Poly(propylene fumarate)/hydroxyapatite nanocomposite/black phosphorus nanosheet phosphate composites for enhanced bone repair

Poly(propylene fumarate)/hydroxyapatite nanocomposite/black phosphorus nanosheet phosphate composites for enhanced bone repair | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Poly(propylene fumarate)/hydroxyapatite nanocomposite/black phosphorus nanosheet phosphate composites for enhanced bone repair Xiaoxia Huang, Jiahan Chen, Rui Ma, Jianghua Wang, Yong Teng This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6050660/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 30 Jul, 2025 Read the published version in Journal of Orthopaedic Surgery and Research → Version 1 posted 12 You are reading this latest preprint version Abstract Background Bone defects due to trauma, infections, congenital malformations, and tumor resection remain significant health challenges. The polymethyl methacrylate (PMMA) bone cement’s limitations in orthopedic applications arise from its lack of bioactivity and the toxicity of its monomers. Hydroxyapatite (HA) cement is widely used for bone reconstruction despite its inherent brittleness. Biodegradable poly(propylene fumarate) (PPF) is recognized for its exceptional performance in addressing these bone defects and providing adequate mechanical support. Black phosphorus (BP) nanosheets (BPNs) have attracted attention due to their unique puckered honeycomb lattice, broad absorption spectrum, high specific surface area, excellent antibacterial properties, and tunable bandgap. In this study, we developed a novel bone cement, PPF/HA/BP, via thermal crosslinking and conducted in vitro evaluation. Methods PPF was synthesized using a two-step approach, whereas BPNs were produced through a liquid-phase exfoliation technique. Then, PPF/HA/BP composite materials were created using a thermal crosslinking process, followed by a thorough examination of their mechanical characteristics, compatibility with cells, osteogenic activity, and degradation properties. Results PPF/HA/BP bone cement was designed by optimizing formulation to possess mechanical properties comparable to bone tissue. PPF bone cement had a polymerization time of 8.16 ± 0.35 min and a temperature of 62.67 ± 0.67°C. HA reduced polymerization time and temperature. PPF/HA/BP exhibited a polymerization time of 6.70 ± 0.10 min and a maximum temperature of 52.5°C ( P < 0.05). PPF/HA/BP enhanced the adhesion, proliferation, and mineralization of preosteoblasts on its surface and demonstrated photothermal properties. When exposed to an 808 nm laser, the bone cement’s temperature rose to 50ºC. After co-culturing with MC3T3-E1 for 14 days, the PPF/HA/BP group exhibited significantly higher expression of ALP , COL I , and RUNX2 compared to the PPF and PPF/HA groups ( P 0.05). Conclusion Biodegradable PPF/HA/BP demonstrated satisfactory mechanical properties, degradation behavior, outstanding photothermal characteristics, excellent biocompatibility, and osteogenic activity. It also promoted bone regeneration by enhancing the proliferation and differentiation of MC3T3-E1 cells in vitro and upregulating the related genes’ expression. Poly(propylene fumarate) Black phosphorus nanomaterials Hydroxyapatite nanocomposite Bone regeneration Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 Background Advancements in construction and transportation have led to an increase in fractures and bone defects due to high-energy traumas. Large bone defects resulting from ischemic necrosis, tumors, joint prosthesis replacements, and osteoporosis pose significant challenges for orthopedic surgeons. Although autogenous bone grafts are effective in bone defect reconstruction, they are limited in repairing large defects and might cause complications at the donor site. Fresh allogeneic bone, though useful, carries risks of immune rejection and disease transmission [ 1 – 3 ]. Currently, commonly used artificial bone tissue repair materials include PMMA bone cement, tricalcium phosphate ceramics, hydroxyapatite ceramics, and calcium phosphate-based biological bone cements. However, PMMA cement has significant limitations. The high exothermic temperature during polymerization can potentially damage surrounding cells. The excessive hardness of PMMA cement is incompatible with vertebral biomechanics, increasing the risk of proximal vertebral fractures [ 4 ]. Recent advancements in tissue engineering have led to the development of promising materials. However, bone substitutes based on natural polymers often exhibit poor biomechanical properties, as they are more suited to soft tissues and fail to meet the mechanical strength, biodegradation rate, and stability necessary for bone tissue [ 5 ]. Compared to natural polymer materials, the physical and chemical properties and the microstructure of synthetic polymer materials are more easily modifiable. The desired properties can be achieved by strategically designing polymer functional groups or adjusting the preparation process parameters [ 6 ]. Versatility is their most significant advantage. PPF is one of the most attractive polymers, with many potential applications in orthopedics. Due to its biodegradability [ 7 ], expandability, certain mechanical properties [ 8 ], and biocompatibility, PPF can gradually replace bone trabecular tissue [ 9 ]. However, PPF cannot provide material properties that closely match those of cortical bone. HA is a bioceramic with a chemical composition and morphology similar to that of bone, with biocompatibility and excellent bone conductivity [ 10 , 11 ]. Incorporating HA into the PPF material increases its strength and enhances its osteoinductive capability. In addition, implant-related infections pose a persistent risk, potentially resulting in severe clinical complications such as implant failure, chronic conditions, and the need for repeated surgical interventions. Zheng et al. [ 12 ] developed composite nanosheets (BPNs@phy) loaded with phytoextracts (Phy) with antibacterial properties. The material exhibited excellent antibacterial effects upon exposure to 808 nm near-infrared light. Recent studies suggest that phosphates may contribute to the mineralization of biomimetic calcium phosphate, enhancing the adhesion, differentiation, and proliferation of bone cells [ 13 ]. After degradation, BPNs can be converted into calcium phosphate, which promotes bone regeneration in physiological environments. BPNs comprise corrugated planes of phosphorus atoms held together by weak van der Waals forces, allowing easy exfoliation into thin BPNs [ 14 ]. In addition, BP is highly reactive toward oxygen and water due to the lone electron pairs on each phosphorus atom in the layered structure [ 15 ]. Such a characteristic limits its biomedical applications and further research [ 16 ]. In previous work, our research group prepared BP/PPF bone cement through thermal crosslinking [ 17 ]. In vitro studies demonstrated its ability to release phosphate ions slowly and continuously. Based on these findings, we hypothesize that introducing BPNs onto the surface of HA significantly enhances its stability and bioactivity. This study constructed PPF/HA/BP bone cement and systematically investigated the impact of different component ratios on the material’s mechanical properties. Key properties were evaluated, including compressive strength, elastic modulus, exothermic temperature, and crosslinking time. Additionally, biocompatibility, in vitro degradation, and safety were assessed through molecular, synthesis, and cell experiments to provide a solid foundation for clinical applications (Fig. 1 ). Materials and methods Preparation of PPF In a light-protected environment, 329.6 mL of diethyl fumarate and 440.65 mL of 1,2-propanediol were transferred into a three-neck round-bottom flask, followed by adding 0.45 g of zinc chloride and 2.75 g of hydroquinone and stirring the mixture. The apparatus was connected to a condenser, and the stirring speed was gradually increased to 300 rpm under nitrogen protection. The temperature of the silicone oil bath was raised from 110°C to 150°C, maintaining it for several hours until approximately 90% of the ethanol was recovered. The vacuum system was connected to a liquid nitrogen condenser, and the pressure was maintained at < 1 mmHg. The mixture was gradually heated to 150°C, avoiding bumping. Once the desired molecular weight was reached, heat was stopped, the air inlet valve was opened slowly, and the vacuum pump was stopped. Equal volumes of 1.85% HCl and saturated NaCl solutions were added to purify the PPF. The mixture was dried with anhydrous Na₂SO₄ overnight and filtered to remove Na₂SO₄ to obtain a clear, pale yellow polymer. Finally, rotary evaporation at 45°C was used under reduced pressure to remove dichloromethane. Preparation of BPNs Fifty mg of BPNs was weighed and added to an N-vinyl-2-pyrrolidone(NVP) solution in a mortar. The mixture was transferred into a brown bottle, and NVP was added to increase the volume to 100 mL. Argon was introduced, and the bottle was sealed. The solution was sonicated in an ice–water bath at 250 W for 12 hours, followed by processing with an ultrasonic cell crusher at 180 W (5 s on/5 s off) for another 12 hours, with ice added to prevent overheating. The mixture was centrifuged at 1300 rpm for 15 min to collect the supernatant, which was then centrifuged at 9660×g (4 () for 20 min to obtain BPNs. The BPNs were washed three times with NVP and resuspended in 100 mL of NVP. Argon was bubbled through, and the suspension was stored at 4°C for future use. Preparation of PPF/HA/BP This study builds upon previous experimental findings to propose an orthogonal experimental design featuring four factors, each with three levels[ 17 , 18 ]. The investigated factors included the heat crosslinking initiator benzoyl peroxide (BPO), BPNs, the crosslinking agent NVP, and HA. These selections aimed to elucidate the effects of various materials and conditions on the experimental outcomes and provide essential data for optimizing the performance of bone cement. The overall experimental design incorporated nine experimental groups, with the sequence and combinations organized according to a standardized orthogonal experimental design table. By assessing the mechanical properties of bone cement formulations, specifically the compressive strength and elastic modulus, this study sought to identify the optimal experimental scheme (Table 1 ). Table 1 Horizontal design table of orthogonal factors A.BPO(mg) B.BP(ug) C.NVP(ml) D.HA(g) 1 5 100 0.25 0.1 2 7.5 200 0.375 0.2 3 10 300 0.5 0.3 After the samples were centrifuged to collect sufficient BPNs, they were resuspended in NVP, and an appropriate amount of hydroxyapatite was added. A certain proportion of BPO was subsequently added. The resulting mixture was then introduced into the pre-prepared PPF and thoroughly stirred, and an initiator, dimethyl-P-toluidine (DMPT), was added. The mixture was stirred until fully homogeneous. Finally, a syringe was used to inject the mixture into a custom-made polytetrafluoroethylene (PTFE) mold measuring 10 mm in diameter and 15 mm in height. Characterization The PPF polymer was dissolved in CDCl 3 and analyzed via a 600-MHz NMR spectrometer to obtain the 1 H-NMR spectrum. The relative molecular weight and polydispersity index of PPF were determined using an Agilent 1260 Infinity II GPC, with chloroform as the eluent and polystyrene as the standard. PPF was mixed with KBr, compressed into pellets, and analyzed via a Thermo Scientific Nicolet iS20 FTIR spectrometer from 4000 to 500 cm − 1 with a 4-cm − 1 resolution. The BPNs were washed three times with anhydrous ethanol, deposited onto a copper grid, and allowed to dry at room temperature, followed by examining their morphology and structure via transmission electron microscopy (FEI Talos F200X, USA) at an operating voltage of 200 kV. BPNs and dried KBr were ground together to form thin pellets, and the functional groups were analyzed via an FTIR spectrometer (FTIR-650, Tianjin Guangdong). After being centrifugally dispersed in deionized water, the particle size and zeta potential of BPNs were measured in the aqueous medium via a Zetasizer Ultra (USA). The morphology and thickness of BPNs were assessed using atomic force microscopy (AFM) (Bruker Dimension Icon, Germany). The samples were initially cured using a PTFE mold measuring 10 mm in diameter and 2 mm in height. The samples were subsequently pre-frozen at -60°C for 6 hours in a freeze-dryer, followed by vacuum freeze-drying for 24 hours. After drying, the samples were sealed and stored in a refrigerator at 4°C. Before imaging, the samples were gold-coated under vacuum and analyzed under a scanning electron microscope (JSM7610FPlus, Japan). Mechanical properties The samples, measuring 10 mm in diameter and 15 mm in height, were fabricated for compression tests under the ISO 604 standard[ 19 , 20 ]. A universal electromechanical testing machine equipped with a 30-kN load cell (E44.304, MTS Systems (China) LTD, CN) was used to conduct the compression tests. The compression rate was set at the standard value of 2 mm/min. Before the tests, the upper and lower surfaces of the samples were meticulously polished with sandpaper to ensure their parallelism. Setting properties The final curing time was recorded using a Vicat apparatus (WKY-1000, Tianjian Instrument Co., Ltd., CN), which was calculated as the duration from the initiation of crosslinking to the formation of a solidified structure. Hydrophilic experiment of bone cement The bone cement’s hydrophilic properties were investigated by selecting PPF/HA/BP, PPF/HA, and PPF materials known for their optimal mechanical properties following formulation optimization. The contact angle, representing the angle formed between the liquid and the solid material, was measured to assess the hydrophilic characteristics. Five parallel samples were prepared for each group, with the bone cement molded in a polytetrafluoroethylene mold measuring 10 mm in diameter and 2 mm in height. The materials were placed on a test platform to measure the contact using a JY-82 contact angle meter. The platform was adjusted to a horizontal position, and an automatic titration system was used to dispense water droplets. Images were captured, and the contact angle was determined by fitting the data according to the Young–Laplace equation. In vitro bone cement degradation Using a PTFE mold, a cylindrical sample was fabricated for the in vitro degradation study, measuring 6 mm in diameter and 10 mm in height. Each sample’s initial weight (M 0 ) was recorded after it had been dried in a vacuum drying oven at 37°C. The prepared sample was subsequently fully immersed in a plastic bottle containing a PBS solution (pH = 7.3), with the solution being replaced weekly. The bone cement-to-PBS buffer ratio was 0.1 g: 10 mL, and the mixture was then continuously incubated in a constant-temperature water bath shaker at 37°C. The samples were retrieved from the solution every week. The surface water was wiped off with filter paper, and the weight was recorded as M 1 . The samples were subsequently dried in a vacuum drying oven at 37°C until completely dehydrated, and the weight was recorded as M 2 . This process continued for four weeks. The initial weight of the sample (M 0 ) was then compared to the sample’s weight after degradation (M t ) to determine the bone cement’s rate of mass loss. In vitro photothermal conversion efficiency The pre-prepared PPF, PPF/HA, and PPF/HA/BP discs measuring 6 mm in diameter and 1 cm in height were submerged in 0.5 mL of PBS. The samples were subsequently irradiated with an 808 nm laser with a consistent power density of 1.0 W/cm². The irradiation time was 10 min, and the distance between the light source and the sample was 12 cm. The samples’ heating curves were recorded using a temperature recorder. To evaluate the stability of the photothermal effect, the PPF/HA/BP bone cement was subsequently irradiated five times. The distance between the light source and the sample was 12 cm, and the heating curves of the samples were recorded using a temperature recorder. Cell culture MC3T3-E1 cells were grown and revived in α-MEM supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. To conduct the osteogenic differentiation assay, the α-MEM was replaced with OriCell osteogenic induction medium containing 10% FBS, 1% penicillin/streptomycin, 100-nM dexamethasone, 50-µg/mL ascorbic acid, and 10-mM β-glycerophosphate. Before being cultured with cells, the cement samples were immersed in 75% alcohol and air-dried under ultraviolet light. Cytotoxicity assay According to GB/T 16886.5–2003/ISO 10993-5-1999, various groups of bone cement materials were initially immersed in 75% alcohol for 30 min and then transferred to a complete medium for 72 hours. The extract was subsequently filtered through a 0.22-µm sterile filter. The cells were counted in a cell counting chamber and then seeded at a density of 5,000 cells/well in 96-well plates. The plates were cultured overnight to allow the cells to fully adhere. The old culture medium was discarded, and the corresponding bone cement extracts were added to each group, with a blank control group lacking material extract. The cells were then cultured for 1, 3, or 5 days. Following the specified incubation period, the extract was removed, and 100 µL of complete culture medium containing 10% CCK8 (Cell Counting Kit-8) was added to each well. The plates were incubated in the dark at 37°C under 5% CO₂ for 1 hour. After incubation, each sample’s optical density (OD) was measured at a wavelength of 450 nm using a microplate reader. The effects of pure PPF, PPF/HA, and PPF/HA/BP extracts on cell morphology were assessed using live/dead (Yeasen 40747ES76, Shanghai, China) and cytoskeletal staining, with the cytoskeleton labeled with phalloidin and the nuclei labeled with DAPI (Servicebio G1012, Servicebio G1041, Wuhan, China). The stained cells were observed under a confocal laser scanning microscope (CLSM; Leica TCS SP2, Heidelberg, Germany) to evaluate cell survival and morphological spread. Cytocompatibility tests The PPF, PPF/HA, and PPF/HA/BP bone cements were meticulously prepared using polytetrafluoroethylene molds measuring 10 mm in diameter and 2 mm in height, with five samples in each group. The prepared bone cements were carefully placed in a biosafety cabinet, soaked in 75% alcohol for 30 min, and gently air-dried. A suspension of MC3T3-E1 cells (3×10 4 cells/mL) was seeded onto the material’s surface and cultured in an incubator for 48 hours. Then, the samples were fixed with 4% paraformaldehyde for 30 min, followed by gradient dehydration with alcohol. The samples were then freeze-dried, gold-coated, and analyzed by scanning electron microscopy (SEM). To each well of a 6-well plate, 1 mL of 0.1% gelatin was added, and the mixture was incubated for 30 min at 37°C. Resuspended MC3T3-E1 cells were then seeded into 6-well plates at a density of 1×10 5 cells/mL, with 2 mL of the cellular suspension added to each well. The plates were incubated at 37ºC under 5% carbon dioxide and maintained under saturated humidity. Upon reaching approximately 70% confluence after incubating for 24‒48 hours, the medium was replaced with an osteogenic medium composed of PPF, PPF/HA, and PPF/HA/BP extracts. The cells were induced to differentiate via the aforementioned extraction medium for 7, 14, and 21 days, with three replicate wells for each time interval. The culture medium was replaced every 48 hours to ensure optimal cell culture conditions. To quantify the material-induced capability at different time points (7, 14, and 21 days), after culturing for the respective durations, the medium was removed, followed by two washes with PBS. Alkaline phosphatase(Beyotime C3206, Shanghai, China) and Alizarin Red dye((OriCell ALIR-10001, Guangzhou, China)) were then added. After incubating for 20 minutes, the samples were washed twice with PBS and observed under a microscope to capture images. Osteogenic-related gene expression Real-time quantitative polymerase chain reaction (QPCR) was used to quantify the expression of osteogenic genes in MC3T3-E1 cells (n = 9). The MC3T3-E1 cells in suspension were adjusted to the required concentration and plated into 6-well plates at a density of 2×10 4 cells/mL, with 2 mL/well. These cells were subsequently incubated at 37°C under 5% carbon dioxide and high humidity. When the cell confluence reached approximately 70%, osteogenic differentiation was generally triggered within 24‒48 hours of incubation. Bone formation-associated gene expression was evaluated through co-culture with the extract. The cells were then treated with an osteogenic medium for 14 days, and the medium was changed every 48 hours. Total RNA was extracted from the cells using an RNeasy Mini Kit (Qiagen 74104, Hilden, Germany). QPCR was performed to measure the mRNA expression of marker genes, including alkaline phosphatase ( ALP ), runt-related transcription factor 2 ( RUNX2 ), and collagen I ( Col-I ). Beta-actin was used as the internal control. Table S1 presents the primer sequences. Reverse transcription was performed on 1 mg of RNA using the reverse transcription kit SuperScriptTM III Reverse Transcriptase. The qRT‒PCR assays were conducted using SYBR Premix Ex Taq II (×2) as a PCR reagent on an ABI 7500 rapid machine (Applied Biosystems StepOne, MA, US). The results were calculated using the delta‒delta Ct method (2 −△△ Ct). Statistical analysis Quantitative variables were reported as means ± SDs. The normality of the data was assessed using the Shapiro‒Wilk test. The Mann‒Whitney U test was used to compare skewed data and determine significant differences. For non-skewed data, one-way analysis of variance (ANOVA) was conducted with post hoc pairwise comparisons. GraphPad Prism 9.0 software (San Diego, CA, US) was used for all analyses and graphic representations. A P-value of < 0.05 was considered statistically significant. Results Characterization of PPF and BPNs In the 1 H-NMR spectrum of the PPF polymer, the peak at 7.26 ppm corresponded to the solvent, deuterated chloroform. The signal at 6.86 ppm was attributed to the double bond present in the PPF molecular chain. The methylidene signals in the PPF were observed at 5.28 and 4.24 ppm, whereas the methyl signal was detected at 1.23 ppm (Fig. 2 ). As shown in Fig. 3 a and b, the BPNs exhibited an irregular morphology with an approximate size of 300 nm in the TEM images. AFM characterization of the BPNs revealed a thickness of approximately 5 nm (Fig. 3 c). Figure 3 d shows that the average particle size of the BPNs synthesized in this study was 314.7 nm. Characterization of PPF/HA/BP The scanning electron microscopy (SEM) images (Fig. 4 ) illustrated the smooth surface of the PPF bone cement material. Hydroxyapatite particles and black phosphorus nanosheets were partially deposited on the surface of the PPF, although they did not completely cover the material. Additionally, the results of the hydrophilic angle tests indicated that hydroxyapatite particles and black phosphorus nanosheets significantly influenced the surface hydrophilicity of the bone cement, suggesting that the changes in the hydrophilic angle were statistically significant ( P < 0.001). Mechanical properties PPF bone cement with various formulations exhibited remarkable mechanical strength, with a compressive strength range of 70–150 MPa and an elastic modulus range of 600‒900 MPa, suggesting a close resemblance to human bone tissue regarding its mechanical properties (Table 2 ). Table 2 Results of bone cement orthogonal test Test number Factor Result A B C D Compressive strength (Mpa) Modulus of elasticity (Mpa) 1 1 1 1 1 110.2 682 2 1 2 2 2 122.3 733.4 3 1 3 3 3 73.7 710.6 4 2 1 2 3 134 903.7 5 2 2 3 1 132 829.2 6 2 3 1 2 145.2 758.8 7 3 1 3 2 147.9 844.8 8 3 2 1 3 152.3 868.9 9 3 3 2 1 127.9 846.9 Based on the results of the range analysis, the influence of factors A, B, C, and D on the compressive strength was as follows: A (BPO) > B (BPNs) > D (HA) > C (NVP). Among them, BPO and BPNs positively impacted the compressive strength of bone cement, indicating that the higher their content, the greater the compressive strength of the bone cement. The optimal level of HA for compressive strength was at the second level. In contrast, NVP had an adverse effect on compressive strength, indicating that the higher its content, the lower the compressive strength of the bone cement. Therefore, based on the range analysis, the optimal formulation for compressive strength is A3B2D2C1. Additionally, based on the range analysis, the influence of factors A, B, C, and D on the elastic modulus was as follows: A (BPO) > C (NVP) > D (HA) > B (BPNs). BPO and HA positively affected the elastic modulus, indicating that the higher their content, the greater the elastic modulus of the bone cement. BPNs and NVP had an optimal formulation at the second level. Therefore, based on the range analysis, the optimal formulation for the elastic modulus is A3C2D3B2. Finally, based on the comprehensive balance method, the best bone cement formulation is A3B2C1D2 (Fig. 5 and Table 3 ). Table 3 Analysis of compressive strength and elastic modulus of bone cement Compressive strength Modulus of elasticity A B C D A B C D K1 102.06 130.7 135.9 123.36 708.67 810.16 769.9 786.03 K2 137.06 135.53 128.06 138.46 830.56 810.5 828 779 K3 142.7 115.6 117.86 120 853.53 772.1 794.86 827.73 R 40.63 19.93 18.03 18.46 144.86 27.13 58.1 48.73 Best index 3 2 1 2 3 2 2 3 Setting properties The polymerization time of PPF bone cement was 8.16 ± 0.35 min, and the polymerization temperature was 62.67 ± 0.67°C. Incorporating hydroxyapatite (HA) shortened the polymerization time of PPF/HA bone cement and decreased the polymerization temperature. Significant differences in polymerization time and temperature were observed between the two groups ( P < 0.01). When BPNs were added along with hydroxyapatite, the maximum temperature of the PPF/HA/BP bone cement reached 52.5°C, with a significant difference between the two groups ( P < 0.01). The polymerization time of PPF/HA/BP was 6.70 ± 0.10 min, which was significantly shorter than that of PPF/HA (7.33 ± 0.15), with a significant difference between the two groups ( P < 0.001) (Fig. 6 ). In vitro Degradation Behavior and Photothermal Conversion Efficiency of PPF, PPF/HA, and PPF/HA/BP Figure 7 illustrates the degradation process of various materials in a human-simulated solution in vitro. The pH of these materials consistently remained close to 7.4. The degradation rate of the PPF-based bone cement was relatively rapid during the initial two weeks, stabilizing thereafter. The scaffold composite exhibited excellent adaptability when subjected to a dynamic in vivo environment. After irradiation at 808 nm with a power density of 1.0 W/cm 2 for 10 min, the temperature of the PPF/HA/BP group was 17°C higher than that of the PPF and PPF/HA groups. Corresponding experiments were conducted to evaluate the repeatability of the photothermal properties of the bone cement in the PPF/HA/BP group. After completing five photothermal cycles, the temperature change curve of the bone cement in the BP group exhibited minimal variations, with the maximum temperature stabilizing at approximately 50ºC, indicating that its photothermal conversion performance remained consistent across multiple cycles. Following irradiation at a wavelength of 808 nm and a power density of 1.0 W/cm 2 for 10 min, the PPF/HA/BP group presented a temperature increase of 17ºC compared with those of the PPF and PPF/HA groups. Relevant experiments were conducted.to evaluate the consistency of the photothermal characteristics of the bone cement within the PPF/HA/BP group. After completing five photothermal cycles, the temperature variation curve for the BP group demonstrated minimal changes, with the peak temperature stabilizing at approximately 50ºC, indicating that its photothermal conversion efficiency remained stable across multiple cycles (Fig. 8 ). Cell adhesion and cytotoxicity assay MC3T3-E1 cells were co-cultured with the extracts of various bone cements, including pure PPF, PPF/HA, and PPF/HA/BP, for 1, 3, and 5 days to evaluate their in vitro cytocompatibility (Fig. 9 ). The results demonstrated no significant difference in cell viability between the pure PPF bone cement extract-treated and control groups ( P > 0.05). In contrast, cell viability in the PPF/HA and PPF/HA/BP bone cement extract-treated groups was significantly higher than the control and pure PPF bone cement-treated groups ( P < 0.05). Additionally, statistically significant differences were observed in cell proliferation between the PPF/HA/BP and PPF/HA bone cement extract co-cultured groups, indicating that both PPF/HA/BP and PPF/HA bone cement materials promoted cell proliferation. However, PPF/HA/BP exhibited a more pronounced effect on osteoblast proliferation. Figure 10 shows the adhesion of MC3T-E1 cells to the surface of the bone cement. The results indicated significantly less cell adhesion on the PPF cement surface than on the PPF/HA and PPF/HA/BP cement surfaces. Additionally, the pseudopodia of the cells actively adhered to the PPF/HA bone cement, particularly on the hydroxyapatite particles, suggesting a strong interaction between the cells and the material. The cells exhibited a clear overlapping growth pattern in cultures with PPF/HA/BP bone cement. Live/dead fluorescence staining was used to assess cell survival. The results revealed a significantly higher cell survival rate in the experimental group compared to the control group (Fig. 11 ). Notably, the PPF/HA/BP group demonstrated outstanding performance, with the highest number of living cells, highlighting its effectiveness in promoting cell survival. Additionally, as the cell growth period increased, the number of live cells gradually increased, and the area covered by fluorescence staining significantly expanded, further confirming the positive impact of the culture conditions on cell survival. Under the effect of different types of bone cement extracts, MC3T3 cells presented a polygonal or spindle shape, with the cytoskeleton exhibiting a bundle arrangement parallel to the vertical axis of the cell, forming a network structure. This further indicates that PPF-based materials have no apparent cytotoxicity. The PPF/HA/BP group’s cytoskeletal structure was clearer, the overall spreading state was good, and the cytoskeletal structure and spreading state were better than those of the control group (Fig. 12 ). Alkaline phosphatase staining and alizarin red staining MC3T3-E1 cells were induced with an osteogenic induction medium and co-cultured with various extracts, with observations at 7 and 14 days. As shown in Fig. 13 , no significant abnormalities were noted in the staining of the PPF or positive control groups during the 7-day staining experiment, indicating that the pure PPF material exhibited no significant toxicity and did not interfere with the MC3T3-E1 cells’ early mineralization. The ALP staining results for the PPF/HA and PPF/HA/BP material groups were more intense compared to the positive control group, with the PPF/HA/BP group displaying the most pronounced staining. As the co-culture period was extended, the differences between the groups became increasingly evident. After 21 days of induction, the number of calcium nodules in the PPF/HA and PPF/HA/BP groups was higher than that in the positive control and PPF groups. Moreover, the calcium nodules in the PPF/HA/BP material group were larger, confirming that both the PPF/HA and PPF/HA/BP materials promoted osteogenic differentiation, with the PPF/HA/BP group showing the most pronounced effect. Osteogenesis-related gene expression The expression of ALP , COL I , and RUNX2 in the PPF/HA/BP and PPF/HA groups was significantly higher than the control group after 14 days of co-culture ( P < 0.001). Moreover, the PPF/HA/BP group showed significantly higher expression levels compared to the PPF/HA group( P 0.05), suggesting that PPF/HA/BP had favorable osteogenic differentiation activity in vitro (Fig. 14 ). Discussion The prevalence of bone-related diseases is increasing due to factors such as an aging population and sports injuries. In recent years, the number of patients presenting with bone injuries or defects has notably increased, posing significant clinical challenges for effective bone repair. Relevant studies indicate that the global number of fracture cases increased from 133.4 million in 1990 to 178 million in 2019, reflecting an increase of over one-third [ 21 ]. However, when the size of a bone defect exceeds a certain critical threshold, the bone’s self-repair ability is significantly impaired, hindering the restoration of its normal function and structure. When a bone defect exceeds a specific size, its self-repair ability is insufficient, necessitating bone healing promotion through surgical intervention. Although autologous bone transplantation is the“gold standard” [ 22 ], it is associated with problems such as donor shortage, postoperative infection, and pain, limiting its application. Although allogeneic bone transplantation can provide an alternative, there is a risk of immune rejection and disease transmission. Therefore, safer and more effective bone repair programs are urgently needed to meet clinical needs and improve patient outcomes [ 23 ]. Natural proteins and peptides, particularly those derived from collagen, silk fibroin, fibrin, and gelatin, have shown considerable promise in bone tissue engineering. Despite collagen’s several advantages, such as low antigenicity, favorable hydrophilicity, and ease of modification, its stability is diminished under standard physiological conditions [ 24 ]. Thus, enhancing the biomechanical properties of polymers and improving their compatibility with bone tissue is a critical avenue for future research. PPF has attracted significant attention because of its outstanding properties and has progressively emerged as a promising material in biomedical research. It offers many potential applications, particularly in replacing bone trabecular tissue. Its performance advantages make it an ideal candidate material [ 25 ]. Kimicata et al. [ 26 ] integrated extracellular matrix components from tissues with PPF-based materials to develop a biodegradable biological scaffold suitable for use as a vascular graft. This approach uses the advantages of both components, providing a platform for cell growth due to favorable material properties. Although PPF has excellent mechanical properties and biocompatibility as a bone cement, it lacks bone-inductive capabilities and cannot directly promote bone regeneration. The HA/polymer composite scaffold exhibited exceptional mechanical properties, osteogenic potential, and slow release. As a scaffold for bone regeneration, significant advancements have been made in applying growth factors, cells, and drug delivery systems for bone reconstruction [ 27 ]. Teng et al. [ 18 ] developed PPF/HA cervical cages using ASTM standard molds to evaluate the compression, bending, tensile, and hardness properties of PPF:HA bulk materials. The findings indicated that this polymer composite could be a promising candidate for cervical cages. Additionally, a cylindrical disk of poly-ε-caprolactone was created through melt deposition and modified with nanohydroxyapatite and polypropyl fumarate. Tissue regeneration enhancement was assessed via bone density measurements and micro-CT imaging [ 28 ]. However, some studies have shown that hydroxyapatite has the lowest solubility among various calcium phosphate salts [ 29 ], limiting its application in bone tissue engineering repair materials. In recent years, black phosphorus nanosheets have demonstrated significant advantages in bone regeneration, and their unique properties have made them promising candidate materials for bone tissue repair. As the most stable allotrope of elemental phosphorus, black phosphorus has a high degree of homology with the inorganic phosphorus found in human bone. Therefore, it provides a robust scientific basis for its application in bone mineralization and healing processes. Furthermore, black phosphorus exhibits excellent biocompatibility and superior degradation performance. In this study, the optimal bone cement, with mechanical properties most similar to human bone, was prepared using orthogonal design optimization. Additionally, hydrophilicity experiments demonstrated that incorporating BPNs and HA significantly enhanced the surface hydrophilicity of the PPF-based materials. Metal implants are extensively used in orthopedics, providing several advantages. However, they have significant drawbacks [ 30 ], including high costs, manufacturing complexities, and the potential for irregular stress distribution, which can impact long-term stability. Furthermore, metal implants may produce artifacts during X-ray or CT scan examinations [ 31 ], compromising diagnostic accuracy. Additionally, the nonbiodegradable nature of metal materials often necessitates removal through secondary surgery, increasing patient risks, possible allergic reactions, or other complications. The unsaturated double bond feature of PPF enables it to react with various molecules, facilitating the formation of crosslinked products with compounds such as N-vinylpyrrolidone, bioceramics, and natural organic macromolecules. As a biodegradable material, PPF breaks down into fumaric acid and propylene glycol during degradation. These byproducts are metabolized and excreted from the body, ensuring the safety and biocompatibility of the material within the organism [ 32 ]. The present study demonstrated that the pH of the PPF-based materials remained stable at 7.4 throughout the degradation process, further confirming that the PPF did not disrupt the internal environment. The SEM images indicated proper adhesion of MC3T3 cells to the surface, indicating a healthy growth pattern. This interaction suggests that the PPF/HA/BP composite material created an environment conducive to cell attachment and proliferation. Implant-associated infections have long been a significant factor affecting the success of implant procedures. These infections result in physical pain for patients and increase medical costs [ 33 – 35 ]. Clinical trials have demonstrated that bacterial proliferation in the surrounding microenvironment can lead to implant failure. The hydroxyapatite composite lacks intrinsic antibacterial properties and fails to exert an antibacterial effect when implanted in the body. In contrast, as a novel antibacterial material, black phosphorus possesses a unique two-dimensional layered structure that facilitates effective interactions with bacterial cell membranes through physical mechanisms [ 12 , 36 ]. Naskar et al. [ 37 ] demonstrated the remarkable photothermal antibacterial properties of Au-ZnO-BP nanocomposites against multidrug-resistant Staphylococcus aureus strains. A thin layer of black phosphorus nanosheets, which measured 300 nm and featured a controlled particle size, was successfully prepared through liquid-phase exfoliation in this study. Compared with the PPF and PPF/HA groups, the temperature of the PPF/HA/BP group increased by 17°C after irradiation at a wavelength of 808 nm with a power density of 1.0 W/cm² for 10 min. To assess the consistency of the photothermal properties of the bone cement in the PPF/HA/BP group, the temperature variation curve following five photothermal cycles demonstrated the slightest change in the BP group, with the peak temperature stabilizing at approximately 50°C. This stability indicates that its photothermal conversion efficiency remained consistent across multiple cycles. MC3T3-E1 cells are widely used as osteoblast precursors, as they differentiate into mature osteoblasts over time under osteogenic induction. Approximately 14 days after induction, the cells reach a critical stage of osteogenic differentiation, with significant changes in the expression of key osteogenesis-related genes such as ALP, RUNX2 , and COL-1 . PCR analysis of these genes accurately reflects the osteogenic differentiation status. ALP is involved in bone matrix mineralization [ 38 ], RUNX2 acts as a key transcription factor for osteoblast differentiation, and COL I , the primary organic component of the bone matrix [ 39 ], is essential for maintaining bone structure and mechanical properties. According to several previous studies [ 40 – 42 ], black phosphorus nanosheets can upregulate the expression of osteogenesis-related genes upon near-infrared light irradiation and show promise in preventing clinical bacterial infections. They also enhance the mechanical strength of dynamic self-healing hydrogels and enable the gel scaffold to retain protein binding [ 43 ]. Black phosphorus is degraded in vivo and transforms into nontoxic phosphate ions that can effectively bind with calcium ions. This process is crucial in enhancing bone hardness and promoting bone regeneration. This study demonstrated that the degradation of black phosphorus, in conjunction with HA, synergistically upregulated osteogenic genes such as ALP , COL I , and RUNX2 , further inducing bone mineralization [ 44 , 45 ]. This study indicated a significant increase in alkaline phosphatase activity in the PPF/HA/BP group. Furthermore, the expression of osteogenic-related genes, including ALP, COL I , and RUNX2 , was notably upregulated compared to other groups. When designing orthopedic implants, factors other than only mechanical properties are essential. Achieving a balance between mechanical strength, biocompatibility, and degradation rate is vital for ensuring optimal performance and long-term success. However, natural polymer-based bone substitutes often pose specific challenges, particularly regarding their poor biomechanical properties. A primary concern is that the inherent characteristics of natural polymers are generally more compatible with soft tissues, which complicates their ability to fulfill the mechanical strength, biodegradation rate, and stability requirements essential for bone tissue [ 10 , 46 , 47 ]. The PPF-based bone cement rapidly degraded during the initial two weeks, after which the degradation rate diminished, allowing for an extended period and space for bone healing. As the material progressively degrades, bone tissue begins to infiltrate the area, ensuring the stability of the bone defect site and mitigating the risk of losing support due to the rapid degradation of the material. Consequently, it ensures that the bone defect site is consistently and stably supported throughout the healing process, ultimately facilitating effective integration with the bone tissue. This study conducted experiments on the materials’ mechanical properties, cellular behavior, degradation rate, biocompatibility, and osteogenic activity. Among these, PPF/HA/BP exhibited mild degradation behavior, good biocompatibility, and high osteogenic induction activity, making it particularly suitable for meeting the clinical demands of biodegradable bone implants. However, several aspects of the results warrant further investigation. First, the material characterization tests did not consider dynamic fluid flow, stress effects, or the frictional resistance of the porous scaffolds. Additionally, the impact of the porous structure design with varying porosities on the mechanical properties and biodegradable characteristics should be further clarified. Finally, future research should focus on designing degradable scaffolds integrated with antibiotic-loaded materials via 3D printing technology. Conclusion The biodegradable PPF/HA/BP composite demonstrated exceptional mechanical properties, favorable degradation behavior, outstanding photothermal characteristics, excellent biocompatibility, and significant osteoinductive activity. It promoted the proliferation and differentiation of MC3T3-E1 cells in vitro, upregulated the expression of key genes, and facilitated bone regeneration. This composite provided an environment conducive to osteoblasts and effectively stimulated the cellular processes involved in bone formation, making it a highly promising material for bone tissue engineering. An in vivo study will be carried out in later stages to further confirm the osteogenic effects of the material. Moreover, incorporating BPNs significantly enhanced the photothermal response of the material, offering the potential for more precise and localized treatment strategies in regenerative medicine. Additionally, the synergistic interaction between PPF, HA, and BPNs ensured the mechanical stability of the scaffold and accelerated the healing process by supporting the regeneration of bone tissue under physiological conditions. In the future, we aim to fabricate porous PPF/HA/BP scaffolds that incorporate drugs to enable an integrated system for controlled drug release and structural-osteogenic functionality. Abbreviations AFM atomic force microscopy ALP alkaline phosphatase BP black phosphorus ARS alizarin red staining BPNs black phosphorus nanosheets BPO benzoyl peroxide β-GP β-glycerophosphate CCK8 cell counting kit-8 Col I collagen I DMPT dimethyl-P-toluidine HA hydroxyapatite NVP N-vinyl-2-pyrrolidone OD optical density Phy phytoextracts PMMA polymethylmethacrylate PPF poly(propylene fumarate) PTFE polytetrafuoroethylene RUNX2 Runt-related transcription factor 2 SEM scanning electron microscope Declarations Consent for publication Written informed consent was obtained from the patient for publication of this case report and any accompanying images. A copy of the written consent is available for review by the Editor of this journal. Competing interests The authors declare that they have no conflict of interest. Author details 1 Graduate School of Xinjiang Medical University, Urumqi, Xinjiang, China. 2 Department of Orthopedics, General Hospital of Xinjiang Military Region, Urumqi, Xinjiang, China. 3 Department of Orthopedics, the First Affiliated Hospital of Chengdu Medical College.4 Department of Pharmacy, General Hospital of Xinjiang Military Region, Urumqi, Xinjiang, China. Funding The research was funded by the Natural Science Foundation of Xinjiang Uygur Autonomous Region (2023D01C93) and (CYFY-GQ73). Author Contribution HXX conducted the study; collected, analyzed, and interpreted the data; and wrote the manuscript. CJ interpreted the data and edited the manuscript. JW reviewed the manuscript. RM planned the project YT edited the manuscript and reviewed the manuscript. XH and CJ have equally to equally to this study. Acknowledgements Not applicable. 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Supplementary Files TableS1.docx Cite Share Download PDF Status: Published Journal Publication published 30 Jul, 2025 Read the published version in Journal of Orthopaedic Surgery and Research → Version 1 posted Editorial decision: Revision requested 09 May, 2025 Reviews received at journal 03 May, 2025 Reviews received at journal 01 May, 2025 Reviews received at journal 28 Apr, 2025 Reviewers agreed at journal 28 Apr, 2025 Reviews received at journal 27 Apr, 2025 Reviewers agreed at journal 26 Apr, 2025 Reviewers agreed at journal 24 Apr, 2025 Reviewers agreed at journal 24 Apr, 2025 Reviewers invited by journal 24 Apr, 2025 Submission checks completed at journal 21 Apr, 2025 First submitted to journal 17 Apr, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6050660","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":449207569,"identity":"ed7b25c8-c899-45a1-8544-db3e0359526b","order_by":0,"name":"Xiaoxia Huang","email":"","orcid":"","institution":"Xinjiang Medical University","correspondingAuthor":false,"prefix":"","firstName":"Xiaoxia","middleName":"","lastName":"Huang","suffix":""},{"id":449207572,"identity":"b72b51b0-5160-4dc6-b1d4-4a275957cf53","order_by":1,"name":"Jiahan Chen","email":"","orcid":"","institution":"the First Affiliated Hospital of Chengdu Medical College","correspondingAuthor":false,"prefix":"","firstName":"Jiahan","middleName":"","lastName":"Chen","suffix":""},{"id":449207574,"identity":"707b1d20-3161-4d18-b167-413d55bd3382","order_by":2,"name":"Rui Ma","email":"","orcid":"","institution":"Xinjiang Medical University","correspondingAuthor":false,"prefix":"","firstName":"Rui","middleName":"","lastName":"Ma","suffix":""},{"id":449207575,"identity":"18653cd9-482a-4623-b552-a87d3b298b88","order_by":3,"name":"Jianghua Wang","email":"","orcid":"","institution":"General Hospital of Xinjiang Military Region","correspondingAuthor":false,"prefix":"","firstName":"Jianghua","middleName":"","lastName":"Wang","suffix":""},{"id":449207576,"identity":"d5ab4dde-3236-4079-b32b-ae35c574ff29","order_by":4,"name":"Yong Teng","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA4UlEQVRIiWNgGAWjYHACNgTzg4GNHRE6mNkYDkCZjDMK0pJJ08LM8+EQYwMhDfzS5489/lBxx27D8d7Dr20MDjAzsB8+ugGfFsm+ZHaDA2eeJW84cy7NOsfgDh8DT1raDXxaDM4ws0kcbDucbHAjx8w4x+AZM4MEjxkRWv5BtVgYHGZsIE5Lw2E7oBbjxwzEaJHsYTaTOHPscILkmTNmjD0GaclshPzCz8P4TKKi5rA93/Ee4w8//tjY8bMfPoZXCwwkLjjAwCYBYrERUgoD9vINDMwfiFU9CkbBKBgFIwsAAPvKTil5A/k0AAAAAElFTkSuQmCC","orcid":"","institution":"General Hospital of Xinjiang Military Region","correspondingAuthor":true,"prefix":"","firstName":"Yong","middleName":"","lastName":"Teng","suffix":""}],"badges":[],"createdAt":"2025-02-17 19:53:29","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6050660/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6050660/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s13018-025-06028-z","type":"published","date":"2025-07-30T16:05:31+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":81607717,"identity":"4ab4f680-6f62-471f-ac66-25dbd855818e","added_by":"auto","created_at":"2025-04-29 06:17:47","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":84427,"visible":true,"origin":"","legend":"\u003cp\u003ePreparation of PPF/HA/BP bone cement and its application in bone tissue\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/02cae2dfdb5977a87791c69e.jpg"},{"id":81608205,"identity":"4535d916-848a-4536-bbe6-f55eacbca055","added_by":"auto","created_at":"2025-04-29 06:25:46","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1148592,"visible":true,"origin":"","legend":"\u003cp\u003e1H-NMR spectra of PPF copolymer\u003c/p\u003e","description":"","filename":"Fig.2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/825c825b639dcf93f8f6939c.jpg"},{"id":81608222,"identity":"fa0239ad-a69b-41ae-9915-093daeda1dd0","added_by":"auto","created_at":"2025-04-29 06:25:48","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":779542,"visible":true,"origin":"","legend":"\u003cp\u003eA and B. Transmission electron microscopy images of BPNs; C. AFM selected area thickness analysis image; D. Average particle size distribution of BPNs.\u003c/p\u003e","description":"","filename":"Fig.3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/481ccde819b076e4aa6b86f3.jpg"},{"id":81608218,"identity":"f6e3080f-83b4-4b6c-a389-cb629a8179ed","added_by":"auto","created_at":"2025-04-29 06:25:47","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":517107,"visible":true,"origin":"","legend":"\u003cp\u003eSEM characterization and hydrophilicity experiments of different materials. A-C. SEM surface morphology of PPF, PPF/HA, and PPF/HA/BP bone cement materials; D. Water contact angles of PPF, PPF/HA, and PPF/HA/BP bone cement materials; E. Comparison of hydrophilicity angles of the three materials.(***\u003cem\u003eP\u003c/em\u003e＜0.001 VS PPF;\u003csup\u003e###\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e＜0.001 VS PPF/HA)\u003c/p\u003e","description":"","filename":"Fig.4R1.png","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/7d891327e606f37a1e6b5391.png"},{"id":81609497,"identity":"1a9804e7-c168-40bd-abaf-12b78e2f117e","added_by":"auto","created_at":"2025-04-29 06:41:47","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":667549,"visible":true,"origin":"","legend":"\u003cp\u003eStress-strain curves for different ratios.\u003c/p\u003e","description":"","filename":"Fig.5R1.tif.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/feaa620ad0518a2d2b0a7794.jpg"},{"id":81607742,"identity":"308152ca-6f09-4526-86a9-c65922735b20","added_by":"auto","created_at":"2025-04-29 06:17:48","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":216161,"visible":true,"origin":"","legend":"\u003cp\u003ePolymerization temperature and time of bone cement: (A) Falling polymerization temperature of different bone cements; (B) Different polymerization time of bone cement.(**\u003cem\u003eP\u003c/em\u003e＜0.01 VS PPF;***\u003cem\u003eP\u003c/em\u003e＜0.001 VS PPF;\u003csup\u003e##\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e＜0.01 VS PPF/HA;\u003csup\u003e###\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e＜0.001 VS PPF/HA)\u003c/p\u003e","description":"","filename":"Fig.6R1.tif.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/69e3b7315d28b5997276afed.jpg"},{"id":81607758,"identity":"628d2717-f9e9-4311-8711-35e9e6a20d83","added_by":"auto","created_at":"2025-04-29 06:17:49","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":5879579,"visible":true,"origin":"","legend":"\u003cp\u003eIn vitro degradation experiment of bone cement: (A) PH changes during the degradation of bone cement; (B) Weight loss during the degradation of bone cement;(C) Changes in water absorption of bone cement; (D) Expansion during the degradation process of bone cement.\u003c/p\u003e","description":"","filename":"Fig.7R1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/bc0bccfe62cf731ce09f48ed.jpg"},{"id":81607750,"identity":"024fbbf1-9df4-483d-b246-33ad5e1ead6a","added_by":"auto","created_at":"2025-04-29 06:17:49","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":1031112,"visible":true,"origin":"","legend":"\u003cp\u003e808nm near-infrared light (1W/cm²) irradiation on bone cement: A.Temperature variation curve; B. Repeated cycle irradiation temperature variation curve.\u003c/p\u003e","description":"","filename":"Fig.8R1.png","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/e50155b1aaeb3aea60f5dc20.png"},{"id":81608208,"identity":"6c7ab516-7c48-4fc5-a51b-10e4790d3d72","added_by":"auto","created_at":"2025-04-29 06:25:47","extension":"jpg","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":93038,"visible":true,"origin":"","legend":"\u003cp\u003eThe activity of MC3T3-E1 cells co-cultured in PPF, PPF/HA, and PPF/HA/BP bone cement extracts was assessed at 1, 3, and 5 days. A blank control group with no material extract was also included.(\u003csup\u003ens\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e＞0.05 VS Control;**\u003cem\u003eP\u003c/em\u003e＜0.01 VS Control;***\u003cem\u003eP\u003c/em\u003e＜0.001 VS Control;\u003csup\u003e##\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e＜0.01 VS PPF/HA;\u003csup\u003e###\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e＜0.001 VS PPF/HA)\u003c/p\u003e","description":"","filename":"Fig.9R1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/54c1988dc8a0b64dba395863.jpg"},{"id":81607737,"identity":"87f4dc03-e522-4883-b856-0945b8d71762","added_by":"auto","created_at":"2025-04-29 06:17:48","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":1085163,"visible":true,"origin":"","legend":"\u003cp\u003eCell growth on the surface of bone cement under SEM.\u003c/p\u003e","description":"","filename":"Fig.10R1.png","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/4327bd5b7d7fb37db3588c85.png"},{"id":81608219,"identity":"0036831a-08b2-4f0d-a238-80c60085c2a4","added_by":"auto","created_at":"2025-04-29 06:25:47","extension":"jpg","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":14910565,"visible":true,"origin":"","legend":"\u003cp\u003eStaining experiment of living and dead cells (Live cells were stained with green ffuorescence, dead cells with red ffuorescence).\u003c/p\u003e","description":"","filename":"Fig.11R1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/9b06b750bd5142df3ff907be.jpg"},{"id":81608233,"identity":"afa02693-cf88-4788-af39-cd6146dec195","added_by":"auto","created_at":"2025-04-29 06:25:49","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":709135,"visible":true,"origin":"","legend":"\u003cp\u003ePhalloidin experiment.Atin fflaments with ffuorescein (red), and nuclei with DAPI (blue)\u003c/p\u003e","description":"","filename":"Fig.12R1.png","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/50e8684359ce62c43b30f663.png"},{"id":81608243,"identity":"8c32dc28-69c0-4ab5-8a71-410bc53f2be1","added_by":"auto","created_at":"2025-04-29 06:25:50","extension":"png","order_by":13,"title":"Figure 13","display":"","copyAsset":false,"role":"figure","size":141715832,"visible":true,"origin":"","legend":"\u003cp\u003eALP staining at 7 and 14 days, and Alizarin Red staining at 21 days for each group.\u003c/p\u003e","description":"","filename":"Fig.13R1.png","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/683ffbe07ddac5efb47d1c24.png"},{"id":81608600,"identity":"5f71e5e6-187c-4345-b85f-06efd0718b6c","added_by":"auto","created_at":"2025-04-29 06:33:47","extension":"png","order_by":14,"title":"Figure 14","display":"","copyAsset":false,"role":"figure","size":451913,"visible":true,"origin":"","legend":"\u003cp\u003eExpression of osteogenic-related genes in MC3T3-E1 cells treated with different materials.(\u003csup\u003ens\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e＞0.05 VS Control;***\u003cem\u003eP\u003c/em\u003e＜0.001 VS Control;\u003csup\u003e##\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e＜0.01 VS PPF/HA;\u003csup\u003e###\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e＜0.001 VS PPF/HA)\u003c/p\u003e","description":"","filename":"Fig.14R1.png","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/130e4dcc460a75a145586572.png"},{"id":88268315,"identity":"fb826824-f9d3-44c4-95ec-e24b777d9aae","added_by":"auto","created_at":"2025-08-04 16:50:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":100877299,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/8dcbc0cc-1395-411f-a498-fe7b2ea68dfc.pdf"},{"id":81607708,"identity":"ddceefe5-f717-441d-887a-a755b6b52fd0","added_by":"auto","created_at":"2025-04-29 06:17:46","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":17473,"visible":true,"origin":"","legend":"","description":"","filename":"TableS1.docx","url":"https://assets-eu.researchsquare.com/files/rs-6050660/v1/11695959afd29caf9053d4cf.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Poly(propylene fumarate)/hydroxyapatite nanocomposite/black phosphorus nanosheet phosphate composites for enhanced bone repair","fulltext":[{"header":"Background","content":"\u003cp\u003eAdvancements in construction and transportation have led to an increase in fractures and bone defects due to high-energy traumas. Large bone defects resulting from ischemic necrosis, tumors, joint prosthesis replacements, and osteoporosis pose significant challenges for orthopedic surgeons. Although autogenous bone grafts are effective in bone defect reconstruction, they are limited in repairing large defects and might cause complications at the donor site. Fresh allogeneic bone, though useful, carries risks of immune rejection and disease transmission [\u003cspan additionalcitationids=\"CR2\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Currently, commonly used artificial bone tissue repair materials include PMMA bone cement, tricalcium phosphate ceramics, hydroxyapatite ceramics, and calcium phosphate-based biological bone cements. However, PMMA cement has significant limitations. The high exothermic temperature during polymerization can potentially damage surrounding cells. The excessive hardness of PMMA cement is incompatible with vertebral biomechanics, increasing the risk of proximal vertebral fractures [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eRecent advancements in tissue engineering have led to the development of promising materials. However, bone substitutes based on natural polymers often exhibit poor biomechanical properties, as they are more suited to soft tissues and fail to meet the mechanical strength, biodegradation rate, and stability necessary for bone tissue [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Compared to natural polymer materials, the physical and chemical properties and the microstructure of synthetic polymer materials are more easily modifiable. The desired properties can be achieved by strategically designing polymer functional groups or adjusting the preparation process parameters [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Versatility is their most significant advantage. PPF is one of the most attractive polymers, with many potential applications in orthopedics. Due to its biodegradability [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e], expandability, certain mechanical properties [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e], and biocompatibility, PPF can gradually replace bone trabecular tissue [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. However, PPF cannot provide material properties that closely match those of cortical bone. HA is a bioceramic with a chemical composition and morphology similar to that of bone, with biocompatibility and excellent bone conductivity [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Incorporating HA into the PPF material increases its strength and enhances its osteoinductive capability.\u003c/p\u003e \u003cp\u003eIn addition, implant-related infections pose a persistent risk, potentially resulting in severe clinical complications such as implant failure, chronic conditions, and the need for repeated surgical interventions. Zheng et al. [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e] developed composite nanosheets (BPNs@phy) loaded with phytoextracts (Phy) with antibacterial properties. The material exhibited excellent antibacterial effects upon exposure to 808 nm near-infrared light. Recent studies suggest that phosphates may contribute to the mineralization of biomimetic calcium phosphate, enhancing the adhesion, differentiation, and proliferation of bone cells [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. After degradation, BPNs can be converted into calcium phosphate, which promotes bone regeneration in physiological environments. BPNs comprise corrugated planes of phosphorus atoms held together by weak van der Waals forces, allowing easy exfoliation into thin BPNs [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. In addition, BP is highly reactive toward oxygen and water due to the lone electron pairs on each phosphorus atom in the layered structure [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Such a characteristic limits its biomedical applications and further research [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. In previous work, our research group prepared BP/PPF bone cement through thermal crosslinking [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. In vitro studies demonstrated its ability to release phosphate ions slowly and continuously. Based on these findings, we hypothesize that introducing BPNs onto the surface of HA significantly enhances its stability and bioactivity.\u003c/p\u003e \u003cp\u003eThis study constructed PPF/HA/BP bone cement and systematically investigated the impact of different component ratios on the material\u0026rsquo;s mechanical properties. Key properties were evaluated, including compressive strength, elastic modulus, exothermic temperature, and crosslinking time. Additionally, biocompatibility, in vitro degradation, and safety were assessed through molecular, synthesis, and cell experiments to provide a solid foundation for clinical applications (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePreparation of PPF\u003c/h2\u003e \u003cp\u003eIn a light-protected environment, 329.6 mL of diethyl fumarate and 440.65 mL of 1,2-propanediol were transferred into a three-neck round-bottom flask, followed by adding 0.45 g of zinc chloride and 2.75 g of hydroquinone and stirring the mixture. The apparatus was connected to a condenser, and the stirring speed was gradually increased to 300 rpm under nitrogen protection. The temperature of the silicone oil bath was raised from 110\u0026deg;C to 150\u0026deg;C, maintaining it for several hours until approximately 90% of the ethanol was recovered. The vacuum system was connected to a liquid nitrogen condenser, and the pressure was maintained at \u0026lt;\u0026thinsp;1 mmHg. The mixture was gradually heated to 150\u0026deg;C, avoiding bumping. Once the desired molecular weight was reached, heat was stopped, the air inlet valve was opened slowly, and the vacuum pump was stopped. Equal volumes of 1.85% HCl and saturated NaCl solutions were added to purify the PPF. The mixture was dried with anhydrous Na₂SO₄ overnight and filtered to remove Na₂SO₄ to obtain a clear, pale yellow polymer. Finally, rotary evaporation at 45\u0026deg;C was used under reduced pressure to remove dichloromethane.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003ePreparation of BPNs\u003c/h3\u003e\n\u003cp\u003eFifty mg of BPNs was weighed and added to an N-vinyl-2-pyrrolidone(NVP) solution in a mortar. The mixture was transferred into a brown bottle, and NVP was added to increase the volume to 100 mL. Argon was introduced, and the bottle was sealed. The solution was sonicated in an ice\u0026ndash;water bath at 250 W for 12 hours, followed by processing with an ultrasonic cell crusher at 180 W (5 s on/5 s off) for another 12 hours, with ice added to prevent overheating. The mixture was centrifuged at 1300 rpm for 15 min to collect the supernatant, which was then centrifuged at 9660\u0026times;g (4 () for 20 min to obtain BPNs. The BPNs were washed three times with NVP and resuspended in 100 mL of NVP. Argon was bubbled through, and the suspension was stored at 4\u0026deg;C for future use.\u003c/p\u003e\n\u003ch3\u003ePreparation of PPF/HA/BP\u003c/h3\u003e\n\u003cp\u003eThis study builds upon previous experimental findings to propose an orthogonal experimental design featuring four factors, each with three levels[\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. The investigated factors included the heat crosslinking initiator benzoyl peroxide (BPO), BPNs, the crosslinking agent NVP, and HA. These selections aimed to elucidate the effects of various materials and conditions on the experimental outcomes and provide essential data for optimizing the performance of bone cement. The overall experimental design incorporated nine experimental groups, with the sequence and combinations organized according to a standardized orthogonal experimental design table. By assessing the mechanical properties of bone cement formulations, specifically the compressive strength and elastic modulus, this study sought to identify the optimal experimental scheme (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eHorizontal design table of orthogonal factors\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA.BPO(mg)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eB.BP(ug)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC.NVP(ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eD.HA(g)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.375\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e300\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eAfter the samples were centrifuged to collect sufficient BPNs, they were resuspended in NVP, and an appropriate amount of hydroxyapatite was added. A certain proportion of BPO was subsequently added. The resulting mixture was then introduced into the pre-prepared PPF and thoroughly stirred, and an initiator, dimethyl-P-toluidine (DMPT), was added. The mixture was stirred until fully homogeneous. Finally, a syringe was used to inject the mixture into a custom-made polytetrafluoroethylene (PTFE) mold measuring 10 mm in diameter and 15 mm in height.\u003c/p\u003e\n\u003ch3\u003eCharacterization\u003c/h3\u003e\n\u003cp\u003eThe PPF polymer was dissolved in CDCl\u003csub\u003e3\u003c/sub\u003e and analyzed via a 600-MHz NMR spectrometer to obtain the \u003csup\u003e1\u003c/sup\u003eH-NMR spectrum. The relative molecular weight and polydispersity index of PPF were determined using an Agilent 1260 Infinity II GPC, with chloroform as the eluent and polystyrene as the standard. PPF was mixed with KBr, compressed into pellets, and analyzed via a Thermo Scientific Nicolet iS20 FTIR spectrometer from 4000 to 500 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e with a 4-cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e resolution. The BPNs were washed three times with anhydrous ethanol, deposited onto a copper grid, and allowed to dry at room temperature, followed by examining their morphology and structure via transmission electron microscopy (FEI Talos F200X, USA) at an operating voltage of 200 kV. BPNs and dried KBr were ground together to form thin pellets, and the functional groups were analyzed via an FTIR spectrometer (FTIR-650, Tianjin Guangdong). After being centrifugally dispersed in deionized water, the particle size and zeta potential of BPNs were measured in the aqueous medium via a Zetasizer Ultra (USA). The morphology and thickness of BPNs were assessed using atomic force microscopy (AFM) (Bruker Dimension Icon, Germany). The samples were initially cured using a PTFE mold measuring 10 mm in diameter and 2 mm in height. The samples were subsequently pre-frozen at -60\u0026deg;C for 6 hours in a freeze-dryer, followed by vacuum freeze-drying for 24 hours. After drying, the samples were sealed and stored in a refrigerator at 4\u0026deg;C. Before imaging, the samples were gold-coated under vacuum and analyzed under a scanning electron microscope (JSM7610FPlus, Japan).\u003c/p\u003e\n\u003ch3\u003eMechanical properties\u003c/h3\u003e\n\u003cp\u003eThe samples, measuring 10 mm in diameter and 15 mm in height, were fabricated for compression tests under the ISO 604 standard[\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. A universal electromechanical testing machine equipped with a 30-kN load cell (E44.304, MTS Systems (China) LTD, CN) was used to conduct the compression tests. The compression rate was set at the standard value of 2 mm/min. Before the tests, the upper and lower surfaces of the samples were meticulously polished with sandpaper to ensure their parallelism.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eSetting properties\u003c/h2\u003e \u003cp\u003eThe final curing time was recorded using a Vicat apparatus (WKY-1000, Tianjian Instrument Co., Ltd., CN), which was calculated as the duration from the initiation of crosslinking to the formation of a solidified structure.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eHydrophilic experiment of bone cement\u003c/h3\u003e\n\u003cp\u003eThe bone cement\u0026rsquo;s hydrophilic properties were investigated by selecting PPF/HA/BP, PPF/HA, and PPF materials known for their optimal mechanical properties following formulation optimization. The contact angle, representing the angle formed between the liquid and the solid material, was measured to assess the hydrophilic characteristics. Five parallel samples were prepared for each group, with the bone cement molded in a polytetrafluoroethylene mold measuring 10 mm in diameter and 2 mm in height. The materials were placed on a test platform to measure the contact using a JY-82 contact angle meter. The platform was adjusted to a horizontal position, and an automatic titration system was used to dispense water droplets. Images were captured, and the contact angle was determined by fitting the data according to the Young\u0026ndash;Laplace equation.\u003c/p\u003e\n\u003ch3\u003eIn vitro bone cement degradation\u003c/h3\u003e\n\u003cp\u003eUsing a PTFE mold, a cylindrical sample was fabricated for the in vitro degradation study, measuring 6 mm in diameter and 10 mm in height. Each sample\u0026rsquo;s initial weight (M\u003csub\u003e0\u003c/sub\u003e) was recorded after it had been dried in a vacuum drying oven at 37\u0026deg;C. The prepared sample was subsequently fully immersed in a plastic bottle containing a PBS solution (pH\u0026thinsp;=\u0026thinsp;7.3), with the solution being replaced weekly. The bone cement-to-PBS buffer ratio was 0.1 g: 10 mL, and the mixture was then continuously incubated in a constant-temperature water bath shaker at 37\u0026deg;C. The samples were retrieved from the solution every week. The surface water was wiped off with filter paper, and the weight was recorded as M\u003csub\u003e1\u003c/sub\u003e. The samples were subsequently dried in a vacuum drying oven at 37\u0026deg;C until completely dehydrated, and the weight was recorded as M\u003csub\u003e2\u003c/sub\u003e. This process continued for four weeks. The initial weight of the sample (M\u003csub\u003e0\u003c/sub\u003e) was then compared to the sample\u0026rsquo;s weight after degradation (M\u003csub\u003et\u003c/sub\u003e) to determine the bone cement\u0026rsquo;s rate of mass loss.\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eIn vitro photothermal conversion efficiency\u003c/h2\u003e \u003cp\u003eThe pre-prepared PPF, PPF/HA, and PPF/HA/BP discs measuring 6 mm in diameter and 1 cm in height were submerged in 0.5 mL of PBS. The samples were subsequently irradiated with an 808 nm laser with a consistent power density of 1.0 W/cm\u0026sup2;. The irradiation time was 10 min, and the distance between the light source and the sample was 12 cm. The samples\u0026rsquo; heating curves were recorded using a temperature recorder. To evaluate the stability of the photothermal effect, the PPF/HA/BP bone cement was subsequently irradiated five times. The distance between the light source and the sample was 12 cm, and the heating curves of the samples were recorded using a temperature recorder.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eCell culture\u003c/h2\u003e \u003cp\u003eMC3T3-E1 cells were grown and revived in α-MEM supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. To conduct the osteogenic differentiation assay, the α-MEM was replaced with OriCell osteogenic induction medium containing 10% FBS, 1% penicillin/streptomycin, 100-nM dexamethasone, 50-\u0026micro;g/mL ascorbic acid, and 10-mM β-glycerophosphate. Before being cultured with cells, the cement samples were immersed in 75% alcohol and air-dried under ultraviolet light.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eCytotoxicity assay\u003c/h2\u003e \u003cp\u003eAccording to GB/T 16886.5\u0026ndash;2003/ISO 10993-5-1999, various groups of bone cement materials were initially immersed in 75% alcohol for 30 min and then transferred to a complete medium for 72 hours. The extract was subsequently filtered through a 0.22-\u0026micro;m sterile filter. The cells were counted in a cell counting chamber and then seeded at a density of 5,000 cells/well in 96-well plates. The plates were cultured overnight to allow the cells to fully adhere. The old culture medium was discarded, and the corresponding bone cement extracts were added to each group, with a blank control group lacking material extract. The cells were then cultured for 1, 3, or 5 days. Following the specified incubation period, the extract was removed, and 100 \u0026micro;L of complete culture medium containing 10% CCK8 (Cell Counting Kit-8) was added to each well. The plates were incubated in the dark at 37\u0026deg;C under 5% CO₂ for 1 hour. After incubation, each sample\u0026rsquo;s optical density (OD) was measured at a wavelength of 450 nm using a microplate reader.\u003c/p\u003e \u003cp\u003eThe effects of pure PPF, PPF/HA, and PPF/HA/BP extracts on cell morphology were assessed using live/dead (Yeasen 40747ES76, Shanghai, China) and cytoskeletal staining, with the cytoskeleton labeled with phalloidin and the nuclei labeled with DAPI (Servicebio G1012, Servicebio G1041, Wuhan, China). The stained cells were observed under a confocal laser scanning microscope (CLSM; Leica TCS SP2, Heidelberg, Germany) to evaluate cell survival and morphological spread.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eCytocompatibility tests\u003c/h2\u003e \u003cp\u003eThe PPF, PPF/HA, and PPF/HA/BP bone cements were meticulously prepared using polytetrafluoroethylene molds measuring 10 mm in diameter and 2 mm in height, with five samples in each group. The prepared bone cements were carefully placed in a biosafety cabinet, soaked in 75% alcohol for 30 min, and gently air-dried. A suspension of MC3T3-E1 cells (3\u0026times;10\u003csup\u003e4\u003c/sup\u003e cells/mL) was seeded onto the material\u0026rsquo;s surface and cultured in an incubator for 48 hours. Then, the samples were fixed with 4% paraformaldehyde for 30 min, followed by gradient dehydration with alcohol. The samples were then freeze-dried, gold-coated, and analyzed by scanning electron microscopy (SEM).\u003c/p\u003e \u003cp\u003eTo each well of a 6-well plate, 1 mL of 0.1% gelatin was added, and the mixture was incubated for 30 min at 37\u0026deg;C. Resuspended MC3T3-E1 cells were then seeded into 6-well plates at a density of 1\u0026times;10\u003csup\u003e5\u003c/sup\u003e cells/mL, with 2 mL of the cellular suspension added to each well. The plates were incubated at 37\u0026ordm;C under 5% carbon dioxide and maintained under saturated humidity. Upon reaching approximately 70% confluence after incubating for 24‒48 hours, the medium was replaced with an osteogenic medium composed of PPF, PPF/HA, and PPF/HA/BP extracts. The cells were induced to differentiate via the aforementioned extraction medium for 7, 14, and 21 days, with three replicate wells for each time interval. The culture medium was replaced every 48 hours to ensure optimal cell culture conditions. To quantify the material-induced capability at different time points (7, 14, and 21 days), after culturing for the respective durations, the medium was removed, followed by two washes with PBS. Alkaline phosphatase(Beyotime C3206, Shanghai, China) and Alizarin Red dye((OriCell ALIR-10001, Guangzhou, China)) were then added. After incubating for 20 minutes, the samples were washed twice with PBS and observed under a microscope to capture images.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eOsteogenic-related gene expression\u003c/h2\u003e \u003cp\u003eReal-time quantitative polymerase chain reaction (QPCR) was used to quantify the expression of osteogenic genes in MC3T3-E1 cells (n\u0026thinsp;=\u0026thinsp;9). The MC3T3-E1 cells in suspension were adjusted to the required concentration and plated into 6-well plates at a density of 2\u0026times;10\u003csup\u003e4\u003c/sup\u003e cells/mL, with 2 mL/well. These cells were subsequently incubated at 37\u0026deg;C under 5% carbon dioxide and high humidity. When the cell confluence reached approximately 70%, osteogenic differentiation was generally triggered within 24‒48 hours of incubation. Bone formation-associated gene expression was evaluated through co-culture with the extract. The cells were then treated with an osteogenic medium for 14 days, and the medium was changed every 48 hours. Total RNA was extracted from the cells using an RNeasy Mini Kit (Qiagen 74104, Hilden, Germany). QPCR was performed to measure the mRNA expression of marker genes, including alkaline phosphatase (\u003cem\u003eALP\u003c/em\u003e), runt-related transcription factor 2 (\u003cem\u003eRUNX2\u003c/em\u003e), and collagen I (\u003cem\u003eCol-I\u003c/em\u003e). Beta-actin was used as the internal control. \u003cb\u003eTable S1\u003c/b\u003e presents the primer sequences. Reverse transcription was performed on 1 mg of RNA using the reverse transcription kit SuperScriptTM III Reverse Transcriptase. The qRT‒PCR assays were conducted using SYBR Premix Ex Taq II (\u0026times;2) as a PCR reagent on an ABI 7500 rapid machine (Applied Biosystems StepOne, MA, US). The results were calculated using the delta‒delta Ct method (2\u003csup\u003e\u0026minus;△△\u003c/sup\u003eCt).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eQuantitative variables were reported as means\u0026thinsp;\u0026plusmn;\u0026thinsp;SDs. The normality of the data was assessed using the Shapiro‒Wilk test. The Mann‒Whitney U test was used to compare skewed data and determine significant differences. For non-skewed data, one-way analysis of variance (ANOVA) was conducted with post hoc pairwise comparisons. GraphPad Prism 9.0 software (San Diego, CA, US) was used for all analyses and graphic representations. A P-value of \u0026lt;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eCharacterization of PPF and BPNs\u003c/h2\u003e \u003cp\u003eIn the \u003csup\u003e1\u003c/sup\u003eH-NMR spectrum of the PPF polymer, the peak at 7.26 ppm corresponded to the solvent, deuterated chloroform. The signal at 6.86 ppm was attributed to the double bond present in the PPF molecular chain. The methylidene signals in the PPF were observed at 5.28 and 4.24 ppm, whereas the methyl signal was detected at 1.23 ppm (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eAs shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea and b, the BPNs exhibited an irregular morphology with an approximate size of 300 nm in the TEM images. AFM characterization of the BPNs revealed a thickness of approximately 5 nm (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ec). Figure\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ed shows that the average particle size of the BPNs synthesized in this study was 314.7 nm.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003eCharacterization of PPF/HA/BP\u003c/h2\u003e \u003cp\u003eThe scanning electron microscopy (SEM) images (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e) illustrated the smooth surface of the PPF bone cement material. Hydroxyapatite particles and black phosphorus nanosheets were partially deposited on the surface of the PPF, although they did not completely cover the material. Additionally, the results of the hydrophilic angle tests indicated that hydroxyapatite particles and black phosphorus nanosheets significantly influenced the surface hydrophilicity of the bone cement, suggesting that the changes in the hydrophilic angle were statistically significant (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eMechanical properties\u003c/h2\u003e \u003cp\u003ePPF bone cement with various formulations exhibited remarkable mechanical strength, with a compressive strength range of 70\u0026ndash;150 MPa and an elastic modulus range of 600‒900 MPa, suggesting a close resemblance to human bone tissue regarding its mechanical properties (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eResults of bone cement orthogonal test\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTest number\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFactor\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eResult\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCompressive strength (Mpa)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eModulus of elasticity (Mpa)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e110.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e682\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e122.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e733.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e73.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e710.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e134\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e903.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e132\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e829.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e145.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e758.8\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e147.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e844.8\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e152.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e868.9\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e127.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e846.9\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eBased on the results of the range analysis, the influence of factors A, B, C, and D on the compressive strength was as follows: A (BPO)\u0026thinsp;\u0026gt;\u0026thinsp;B (BPNs)\u0026thinsp;\u0026gt;\u0026thinsp;D (HA)\u0026thinsp;\u0026gt;\u0026thinsp;C (NVP). Among them, BPO and BPNs positively impacted the compressive strength of bone cement, indicating that the higher their content, the greater the compressive strength of the bone cement. The optimal level of HA for compressive strength was at the second level. In contrast, NVP had an adverse effect on compressive strength, indicating that the higher its content, the lower the compressive strength of the bone cement. Therefore, based on the range analysis, the optimal formulation for compressive strength is A3B2D2C1.\u003c/p\u003e \u003cp\u003eAdditionally, based on the range analysis, the influence of factors A, B, C, and D on the elastic modulus was as follows: A (BPO)\u0026thinsp;\u0026gt;\u0026thinsp;C (NVP)\u0026thinsp;\u0026gt;\u0026thinsp;D (HA)\u0026thinsp;\u0026gt;\u0026thinsp;B (BPNs). BPO and HA positively affected the elastic modulus, indicating that the higher their content, the greater the elastic modulus of the bone cement. BPNs and NVP had an optimal formulation at the second level. Therefore, based on the range analysis, the optimal formulation for the elastic modulus is A3C2D3B2. Finally, based on the comprehensive balance method, the best bone cement formulation is A3B2C1D2 (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e and Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAnalysis of compressive strength and elastic modulus of bone cement\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003eCompressive strength\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c9\" namest=\"c6\"\u003e \u003cp\u003eModulus of elasticity\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK1\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e102.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e130.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e135.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e123.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e708.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e810.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e769.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e786.03\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e137.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e135.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e128.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e138.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e830.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e810.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e828\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e779\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK3\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e142.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e115.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e117.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e120\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e853.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e772.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e794.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e827.73\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eR\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e40.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e19.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e18.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e18.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e144.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e27.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e58.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e48.73\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBest index\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003eSetting properties\u003c/h2\u003e \u003cp\u003eThe polymerization time of PPF bone cement was 8.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35 min, and the polymerization temperature was 62.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.67\u0026deg;C. Incorporating hydroxyapatite (HA) shortened the polymerization time of PPF/HA bone cement and decreased the polymerization temperature. Significant differences in polymerization time and temperature were observed between the two groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01). When BPNs were added along with hydroxyapatite, the maximum temperature of the PPF/HA/BP bone cement reached 52.5\u0026deg;C, with a significant difference between the two groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01). The polymerization time of PPF/HA/BP was 6.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 min, which was significantly shorter than that of PPF/HA (7.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15), with a significant difference between the two groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003eIn vitro Degradation Behavior and Photothermal Conversion Efficiency of PPF, PPF/HA, and PPF/HA/BP\u003c/h2\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e illustrates the degradation process of various materials in a human-simulated solution in vitro. The pH of these materials consistently remained close to 7.4. The degradation rate of the PPF-based bone cement was relatively rapid during the initial two weeks, stabilizing thereafter. The scaffold composite exhibited excellent adaptability when subjected to a dynamic in vivo environment.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eAfter irradiation at 808 nm with a power density of 1.0 W/cm\u003csup\u003e2\u003c/sup\u003e for 10 min, the temperature of the PPF/HA/BP group was 17\u0026deg;C higher than that of the PPF and PPF/HA groups. Corresponding experiments were conducted to evaluate the repeatability of the photothermal properties of the bone cement in the PPF/HA/BP group. After completing five photothermal cycles, the temperature change curve of the bone cement in the BP group exhibited minimal variations, with the maximum temperature stabilizing at approximately 50\u0026ordm;C, indicating that its photothermal conversion performance remained consistent across multiple cycles. Following irradiation at a wavelength of 808 nm and a power density of 1.0 W/cm\u003csup\u003e2\u003c/sup\u003e for 10 min, the PPF/HA/BP group presented a temperature increase of 17\u0026ordm;C compared with those of the PPF and PPF/HA groups. Relevant experiments were conducted.to evaluate the consistency of the photothermal characteristics of the bone cement within the PPF/HA/BP group. After completing five photothermal cycles, the temperature variation curve for the BP group demonstrated minimal changes, with the peak temperature stabilizing at approximately 50\u0026ordm;C, indicating that its photothermal conversion efficiency remained stable across multiple cycles (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e \u003ch2\u003eCell adhesion and cytotoxicity assay\u003c/h2\u003e \u003cp\u003eMC3T3-E1 cells were co-cultured with the extracts of various bone cements, including pure PPF, PPF/HA, and PPF/HA/BP, for 1, 3, and 5 days to evaluate their in vitro cytocompatibility (Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e9\u003c/span\u003e). The results demonstrated no significant difference in cell viability between the pure PPF bone cement extract-treated and control groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). In contrast, cell viability in the PPF/HA and PPF/HA/BP bone cement extract-treated groups was significantly higher than the control and pure PPF bone cement-treated groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Additionally, statistically significant differences were observed in cell proliferation between the PPF/HA/BP and PPF/HA bone cement extract co-cultured groups, indicating that both PPF/HA/BP and PPF/HA bone cement materials promoted cell proliferation. However, PPF/HA/BP exhibited a more pronounced effect on osteoblast proliferation.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e10\u003c/span\u003e shows the adhesion of MC3T-E1 cells to the surface of the bone cement. The results indicated significantly less cell adhesion on the PPF cement surface than on the PPF/HA and PPF/HA/BP cement surfaces. Additionally, the pseudopodia of the cells actively adhered to the PPF/HA bone cement, particularly on the hydroxyapatite particles, suggesting a strong interaction between the cells and the material. The cells exhibited a clear overlapping growth pattern in cultures with PPF/HA/BP bone cement.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eLive/dead fluorescence staining was used to assess cell survival. The results revealed a significantly higher cell survival rate in the experimental group compared to the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e11\u003c/span\u003e). Notably, the PPF/HA/BP group demonstrated outstanding performance, with the highest number of living cells, highlighting its effectiveness in promoting cell survival. Additionally, as the cell growth period increased, the number of live cells gradually increased, and the area covered by fluorescence staining significantly expanded, further confirming the positive impact of the culture conditions on cell survival.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eUnder the effect of different types of bone cement extracts, MC3T3 cells presented a polygonal or spindle shape, with the cytoskeleton exhibiting a bundle arrangement parallel to the vertical axis of the cell, forming a network structure. This further indicates that PPF-based materials have no apparent cytotoxicity. The PPF/HA/BP group\u0026rsquo;s cytoskeletal structure was clearer, the overall spreading state was good, and the cytoskeletal structure and spreading state were better than those of the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig12\" class=\"InternalRef\"\u003e12\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003eAlkaline phosphatase staining and alizarin red staining\u003c/h2\u003e \u003cp\u003eMC3T3-E1 cells were induced with an osteogenic induction medium and co-cultured with various extracts, with observations at 7 and 14 days. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig13\" class=\"InternalRef\"\u003e13\u003c/span\u003e, no significant abnormalities were noted in the staining of the PPF or positive control groups during the 7-day staining experiment, indicating that the pure PPF material exhibited no significant toxicity and did not interfere with the MC3T3-E1 cells\u0026rsquo; early mineralization. The ALP staining results for the PPF/HA and PPF/HA/BP material groups were more intense compared to the positive control group, with the PPF/HA/BP group displaying the most pronounced staining. As the co-culture period was extended, the differences between the groups became increasingly evident. After 21 days of induction, the number of calcium nodules in the PPF/HA and PPF/HA/BP groups was higher than that in the positive control and PPF groups. Moreover, the calcium nodules in the PPF/HA/BP material group were larger, confirming that both the PPF/HA and PPF/HA/BP materials promoted osteogenic differentiation, with the PPF/HA/BP group showing the most pronounced effect.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec25\" class=\"Section3\"\u003e \u003ch2\u003eOsteogenesis-related gene expression\u003c/h2\u003e \u003cp\u003eThe expression of \u003cem\u003eALP\u003c/em\u003e, \u003cem\u003eCOL I\u003c/em\u003e, and \u003cem\u003eRUNX2\u003c/em\u003e in the PPF/HA/BP and PPF/HA groups was significantly higher than the control group after 14 days of co-culture (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). Moreover, the PPF/HA/BP group showed significantly higher expression levels compared to the PPF/HA group(\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). There was no significant difference in the expression of \u003cem\u003eALP, COL I, or RUNX2\u003c/em\u003e between the PPF and positive control groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05), suggesting that PPF/HA/BP had favorable osteogenic differentiation activity in vitro (Fig.\u0026nbsp;\u003cspan refid=\"Fig14\" class=\"InternalRef\"\u003e14\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe prevalence of bone-related diseases is increasing due to factors such as an aging population and sports injuries. In recent years, the number of patients presenting with bone injuries or defects has notably increased, posing significant clinical challenges for effective bone repair. Relevant studies indicate that the global number of fracture cases increased from 133.4\u0026nbsp;million in 1990 to 178\u0026nbsp;million in 2019, reflecting an increase of over one-third [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. However, when the size of a bone defect exceeds a certain critical threshold, the bone\u0026rsquo;s self-repair ability is significantly impaired, hindering the restoration of its normal function and structure. When a bone defect exceeds a specific size, its self-repair ability is insufficient, necessitating bone healing promotion through surgical intervention. Although autologous bone transplantation is the\u0026ldquo;gold standard\u0026rdquo; [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], it is associated with problems such as donor shortage, postoperative infection, and pain, limiting its application. Although allogeneic bone transplantation can provide an alternative, there is a risk of immune rejection and disease transmission. Therefore, safer and more effective bone repair programs are urgently needed to meet clinical needs and improve patient outcomes [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eNatural proteins and peptides, particularly those derived from collagen, silk fibroin, fibrin, and gelatin, have shown considerable promise in bone tissue engineering. Despite collagen\u0026rsquo;s several advantages, such as low antigenicity, favorable hydrophilicity, and ease of modification, its stability is diminished under standard physiological conditions [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Thus, enhancing the biomechanical properties of polymers and improving their compatibility with bone tissue is a critical avenue for future research. PPF has attracted significant attention because of its outstanding properties and has progressively emerged as a promising material in biomedical research. It offers many potential applications, particularly in replacing bone trabecular tissue. Its performance advantages make it an ideal candidate material [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Kimicata et al. [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e] integrated extracellular matrix components from tissues with PPF-based materials to develop a biodegradable biological scaffold suitable for use as a vascular graft. This approach uses the advantages of both components, providing a platform for cell growth due to favorable material properties. Although PPF has excellent mechanical properties and biocompatibility as a bone cement, it lacks bone-inductive capabilities and cannot directly promote bone regeneration.\u003c/p\u003e \u003cp\u003eThe HA/polymer composite scaffold exhibited exceptional mechanical properties, osteogenic potential, and slow release. As a scaffold for bone regeneration, significant advancements have been made in applying growth factors, cells, and drug delivery systems for bone reconstruction [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Teng et al. [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e] developed PPF/HA cervical cages using ASTM standard molds to evaluate the compression, bending, tensile, and hardness properties of PPF:HA bulk materials. The findings indicated that this polymer composite could be a promising candidate for cervical cages. Additionally, a cylindrical disk of poly-ε-caprolactone was created through melt deposition and modified with nanohydroxyapatite and polypropyl fumarate. Tissue regeneration enhancement was assessed via bone density measurements and micro-CT imaging [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. However, some studies have shown that hydroxyapatite has the lowest solubility among various calcium phosphate salts [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e], limiting its application in bone tissue engineering repair materials. In recent years, black phosphorus nanosheets have demonstrated significant advantages in bone regeneration, and their unique properties have made them promising candidate materials for bone tissue repair. As the most stable allotrope of elemental phosphorus, black phosphorus has a high degree of homology with the inorganic phosphorus found in human bone. Therefore, it provides a robust scientific basis for its application in bone mineralization and healing processes. Furthermore, black phosphorus exhibits excellent biocompatibility and superior degradation performance. In this study, the optimal bone cement, with mechanical properties most similar to human bone, was prepared using orthogonal design optimization. Additionally, hydrophilicity experiments demonstrated that incorporating BPNs and HA significantly enhanced the surface hydrophilicity of the PPF-based materials.\u003c/p\u003e \u003cp\u003eMetal implants are extensively used in orthopedics, providing several advantages. However, they have significant drawbacks [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e], including high costs, manufacturing complexities, and the potential for irregular stress distribution, which can impact long-term stability. Furthermore, metal implants may produce artifacts during X-ray or CT scan examinations [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e], compromising diagnostic accuracy. Additionally, the nonbiodegradable nature of metal materials often necessitates removal through secondary surgery, increasing patient risks, possible allergic reactions, or other complications. The unsaturated double bond feature of PPF enables it to react with various molecules, facilitating the formation of crosslinked products with compounds such as N-vinylpyrrolidone, bioceramics, and natural organic macromolecules. As a biodegradable material, PPF breaks down into fumaric acid and propylene glycol during degradation. These byproducts are metabolized and excreted from the body, ensuring the safety and biocompatibility of the material within the organism [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. The present study demonstrated that the pH of the PPF-based materials remained stable at 7.4 throughout the degradation process, further confirming that the PPF did not disrupt the internal environment. The SEM images indicated proper adhesion of MC3T3 cells to the surface, indicating a healthy growth pattern. This interaction suggests that the PPF/HA/BP composite material created an environment conducive to cell attachment and proliferation.\u003c/p\u003e \u003cp\u003eImplant-associated infections have long been a significant factor affecting the success of implant procedures. These infections result in physical pain for patients and increase medical costs [\u003cspan additionalcitationids=\"CR34\" citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Clinical trials have demonstrated that bacterial proliferation in the surrounding microenvironment can lead to implant failure. The hydroxyapatite composite lacks intrinsic antibacterial properties and fails to exert an antibacterial effect when implanted in the body. In contrast, as a novel antibacterial material, black phosphorus possesses a unique two-dimensional layered structure that facilitates effective interactions with bacterial cell membranes through physical mechanisms [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Naskar et al. [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] demonstrated the remarkable photothermal antibacterial properties of Au-ZnO-BP nanocomposites against multidrug-resistant \u003cem\u003eStaphylococcus aureus\u003c/em\u003e strains. A thin layer of black phosphorus nanosheets, which measured 300 nm and featured a controlled particle size, was successfully prepared through liquid-phase exfoliation in this study. Compared with the PPF and PPF/HA groups, the temperature of the PPF/HA/BP group increased by 17\u0026deg;C after irradiation at a wavelength of 808 nm with a power density of 1.0 W/cm\u0026sup2; for 10 min. To assess the consistency of the photothermal properties of the bone cement in the PPF/HA/BP group, the temperature variation curve following five photothermal cycles demonstrated the slightest change in the BP group, with the peak temperature stabilizing at approximately 50\u0026deg;C. This stability indicates that its photothermal conversion efficiency remained consistent across multiple cycles.\u003c/p\u003e \u003cp\u003eMC3T3-E1 cells are widely used as osteoblast precursors, as they differentiate into mature osteoblasts over time under osteogenic induction. Approximately 14 days after induction, the cells reach a critical stage of osteogenic differentiation, with significant changes in the expression of key osteogenesis-related genes such as \u003cem\u003eALP, RUNX2\u003c/em\u003e, and \u003cem\u003eCOL-1\u003c/em\u003e. PCR analysis of these genes accurately reflects the osteogenic differentiation status. \u003cem\u003eALP\u003c/em\u003e is involved in bone matrix mineralization [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e], \u003cem\u003eRUNX2\u003c/em\u003e acts as a key transcription factor for osteoblast differentiation, and \u003cem\u003eCOL I\u003c/em\u003e, the primary organic component of the bone matrix [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e], is essential for maintaining bone structure and mechanical properties. According to several previous studies [\u003cspan additionalcitationids=\"CR41\" citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e], black phosphorus nanosheets can upregulate the expression of osteogenesis-related genes upon near-infrared light irradiation and show promise in preventing clinical bacterial infections. They also enhance the mechanical strength of dynamic self-healing hydrogels and enable the gel scaffold to retain protein binding [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. Black phosphorus is degraded in vivo and transforms into nontoxic phosphate ions that can effectively bind with calcium ions. This process is crucial in enhancing bone hardness and promoting bone regeneration. This study demonstrated that the degradation of black phosphorus, in conjunction with HA, synergistically upregulated osteogenic genes such as \u003cem\u003eALP\u003c/em\u003e, \u003cem\u003eCOL I\u003c/em\u003e, and \u003cem\u003eRUNX2\u003c/em\u003e, further inducing bone mineralization [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. This study indicated a significant increase in alkaline phosphatase activity in the PPF/HA/BP group. Furthermore, the expression of osteogenic-related genes, including \u003cem\u003eALP, COL I\u003c/em\u003e, and \u003cem\u003eRUNX2\u003c/em\u003e, was notably upregulated compared to other groups.\u003c/p\u003e \u003cp\u003eWhen designing orthopedic implants, factors other than only mechanical properties are essential. Achieving a balance between mechanical strength, biocompatibility, and degradation rate is vital for ensuring optimal performance and long-term success. However, natural polymer-based bone substitutes often pose specific challenges, particularly regarding their poor biomechanical properties. A primary concern is that the inherent characteristics of natural polymers are generally more compatible with soft tissues, which complicates their ability to fulfill the mechanical strength, biodegradation rate, and stability requirements essential for bone tissue [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. The PPF-based bone cement rapidly degraded during the initial two weeks, after which the degradation rate diminished, allowing for an extended period and space for bone healing. As the material progressively degrades, bone tissue begins to infiltrate the area, ensuring the stability of the bone defect site and mitigating the risk of losing support due to the rapid degradation of the material. Consequently, it ensures that the bone defect site is consistently and stably supported throughout the healing process, ultimately facilitating effective integration with the bone tissue.\u003c/p\u003e \u003cp\u003eThis study conducted experiments on the materials\u0026rsquo; mechanical properties, cellular behavior, degradation rate, biocompatibility, and osteogenic activity. Among these, PPF/HA/BP exhibited mild degradation behavior, good biocompatibility, and high osteogenic induction activity, making it particularly suitable for meeting the clinical demands of biodegradable bone implants. However, several aspects of the results warrant further investigation. First, the material characterization tests did not consider dynamic fluid flow, stress effects, or the frictional resistance of the porous scaffolds. Additionally, the impact of the porous structure design with varying porosities on the mechanical properties and biodegradable characteristics should be further clarified. Finally, future research should focus on designing degradable scaffolds integrated with antibiotic-loaded materials via 3D printing technology.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe biodegradable PPF/HA/BP composite demonstrated exceptional mechanical properties, favorable degradation behavior, outstanding photothermal characteristics, excellent biocompatibility, and significant osteoinductive activity. It promoted the proliferation and differentiation of MC3T3-E1 cells in vitro, upregulated the expression of key genes, and facilitated bone regeneration. This composite provided an environment conducive to osteoblasts and effectively stimulated the cellular processes involved in bone formation, making it a highly promising material for bone tissue engineering. An in vivo study will be carried out in later stages to further confirm the osteogenic effects of the material. Moreover, incorporating BPNs significantly enhanced the photothermal response of the material, offering the potential for more precise and localized treatment strategies in regenerative medicine. Additionally, the synergistic interaction between PPF, HA, and BPNs ensured the mechanical stability of the scaffold and accelerated the healing process by supporting the regeneration of bone tissue under physiological conditions. In the future, we aim to fabricate porous PPF/HA/BP scaffolds that incorporate drugs to enable an integrated system for controlled drug release and structural-osteogenic functionality.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eAFM\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eatomic force microscopy\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eALP\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ealkaline phosphatase\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eBP\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eblack phosphorus\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eARS\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ealizarin red staining\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eBPNs\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eblack phosphorus nanosheets\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eBPO\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ebenzoyl peroxide\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eβ-GP\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eβ-glycerophosphate\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCCK8\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ecell counting kit-8\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCol I\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ecollagen I\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eDMPT\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003edimethyl-P-toluidine\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eHA\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ehydroxyapatite\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eNVP\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eN-vinyl-2-pyrrolidone\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eOD\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eoptical density\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003ePhy\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ephytoextracts\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003ePMMA\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003epolymethylmethacrylate\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003ePPF\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003epoly(propylene fumarate)\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003ePTFE\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003epolytetrafuoroethylene\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eRUNX2\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eRunt-related transcription factor 2\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eSEM\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003escanning electron microscope\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eConsent for publication\u003c/h2\u003e \u003cp\u003eWritten informed consent was obtained from the patient for publication of this case report and any accompanying images. A copy of the written consent is available for review by the Editor of this journal.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eCompeting interests\u003c/strong\u003e \u003cp\u003eThe authors declare that they have no conflict of interest.\u003c/p\u003e \u003c/p\u003e\u003cp\u003e \u003ch2\u003eAuthor details\u003c/h2\u003e \u003cp\u003e1 Graduate School of Xinjiang Medical University, Urumqi, Xinjiang, China. 2 Department of Orthopedics, General Hospital of Xinjiang Military Region, Urumqi, Xinjiang, China. 3 Department of Orthopedics, the First Affiliated Hospital of Chengdu Medical College.4 Department of Pharmacy, General Hospital of Xinjiang Military Region, Urumqi, Xinjiang, China.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThe research was funded by the Natural Science Foundation of Xinjiang Uygur Autonomous Region (2023D01C93) and (CYFY-GQ73).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eHXX conducted the study; collected, analyzed, and interpreted the data; and wrote the manuscript. CJ interpreted the data and edited the manuscript. JW reviewed the manuscript. RM planned the project YT edited the manuscript and reviewed the manuscript. XH and CJ have equally to equally to this study.\u003c/p\u003e\u003ch2\u003eAcknowledgements\u003c/h2\u003e \u003cp\u003eNot applicable.\u003c/p\u003e\u003ch2\u003eAvailability of data and materials\u003c/h2\u003e \u003cp\u003eThe datasets analyzed during the current study are available from the corresponding author upon reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eZhou S, Xiao C, Fan L, Yang J, Ge R, Cai M, Yuan K, Li C, Crawford RW, Xiao Y, et al. Injectable ultrasound-powered bone-adhesive nanocomposite hydrogel for electrically accelerated irregular bone defect healing. 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Adv Healthc Mater. 2020;9(10):e2000265.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMiao Y, Chen Y, Luo J, Liu X, Yang Q, Shi X, Wang Y. Black phosphorus nanosheets-enabled DNA hydrogel integrating 3D-printed scaffold for promoting vascularized bone regeneration. Bioact Mater. 2023;21:97\u0026ndash;109.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eXu H, Liu X, George MN, Miller AL 2nd, Park S, Xu H, Terzic A, Lu L. Black phosphorus incorporation modulates nanocomposite hydrogel properties and subsequent MC3T3 cell attachment, proliferation, and differentiation. J Biomed Mater Res A. 2021;109(9):1633\u0026ndash;45.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi L, Yu M, Li Y, Li Q, Yang H, Zheng M, Han Y, Lu D, Lu S, Gui L. Synergistic anti-inflammatory and osteogenic n-HA/resveratrol/chitosan composite microspheres for osteoporotic bone regeneration. Bioact Mater. 2021;6(5):1255\u0026ndash;66.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFan J, Abedi-Dorcheh K, Sadat Vaziri A, Kazemi-Aghdam F, Rafieyan S, Sohrabinejad M, Ghorbani M, Rastegar Adib F, Ghasemi Z, Klavins K et al. A Review of Recent Advances in Natural Polymer-Based Scaffolds for Musculoskeletal Tissue Engineering. Polym (Basel) 2022, 14(10).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharma S, Sudhakara P, Singh J, Ilyas RA, Asyraf MRM, Razman MR. Critical Review of Biodegradable and Bioactive Polymer Composites for Bone Tissue Engineering and Drug Delivery Applications. \u003cem\u003ePolymers (Basel)\u003c/em\u003e 2021, 13(16).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"journal-of-orthopaedic-surgery-and-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"josr","sideBox":"Learn more about [Journal of Orthopaedic Surgery and Research](http://josr-online.biomedcentral.com)","snPcode":"13018","submissionUrl":"https://submission.nature.com/new-submission/13018/3","title":"Journal of Orthopaedic Surgery and Research","twitterHandle":"@MSKmedBMC","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Poly(propylene fumarate), Black phosphorus nanomaterials, Hydroxyapatite nanocomposite, Bone regeneration","lastPublishedDoi":"10.21203/rs.3.rs-6050660/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6050660/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eBone defects due to trauma, infections, congenital malformations, and tumor resection remain significant health challenges. The polymethyl methacrylate (PMMA) bone cement\u0026rsquo;s limitations in orthopedic applications arise from its lack of bioactivity and the toxicity of its monomers. Hydroxyapatite (HA) cement is widely used for bone reconstruction despite its inherent brittleness. Biodegradable poly(propylene fumarate) (PPF) is recognized for its exceptional performance in addressing these bone defects and providing adequate mechanical support. Black phosphorus (BP) nanosheets (BPNs) have attracted attention due to their unique puckered honeycomb lattice, broad absorption spectrum, high specific surface area, excellent antibacterial properties, and tunable bandgap. In this study, we developed a novel bone cement, PPF/HA/BP, via thermal crosslinking and conducted in vitro evaluation.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003ePPF was synthesized using a two-step approach, whereas BPNs were produced through a liquid-phase exfoliation technique. Then, PPF/HA/BP composite materials were created using a thermal crosslinking process, followed by a thorough examination of their mechanical characteristics, compatibility with cells, osteogenic activity, and degradation properties.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003ePPF/HA/BP bone cement was designed by optimizing formulation to possess mechanical properties comparable to bone tissue. PPF bone cement had a polymerization time of 8.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35 min and a temperature of 62.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.67\u0026deg;C. HA reduced polymerization time and temperature. PPF/HA/BP exhibited a polymerization time of 6.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 min and a maximum temperature of 52.5\u0026deg;C (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). PPF/HA/BP enhanced the adhesion, proliferation, and mineralization of preosteoblasts on its surface and demonstrated photothermal properties. When exposed to an 808 nm laser, the bone cement\u0026rsquo;s temperature rose to 50\u0026ordm;C. After co-culturing with MC3T3-E1 for 14 days, the PPF/HA/BP group exhibited significantly higher expression of \u003cem\u003eALP\u003c/em\u003e, \u003cem\u003eCOL I\u003c/em\u003e, and \u003cem\u003eRUNX2\u003c/em\u003e compared to the PPF and PPF/HA groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). However, no statistically significant difference was observed in the expression of \u003cem\u003eALP, COL I\u003c/em\u003e, and \u003cem\u003eRUNX2\u003c/em\u003e between the PPF and positive control groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eBiodegradable PPF/HA/BP demonstrated satisfactory mechanical properties, degradation behavior, outstanding photothermal characteristics, excellent biocompatibility, and osteogenic activity. It also promoted bone regeneration by enhancing the proliferation and differentiation of MC3T3-E1 cells in vitro and upregulating the related genes\u0026rsquo; expression.\u003c/p\u003e","manuscriptTitle":"Poly(propylene fumarate)/hydroxyapatite nanocomposite/black phosphorus nanosheet phosphate composites for enhanced bone repair","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-29 06:17:41","doi":"10.21203/rs.3.rs-6050660/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-05-09T04:59:28+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-05-04T03:03:59+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-05-01T21:28:56+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-04-28T15:15:15+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"113440741723822521678748907386728772689","date":"2025-04-28T14:37:37+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-04-28T02:16:57+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"50306322860590632984872127763301612158","date":"2025-04-27T00:38:57+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"92422520495067934514646557573432941914","date":"2025-04-24T21:25:12+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"136705352623468878087856470759285974587","date":"2025-04-24T19:02:07+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-04-24T11:12:31+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-04-21T07:13:50+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Orthopaedic Surgery and Research","date":"2025-04-18T03:53:00+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-orthopaedic-surgery-and-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"josr","sideBox":"Learn more about [Journal of Orthopaedic Surgery and Research](http://josr-online.biomedcentral.com)","snPcode":"13018","submissionUrl":"https://submission.nature.com/new-submission/13018/3","title":"Journal of Orthopaedic Surgery and Research","twitterHandle":"@MSKmedBMC","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"9b202532-1660-4341-8c02-3b19f5bcdb06","owner":[],"postedDate":"April 29th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-08-04T16:42:31+00:00","versionOfRecord":{"articleIdentity":"rs-6050660","link":"https://doi.org/10.1186/s13018-025-06028-z","journal":{"identity":"journal-of-orthopaedic-surgery-and-research","isVorOnly":false,"title":"Journal of Orthopaedic Surgery and Research"},"publishedOn":"2025-07-30 16:05:31","publishedOnDateReadable":"July 30th, 2025"},"versionCreatedAt":"2025-04-29 06:17:41","video":"","vorDoi":"10.1186/s13018-025-06028-z","vorDoiUrl":"https://doi.org/10.1186/s13018-025-06028-z","workflowStages":[]},"version":"v1","identity":"rs-6050660","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6050660","identity":"rs-6050660","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}