Investigation of bone defect repair with nanocomposite containing octacalcium phosphate (OCP) and titanium oxide on rat femur bone

Investigation of bone defect repair with nanocomposite containing octacalcium phosphate (OCP) and titanium oxide on rat femur bone | 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 Investigation of bone defect repair with nanocomposite containing octacalcium phosphate (OCP) and titanium oxide on rat femur bone Nima Ilbeygi, Alireza Jahandideh, Abolfazl Akbarzadeh, Mohammad Nasrollahzadeh Masouleh, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7215028/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Bone defect repair remains a major clinical challenge, requiring the development of effective biomaterials to address the limitations of autografts and allografts. This study examined the regenerative potential of a nanocomposite composed of octacalcium phosphate (OCP) and titanium dioxide (TiO₂) in a rat femoral defect model. The OCP-TiO₂ nanocomposite was produced through electrospinning and analyzed using Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM), confirming its structural integrity and osteoconductive properties. Rats were divided into three groups: control (untreated defect), autograft, and OCP-TiO₂ nanocomposite. Histopathological, immunohistochemical (VEGF, Osteocalcin, Ki67), and radiological assessments were performed at 15-, 30-, and 60-days post-implantation. Findings showed that the OCP-TiO₂ group experienced enhanced bone regeneration, with earlier trabecular bone formation, less inflammation, and improved osteointegration compared to autografts and controls. Radiological evaluations indicated faster defect remodeling in the OCP-TiO₂ group, while autografts displayed ongoing inflammation. These results indicate that the OCP-TiO₂ nanocomposite is a promising alternative to autografts, offering improved osteoconductivity, mechanical stability, and biocompatibility for bone defect repair. Future research should focus on long-term outcomes and clinical applications. Bone regeneration Octacalcium phosphate (OCP) Titanium oxide (TiO₂) Nanocomposite Osteoconductivity Femoral defect Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Bone defect repair is the process of bone tissue regeneration, which is a multidimensional and multifactorial process. A series of new cellular and molecular events occurs to re-form bone tissue. Subsequently, we need systemic and local factors for regular bone formation, and any disruption in this process disrupts repair [ 1 , 2 ] Many studies have been conducted on types of autologous and non-autologous substitutes in various animal models to determine which compound can be used as a bone tissue substitute and is preferable. In this regard, two issues can be examined: the ability to induce bone formation and the ability to disperse and scaffold the material placed in the bone defect for repair, both of which are equally important [ 2 , 3 ]. The remarkable progress achieved in tissue engineering over the past decades has motivated researchers to develop biomaterials capable of mimicking the structure and function of the collagen-rich extracellular matrix, supporting cell adhesion, proliferation, and differentiation. Annually, bone defects affect over 20 million patients worldwide, often caused by trauma, tumors, infections, or congenital disorders, which severely impair patients’ health and quality of life [ 4 ]. Addressing these defects is a crucial medical goal. Traditional methods like autografts and allografts are still not perfect: autografts, although providing a good environment for bone healing, have issues such as donor site complications, limited supply, and postoperative pain [ 5 ], while allografts pose risks of immune rejection and disease transmission [ 6 , 7 ]. To overcome these challenges, research has increasingly focused on biomimetic biomaterials as innovative options for bone regeneration, seeking to surpass the limitations of conventional treatments [ 7 – 9 ]. The advanced biomaterials are fabricated to promote the proliferation of mineralized collagen extracellular matrix, aiming to enhance the biological functionality and compatibility of human tissues. Because of its acceptable biocompatibility and mechanical stability, mineralized collagen can stimulate osteogenesis, making it a key material in bone tissue engineering [ 10 – 12 ]. Bone fracture regeneration is a multiple-stage process, ranging from initial inflammation to later remodeling. Biomaterials with a calcium phosphate structure regulate these stages because of their compositional similarity to natural bone [ 12 , 13 ]. Octacalcium phosphate (OCP) is a promising candidate to assist to biological apatite crystals within bone matrices as a precursor [ 10 , 13 ]. OCP, with its chemical formula Ca8H2(PO4)6·5H2O and a Ca/P molar ratio of 1.33, significantly enhances osteoconductivity and exhibits notable osteoinductive potential, making it highly effective for supportive bone replacement [ 13 ]. However, biological grafts are the gold standard among traditional methods; they have various limitations. Researchers have developed synthetic alternatives such as calcium phosphate ceramics, calcium sulfate, and bioactive glasses. Nevertheless, these alternatives face significant challenges: most only exhibit two of the four essential characteristics for effective bone repair—osteointegration, osteoconduction, osteoinduction, and osteogenesis [ 14 ]. For instance, calcium phosphate ceramics, while effective at guiding bone formation, lack other essential functionalities when used alone [ 15 ]. Their brittleness and poor mechanical strength further limit their use in load-bearing areas [ 16 ]. Additionally, inconsistent degradation rates of these ceramics can hinder optimal bone regeneration and compromise structural stability during healing. Calcium sulfate, a commonly used synthetic material, breaks down quickly and does not have enough mechanical strength for bone repair uses [ 17 ] .Some synthetic alternatives have also caused serious complications. For instance, geneX®, a calcium sulfate-based implant, was associated with inflammation, delayed wound healing, and cyst formation in clinical cases, leading to early termination of a trial [ 17 ]. Bioactive glasses, although able to form strong bonds with bone through hydroxyapatite crystallization, lack sufficient mechanical strength for a standalone application [ 17 ] .Likewise, synthetic hydroxyapatite misses trace elements (such., Na+, Mg2+, K+, Sr+) found in natural bone minerals, which reduces its biocompatibility and slows its resorption in the body [ 18 ]. Equipment such as bone grafts and synthetic materials that are typically used to repair bone defects can be used in conjunction with standard surgical methods. Numerous materials have been developed to regenerate bone tissue to provide an alternative to autologous bone grafting, which has certain disadvantages such as limited tissue availability and morbidity associated with harvesting the bone from a second operative site [ 19 ] .Calcium phosphates, such ashydroxyapatite (HA: Ca10(PO4)6(OH)2) and β-tricalcium phosphate (β‐TCP: Ca3(PO4)2), are standard materials that are typically used for bone regeneration. Their synthetic forms are widely employed as bone substitutes in clinical settings [ 20 , 21 ]. Materials Octacalcium phosphate was purchased from Merck (Germany). All other reagents and solvents were of analytical grade and used without further purification. TiO2 (rutile) was obtained from Strem Chemicals Inc. (USA). Fabrication and characterization To prepare a 15% weight/volume solution for the experimental groups, an Octacalcium phosphate (OCP) and polyurethane mixture is dissolved in acetic acid and dichloromethane (DCM), respectively. The compounds are used at a molar ratio of 4:1. In the experimental group, the synthesis and manufacturing of Octacalcium phosphate (OCP), polyurethane, and titanium dioxide involved the use of 0.05 grams of titanium dioxide. A solution with a concentration of 15% by weight or volume was supplemented with Octacalcium phosphate (OCP). After preparing the solutions, they were stirred with a magnetic stirrer at 25 degrees Celsius for 24 hours. The experimental groups used the electrospinning process to produce nanodimension scaffolds and nanofibers. The electrospinning machine was set to a speed of 130 revolutions per minute (rpm), a voltage of 20–25 kilovolts (kV), a temperature of 25 degrees Celsius, a distance of 15 centimeters, and an injection rate of 2 milliliters per hour. FTIR Characterization The structural composition of the experimental group (octacalcium phosphate/polyurethane/Titanium Oxide) was assessed using Fourier Transform Infrared Spectroscopy (FTIR; Bruker IFS 48 instrument). All spectra were recorded in the range of 650–4000 cm − 1 at a scan speed of 16 scans per minute with a resolution of 1 cm − 1 in a KBr-diluted medium (Fig. 1 ). Scanning Electron Microscope (SEM) Characterization To analyze the microstructure of the nanofibers, samples were lyophilized in a freeze-dryer (Alpha 1–4 LD plus; Martin Christ) for 48 hours at − 50°C. The dried nanofibers were then coated with gold in a vacuum and examined using a scanning electron microscope (SEM; TESCAN Vega II) (Fig. 2 ). Experimental groups The study groups consist of the first group with 12 participants (control group), the second group treated with an octacalcium phosphate and titanium oxide nanocomposite, polyurethane, and the third group treated with an autograft. Bone damage model and nanocomposite implantation To implant the nanocomposite, animals are initially anesthetized with a combination of ketamine (10% 50 mg/kg) and xylazine (2% 5 mg/kg). After preparing the area aseptically, an incision is made in the animal's thigh. The muscles in the area are removed, and a 3.5 mm defect is created in the femoral shaft using an orthopedic low-speed drill and a special drill. In the first group, the defect is left without implanting the nanocomposite, and the area is closed. In the second group, the defect is closed by implanting octacalcium phosphate and titanium oxide nanocomposite, while in the third group, the defect is filled and closed using bone fragments from the drilled area. After implanting the nanocomposite, the muscles in the area are sutured with 0–4 Vicryl thread, and finally, the skin is closed with 0–4 nylon thread. To prevent possible infections, all test animals will be injected intramuscularly with cefazolin at a dosage of 20 mg per kilogram of body weight and provided with painkillers for 3 days. Once a day, the area will be checked for possible swelling or inflammation, suture openings, discharge, or signs of infection. The skin sutures will be removed 10 days after the operation. First, radiographs will be taken of the desired area at intervals of 15, 30, 45, and 60 days for radiological evaluations. Histology assessment To perform histopathological and immunohistochemical tests, bone samples will be taken from every 3 mice in each group at 15, 30, 45, and 60 days. For this purpose, the animals will be euthanized by injecting lethal doses of intravenous anesthetic Sodium Thiopental (intracardiac injection), and then the femur will be removed for histopathological examination. The samples will be transported to the laboratory in a 10% formalin stabilizing solution, and the formalin solution will be replaced 24 hours later. Histopathological evaluation of the healing process will be performed with trichrome staining. Table 1 lists the scoring degree to which healing is assessed [ 22 , 23 ]. Table 1 Grading of the histopathological lesions of the defects Phenomena Description Phenomena Score Union Failure to fill the defect 0 Only fibrotic tissue 1 Equal amounts of fibrotic and cartilage tissue 2 High amounts of cartilage and little fibrosis 3 Only cartilage 4 Lots of cartilage with little immature bone 5 Immature cartilage and bone in equal proportion 6 Prominent amount of immature bone and little cartilage 7 Healing with immature bone 8 Healing with mature bone 9 Inflammation Absence of inflammatory cells 0 Presence of inflammatory cells less than 25% 1 Presence of inflammatory cells 25–50% 2 Presence of inflammatory cells 50–75% 3 Presence of inflammatory cells more than 75% 4 Remodeling No Remodeling 0 Remodeling less than 25% 1 Remodeling 25%-50% 2 Remodeling 50%-75% 3 More than 75% Remodeling 4 Absorption of scaffolds No Absorption 0 Absorption 25–50% 1 Absorption 50%-75% 2 Complete Absorption 3 20 Radiological assessment Radiological methods have been employed with remarkable success in the assessment of bone repair procedures. In experimental settings, the presurgical assessment allows the evaluation of exact anatomy and bone dimensions, aiding in the selection of fixation devices and implants. Radiological assessment as a basic postoperative method enables to evaluation of the quality of replacement and detection of the fracture. Sample radiographic images of the rabbit calvarial bone, with or without a grafted material, are shown in Fig. 3 . In this study, we applied a computed tomography scan (CT scan) and a computer to create cross-sectional images, also known as slices, of the bones, blood vessels, and soft tissues inside the body. The skulls of the rabbits were CT scanned on the 30th and 60th days after surgery to evaluate the amount of bone repair and hard tissue formation in the area. A CT scan of the calvaria region was performed after euthanasia and skull removal in the animals. The grading of the CT scan was done in the following Table 2 : Table 2 Grading of the sonographic evaluations of the defects Standard unit variable description changeable scale % The volume of newly formed bone/the total volume of the desired area Bone volume fraction 1 % The volume of the remaining grafted material / the total volume of the desired area volume fraction of the remaining material 2 1/mm Measurement of the average number of trabeculae per unit length Bone trabeculae count 3 mm Average thickness of trabeculae (assessed using direct 3D methods) The thickness of bone trabeculae 4 mm Average distance between trabeculae (assessed using direct 3D methods) The distance between bony trabeculae 5 Statistical analysis Results were expressed as the mean ± standard deviation. We used SPSS (version 16.1, IBM, USA). One-way ANOVA analysis of variance was used to compare the means from multiple experimental groups, followed by a post hoc Tukey test. P < 0.05 was considered statistically significant. Results Nanofibers Characterizations: ATR-IR analysis The ATR-IR technique was used to investigate the presence of chemical functional groups within the structure of the polyurethane/octacalcium phosphate composite with titanium oxide; the results are reported in the form of a graph based on the frequency of the elements present in the chemical structure. The vibrations in the graph corresponding to frequencies between 3961-3604 and 1523-cm-1 indicate the presence of N-H and C-N functional groups in the polyurethane polymer, respectively. Also, the frequency between 1100-1072-cm-1 indicates the presence of C-O-C functional groups in the polyurethane, and the wavelengths between 1001-920, 1260-1072, and 500-700 cm-1 indicate the functional groups of octacalcium phosphate Ca, HP, PO4-. The vibrations observed at around 2965 cm-1 in the diagrams refer to the aliphatic C–H functional group in the nanocomposite. On the other hand, the vibrations at frequencies of 1770–1726, 1449–1072, and 800–690 cm-1 indicate the Ti-o-o functional groups of titanium oxide that have reacted with the chemical structure of the composite and are present (Diagram 1). SEM Analysis The results of examining the morphology of the prepared nanofibers for the polyurethane/octacalcium phosphate composite with titanium using the SEM scanning electron microscope method showed that the prepared nanofibers can maintain a three-dimensional structure with interconnected porosity. This porosity and interconnection of the nanofibers can be a reason for the presence of titanium dioxide, which creates a more suitable morphology and topography in the scaffold and can indicate the presence of suitable sites for surface adhesion in the prepared nanofiber strands( Figure1). Histopathological Results : The results of the test after 15,30, and 60 days after treatment indicated that Dense fibrous connective tissue with fibroblasts and fibrocytes, relatively low infiltration of mononuclear inflammatory cells, covers the outer surface of the defect. In deeper areas, immature and scattered trabeculae of newly formed bone are observed. Extensive fibrous connective tissue with fibroblasts and fibrocytes, and moderate to severe infiltration of mononuclear inflammatory cells surrounding the autograft particles placed in the defect. Limited bone formation is seen in the center and around the autograft, as well as at the margins of the defect and near the old bone. Limited fibrous connective tissue with severe infiltration of neutrophilic inflammatory cells surrounding the scaffold particles placed in the defect. Bone formation appears as scattered islands among the formed connective tissue Moreover, the results 30 days after implantation in all experimental groups indicated that in the control group, Dense fibrous connective tissue with fibroblasts, collagen Fibers, with very mild to negligible infiltration of inflammatory cells. Immature and scattered trabeculae of newly formed bone are observed. Fibrous connective tissue, containing fibroblasts, fibrocytes, and collagen fibers, with moderate to severe inflammatory cell infiltration, surrounds the autograft pieces placed in the defect. Newly formed bone trabeculae adjacent to the connective tissue around the autograft are evident( Figure2). The TiO₂-OCP group showed significantly more bone formation than the others. Severe inflammation was localized around scaffold remnants. However, in the fibrous connective tissue, the inflammation was negligible to mild. Among the treatment groups, the TiO₂-OCP group showed the highest bone formation with the least inflammation . At 60 days, the control group showed integrated trabeculae of newly formed bone adjacent to the old cortical bone . Despite bone formation in the connective tissue filling the defect, severe inflammation was observed. The TiO₂-OCP group showed the most prominent bone formation. Newly formed trabeculae had fused, forming relatively mature cortical bone plates. Comparison of Groups at Day 15 : The control group shows moderate bone formation with the least inflammation. The TiO₂-OCP group shows the highest bone formation with the lowest inflammation(Table 3) . The process of scoring Histopathological Results will be as follows : 1. Bone Formation: o 0: No formation o 1: Minimal formation o 2: Moderate formation o 3: Marked formation (bridging of the defect) 2. Integration with Host Bone: o 0: No integration o 1: Partial integration o 2: Almost complete integration o 3: Complete integration 3. Inflammation: o 0: No inflammation o 1: Mild inflammation o 2: Moderate inflammation o 3: Severe inflammation Table 3. Histopathological Result. The results of the test after 15 days after treatment. Bone Formation Integration with Host Bone Inflammation Group 1 0 1 Control 2 1 2 Control 1 0 2 Control 1 1 1 Control 1 0 3 Autograft 1 0 3 Autograft 1 1 3 Autograft 1 1 2 Autograft 2 1 1 OCP-TIO2 2 1 1 OCP-TIO2 2 1 1 OCP-TIO2 2 1 1 OCP-TIO2 Comparison of Groups at Day 30 : Among the treatment groups, the TiO₂-OCP group showed the highest bone formation with the least inflammation (Table 4) . Table 4. Histopathological Result. The results of the test after 30 days after treatment. Bone Formation Integration with Host Bone Inflammation Group 1 0 2 Control 2 1 1 Control 1 0 1 Control 2 1 2 Control 1 1 2 Autograft 2 0 3 Autograft 1 1 3 Autograft 2 1 2 Autograft 2 1 1 OCP-TIO2 3 2 1 OCP-TIO2 3 2 1 OCP-TIO2 2 2 1 OCP-TIO2 Table 5. Histopathological Result. The results of the test after 60 days after treatment. Bone Formation Integration with Host Bone Inflammation Group 3 2 1 Control 2 2 0 Control 2 1 1 Control 2 1 1 Control 2 1 2 Autograft 2 1 3 Autograft 1 1 3 Autograft 2 1 2 Autograft 3 3 1 OCP-TIO2 3 2 1 OCP-TIO2 3 2 0 OCP-TIO2 3 3 1 OCP-TIO2 Immunohistochemistry Results Immunohistochemical analysis was conducted for VEGF, Osteocalcin, and Ki67 on Day 15, and Osteocalcin as an osteogenesis marker at 15-, 30-, and 60-days post-treatment in each experimental group using the Allred scoring method (Figure 3). The staining outcomes were examined and compared on days 15, 30, and 60 after transplantation, focusing on new bone formation, integration with host tissue, and inflammation. These results are detailed in separate scoring tables. The findings showed that at 60 days after defect creation, mature bone trabeculae, well-integrated with the host bone, had formed. Meanwhile, moderate bone formation with minimal inflammation occurred at 15- and 30-days post-defect creation, respectively. The results indicate that the TiO₂-OCP experimental groups exhibited the highest bone formation, low inflammation, and scattered bone islands compared to the autograft and control groups. Moreover, in the autograft group, persistent inflammation and moderate bone formation near graft fragments were observed, with severe inflammation at 30- and 60-days post-treatment. In contrast, the TiO₂-OCP group showed the best results: mature cortical bone with Haversian canals, complete integration, and minimal inflammation at 60 days post-implantation, suggesting that the TiO₂-OCP composite is a superior alternative to autografts. It enhances bone repair while reducing inflammatory complications. Autografts, on the other hand, caused persistent inflammation, delaying healing despite some bone formation. The control group displayed slower but steady healing with less inflammation than autografts. Microscopy Method for Allred scoring for VEGF and Osteocalcin and Ki67 Proportion score 0—No cells are immunoreactive,1—≤ 1% of cells are immunoreactive 2—1–10% of cells are immunoreactive 3—11–33% of cells are immunoreactive 4—34–66% of cells are immunoreactive 5—67–100% of cells are immunoreactive Intensity score 0—Negative. 1—Weak. 2—Intermediate. 3—Strong. Allred score (P+I) 0–1—Negative. 2–3—Weak 4–6—Moderate 7–8—Strong Table 6. Day 15 Results VEGF Ki67 Osteocalcin Group 2 2 0 Control 1 2 0 Control 1 2 0 Control 1 1 0 Control 3 3 3 Autograft 1 2 2 Autograft 1 2 2 Autograft 2 3 2 Autograft 3 3 4 OCP-TiO2 2 2 3 OCP-TiO2 2 3 3 OCP-TiO2 3 2 3 OCP-TiO2 Radiological Results Radiological Evaluation results indicated that, TiO₂-OCP scaffold showed the fastest and most complete bone regeneration at all-time points. And autografts performed better than controls but lagged behind TiO₂-OCP in remodeling, control was slowest, with persistent defects even at Day 60.so, TiO₂-OCP is superior to autografts in accelerating bone repair and achieving structural restoration with earlier remodeling(Figur5-7). Autografts, while effective, delay remodeling and may cause complications (e.g., persistent radiolucency). Control healing is insufficient for critical-sized defects, emphasizing the need for bone graft substitutes.The process of scoring radiographs will be as follows : Table 6. Day 15 Radiological Scores Group Bone formation Proximal/Distal union Remodeling Total score Control 1 1/1 0 3 Control 0 0/0 0 0 Control 1 0/1 0 2 Control 1 0/1 0 2 Autograft 2 1/2 0 5 Autograft 1 1/1 0 3 Autograft 2 0/1 0 3 Autograft 1 1/1 0 3 OCP-TiO2 3 1/2 0 6 OCP-TiO2 3 2/2 0 6 OCP-TiO2 2 1/2 0 5 OCP-TiO2 3 1/1 0 5 Table 7. Day 30 Radiological Scores Total score Remodeling Proximal/Distal union Bone formation Group 3 0 1/1 1 Control 3 0 1/1 1 Control 4 0 1/1 2 Control 3 0 1/1 1 Control 5 0 1/2 2 Control 4 0 1/1 2 Autograft 4 0 1/1 2 Autograft 4 0 1/1 2 Autograft 9 1 2/2 4 OCP-TiO2 8 1 2/2 3 OCP-TiO2 8 1 1/2 4 OCP-TiO2 9 1 2/2 4 OCP-TiO2 Discussion Since ancient times, finding effective treatments to regenerate damaged or dysfunctional bone tissues has been the most challenging and persistent issue in medicine. The conventional approach, like tissue or organ transplantation, comes with significant drawbacks, including donor shortages, high costs, immune rejection, and complications from foreign tissue implantation. These limitations have motivated the exploration of novel strategies that replace current treatments and are designed to closely mimic natural tissue [ 24 , 25 ]. Bone tissue engineering, through the fabrication of bone substitutes, currently provides innovative solutions for managing bone injuries. A promising opportunity is using bone substitutes that can boost bone formation. Advances in nanotechnology have enabled the development of polymer-based scaffolds that incorporate nanoparticles and nanofibers [ 26 ]. These nano-engineered fibers offer a larger surface area than traditional fibers, allowing for better material integration and enhanced mechanical strength. According to the latest studies, the composite of semiconductive biomaterials combined with nanoscale metal-based materials like octacalcium phosphate (OCP) and titanium oxide (TiO₂) could revolutionize the treatment of bone injuries in humans. Therefore, TiO₂’s effectiveness is improved when incorporated into OCP, supporting its inclusion in bone regeneration research [ 12 ]. The growing body of research on octacalcium phosphate (OCP) nanocomposites shows their remarkable potential in bone regeneration. Building on early findings of OCP's osteoconductive properties [ 27 ], subsequent studies have systematically explored and expanded its applications in tissue engineering. Notably, Heydari et al. advanced the field by successfully incorporating OCP into polycaprolactone scaffolds, demonstrating enhanced osteoblast proliferation—a critical step toward clinical applications [ 18 ]. The regenerative mechanisms of OCP seem complex. Kurobane et al. offered important insights into its ability to promote blood vessel growth, showing that OCP/gelatin composites not only support bone formation but also enhance vascularization, which is a major challenge in repairing large bone defects. This dual role was further backed by Kawai et al., whose clinical studies in oral surgery confirmed that OCP-based materials are both safe and effective, while also showing their potential as alternatives to autologous grafts [ 21 , 28 , 29 ]. Recent work by Hamaei et al. has advanced our understanding by establishing a dose-response relationship between OCP crystal density and osteogenic outcomes. This finding indicates that optimizing the material could lead to even greater clinical effectiveness. Combined with the antimicrobial properties of TiO₂ nanocomposites, these developments provide a comprehensive solution that addresses multiple aspects of bone regeneration: structural support, biological activity, and infection prevention. Collectively, these studies represent significant progress toward overcoming the limitations of traditional transplantation methods. The consistent findings across in vitro, animal, and clinical studies strongly support the translational potential of OCP-based nanocomposites. Future research should focus on scaling up production, optimizing composite formulations for specific clinical applications, and conducting larger-scale clinical trials to fully realize this technology's potential in regenerative medicine. Declarations Acknowledgments The authors would like to thank the Department of Clinical Science, Science and Research Branch, Islamic Azad University, and Department of Medical Nanotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences for financial support of this project (Grant NO: 62379). Funding This study was funded by the Department of Medical Nanotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences (Grant NO: 62379). Conflict of interest The authors declare that they have no conflict of interest. Ethical Approval The ethical approval for this paper was obtained from the research ethics committee of Tabriz University of Medical Sciences (IR.IAU.SRB.REC.1402.346 ( . Author contributions Nima Ilbeygi: Conceptualization, Methodology, Investigation, Formal analysis, Writing - Review & Editing Alireza Jahandideh, Abolfazl Akbarzadeh: Writing-original draft, Writing - Review & Editing Saeed Hesaraki: Methodology, Investigation Mohammad Nasrollahzadeh Masouleh, and Abolfazl Akbarzadeh: Project administration, Conceptualization, Supervision, Resources, Funding acquisition Data Availability Statement (DAS) Not applicable. References Hench, L. L., et al. (1971). Bonding mechanisms at the interface of ceramic prosthetic materials. 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(2015). Effect of octacalcium-phosphate-modified micro/nanostructured titania surfaces on osteoblast response. ACS applied materials & interfaces , 7 (26), 14384–14396. Kawai, T., et al. (2020). Clinical study of octacalcium phosphate and collagen composite in oral and maxillofacial surgery. Journal of tissue engineering , 11 , 2041731419896449. Hamai, R., et al. (2022). Octacalcium phosphate crystals including a higher density dislocation improve its materials osteogenecity. Applied Materials Today , 26 , 101279. Charts Chart 1 is available in the Supplementary Files section Additional Declarations No competing interests reported. Supplementary Files image1.png Chart 1: ATR-IR test analysis results Cite Share Download PDF Status: Posted Version 1 posted 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-7215028","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":496599180,"identity":"fc00f989-0b2e-41a9-948d-b3c35ad2ee79","order_by":0,"name":"Nima Ilbeygi","email":"","orcid":"","institution":"Islamic Azad university","correspondingAuthor":false,"prefix":"","firstName":"Nima","middleName":"","lastName":"Ilbeygi","suffix":""},{"id":496599183,"identity":"4ba041e6-b548-4958-9740-7497bc21ba57","order_by":1,"name":"Alireza Jahandideh","email":"","orcid":"","institution":"Islamic Azad university","correspondingAuthor":false,"prefix":"","firstName":"Alireza","middleName":"","lastName":"Jahandideh","suffix":""},{"id":496599185,"identity":"eafd530b-af4b-41e0-b7dc-c3e82bbe9199","order_by":2,"name":"Abolfazl Akbarzadeh","email":"data:image/png;base64,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","orcid":"","institution":"Tabriz university of medical sciences","correspondingAuthor":true,"prefix":"","firstName":"Abolfazl","middleName":"","lastName":"Akbarzadeh","suffix":""},{"id":496599187,"identity":"c27f0e35-1fae-4acf-98a4-301ca4699050","order_by":3,"name":"Mohammad Nasrollahzadeh Masouleh","email":"","orcid":"","institution":"Islamic Azad university","correspondingAuthor":false,"prefix":"","firstName":"Mohammad","middleName":"Nasrollahzadeh","lastName":"Masouleh","suffix":""},{"id":496599190,"identity":"34fa0d33-013e-4e6d-93a1-1aed71260b90","order_by":4,"name":"Saeed hesaraki","email":"","orcid":"","institution":"Islamic Azad university","correspondingAuthor":false,"prefix":"","firstName":"Saeed","middleName":"","lastName":"hesaraki","suffix":""}],"badges":[],"createdAt":"2025-07-25 14:08:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7215028/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7215028/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":88812771,"identity":"3d3ed6d2-81a3-42c7-be7d-a0d2d523549c","added_by":"auto","created_at":"2025-08-11 15:51:50","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":183049,"visible":true,"origin":"","legend":"\u003cp\u003eResults of morphological investigation of prepared nanofibers for polyurethane/octacalcium phosphate composite with titanium (a-c).\u003c/p\u003e","description":"","filename":"image2.png","url":"https://assets-eu.researchsquare.com/files/rs-7215028/v1/e64e70c3a7edf775f53d3f24.png"},{"id":88812772,"identity":"4886ced2-cb0a-4d5a-a2c8-f0f28446917e","added_by":"auto","created_at":"2025-08-11 15:51:50","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":788576,"visible":true,"origin":"","legend":"\u003cp\u003eHistologic assessments 15,30, and 60 days after the defect surgery. (a,d,g) histological staining (H\u0026amp;E) of the defect area 15 day after surgery, (b,e,h) histological staining (H\u0026amp;E) of the defect area 30 days after surgery staining and (c,f,i) histological staining (H\u0026amp;E) of the defect area 60 days after surgery staining to visualize (magnification: 100x).\u003c/p\u003e","description":"","filename":"image3.png","url":"https://assets-eu.researchsquare.com/files/rs-7215028/v1/9afe812bb37c9af3bb77e6df.png"},{"id":88813972,"identity":"40eaf2af-1121-4b31-9855-f0b485d8c43e","added_by":"auto","created_at":"2025-08-11 15:59:50","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":834146,"visible":true,"origin":"","legend":"\u003cp\u003eImmunological assessments for Osteocalcin marker in 15,30 and 60 days after the defect surgery. (a,d,g) Osteocalcin marker of the defect area 15 day after surgery, (b,e,h) Osteocalcin marker of the defect area 30 days after surgery staining and (c,f,i) Osteocalcin marker of the defect area 60 days after surgery staining to visualize (magnification: 100x)\u003c/p\u003e","description":"","filename":"image4.png","url":"https://assets-eu.researchsquare.com/files/rs-7215028/v1/bc228f5b62924c245bac9ec9.png"},{"id":88813971,"identity":"3e2a7329-dc6c-4b64-a2cb-5ebe9edf55c7","added_by":"auto","created_at":"2025-08-11 15:59:50","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":121756,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 5. Radiological\u003c/strong\u003e assessment at 15 days, (A) Control group: minimal new bone formation was observed in the control group. Proximal defect repair was more evident than distal. One control sample showed a complete fracture at the defect site\u003cstrong\u003e (B)Autograph group:\u003c/strong\u003e The defect was more filled compared to the control group, with better proximal repair, (\u003cstrong\u003eC) OCP-TiO2 Implant Group:\u003c/strong\u003e Significant bone formation and defect filling were observed, though remodeling had not started.\u003c/p\u003e","description":"","filename":"image5.png","url":"https://assets-eu.researchsquare.com/files/rs-7215028/v1/d55f7d35f76de66045eab056.png"},{"id":88812786,"identity":"32fe50e1-ce0c-4101-b3cf-6dce96497ab8","added_by":"auto","created_at":"2025-08-11 15:51:50","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":203496,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 6. Radiological\u003c/strong\u003e assessment at 30 days, (A) Control\u003cstrong\u003e \u003c/strong\u003egroup: new bone formation still had a lower rate compared to the other groups. Defect healing was observed in both the proximal and distal parts to a similar extent.\u003cstrong\u003e (B) Autograph group:\u003c/strong\u003ethe extent of defect filling was much more evident compared to the control group, and significant progress was observed compared to day 15. The proximal and distal parts of the defect showed similar healing. (\u003cstrong\u003eC) OCP-TiO2 Implant Group:\u003c/strong\u003e a similar extent in both the proximal and distal parts, close to the maximum. The remodeling process has started in all samples, and in 75% samples, the bone has returned to its original shape.\u003c/p\u003e","description":"","filename":"image7.png","url":"https://assets-eu.researchsquare.com/files/rs-7215028/v1/c169cf023f8c99285997ae56.png"},{"id":88813975,"identity":"845878d5-d508-4165-96bd-939e5fe43949","added_by":"auto","created_at":"2025-08-11 15:59:50","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":231677,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 7. Radiological\u003c/strong\u003e assessment at 60 days, (A) Control\u003cstrong\u003e \u003c/strong\u003egroup: new bone formation had increased significantly compared to previous stages, and defect healing was observed in both proximal and distal parts to a similar extent\u003cstrong\u003e. (B) Autograph group:\u003c/strong\u003ehealing has occurred similarly and almost completely in both the proximal and distal parts of the defect. The remodeling process has also been relatively com7-.\u003c/p\u003e","description":"","filename":"image8.png","url":"https://assets-eu.researchsquare.com/files/rs-7215028/v1/78877458a62397e3f2bbd3b9.png"},{"id":89346641,"identity":"b6be133e-75fa-4a08-bb13-bd911abef95f","added_by":"auto","created_at":"2025-08-19 05:01:38","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3751022,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7215028/v1/8497df8c-0759-473d-af95-af627212d559.pdf"},{"id":88812770,"identity":"2d76585b-1ba0-4c3c-bf0b-5a69beb36407","added_by":"auto","created_at":"2025-08-11 15:51:50","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":31423,"visible":true,"origin":"","legend":"\u003cp\u003eChart 1: ATR-IR test analysis results\u003c/p\u003e","description":"","filename":"image1.png","url":"https://assets-eu.researchsquare.com/files/rs-7215028/v1/3e30205eb844a89e9962c4e3.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Investigation of bone defect repair with nanocomposite containing octacalcium phosphate (OCP) and titanium oxide on rat femur bone","fulltext":[{"header":"Introduction","content":"\u003cp\u003eBone defect repair is the process of bone tissue regeneration, which is a multidimensional and multifactorial process. A series of new cellular and molecular events occurs to re-form bone tissue. Subsequently, we need systemic and local factors for regular bone formation, and any disruption in this process disrupts repair [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]\u003c/p\u003e\u003cp\u003eMany studies have been conducted on types of autologous and non-autologous substitutes in various animal models to determine which compound can be used as a bone tissue substitute and is preferable. In this regard, two issues can be examined: the ability to induce bone formation and the ability to disperse and scaffold the material placed in the bone defect for repair, both of which are equally important [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe remarkable progress achieved in tissue engineering over the past decades has motivated researchers to develop biomaterials capable of mimicking the structure and function of the collagen-rich extracellular matrix, supporting cell adhesion, proliferation, and differentiation.\u003c/p\u003e\u003cp\u003eAnnually, bone defects affect over 20\u0026nbsp;million patients worldwide, often caused by trauma, tumors, infections, or congenital disorders, which severely impair patients\u0026rsquo; health and quality of life [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Addressing these defects is a crucial medical goal. Traditional methods like autografts and allografts are still not perfect: autografts, although providing a good environment for bone healing, have issues such as donor site complications, limited supply, and postoperative pain [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], while allografts pose risks of immune rejection and disease transmission [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. To overcome these challenges, research has increasingly focused on biomimetic biomaterials as innovative options for bone regeneration, seeking to surpass the limitations of conventional treatments [\u003cspan additionalcitationids=\"CR8\" citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe advanced biomaterials are fabricated to promote the proliferation of mineralized collagen extracellular matrix, aiming to enhance the biological functionality and compatibility of human tissues. Because of its acceptable biocompatibility and mechanical stability, mineralized collagen can stimulate osteogenesis, making it a key material in bone tissue engineering [\u003cspan additionalcitationids=\"CR11\" citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eBone fracture regeneration is a multiple-stage process, ranging from initial inflammation to later remodeling. Biomaterials with a calcium phosphate structure regulate these stages because of their compositional similarity to natural bone [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eOctacalcium phosphate (OCP) is a promising candidate to assist to biological apatite crystals within bone matrices as a precursor [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eOCP, with its chemical formula Ca8H2(PO4)6\u0026middot;5H2O and a Ca/P molar ratio of 1.33, significantly enhances osteoconductivity and exhibits notable osteoinductive potential, making it highly effective for supportive bone replacement [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eHowever, biological grafts are the gold standard among traditional methods; they have various limitations. Researchers have developed synthetic alternatives such as calcium phosphate ceramics, calcium sulfate, and bioactive glasses. Nevertheless, these alternatives face significant challenges: most only exhibit two of the four essential characteristics for effective bone repair\u0026mdash;osteointegration, osteoconduction, osteoinduction, and osteogenesis [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eFor instance, calcium phosphate ceramics, while effective at guiding bone formation, lack other essential functionalities when used alone [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Their brittleness and poor mechanical strength further limit their use in load-bearing areas [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Additionally, inconsistent degradation rates of these ceramics can hinder optimal bone regeneration and compromise structural stability during healing.\u003c/p\u003e\u003cp\u003eCalcium sulfate, a commonly used synthetic material, breaks down quickly and does not have enough mechanical strength for bone repair uses [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e] .Some synthetic alternatives have also caused serious complications. For instance, geneX\u0026reg;, a calcium sulfate-based implant, was associated with inflammation, delayed wound healing, and cyst formation in clinical cases, leading to early termination of a trial [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Bioactive glasses, although able to form strong bonds with bone through hydroxyapatite crystallization, lack sufficient mechanical strength for a standalone application [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e] .Likewise, synthetic hydroxyapatite misses trace elements (such., Na+, Mg2+, K+, Sr+) found in natural bone minerals, which reduces its biocompatibility and slows its resorption in the body [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eEquipment such as bone grafts and synthetic materials that are typically used to repair bone defects can be used in conjunction with standard surgical methods. Numerous materials have been developed to regenerate bone tissue to provide an alternative to autologous bone grafting, which has certain disadvantages such as limited tissue availability and morbidity associated with harvesting the bone from a second operative site [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e] .Calcium phosphates, such ashydroxyapatite (HA: Ca10(PO4)6(OH)2) and β-tricalcium phosphate (β‐TCP: Ca3(PO4)2), are standard materials that are typically used for bone regeneration. Their synthetic forms are widely employed as bone substitutes in clinical settings [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e"},{"header":"Materials","content":"\u003cp\u003eOctacalcium phosphate was purchased from Merck (Germany). All other reagents and solvents were of analytical grade and used without further purification. TiO2 (rutile) was obtained from Strem Chemicals Inc. (USA).\u003c/p\u003e\u003cp\u003e\u003cb\u003eFabrication and characterization\u003c/b\u003e\u003c/p\u003e\u003cp\u003eTo prepare a 15% weight/volume solution for the experimental groups, an Octacalcium phosphate (OCP) and polyurethane mixture is dissolved in acetic acid and dichloromethane (DCM), respectively. The compounds are used at a molar ratio of 4:1. In the experimental group, the synthesis and manufacturing of Octacalcium phosphate (OCP), polyurethane, and titanium dioxide involved the use of 0.05 grams of titanium dioxide. A solution with a concentration of 15% by weight or volume was supplemented with Octacalcium phosphate (OCP). After preparing the solutions, they were stirred with a magnetic stirrer at 25 degrees Celsius for 24 hours. The experimental groups used the electrospinning process to produce nanodimension scaffolds and nanofibers. The electrospinning machine was set to a speed of 130 revolutions per minute (rpm), a voltage of 20\u0026ndash;25 kilovolts (kV), a temperature of 25 degrees Celsius, a distance of 15 centimeters, and an injection rate of 2 milliliters per hour.\u003c/p\u003e\u003cp\u003e\u003cb\u003eFTIR Characterization\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe structural composition of the experimental group (octacalcium phosphate/polyurethane/Titanium Oxide) was assessed using Fourier Transform Infrared Spectroscopy (FTIR; Bruker IFS 48 instrument). All spectra were recorded in the range of 650\u0026ndash;4000 cm\u0026thinsp;\u0026minus;\u0026thinsp;1 at a scan speed of 16 scans per minute with a resolution of 1 cm\u0026thinsp;\u0026minus;\u0026thinsp;1 in a KBr-diluted medium (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cb\u003eScanning Electron Microscope (SEM) Characterization\u003c/b\u003e\u003c/p\u003e\u003cp\u003eTo analyze the microstructure of the nanofibers, samples were lyophilized in a freeze-dryer (Alpha 1\u0026ndash;4 LD plus; Martin Christ) for 48 hours at \u0026minus;\u0026thinsp;50\u0026deg;C. The dried nanofibers were then coated with gold in a vacuum and examined using a scanning electron microscope (SEM; TESCAN Vega II) (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cb\u003eExperimental groups\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe study groups consist of the first group with 12 participants (control group), the second group treated with an octacalcium phosphate and titanium oxide nanocomposite, polyurethane, and the third group treated with an autograft.\u003c/p\u003e\u003cp\u003e\u003cb\u003eBone damage model and nanocomposite implantation\u003c/b\u003e\u003c/p\u003e\u003cp\u003eTo implant the nanocomposite, animals are initially anesthetized with a combination of ketamine (10% 50 mg/kg) and xylazine (2% 5 mg/kg).\u003c/p\u003e\u003cp\u003eAfter preparing the area aseptically, an incision is made in the animal's thigh. The muscles in the area are removed, and a 3.5 mm defect is created in the femoral shaft using an orthopedic low-speed drill and a special drill. In the first group, the defect is left without implanting the nanocomposite, and the area is closed. In the second group, the defect is closed by implanting octacalcium phosphate and titanium oxide nanocomposite, while in the third group, the defect is filled and closed using bone fragments from the drilled area. After implanting the nanocomposite, the muscles in the area are sutured with 0\u0026ndash;4 Vicryl thread, and finally, the skin is closed with 0\u0026ndash;4 nylon thread. To prevent possible infections, all test animals will be injected intramuscularly with cefazolin at a dosage of 20 mg per kilogram of body weight and provided with painkillers for 3 days. Once a day, the area will be checked for possible swelling or inflammation, suture openings, discharge, or signs of infection. The skin sutures will be removed 10 days after the operation. First, radiographs will be taken of the desired area at intervals of 15, 30, 45, and 60 days for radiological evaluations.\u003c/p\u003e\u003cp\u003e\u003cb\u003eHistology assessment\u003c/b\u003e\u003c/p\u003e\u003cp\u003eTo perform histopathological and immunohistochemical tests, bone samples will be taken from every 3 mice in each group at 15, 30, 45, and 60 days. For this purpose, the animals will be euthanized by injecting lethal doses of intravenous anesthetic Sodium Thiopental (intracardiac injection), and then the femur will be removed for histopathological examination. The samples will be transported to the laboratory in a 10% formalin stabilizing solution, and the formalin solution will be replaced 24 hours later. Histopathological evaluation of the healing process will be performed with trichrome staining. Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e lists the scoring degree to which healing is assessed [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\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\u003eGrading of the histopathological lesions of the defects\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\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\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePhenomena\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eDescription Phenomena\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eScore\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"9\" rowspan=\"10\"\u003e\u003cp\u003eUnion\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eFailure to fill the defect\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eOnly fibrotic tissue\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEqual amounts of fibrotic and cartilage tissue\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHigh amounts of cartilage and little fibrosis\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eOnly cartilage\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLots of cartilage with little immature bone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eImmature cartilage and bone in equal proportion\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eProminent amount of immature bone and little cartilage\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHealing with immature bone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHealing with mature bone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e9\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003eInflammation\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAbsence of inflammatory cells\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePresence of inflammatory cells less than 25%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePresence of inflammatory cells 25\u0026ndash;50%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePresence of inflammatory cells 50\u0026ndash;75%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePresence of inflammatory cells more than 75%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003eRemodeling\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNo Remodeling\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRemodeling less than 25%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRemodeling 25%-50%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRemodeling 50%-75%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMore than 75% Remodeling\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e\u003cp\u003eAbsorption of scaffolds\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNo Absorption\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAbsorption 25\u0026ndash;50%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAbsorption 50%-75%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eComplete Absorption\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eRadiological assessment\u003c/b\u003e\u003c/p\u003e\u003cp\u003eRadiological methods have been employed with remarkable success in the assessment of bone repair procedures. In experimental settings, the presurgical assessment allows the evaluation of exact anatomy and bone dimensions, aiding in the selection of fixation devices and implants. Radiological assessment as a basic postoperative method enables to evaluation of the quality of replacement and detection of the fracture. Sample radiographic images of the rabbit calvarial bone, with or without a grafted material, are shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e\u003cp\u003eIn this study, we applied a computed tomography scan (CT scan) and a computer to create cross-sectional images, also known as slices, of the bones, blood vessels, and soft tissues inside the body.\u003c/p\u003e\u003cp\u003eThe skulls of the rabbits were CT scanned on the 30th and 60th days after surgery to evaluate the amount of bone repair and hard tissue formation in the area. A CT scan of the calvaria region was performed after euthanasia and skull removal in the animals. The grading of the CT scan was done in the following Table \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\u003eGrading of the sonographic evaluations of the defects\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\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=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eStandard unit\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003evariable description\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003echangeable\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003escale\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eThe volume of newly formed bone/the total volume of the desired area\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBone volume fraction\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eThe volume of the remaining grafted material / the total volume of the desired area\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003evolume fraction of the remaining material\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1/mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMeasurement of the average number of trabeculae per unit length\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBone trabeculae count\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003emm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAverage thickness of trabeculae (assessed using direct 3D methods)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eThe thickness of bone trabeculae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003emm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAverage distance between trabeculae (assessed using direct 3D methods)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eThe distance between bony trabeculae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cdiv id=\"Sec2\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003eResults were expressed as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation. We used SPSS (version 16.1, IBM, USA). One-way ANOVA analysis of variance was used to compare the means from multiple experimental groups, followed by a post hoc Tukey test. P\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eNanofibers Characterizations:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eATR-IR analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe ATR-IR technique was used to investigate the presence of chemical functional groups within the structure of the polyurethane/octacalcium phosphate composite with titanium oxide; the results are reported in the form of a graph based on the frequency of the elements present in the chemical structure. The vibrations in the graph corresponding to frequencies between 3961-3604 and 1523-cm-1 indicate the presence of N-H and C-N functional groups in the polyurethane polymer, respectively. Also, the frequency between 1100-1072-cm-1 indicates the presence of C-O-C functional groups in the polyurethane, and the wavelengths between 1001-920, 1260-1072, and 500-700 cm-1 indicate the functional groups of octacalcium phosphate Ca, HP, PO4-. The vibrations observed at around 2965 cm-1 in the diagrams refer to the aliphatic C\u0026ndash;H functional group in the nanocomposite. On the other hand, the vibrations at frequencies of 1770\u0026ndash;1726, 1449\u0026ndash;1072, and 800\u0026ndash;690 cm-1 indicate the Ti-o-o functional groups of titanium oxide that have reacted with the chemical structure of the composite and are present (Diagram 1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSEM Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe results of examining the morphology of the prepared nanofibers for the polyurethane/octacalcium phosphate composite with titanium using the SEM scanning electron microscope method showed that the prepared nanofibers can maintain a three-dimensional structure with interconnected porosity. This porosity and interconnection of the nanofibers can be a reason for the presence of titanium dioxide, which creates a more suitable morphology and topography in the scaffold and can indicate the presence of suitable sites for surface adhesion in the prepared nanofiber strands( Figure1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHistopathological Results\u003cspan dir=\"RTL\"\u003e:\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe results of the test after 15,30, and 60 days after treatment indicated that Dense fibrous connective tissue with fibroblasts and fibrocytes, relatively low infiltration of mononuclear inflammatory cells, covers the outer surface of the defect. In deeper areas, immature and scattered trabeculae of newly formed bone are observed. Extensive fibrous connective tissue with fibroblasts and fibrocytes, and moderate to severe infiltration of mononuclear inflammatory cells surrounding the autograft particles placed in the defect. Limited bone formation is seen in the center and around the autograft, as well as at the margins of the defect and near the old bone. Limited fibrous connective tissue with severe infiltration of neutrophilic inflammatory cells surrounding the scaffold particles placed in the defect. Bone formation appears as scattered islands among the formed connective tissue Moreover, the results 30 days after implantation in all experimental groups indicated that in the control group, Dense fibrous connective tissue with fibroblasts, collagen Fibers, with very mild to negligible infiltration of inflammatory cells. Immature and scattered trabeculae of newly formed bone are observed. Fibrous connective tissue, containing fibroblasts, fibrocytes, and collagen fibers, with moderate to severe inflammatory cell infiltration, surrounds the autograft pieces placed in the defect. Newly formed bone trabeculae adjacent to the connective tissue around the autograft are evident( Figure2).\u003c/p\u003e\n\u003cp\u003eThe TiO₂-OCP group showed significantly more bone formation than the others. Severe inflammation was localized around scaffold remnants. However, in the fibrous connective tissue, the inflammation was negligible to mild.\u003c/p\u003e\n\u003cp\u003eAmong the treatment groups, the TiO₂-OCP group showed the highest bone formation with the least inflammation\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e At 60 days, the control group showed integrated trabeculae of newly formed bone adjacent to the old cortical bone\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eDespite bone formation in the connective tissue filling the defect, severe inflammation was observed. The TiO₂-OCP group showed the most prominent bone formation. Newly formed trabeculae had fused, forming relatively mature cortical bone plates.\u003c/p\u003e\n\u003cp\u003eComparison of Groups at Day 15\u003cspan dir=\"RTL\"\u003e:\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eThe control group shows moderate bone formation with the least inflammation. The TiO₂-OCP group shows the highest bone formation with the lowest inflammation(Table 3)\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eThe process of scoring Histopathological Results will be as follows\u003cspan dir=\"RTL\"\u003e:\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e1. \u003cstrong\u003eBone Formation:\u003c/strong\u003e\u003cbr\u003e\u0026nbsp;o 0: No formation\u003cbr\u003e\u0026nbsp;o 1: Minimal formation\u003cbr\u003e\u0026nbsp;o 2: Moderate formation\u003cbr\u003e\u0026nbsp;o 3: Marked formation (bridging of the defect)\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e2. \u003cstrong\u003eIntegration with Host Bone:\u003c/strong\u003e\u003cbr\u003e\u0026nbsp;o 0: No integration\u003cbr\u003e\u0026nbsp;o 1: Partial integration\u003cbr\u003e\u0026nbsp;o 2: Almost complete integration\u003cbr\u003e\u0026nbsp;o 3: Complete integration\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e3. \u003cstrong\u003eInflammation:\u003c/strong\u003e\u003cbr\u003e\u0026nbsp;o 0: No inflammation\u003cbr\u003e\u0026nbsp;o 1: Mild inflammation\u003cbr\u003e\u0026nbsp;o 2: Moderate inflammation\u003cbr\u003e\u0026nbsp;o 3: Severe inflammation\u003c/p\u003e\n\u003cp\u003eTable 3.\u0026nbsp;Histopathological Result. The results of the test after 15 days after treatment.\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable dir=\"rtl\" border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"601\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eBone Formation\u003cspan dir=\"RTL\"\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eIntegration with Host Bone\u003cspan dir=\"RTL\"\u003e\u0026nbsp;\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eInflammation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cstrong\u003e\u003cspan dir=\"LTR\"\u003eGroup\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e0\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e0\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eControl\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e0\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e3\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eAutograft\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e0\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e3\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eAutograft\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e3\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eAutograft\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eAutograft\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eOCP-TIO2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eOCP-TIO2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eOCP-TIO2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003e\u003cspan dir=\"RTL\"\u003e1\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"LTR\"\u003eOCP-TIO2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eComparison of Groups at Day 30\u003cspan dir=\"RTL\"\u003e:\u0026nbsp;\u003c/span\u003eAmong the treatment groups, the TiO₂-OCP group showed the highest bone formation with the least inflammation (Table 4)\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eTable 4. Histopathological Result. The results of the test after 30 days after treatment.\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable dir=\"rtl\" border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"599\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eBone Formation \u0026nbsp;\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eIntegration with Host Bone\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eInflammation\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cstrong\u003e\u003cspan dir=\"LTR\"\u003eGroup\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TIO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TIO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TIO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 131px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 138px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 201px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TIO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 5. Histopathological Result. The results of the test after 60 days after treatment.\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable dir=\"rtl\" border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"620\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eBone Formation \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eIntegration with Host Bone\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eInflammation\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cstrong\u003e\u003cspan dir=\"LTR\"\u003eGroup\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TIO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TIO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TIO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 161px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 163px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 172px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TIO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eImmunohistochemistry Results\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eImmunohistochemical analysis was conducted for VEGF, Osteocalcin, and Ki67 on Day 15, and Osteocalcin as an osteogenesis marker at 15-, 30-, and 60-days post-treatment in each experimental group using the Allred scoring method (Figure 3). The staining outcomes were examined and compared on days 15, 30, and 60 after transplantation, focusing on new bone formation, integration with host tissue, and inflammation. These results are detailed in separate scoring tables. The findings showed that at 60 days after defect creation, mature bone trabeculae, well-integrated with the host bone, had formed. Meanwhile, moderate bone formation with minimal inflammation occurred at 15- and 30-days post-defect creation, respectively. The results indicate that the TiO₂-OCP experimental groups exhibited the highest bone formation, low inflammation, and scattered bone islands compared to the autograft and control groups. Moreover, in the autograft group, persistent inflammation and moderate bone formation near graft fragments were observed, with severe inflammation at 30- and 60-days post-treatment. In contrast, the TiO₂-OCP group showed the best results: mature cortical bone with Haversian canals,\u0026nbsp;complete integration, and\u0026nbsp;minimal inflammation at 60 days post-implantation, suggesting that the TiO₂-OCP composite\u0026nbsp;is a\u0026nbsp;superior alternative to autografts. It enhances bone repair while reducing inflammatory complications. Autografts, on the other hand, caused persistent inflammation, delaying healing despite some bone formation. The control group displayed slower but steady healing with less inflammation than autografts.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMicroscopy Method for Allred\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003escoring for VEGF and\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eOsteocalcin and Ki67\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eProportion\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003escore\u003c/strong\u003e\u003cbr\u003e0\u0026mdash;No cells are immunoreactive,1\u0026mdash;\u0026le; 1% of cells are immunoreactive\u003cbr\u003e2\u0026mdash;1\u0026ndash;10% of cells are immunoreactive\u003cbr\u003e3\u0026mdash;11\u0026ndash;33% of cells are immunoreactive\u003c/p\u003e\n\u003cp\u003e4\u0026mdash;34\u0026ndash;66% of cells are immunoreactive\u003cbr\u003e5\u0026mdash;67\u0026ndash;100% of cells are immunoreactive\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIntensity\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003escore\u003c/strong\u003e\u003cbr\u003e0\u0026mdash;Negative.\u003cbr\u003e1\u0026mdash;Weak.\u003cbr\u003e2\u0026mdash;Intermediate.\u003cbr\u003e3\u0026mdash;Strong.\u003cbr\u003e\u0026nbsp;\u003cbr\u003e\u003cstrong\u003eAllred\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003escore (P+I)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e0\u0026ndash;1\u0026mdash;Negative.\u003cbr\u003e2\u0026ndash;3\u0026mdash;Weak\u003cbr\u003e4\u0026ndash;6\u0026mdash;Moderate\u003cbr\u003e7\u0026ndash;8\u0026mdash;Strong\u003c/p\u003e\n\u003cp\u003eTable 6. \u003cstrong\u003eDay 15 Results\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable dir=\"rtl\" border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"601\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eVEGF\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eKi67\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOsteocalcin\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cstrong\u003e\u003cspan dir=\"LTR\"\u003eGroup\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TiO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TiO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TiO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 150px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TiO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRadiological Results\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eRadiological Evaluation results indicated that, TiO₂-OCP scaffold\u0026nbsp;showed the\u0026nbsp;fastest and most complete bone regeneration\u0026nbsp;at all-time points. And autografts\u0026nbsp;performed better than controls but\u0026nbsp;lagged behind TiO₂-OCP in remodeling, control\u0026nbsp;was\u0026nbsp;slowest, with persistent defects even at Day 60.so, TiO₂-OCP\u0026nbsp;is\u0026nbsp;superior to autografts\u0026nbsp;in\u0026nbsp;accelerating bone repair\u0026nbsp;and achieving\u0026nbsp;structural restoration\u0026nbsp;with\u0026nbsp;earlier remodeling(Figur5-7). Autografts, while effective,\u0026nbsp;delay remodeling\u0026nbsp;and may cause complications (e.g., persistent radiolucency). Control healing\u0026nbsp;is insufficient for critical-sized defects, emphasizing the need for\u0026nbsp;bone graft substitutes.The process of scoring radiographs will be as follows\u003cspan dir=\"RTL\"\u003e:\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eTable 6. \u003cstrong\u003eDay 15 Radiological Scores\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable dir=\"rtl\" border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cstrong\u003e\u003cspan dir=\"LTR\"\u003eGroup\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eBone formation\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eProximal/Distal union\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eRemodeling\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eTotal score\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0/0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TiO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TiO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2/2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TiO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 116px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TiO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 133px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003eTable 7. \u003cstrong\u003eDay 30 Radiological Scores\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable dir=\"rtl\" border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"602\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eTotal score\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eRemodeling\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eProximal/Distal union\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eBone formation\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cstrong\u003e\u003cspan dir=\"LTR\"\u003eGroup\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eAutograft\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2/2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TiO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2/2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TiO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1/2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TiO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 117px;\"\u003e\n \u003cp dir=\"RTL\"\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp dir=\"RTL\"\u003e2/2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 119px;\"\u003e\n \u003cp dir=\"RTL\"\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp dir=\"RTL\"\u003e\u003cspan dir=\"LTR\"\u003eOCP-TiO2\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eSince ancient times, finding effective treatments to regenerate damaged or dysfunctional bone tissues has been the most challenging and persistent issue in medicine. The conventional approach, like tissue or organ transplantation, comes with significant drawbacks, including donor shortages, high costs, immune rejection, and complications from foreign tissue implantation. These limitations have motivated the exploration of novel strategies that replace current treatments and are designed to closely mimic natural tissue [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eBone tissue engineering, through the fabrication of bone substitutes, currently provides innovative solutions for managing bone injuries. A promising opportunity is using bone substitutes that can boost bone formation. Advances in nanotechnology have enabled the development of polymer-based scaffolds that incorporate nanoparticles and nanofibers [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. These nano-engineered fibers offer a larger surface area than traditional fibers, allowing for better material integration and enhanced mechanical strength. According to the latest studies, the composite of semiconductive biomaterials combined with nanoscale metal-based materials like octacalcium phosphate (OCP) and titanium oxide (TiO₂) could revolutionize the treatment of bone injuries in humans. Therefore, TiO₂\u0026rsquo;s effectiveness is improved when incorporated into OCP, supporting its inclusion in bone regeneration research [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe growing body of research on octacalcium phosphate (OCP) nanocomposites shows their remarkable potential in bone regeneration. Building on early findings of OCP's osteoconductive properties [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e], subsequent studies have systematically explored and expanded its applications in tissue engineering. Notably, Heydari et al. advanced the field by successfully incorporating OCP into polycaprolactone scaffolds, demonstrating enhanced osteoblast proliferation\u0026mdash;a critical step toward clinical applications [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe regenerative mechanisms of OCP seem complex. Kurobane et al. offered important insights into its ability to promote blood vessel growth, showing that OCP/gelatin composites not only support bone formation but also enhance vascularization, which is a major challenge in repairing large bone defects. This dual role was further backed by Kawai et al., whose clinical studies in oral surgery confirmed that OCP-based materials are both safe and effective, while also showing their potential as alternatives to autologous grafts [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eRecent work by Hamaei et al. has advanced our understanding by establishing a dose-response relationship between OCP crystal density and osteogenic outcomes. This finding indicates that optimizing the material could lead to even greater clinical effectiveness. Combined with the antimicrobial properties of TiO₂ nanocomposites, these developments provide a comprehensive solution that addresses multiple aspects of bone regeneration: structural support, biological activity, and infection prevention.\u003c/p\u003e\u003cp\u003eCollectively, these studies represent significant progress toward overcoming the limitations of traditional transplantation methods. The consistent findings across in vitro, animal, and clinical studies strongly support the translational potential of OCP-based nanocomposites. Future research should focus on scaling up production, optimizing composite formulations for specific clinical applications, and conducting larger-scale clinical trials to fully realize this technology's potential in regenerative medicine.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank the\u003csup\u003e\u0026nbsp;\u003c/sup\u003eDepartment of Clinical Science, Science and Research Branch, Islamic Azad University, and Department of Medical Nanotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences for financial support of this project (Grant NO: 62379).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was funded by the Department of Medical Nanotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences (Grant NO: 62379).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical Approval\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe ethical approval for this paper was obtained from the research ethics committee of Tabriz University of Medical Sciences (IR.IAU.SRB.REC.1402.346\u003cspan dir=\"RTL\"\u003e\u0026nbsp;(\u003c/span\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNima Ilbeygi:\u003c/strong\u003e Conceptualization, Methodology, Investigation,\u0026nbsp;Formal analysis, Writing - Review \u0026amp; Editing\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAlireza Jahandideh, Abolfazl Akbarzadeh:\u003c/strong\u003e\u003csup\u003e\u0026nbsp;\u003c/sup\u003eWriting-original draft, Writing - Review \u0026amp; Editing\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSaeed Hesaraki:\u0026nbsp;\u003c/strong\u003eMethodology, Investigation\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMohammad Nasrollahzadeh Masouleh, and Abolfazl Akbarzadeh:\u003c/strong\u003e Project administration, Conceptualization, Supervision, Resources, Funding acquisition \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement (DAS)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eHench, L. 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(2018). \u003cem\u003eAssessment of tricalcium phosphate/collagen (TCP/collagene)nanocomposite scaffold compared with hydroxyapatite (HA) on healing of segmental femur bone defect in rabbits.\u003c/em\u003e Artificial Cells, Nanomedicine, and Biotechnology, 46(2): pp. 242\u0026ndash;249.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSuzuki, O., et al. (2006). Bone formation enhanced by implanted octacalcium phosphate involving conversion into Ca-deficient hydroxyapatite. \u003cem\u003eBiomaterials\u003c/em\u003e, \u003cem\u003e27\u003c/em\u003e(13), 2671\u0026ndash;2681.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKamakura, S., et al. (1999). Implantation of octacalcium phosphate (OCP) in rat skull defects enhances bone repair. \u003cem\u003eJournal of dental research\u003c/em\u003e, \u003cem\u003e78\u003c/em\u003e(11), 1682\u0026ndash;1687.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMiyatake, N., et al. (2009). Effect of partial hydrolysis of octacalcium phosphate on its osteoconductive characteristics. \u003cem\u003eBiomaterials\u003c/em\u003e, \u003cem\u003e30\u003c/em\u003e(6), 1005\u0026ndash;1014.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eJiang, P., et al. (2015). Effect of octacalcium-phosphate-modified micro/nanostructured titania surfaces on osteoblast response. \u003cem\u003eACS applied materials \u0026amp; interfaces\u003c/em\u003e, \u003cem\u003e7\u003c/em\u003e(26), 14384\u0026ndash;14396.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKawai, T., et al. (2020). Clinical study of octacalcium phosphate and collagen composite in oral and maxillofacial surgery. \u003cem\u003eJournal of tissue engineering\u003c/em\u003e, \u003cem\u003e11\u003c/em\u003e, 2041731419896449.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHamai, R., et al. (2022). Octacalcium phosphate crystals including a higher density dislocation improve its materials osteogenecity. \u003cem\u003eApplied Materials Today\u003c/em\u003e, \u003cem\u003e26\u003c/em\u003e, 101279.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Charts","content":"\u003cp\u003eChart 1 is available in the Supplementary Files section\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Bone regeneration, Octacalcium phosphate (OCP), Titanium oxide (TiO₂), Nanocomposite, Osteoconductivity, Femoral defect","lastPublishedDoi":"10.21203/rs.3.rs-7215028/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7215028/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBone defect repair remains a major clinical challenge, requiring the development of effective biomaterials to address the limitations of autografts and allografts. This study examined the regenerative potential of a nanocomposite composed of octacalcium phosphate (OCP) and titanium dioxide (TiO₂) in a rat femoral defect model. The OCP-TiO₂ nanocomposite was produced through electrospinning and analyzed using Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM), confirming its structural integrity and osteoconductive properties. Rats were divided into three groups: control (untreated defect), autograft, and OCP-TiO₂ nanocomposite. Histopathological, immunohistochemical (VEGF, Osteocalcin, Ki67), and radiological assessments were performed at 15-, 30-, and 60-days post-implantation. Findings showed that the OCP-TiO₂ group experienced enhanced bone regeneration, with earlier trabecular bone formation, less inflammation, and improved osteointegration compared to autografts and controls. Radiological evaluations indicated faster defect remodeling in the OCP-TiO₂ group, while autografts displayed ongoing inflammation.\u003c/p\u003e\u003cp\u003eThese results indicate that the OCP-TiO₂ nanocomposite is a promising alternative to autografts, offering improved osteoconductivity, mechanical stability, and biocompatibility for bone defect repair. Future research should focus on long-term outcomes and clinical applications.\u003c/p\u003e","manuscriptTitle":"Investigation of bone defect repair with nanocomposite containing octacalcium phosphate (OCP) and titanium oxide on rat femur bone","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-11 15:51:45","doi":"10.21203/rs.3.rs-7215028/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"fc9f022d-a0ff-4fbd-bbbd-1589f70b764b","owner":[],"postedDate":"August 11th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-11-23T17:38:23+00:00","versionOfRecord":[],"versionCreatedAt":"2025-08-11 15:51:45","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7215028","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7215028","identity":"rs-7215028","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}