Antibiofilm, regenerative and bone homeostasis potential of the synergistic association of synoeca-MP peptide with chlorhexidine in oral cavity opportunistic infections

Antibiofilm, regenerative and bone homeostasis potential of the synergistic association of synoeca-MP peptide with chlorhexidine in oral cavity opportunistic infections | 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 Antibiofilm, regenerative and bone homeostasis potential of the synergistic association of synoeca-MP peptide with chlorhexidine in oral cavity opportunistic infections Ingrid Aquino Reichert Barin, Johnny Carvalho da Silva, Raquel Figuerêdo Ramos, and 11 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4595835/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 Objective To evaluate in vitro the association between synoeca-MP peptide and chlorhexidine, regarding their antimicrobial and antibiofilm activities, saliva stability, effect on tissue repair, bone resorption processes, and mineralized matrix formation. Methodology Initially, the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and antibiofilm concentration were determined. The synergism and degradation of synoeca-MP and chlorhexidine in human saliva were assessed. Furthermore, biocompatibility was evaluated using MTT assays, hemolytic assays, and proliferation and migration assays of periodontal ligament cells. Finally, bone homeostasis was evaluated through osteoclastogenesis assays, alkaline phosphatase determination, and mineralized matrix formation assay with SaOs-2 and ligament cells. Results The antimicrobial and antibiofilm activity against the tested microorganisms was confirmed. Low synergistic concentrations of the synoeca-MP and chlorhexidine combination inhibited tested microorganisms. The association of these molecules remained stable in healthy saliva. Nevertheless, it degraded as the severity of periodontal disease increased. Additionally, lower synergistic concentrations of the combination were not cytotoxic to human cells, promoted the proliferation and migration of ligament cells, inhibited osteoclastogenesis, and increased mineral matrix formation of ligament cells and SaOs-2. Conclusion Synoeca-MP and chlorhexidine combination shows potential for oral diseases treatment, as evidenced by its antimicrobial activity, regenerative potential, saliva stability, and bone homeostasis. It may be particularly effective for opportunistic oral infections and in conjunction with mechanical therapy. Clinical relevance The results obtained allow us to confirm the safety and expected effects of this combination, making it a promising alternative for hospitalized patients. Synergism Host defense peptide Periodontal disease Opportunistic infections Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction The control of bacterial biofilm from the oral cavity and respiratory support equipment can avoid nosocomial pneumonia infections in hospital settings [ 1 ]. It is known that patients admitted to intensive care units (ICU), subject to low immunity, may have respiratory infections, most often caused by nosocomial pneumonia and responsible for up to 50% of deaths in the ICU environment, making them a worldwide public health problem [ 2 ]. Oral immunoinflammatory imbalance can alter oral microbiota, leading to the emergence of opportunistic microbial species [ 3 ], such as Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus. aureus and Candida albicans . In this context, the ecological diversity of the oral and periodontal microenvironment may provide suitable conditions for colonization by microorganisms that are not usually considered members of the oral microbiota in subgingival areas [ 4 ]. Such microorganisms have the ability to organize themselves in biofilm and to adhere to dental calculus [ 5 ]. The standard treatment for opportunistic oral infections consists of mechanical microorganisms and biofilm removal [ 6 ]. It is important to evaluate anatomic involvement and the management of any underlying comorbidities and, if necessary, antibiotic, antifungal, antiviral, or antiparasitic adjuvant drugs can be used [ 7 ]. However, most conventional antibiotics are uneffective at clearing biofilm-associated infections [ 8 ]. In a clinical and hospital context, where patients maintain relatively poor oral hygiene conditions, the use of 0.12% chlorhexidine is recommended for oral biofilm control [ 9 ]. This form of control especially occurs in the presence of ventilation equipment, motor difficulties normally in cases in which patients are in an unconscious or sedated state [ 10 ]. Among the most common adverse effects, the following stand out tooth pigmentation, altered taste perception, increased formation of dental calculus, dry mouth, soft tissue burns, and edema of the lips and glands [ 11 ]. For this reason, there is a need to develop new agents with antimicrobial, immunomodulatory, and antibiofilm action. Host defense peptides (HDPs) are a class of biomolecules that can have both antimicrobial acivity against bacteria, viruses and fungi, as well as performing important immunomodulatory functions in angiogenesis, wound healing, and chemotaxis [ 12 ]. In addition, it has been shown that some peptides increase antibiotic action to prevent biofilm formation and eradicate mature biofilms [ 13 ]. One example is the synergistic combination of peptide IDR 1018 with low levels of the antibiotics ceftazidime, ciprofloxacin, imipenem, or tobramycin, which was able to reduce the concentration of antibiotic required to eradicate biofilms [ 12 , 13 ]. In this condition, the synoeca-MP peptide, derived from the venom of Synoeca surinama , a species of South American wasp, has demonstrated important antimicrobial results against Gram-positive and -negative strains. Synoeca-MP is composed of 14 amino acid residues and is effective mainly against antibiotic-resistant bacteria, such as S. aureus, Escherichia coli and E. faecalis [ 14 ]. Likewise, previous studies have also shown this peptide’s synergistic antimicrobial activity against P. aeruginosa and its antibiofilm effect, as well as immunomodulatory activity when associated with chlorhexidine, at low concentrations of both. This association can favor the reduction of undesirable effects of chlorhexidine used alone [ 15 ]. However, there is little information regarding this association [ 15 ]. Therefore, the present study aimed to analyze the association between synoeca-MP peptide and chlorhexidine, regarding its antimicrobial, antibiofilm activity, and its stability in saliva. In addition, its effect on tissue repair, in bone resorption processes, and mineralized matrix formation in vitro were also analyzed. The hypothesis was that the association between synoeca-MP peptide and chlorhexidine [ 16 ] is capable of modulating bone homeostasis, inhibiting osteoclastogenesis, and increasing the formation of the mineral matrix in vitro. Moreover, it may have repair potential, given the conditions of tissue destruction caused by opportunistic microorganisms, as well as periodontopathogens of the oral cavity and the periodontal microenvironment [ 17 ]. Material and methods Synoeca-MP peptide and chlorhexidine digluconate preparation Synoeca-MP peptide (INWLKLGKKIIASL-NH 2 ) was synthesized in solid-phase using F-moc methodology, purified (purity > 95%), lyophilized and stored by AminoTech (São Paulo, Brazil) [ 18 ]. To confirm molecular mass and purity, Matrix Assisted Laser Desorption Ionization - Time of Flight (MALDI-ToF) mass spectrometry was used (Supplementary Fig. 1). The peptide was dissolved in ultrapure water and stored at -20°C until use. Chlorhexidine was manipulated for stock solution (20% concentrated) (Via Magistral, Brasília, Distrito Federal, Brazil) and dissolved in ultrapure water [ 19 ]. Determination of minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC) and antibiofilm concentration Bacteriostatic and bactericidal concentrations were determined according to Clinical & Laboratory Standards Institute guidelines, with some adaptations. Analyses were performed to determine their effectiveness against P. aeruginosa (ATCC27853), E. faecalis (ATCC 19433) and S. aureus (ATCC 25923). Growth curves from each microorganism were previously determined to use its log phase during assays. Antibacterial assays were performed with Luria-Bertani media (Invitrogen, Waltham, Massachusetts, USA) with 5x10 5 CFU.mL -1 in a 96-well plate for 18h at 37 ºC incubated in a microplate reader with readings at 600 nm absorbance under medium agitation (BioTek PowerWave HT, Winooski, Vermont, USA). Ampicillin (50 µg.mL -1 – Sigma-Aldrich, San Luis, Missouri, USA) was used as a negative control for antibacterial assays. Chlorhexidine (Via Magistral) was tested up to 512 µg.mL -1 (1013 µM) and MIC was determined by no bacteria growth. After MIC determination, each sample was incubated in an agar plate to assess the presence of viable microorganisms for the definition of MBC. Then, antibiofilm properties of synoeca-MP and chlorhexidine were assessed against preformed biofilms of P. aeruginosa and S. aureus. Bacterium inoculum was diluted in a BM2 minimum medium, consisting of 62 mM potassium phosphate buffer (pH 7.0 - VETEC, Duque de Caxias, Rio de Janeiro, Brazil), 7 mM (NH 4 ) 2 SO 4 (VETEC), 2 mM MgSO 4 mM (Sigma-Aldrich), 10 µM FeSO 4 (VETEC), and 0.5% glucose (Sigma-Aldrich), so that the final concentration reached was 1/100 v/v per well [ 20 ]. Bacteria were plated and kept for 24h at 37°C. Controls were represented by P. aeruginosa and S. aureus bacteria (1/100 v/v) as negative control, ciprofloxacin (Sigma-Aldrich) and the BM2 medium [62 mM potassium phosphate buffer (pH 7.0 - VETEC), 7 mM (NH 4 ) 2 SO 4 (VETEC), 2 mM MgSO 4 mM (Sigma-Aldrich), 10 µM FeSO 4 (VETEC), and 0.5% glucose (Sigma-Aldrich)], as positive control. After 24h, planktonic bacteria were discarded and plates were washed twice with 1x PBS. Serial sample dilutions were performed and added to the preformed biofilm microplates, for 24h. Subsequently, to assess the biofilm’s viability, the MTT assay (Sigma-Aldrich) was performed, according to the manufacturer's standards [ 21 ]. Determination of minimal inhibitory concentration (MIC), minimal fungicidal concentration (MFC) and antibiofilm concentration of Candida albicans Antifungal analyses were performed against C. albicans (ATCC 10231). Experimental assays were performed with RPMI-1640 media (Sigma-Aldrich) with 0.165 mol.L -1 MOPS with 2.5x10 3 CFU.mL -1 in a 96-well plate for 48h at 37 ºC in a microplate reader under medium agitation (BioTek PowerWave HT). Amphotericin B (10 µg.mL -1 – Sigma-Aldrich) was used as a negative control for antifungal assays. Chlorhexidine (Via Magistral) was tested up to 512 µg.mL -1 (1013 µM) and MIC was determined by no fungal growth. After MIC determination, the sample was incubated in an agar medium for determination of MFC. Fungal antibiofilm properties of synoeca-MP and chlorhexidine were assessed against preformed biofilms of C. albicans . Then, cells were centrifuged, after 48 h growth, at 3000 g, 5 min, at 4 ºC, washed twice with sterile PBS and then resuspended in BM2 media [62 mM potassium phosphate buffer (pH 7.0 - VETEC), 7 mM (NH 4 ) 2 SO 4 (VETEC), 2 mM MgSO 4 mM (Sigma-Aldrich), 10 µM FeSO 4 (VETEC), and 0.5% glucose (Sigma-Aldrich)]. Cells were seeded at 1x10 7 CFU per well in 96-well plates and incubated for 48h at 37 ºC. Antimicrobial agents were added to plates and incubated for 24h more. All agents were tested according to the following concentrations: chlorhexidine up to 128 µg.mL -1 (253 µM) and synoeca-MP up to 128 µg.mL -1 (80 µM). Amphotericin B (Sigma-Aldrich) at 16 µg.mL -1 was used as a positive control. The effect on biofilms was evaluated by XTT cell viability kit assays (Biotium, Fremont, California, USA) and plates were read at 470 nm absorbance [ 22 ]. Synergism assay After the determination of minimum inhibitory concentrations, the synergism assay was assessed for the combination of chlorhexidine and synoeca-MP using the microdilution growth inhibition assay protocol, according to the Clinical and Laboratory Standards Institute (CLSI) [ 23 ]. Thus, concentrations below its MICs were tested according to the checkerboard method with adaptations [ 24 ]. Assays were performed against P. aeruginosa , S. aureus and C. albicans as previously described to determine MIC. The fractional inhibitory concentration (FIC) index determined synergistic interactions according to the following calculation: [(MIC of 1 combined to 2) / (MIC of 1)] + [(MIC of 2 combined to 1) / (MIC of 2)]. Values < 0.5 were considered as a synergic interaction. Degradation/integrity of synoeca-MP and chlorhexidine in human saliva The protocol of this study was approved by the Human Research Ethics Committee (number 90666218.2.0000.0029). Saliva was collected from control patients (without gingival and/or periodontal diseases; n = 7), patients diagnosed with gingivitis (n = 6), and periodontitis (n = 17), prior to clinical dental care, at the graduate dentistry clinic of Universidade Católica de Brasília. Patient data for those with periodontal disease were obtained from medical records, and saliva samples were taken between 8 and 11 am. Unstimulated saliva was collected for 5 min [ 25 ]. Subsequently, 80.26 µM (40.565 µg/mL) of chlorhexidine, 80.26 µM (128 µg/mL) of synoeca-MP, or the same concentration of chlorhexidine and synoeca-MP together were exposed to saliva; the control group was represented by no treatment exposed to saliva control group. To assess the integrity of synoeca-MP and chlorhexidine in saliva, molecular mass, and purity were evaluated using the autoflex speed spectrometer mass spectrometry technique (Bruker Daltonics, Billerica, Massachusetts, USA), using the reflected and positive operating method, adjusted for 400 to 3500 Da, with external calibration [ 26 ]. Synoeca-MP and chlorhexidine were diluted in ultrapure water and deposited on the plate (AnchorchipVar-384 – Bruker Daltonics) together with the matrix in triplicate, awaiting complete crystallization at room temperature after 0, 30, 60, 90, 120, 150, 180, 210 and 240 min. Between the experimental periods, samples were kept at 37°C, to mimic saliva conditions in the body environment. Then, the correlation between the time of degradation of the peptide, chlorhexidine, and their association in the saliva of patients with the different periodontal diagnoses was analyzed. Periodontal ligament cell culture Healthy third molars extracted from patients aged 18 to 30 years were used to obtain periodontal ligament cells. Ethics registration and approval had been obtained from the Human Research Ethics Committee of the Catholic University of Brasília (CAAE: 90666218.2.0000.0029) and all donors signed the understanding and written consent. Right after extraction, extracted teeth were placed in a sterile tube containing Dulbecco Modified Eagle Medium (DMEM; Gibco, Grand Island, New York, USA) without FBS, associated with collagenase type 1 (3 mg.mL -1 ) and dispase (4 mg.mL -1 ), for 1 h at 37 ºC [ 28 ]. After this period, tooth roots were shaved, and tissue structures of the periodontal ligament adhered to the roots were collected in 6-well cell culture plates (Costar Corp., Cambridge, Massachusetts, USA). Cells were cultured in supplemented DMEM culture medium with 10% FBS (Gibco), 1% MEM amino acid solution (Gibco), 0.05% gentamicin (Gibco), 1% L-glutamine (Gibco), 1% penicillin / streptomycin (1000 U.mL -1 ) (Gibco) and kept at 5% CO 2 , 37 ºC and at 95% humidity [ 29 ]. Every three days, the culture medium was changed, until reaching the number of cells required for the experiment. Hemolytic assay The hemolytic activity of synoeca-MP and chlorhexidine was based on the methodology described by Bignami, with adaptations [ 27 ]. Thus, a suspension with 1% cells in PBS was prepared and centrifuged twice (1000 x g, for 2 min). Subsequently, 350 µL of the 1% erythrocyte solution was used to test different concentrations of synoeca-MP and chlorhexidine. Controls were represented by saline solution (0% hemolysis, negative control) and triton X-100 (Sigma-Aldrich) at 0.1% (100% hemolysis, positive control). After 1h, tubes were centrifuged and 100 µL of the supernatant was collected to determine the optical density in a microplate reader at 406 nm. MTT assay Human peripheral blood mononuclear cells (PBMCs), periodontal ligament cells, and SaOs-2 viability were evaluated according to the experimental groups and incubation period of each proposed assay. The positive control was represented by cells in culture medium (100% of cell viability) and the negative control was represented by cells in lysis solution − 10 mM Tris (Sigma-Aldrich), pH 7.4, 1 mM EDTA (Sigma-Aldrich) and 0.1% triton X-100 (Sigma-Aldrich), representing 0% cell viability. At the end of the incubation period, the MTT colorimetric assay (Sigma-Aldrich) was used [ 21 ]. Synoeca-MP and chlorhexidine on cell proliferation and migration assay Cells were plated in 96-well plates (Kasvi, São José dos Pinhais, Paraná, Brazil) and after 24h, synoeca-MP and chlorhexidine stimuli were added. Then, after 24 h, 48 h and 72 h, cell viability was determined by MTT assay. Cell proliferative potential in the presence of tested substances was performed using the Tripan Blue exclusion technique, after 0h, 24h, and 48h of cell incubation [ 30 ]. Confluent periodontal ligament cell cultures were obtained, and their trypsinization and cell resuspension assays were performed in DMEM medium (Gibco) without FBS (same conditions as those used in cell migration assay). Periodontal ligament cells (1x10 5 cells) were added in 1 mL DMEM (Gibco) without FBS to 24-well plates. Experimental groups contained the lowest antimicrobial synergistic concentration of synoeca-MP and chlorhexidine, associated or alone. Then, after completing the experimental incubation, cells were resuspended and the solution was added to 0.4% trypan blue dye (Sigma-Aldrich), for 1 minute. Cells were counted immediately using a Neubauer chamber (Brand GmbH, Wertheim, Baden-Württemberg, Germany). To assess the effect of synoeca-MP, chlorhexidine and their association on cellular proliferation, a scratch assay was performed [ 31 ]. Cells from the periodontal ligament were kept confluent, and through the accomplishment of 3 markings, cell adhesion in the plate was disrupted. After that, the plate was washed with PBS, followed by the addition of supplemented DMEM culture medium, with the addition of all tested substances. Then, after 0 h, 24 h and 48 h, photographs were taken in an inverted electron microscope, with a 10x increase, in order to verify the migratory activity in these periods [ 32 ]. The measurement of cell migration was performed through the analysis of photographs by counting cells using Image J software [ 33 ]. Evaluation of synoeca-MP and chlorhexidine on osteoclastogenesis process Whole blood (about 4 mL) was collected from healthy volunteers. PBMCs were isolated by density gradient centrifugation using Ficoll-Paque (Sigma-Aldrich), following the manufacturer's recommendations. Then, the cell pellet was resuspended in 10 mL of supplemented DMEM (Gibco) and 25 ng.mL -1 of macrophage colony-stimulating factor recombinant (M-CSF; Peprotech, Rocky Hill, Connecticut, USA) [ 34 , 35 ]. After 72h, non-adherent cells were removed, and the adherent cells were plated in 96-well plates (Kasvi). After 24h, synoeca-MP and chlorhexidine stimuli were added. Cellular viability was assessed after 14 days. PBMCs (M-CSF-dependent macrophages) were incubated at 1.6x10 4 cells per well, in 96-well culture plates (Kasvi) with supplemented DMEM (Gibco). Cultures were subjected to stimuli, such as 10 ng.mL -1 of the soluble receptor activator of nuclear factor-κB ligand (sRANKL; Peprotech) and synoeca-MP at its synergistic concentrations with chlorhexidine, associated or alone, for 14 days [ 36 ]. After the fourteenth day, the PBMCs were subjected to tartrate-resistant acid phosphatase (TRAP) staining for later counting of the number of differentiated osteoclasts and the number of nuclei per osteoclast in an inverted microscope. For TRAP staining, the acid phosphatase kit, leukocyte (Sigma-Aldrich) was used, following the manufacturer's recommendations. Osteoclasts were considered TRAP-positive cells (red/orange color), with more than three nuclei inside them [ 37 ]. Evaluation of synoeca-MP and chlorhexidine on mineral matrix formation To evaluate mineral matrix deposition effects, periodontal ligament cells, and SaOs-2 (derived from human osteosarcoma) cells were used [ 38 ]. Periodontal ligament cells and SaOs-2 cells were cultured at a concentration of 1.6x10 4 cells per well, in 6-well culture plates (Kasvi) for 21 days, in supplemented DMEM medium (Gibco), with osteogenic conditions (50 µg.mL -1 ascorbic acid, 100 nM dexamethasone, 1 mM β-glycerophosphate; Sigma-Aldrich), for 21 days. Medium and all stimuli were changed every 3 days. Cultures were subjected to synoeca-MP and chlorhexidine stimuli in synergistic concentrations, after 24h [ 39 , 40 ]. After the incubation period, the concentration of alkaline phosphatase (ALP) and the formation of the mineral matrix were assessed. Alkaline phosphatase. Determination of alkaline phosphatase (ALP) concentration in periodontal ligament and SaOs-2 cell cultures was assessed after 21 days under osteogenic conditions. ALP was measured by the colorimetric method of paranitrophenol, using the Sigmafast p-Nitrophenyl phosphate kit (Sigma-Aldrich). For the ALP assay, the cells were washed with PBS and incubated in 0.05% Triton X-100 for 20 minutes at room temperature, with constant agitation. Cells were transferred, vortexed for 20 seconds, centrifuged for 15 min at 4°C at 2500 RPM, and kept on ice for 20 minutes. Aliquots of cell lysate were incubated with p-Nitrophenol phosphate (p-NF) substrate at 37 ºC for 30 minutes. The reaction was stopped by adding 5 µL 1N NaOH and the absorbance was measured at 405 nm using the microplate reader (Bio-Tek PowerWave HT). A standard p-NF curve was established to determine enzyme activity. The samples were normalized, and protein quantification was determined by Qubit®ฏ [41]. Mineral matrix formation assay. After 21 days of culture, the formation of the mineral matrix of periodontal ligament cells and SaOs-2 cells was determined by alizarin red staining. For this, plates were fixed with 10% (v / v) formaldehyde (VETEC), at room temperature for 15 min. The cell layer was washed twice with distilled water and 1 mL of alizarin red S dye (40 mM; pH 4.1; Sigma-Aldrich) was added per well. The plate was incubated at room temperature for 20 minutes, under constant agitation. After discarding the unincorporated dye, the cell layer was washed 4 times, with 4 mL of distilled water, with a 5-minute stirring between washes. Then, the distilled water was discarded, and the stained cell layer was evaluated under an inverted microscope (Zeiss, Oberkochen, Baden-Württemberg, Germany). To quantify the stained mineral matrix, 800 µL of acetic acid 10% (v/v) (Dinâmica, Indaiatuba, São Paulo, Brazil) was added to each well, followed by incubation for 30 minutes at room temperature, under constant agitation. The cell layer, together with 10% (v/v) acetic acid (Dinâmica), was then scraped from the bottom of the well, vortexing for 30 seconds, covered with 500 µL of mineral oil (VETEC), heated to 85 ºC for 10 minutes and cooled on ice, for 5 minutes. After complete cooling, the suspension was centrifuged at 20,000 x g for 15 minutes and 500 µL of the supernatant transferred to a new tube. For acid neutralization, 200 µL of 10% (v / v) ammonium hydroxide (VETEC) was added. Aliquots (150 µL) of the supernatant were arranged in triplicate in a 96-well plate (Kasvi) for reading in a microplate reader (Bio-Tek PowerWave HT), at 405 nm [ 41 ]. Statistical analysis All experiments were performed in technical and biological triplicates. Data normality was tested by the Shapiro Wilk test, and parametric variables are described as mean and standard deviation or mean and standard error of the mean. Statistically significant differences were considered when p < 0.05. To measure the statistical differences in the continuous variables, the two-way ANOVA was used followed by the Bonferroni post hoc test. The Kaplan-Meier curves were used to show molecule degradation over time (synoeca-MP, chlorhexidine and synoeca-MP plus chlorhexidine), and statistical differences were assessed by Log-Rank (Mantel-Cox) test. Statistical analysis was performed using the GraphPad Prism 5 software (California, USA). Results Antifungal, antibacterial and antibiofilm activity determination Antimicrobial activity of synoeca-MP and chlorhexidine was tested against P. aeruginosa, S. aureus, E. faecalis and C. albicans . Minimal inhibitory concentrations were determined, and all agents were capable of inhibiting yeast and bacteria growth, at synoeca-MP or chlorhexidine concentrations of up to 32 µg.mL -1 , under the assay conditions. However, the minimum bactericidal or fungicidal concentration was up to 64 µg.mL -1 , except when synoeca-MP was evaluated against P. aeruginosa , which did not present MBC until values up to 128 µg.mL -1 (Table 1 ). Further analysis involving antimicrobial properties might increase the potential use of synoeca-MP and chlorhexidine in the oral environment. To this end, synoeca-MP and chlorhexidine were also evaluated for their inhibitory capacity on biofilms created by P. aeruginosa, S. aureus and C. albicans. All tested concentrations of synoeca-MP and chlorhexidine were able to reduce the viability of preformed biofilm of P. aeruginosa . However, synoeca-MP showed the best results at concentrations between 16 and 128 µg.mL -1 (10 to 80 µM), including 50 to 58% of P. aeruginosa biofilm eradication and 128 µg.mL -1 (80 µM) presenting 90% of biofilm eradication of C. albicans and 40% of S. aureus biofilm eradication. On the other hand, chlorhexidine showed its greatest result in the concentration of 128 µg.mL -1 (253 µM), which corresponded to 49% of biofilm reduction against P. aeruginosa and approximately 95% of that of S. aureus . Besides that, 32, 64 and 128 µg.mL -1 (63 to 253 µM) of chlorhexidine can inhibit 80–95% of C. albicans biofilm preformed (Table 1 ). Given the results obtained, we observed the antimicrobial activity as well the activity against pre-formed biofilms by P. aeruginosa , S. aureus , E. faecalis , and C. albicans . In comparative analysis, the peptide synoeca-MP showed an advantage in terms of its microbicidal effects against the colonies of the previously mentioned microorganisms, when compared to chlorhexidine. Thus, an important next point would be to evaluate the possible effects of the interaction of both molecules together. Table 1 Minimal Inhibitory Concentration (MIC), Minimal Bactericidal/Fungicidal Concentration (MBC/MFC) and percentage of Antibiofilm activity of synoeca-MP and chlorhexidine against P. aeruginosa, S. aureus, E. faecalis and C. albicans in vitro . NT* Not tested. Antimicrobial synergistic analysis of synoeca-MP and chlorhexidine Once synoeca-MP and chlorhexidine had demonstrated promising results for antimicrobial activity, antimicrobial synergistic analyses were performed against P. aeruginosa, S. aureus, E. faecalis and C. albicans . Chlorhexidine and synoeca-MP interaction exhibited synergistic antimicrobial activity in different concentrations against P. aeruginosa, S. aureus , and C. albicans (Table 2 ). However, interactions involving synoeca-MP and chlorhexidine against E. faecalis did not demonstrate any synergistic effect. In addition, chlorhexidine association with synoeca-MP reduced 50–99% of MIC concentration when used against P. aeruginosa, S. aureus , and C. albicans . Thus, the synergistic interaction between synoeca-MP and chlorhexidine that demonstrated the best inhibition, at lower concentrations, was 0.125 and 0.25 µg.mL -1 of chlorhexidine and synoeca-MP, respectively, against P. aeruginosa in vitro . The addition of synoeca-MP to chlorhexidine might promote several different effects on the oral environment in inflammatory and infectious conditions. Local and general use of chlorhexidine also encourages the use of this molecule combined with synoeca-MP. Table 2 Synergistic concentrations of synoeca-MP (SYN) and chlorhexidine (CHX). Table with detailed analysis of concentrations was demonstrated according to its combination, fractional inhibitory concentration index (FICI) value and percentage of reduction related to previous inhibitory concentration. Synoeca-MP and chlorhexidine stability in human saliva Once the antimicrobial potential of using chlorhexidine with or without synoeca-MP had been verified, it was necessary to evaluate the stability of its molecules in human saliva. The monoisotopic mass of synoeca-MP, chlorhexidine, matrix control and saliva of all patients was checked to certify their integrity prior to analysis (Supplementary material - Fig. 3 ). For the molecule’s stability in saliva, the last time point at which the ions of the monoisotopic mass of the molecules appeared intact in the spectrum was considered (Supplementary material - Fig. 4 ). Saliva from patients with different periodontal diagnoses were used for these analyses. All data correlating patients' periodontal diagnoses and the time of degradation of all molecules were found in Supplementary table 1 . It was found that the time of degradation of synoeca-MP (Fig. 1 A) and the association of synoeca-MP and chlorhexidine (Fig. 1 C) changed according to the periodontal diagnosis. The degradation time of both molecules was inversely proportional to the severity of periodontal disease; that is, the more severe the periodontal disease, the less time the compounds take to be degraded. On the other hand, chlorhexidine remained stable throughout all evaluated time points (until 240 min) (Fig. 1 B). These results are favorable for a proposal to use these molecules as mouthwashes in the oral cavity. Effect of synoeca-MP, chlorhexidine or both on periodontal ligament cells Once the molecules had shown good integrity in saliva, the next step was to evaluate their effect on periodontal ligament cells. Cell viability of periodontal ligament with stimuli of 0.25 µg.mL -1 (0.16 µM) synoeca-MP and/or 0.125 µg.mL -1 (0.25 µM) chlorhexidine was performed on periodontal ligament cells. No cytotoxicity was observed after 24 h, 48 h, and 72 h, demonstrating security for human periodontal ligament cells. Then, to evaluate the molecules’ possible effects on tissue healing and regeneration, cell cultures of periodontal ligament were subjected to cell proliferation and migration assays. Cultures were stimulated with 0.25 µg.mL -1 (0.16 µM) synoeca-MP and/or 0.125 µg.mL -1 (0.25 µM) chlorhexidine for 24h (Fig. 2 A and C) and 48h (Fig. 2 B and D). The proliferative potential was increased in the presence of synoeca-MP and the synergistic combination of synoeca-MP and chlorhexidine in both experimental periods (p < 0.0001 – Fig. 2 A and B). These results were also similar to the positive control group (periodontal ligament cell in 2% FBS). However, the presence of chlorhexidine led to a similar proliferation to periodontal ligament cells alone, after 24h and 48h. Cell migration is one of the main events during tissue repair and the wound healing process. Thus, periodontal ligament cultures were stimulated with 0.25 µg.mL -1 (0.16 µM) synoeca-MP and/or 0.125 µg.mL -1 (0.25 µM) chlorhexidine for 0h (Fig. 2 E, F, G, H and I), 24 h (Fig. 2 J, K, L, M and N) and 48 h (Fig. 2 O, P, Q, R, and S). It was observed that all tested conditions presented an increase in periodontal ligament cell migration after 24 h, compared to periodontal ligament cells alone (p < 0.0001 - Fig. 2 C). However, after 4h there was an increase in periodontal ligament cell migration in cultures exposed to the synergistic association of chlorhexidine and synoeca-MP (p < 0.0001 - Fig. 2 D and S). It can be inferred that, when comparing synoeca-MP and chlorhexidine, both showed no cytotoxicity. Both synoeca-MP and the synergistic combination of synoeca-MP and chlorhexidine demonstrated increased cell proliferation and migration. These findings are highly valuable for use in the tissue regeneration process. Effect of synoeca-MP, chlorhexidine or both on the osteoclastogenesis process Frist, it was found that none of tested groups showed hemolytic activity. Low concentrations of synoeca-MP and chlorhexidine (0.25 µg.mL -1 (0.16 µM) synoeca-MP and/or 0.125 µg.mL -1 (0.25 µM) chlorhexidine), separated or associated, did not present cytotoxicity to human erythrocytes (Supplementary material - Fig. 1 ). This fact was encouraging for the analysis of the effect of these molecules during the osteoclastogenesis processes. Synoeca-MP and chlorhexidine cytotoxicity was evaluated at synergistic concentrations in peripheral blood mononuclear cells (PBMCs), as well as for its potential for inhibiting osteoclastogenesis. It was shown that cell viability after 14 days remained stable in all tested conditions (Fig. 3 A), demonstrating no cytotoxicity to human PBMCs. It was found that the association between synoeca-MP peptide and chlorhexidine showed the greatest inhibition of the osteoclastogenesis process, reducing it by 86.65% compared to the control group. However, separately, synoeca-MP and chlorhexidine were also able to inhibit osteoclastogenesis by 76.12% and 63.7% respectively, when compared to the sRANKL-stimulated cells (p > 0.0001 - Fig. 3 B). Analyzing the ratio, nuclei/osteoclast, an average of 9.6 nuclei per osteoclast were observed in the sRANKL-stimulated cells, while synoeca-MP, chlorhexidine, and their association showed an average of 3.63, 5.3, and 3.58 nuclei per osteoclast, respectively (Fig. 3 C), presenting a lower osteoclast size with a diminished number of nuclei. In qualitative analysis, it was possible to notice that the multinucleated osteoclasts in sRANKL-stimulated cells were relatively larger compared to other tested groups (Fig. 3 E). The chlorhexidine-stimulated group (Fig. 3 G), synodical-MP-stimulated group (Fig. 3 F) and synoeca-MP with chlorhexidine-stimulated group (Fig. 3 H) presented osteoclasts smaller in size and with similar characteristics to PBMCs (Fig. 3 D), corroborating the results of osteoclast count and number of nuclei per osteoclast. Therefore, these findings are encouraging for the application of these molecules in bone reabsorption process, since the absence of cytotoxicity of both chlorhexidine and Synoeca-MP, both combined and individually, for human cells was demonstrated, something essential for the safety of drugs applied to health. Furthermore, it was found that these molecules, when combined, demonstrated greater efficacy in the osteoclastogenesis downregulation process, including a reduction in the number of nuclei and the size of osteoclasts. Thus, the use of these molecules may be promising for the bone remodeling process. Effect of synoeca-MP, chlorhexidine or both on the mineralized matrix formation Given the promising results of the effect of these molecules during osteoclastogenesis, we began to evaluate the effects of these molecules on the formation of the mineral matrix. To promote the mineralization process, periodontal ligament cells and SaOs-2 cells, derived from human osteosarcoma, were cultured under osteogenic conditions, allowing for osteoblast differentiation (Fig. 4 ). The cell viability of periodontal ligament and SaOs-2 cell cultures was analyzed under osteogenic conditions and stimulated with 0.25 µg.mL -1 (0.16 µM) synoeca-MP and/or 0.125 µg.mL -1 (0.25 µM) chlorhexidine for 24h, and the other analyses then went on to determine the potential of mineralized matrix formation (Fig. 4 A and B). There was no reduction in cell viability with any tested sample when periodontal ligament (Fig. 4 A) and SaOs-2 (Fig. 4 B) cells were stimulated, under osteogenic conditions. To evaluate the effect of synoeca-MP and chlorhexidine on the mineralization process, periodontal ligament, and SaOs-2 cells were cultured under osteogenic conditions for 21 days, and the mineral matrix formation assay was performed by Alizarin red staining. The mineralization potential in periodontal ligament cells (Fig. 4 C) and SaOs-2 cells (Fig. 4 D) stimulated with synoeca-MP and / or chlorhexidine was observed (p < 0.001). However, periodontal ligament cells stimulated with chlorhexidine showed the greatest potential for matrix mineralization, compared to other tested groups (p < 0.0001 - Fig. 4 C and 4C3). Qualitative analysis of mineral matrix formation in cultures of periodontal ligament cells showed the highest intensity of alizarin red staining in the presence of synoeca-MP (Fig. 4 C2), chlorhexidine (Fig. 4 C3) and synoeca-MP and chlorhexidine-associated (Fig. 4 C4), compared to cells in osteogenic medium alone (p < 0.001) (Fig. 4 C1). Similarly, a qualitative analysis of mineral matrix formation based on SaOs-2 cells showed greater intensity of alizarin red staining in cultures stimulated with synoeca-MP (Fig. 4 D2), chlorhexidine (Fig. 4 D3) and synoeca-MP and chlorhexidine associated (Fig. 4 D4), compared to cells in osteogenic medium alone (p < 0.001) (Fig. 4 D1). The alkaline phosphatase production in cultures of periodontal ligament cells, after 21 days, showed no statistically significant differences between any of the tested groups (Fig. 4 E). Additionally, the results showed that synoeca-MP and chlorhexidine did not reduce cell viability, and they promoted the formation of mineral matrix in periodontal ligament cells and SaOs-2 cells. As previously seen, the combination of Synoeca-MP and chlorhexidine also demonstrated efficacy in inhibiting the osteoclastogenesis process. Although synoeca-MP and chlorhexidine showed mineralization potential, periodontal ligament cells stimulated with chlorhexidine exhibited the greatest potential for mineralizing the matrix. Qualitative analyses confirmed these results, showing intense red alizarin staining in the stimulated cultures. Alkaline phosphatase production, an indicator of osteoblastic differentiation, did not significantly differ between the groups. These findings suggest that synoeca-MP and chlorhexidine may play an important role in tissue regeneration, especially in mineralization processes. Discussion The maintenance of oral health in hospitalized patients, who may be immunosuppressed and in precarious oral hygiene conditions, is a major challenge within dentistry [ 1 , 2 ]. In this sense, the oral cavity is an environment prone to the colonization of opportunistic pathogenic microorganisms such as P. aeruginosa, S. aureus, E. faecalis , and C. albicans , which can increase the susceptibility to infections and more serious problems [ 3 ]. Therefore, methods for oral microbial control of hospitalized patients are necessary. Chlorhexidine 0.12% is widely used in the hospital setting to control oral biofilm [ 9 ]. However, prolonged use of this product can result in adverse effects [ 11 ]. In recent decades, the search for new antimicrobial molecules has sparked researchers' interest in antimicrobial peptides [ 12 ]. Consequently, previous studies have isolated several peptides from wasps, which were then classified by their chemical structures and biological activities. Among them, synoeca-MP, derived from the venom of the wasp Synoeca surinama , has shown significant antimicrobial and antibiofilm results against P. aeruginosa and other Gram-positive and Gram-negative strains [ 14 ]. Thus, it may be a promising alternative for controlling pathogenic microorganisms present in the oral microbiota of hospitalized patients. Furthermore, synoeca-MP exhibits synergistic antimicrobial activity when combined with chlorhexidine, allowing for the reduction of the concentration of both molecules to achieve the desired antimicrobial activity. This fact may favor the reduction of undesirable effects of isolated chlorhexidine [ 15 ]. In this scenario, the present study brought new knowledge about the association between the synoeca-MP peptide and chlorhexidine. Herein, its antimicrobial and antibiofilm effect, its stability in human saliva, cytotoxicity, regenerative action, and bone homeostasis in vitro were analyzed. Initially, the antifungal, antibacterial, and antibiofilm activity of synoeca-MP and chlorhexidine against the opportunistic microorganisms involved in oral pathology was determined. The results obtained demonstrate the effectiveness of synoeca-MP and chlorhexidine in inhibiting the growth of P. aeruginosa, S. aureus, E. faecalis , and C. albicans , with minimum inhibitory concentrations of up to 32 µg.mL -1 . Both substances reduced the viability of biofilms formed by these microorganisms, with synoeca-MP demonstrating a more effective eradication of P. aeruginosa and C. albicans biofilms compared to chlorhexidine. The comparative analysis of both molecules suggests that the peptide synoeca-MP may be an encouraging alternative to chlorhexidine in the treatment of oral infections, highlighting the need to investigate a possible synergistic activity between these compounds to develop effective antimicrobial therapeutic strategies. Therefore, this study also verified the potential synergistic antimicrobial activity of these molecules against P. aeruginosa, S. aureus, C. albicans , and E. faecalis [ 42 ]. In this regard, we confirmed the synergism between chlorhexidine and the peptide synoeca-MP against pre-formed biofilms of P. aeruginosa, S. aureus , and C. albicans , but they did not demonstrate any synergistic effect against E. faecalis . The interaction assays exhibited different concentrations of synergistic antimicrobial activity, reducing from 50–99% of the MIC of each molecule in this combination. Additionally, an evaluation of the stability of both chlorhexidine and synoeca-MP molecules in human saliva was necessary, considering the perspective of a possible future mouthwash indicated for patients with different oral conditions. Therefore, our work also investigated the integrity in human saliva after the isolated or combined use of these substances, which revealed fundamental data for their clinical applicability. In this regard, the monoisotopic analysis of these compounds in the saliva of patients with different periodontal diagnoses indicated a relationship between the stability of the molecules and the severity of periodontal disease. It was observed that the greater the periodontal involvement of the individuals, the more rapidly the molecules were degraded. However, chlorhexidine alone maintained the highest degree of stability in the saliva of patients with gingivitis and periodontitis. Furthermore, the combination of synoeca-MP and chlorhexidine was able to increase the stability time of synoeca-MP alone. Such results suggest that chlorhexidine, when combined with synoeca-MP, may be a promising option regarding the development of a new mouthwash. This finding significantly contributes to understanding the stability dynamics of these molecules in human saliva and may guide future therapeutic strategies for the treatment of inflammatory and infectious oral conditions. After infections stemming from oral pathologies, damage and other problems related to tissue integrity can occur. Thus, cell migration is one of the main events during the tissue repair and wound healing process [ 43 ]. In this regard, periodontal ligament cells were exposed to the combination of synoeca-MP and chlorhexidine for the evaluation of cell proliferation and migration. The results obtained demonstrated that the combination of synoeca-MP and chlorhexidine increased cell migration and proliferation at 24h and 48h, with better wound closure after 48h. In addition to synoeca-MP, other host defense peptides have been reported in the literature with regenerative potential. In this context, IDR-1018 has shown promising regenerative properties using 2D and 3D human skin equivalents of fibroblasts, primary keratinocytes, and MeWo melanocyte cell lines [ 43 ]. Furthermore, the antimicrobial peptide LL-37 can promote odonto/osteogenic migration and differentiation of stem cells from the apical papilla by activating the Akt/Wnt/β-catenin signaling pathway [ 44 ]. Therefore, like the other peptides mentioned, synoeca-MP has great potential regarding future clinical applicability, aiming at tissue repair in patients with compromised oral conditions. Moreover, to assess the potential of the combination of synoeca-MP and chlorhexidine, osteoclastogenesis and mineralized matrix formation processes were analyzed. Data obtained demonstrated that the association between synoeca-MP and chlorhexidine showed greater inhibition of the osteoclastogenesis process in vitro . Moreover, the association between both molecules was able to reduce the size and number of nuclei per cell in the few differentiated osteoclasts [ 45 ]. The size and number of nuclei per osteoclast can affect the metabolic activity of osteoclasts. Based on these results, we hypothesize that the increase in metabolic activity may be related to the increase in energy and cell mobility, which can increase areas of bone resorption. Therefore, chlorhexidine and synoeca-MP, when combined, may have promising potential for the treatment of bone resorption. Considering that bone homeostasis results from the balance between bone resorption and deposition, mineral matrix formation is a fundamental process [ 46 ]. Thus, our study evaluated periodontal ligament cells and SaOs-2 cells stimulated with synoeca-MP and/or chlorhexidine, which showed great potential for mineralized matrix formation in the presence of these molecules. However, isolated stimulation with chlorhexidine favored greater formation of mineralized matrix in periodontal ligament cells. Although no damage to osteoblasts and mineralization was observed, probably due to the low concentration of the tested molecules, some studies suggest that chlorhexidine may be harmful to osteoblasts [ 47 , 48 ]. Furthermore, another study concluded that although chlorhexidine has a strong antibacterial capacity against E. faecalis , this molecule is ineffective in promoting mineralization of tissues around the root apex. These findings suggest that synoeca-MP and chlorhexidine may play an important role in tissue regeneration, especially in mineralization processes [ 46 ]. Conclusion Based on the results obtained, it can be concluded that the association of synoeca-MP and chlorhexidine can be applicable to oral diseases. To assess this, both were evaluated for their antimicrobial activity against microorganisms in planktonic and biofilm states, regenerative potential, stability in saliva, and bone homeostasis activity. Overall, the combination of synoeca-MP and chlorhexidine might be effective primarily in cases of opportunistic infections in the oral cavity and could also be associated with mechanical therapy. These molecules with different actions could potentially optimize dental treatments, as they demonstrated effective activity at low concentrations. However, it is important to emphasize that this is a preliminary in vitro study, and further in vivo and clinical analyses must be conducted to confirm these findings. Declarations Author Contribution The present contribution of the study author: All authors contributed to the conception and design of the study. Material preparation, data collection and analysis were carried out by Ingrid A. R. Barin, Johnny C. Silva, Raquel F. Ramos, Stella M. F. Lima, Ana P C. Cantuária, Poliana A. O. Silva, Elaine M. G. L. Dantas, Danilo C. M. Martins, Nelson G. O. Júnior, Osmel F. Martinez, Jesser A. Almeida, Marcelo H. S. Ramada, Octávio L. Franco and Taia M. B. Rezende. The first version of the manuscript was written by Ingrid A. R. Barin, Johnny C. Silva and Raquel F. Ramos, and all authors commented on previous versions of the manuscript. All figures and tables were created by Ingrid A. R. Barin, Johnny C. Silva and Raquel F. Ramos. Bibliographic analysis and corrections were carried out by Stella M. F. Limas and data analysis with the critical resistance of the work was carried out by Taia M. B. Rezende. All authors read and approved the final manuscript. Declarations Section - The present contribution of the study author: The present contribution of the study author: All authors contributed to the conception and design of the study. Material preparation, data collection and analysis were carried out by Ingrid A. R. Barin, Johnny C. Silva, Raquel F. Ramos, Stella M. F. Lima, Ana P C. Cantuária, Poliana A. O. Silva, Elaine M. G. L. Dantas, Danilo C. M. Martins, Nelson G. O. Júnior, Osmel F. Martinez, Jesser A. Almeida, Marcelo H. S. Ramada, Octávio L. Franco and Taia M. B. Rezende. The first version of the manuscript was written by Ingrid A. R. Barin, Johnny C. Silva and Raquel F. Ramos, and all authors commented on previous versions of the manuscript. All figures and tables were created by Ingrid A. R. Barin, Johnny C. Silva and Raquel F. Ramos. Bibliographic analysis and corrections were carried out by Stella M. F. Limas and data analysis with the critical resistance of the work was carried out by Taia M. B. Rezende. All authors read and approved the final manuscript. - Conflict of interest: Not applicable. - Funding: This study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (305242/2022-9), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) (88887.910215/2023-00 and 88887724415/2022-00) and Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF) (00193–00000782/2021-63; 0009-0004-4942-2105 and 00193-00001118/2021-31). - Ethical approval: 90666218.2.0000.0029 - Consent statements: Not applicable. Acknowledgments: This study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (305242/2022-9), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) (88887.910215/2023-00 and 88887724415/2022-00) and Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF) (00193–00000782/2021-63; 0009-0004-4942-2105 and 00193-00001118/2021-31). References Gershonovitch R, Yarom N, Findler M. 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Jul 2015;33(4):359-70. doi:10.1007/s00774-015-0656-4 Liu J, Zhang B, Song S, et al. Bovine collagen peptides compounds promote the proliferation and differentiation of MC3T3-E1 pre-osteoblasts. PloS one. 2014;9(6):e99920. doi:10.1371/journal.pone.0099920 Proksch S, Strobel SL, Vach K, et al. Melatonin as a candidate therapeutic drug for protecting bone cells from chlorhexidine-induced damage. Journal of periodontology. Dec 2014;85(12):e379-89. doi:10.1902/jop.2014.140279 Patel P, Ide M, Coward P, Di Silvio L. The effect of a commercially available chlorhexidine mouthwash product on human osteoblast cells. The European journal of prosthodontics and restorative dentistry. Jun 2006;14(2):67-72. Additional Declarations No competing interests reported. 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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-4595835","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":324802328,"identity":"65d77f1d-b666-43ea-8075-74096f710a4c","order_by":0,"name":"Ingrid Aquino Reichert Barin","email":"","orcid":"","institution":"Universidade Católica de Brasília","correspondingAuthor":false,"prefix":"","firstName":"Ingrid","middleName":"Aquino Reichert","lastName":"Barin","suffix":""},{"id":324802329,"identity":"1868c713-4bc1-4288-915e-0b3d61a8881c","order_by":1,"name":"Johnny Carvalho da Silva","email":"","orcid":"","institution":"University of Brasília","correspondingAuthor":false,"prefix":"","firstName":"Johnny","middleName":"Carvalho da","lastName":"Silva","suffix":""},{"id":324802330,"identity":"7dd70102-fe6b-46ce-9afe-e4f1f7aa4755","order_by":2,"name":"Raquel Figuerêdo Ramos","email":"","orcid":"","institution":"University of Brasília","correspondingAuthor":false,"prefix":"","firstName":"Raquel","middleName":"Figuerêdo","lastName":"Ramos","suffix":""},{"id":324802331,"identity":"1e7a6fbc-3435-4723-8bdc-0bb29eae8084","order_by":3,"name":"Stella Maris de Freitas Lima","email":"","orcid":"","institution":"Universidade Católica de Brasília","correspondingAuthor":false,"prefix":"","firstName":"Stella","middleName":"Maris de Freitas","lastName":"Lima","suffix":""},{"id":324802332,"identity":"c1c857a8-30ba-4bfe-943d-5eb26d140327","order_by":4,"name":"Ana Paula de Castro Cantuária","email":"","orcid":"","institution":"University of Brasília","correspondingAuthor":false,"prefix":"","firstName":"Ana","middleName":"Paula de Castro","lastName":"Cantuária","suffix":""},{"id":324802333,"identity":"c763538f-7b15-4d0c-a9aa-8df21df6a72c","order_by":5,"name":"Poliana Amanda Oliveira Silva","email":"","orcid":"","institution":"University of Brasília","correspondingAuthor":false,"prefix":"","firstName":"Poliana","middleName":"Amanda Oliveira","lastName":"Silva","suffix":""},{"id":324802334,"identity":"ca394359-7d81-4f5c-904c-6ad7b7546e70","order_by":6,"name":"Elaine Maria Guará Lôbo Dantas","email":"","orcid":"","institution":"Universidade Católica de Brasília","correspondingAuthor":false,"prefix":"","firstName":"Elaine","middleName":"Maria Guará Lôbo","lastName":"Dantas","suffix":""},{"id":324802335,"identity":"402bf1e1-0587-48c3-a4a4-e4ea704e1502","order_by":7,"name":"Danilo César Mota Martins","email":"","orcid":"","institution":"University of Brasília","correspondingAuthor":false,"prefix":"","firstName":"Danilo","middleName":"César Mota","lastName":"Martins","suffix":""},{"id":324802336,"identity":"deb99e42-96f5-498c-adf0-c2566f94deb5","order_by":8,"name":"Nelson Gomes de Oliveira Júnior","email":"","orcid":"","institution":"Universidade Católica de Brasília","correspondingAuthor":false,"prefix":"","firstName":"Nelson","middleName":"Gomes de Oliveira","lastName":"Júnior","suffix":""},{"id":324802337,"identity":"d648e866-a88e-415e-b018-8d223dd935a5","order_by":9,"name":"Osmel Fleitas Martinez","email":"","orcid":"","institution":"Universidade Católica de Brasília","correspondingAuthor":false,"prefix":"","firstName":"Osmel","middleName":"Fleitas","lastName":"Martinez","suffix":""},{"id":324802338,"identity":"65ef0b53-4f62-4bef-9ad5-3279c412eef5","order_by":10,"name":"Jeeser Alves de Almeida","email":"","orcid":"","institution":"Federal University of Mato Grosso do Sul","correspondingAuthor":false,"prefix":"","firstName":"Jeeser","middleName":"Alves","lastName":"de Almeida","suffix":""},{"id":324802339,"identity":"e490e5e4-51ee-4639-a1f3-11b94f451118","order_by":11,"name":"Marcelo Henrique Soller Ramada","email":"","orcid":"","institution":"Universidade Católica de Brasília","correspondingAuthor":false,"prefix":"","firstName":"Marcelo","middleName":"Henrique Soller","lastName":"Ramada","suffix":""},{"id":324802340,"identity":"531cf62e-a6a9-4916-ae41-e7d741dfa077","order_by":12,"name":"Octávio Luiz Franco","email":"","orcid":"","institution":"Universidade Católica de Brasília","correspondingAuthor":false,"prefix":"","firstName":"Octávio","middleName":"Luiz","lastName":"Franco","suffix":""},{"id":324802341,"identity":"817e6596-87ca-4680-9304-658896d43896","order_by":13,"name":"Taia Maria Berto Rezende","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAwElEQVRIiWNgGAWjYFACxgYQkgMxDzwgRgMPVIsxWEsCcVogFiU2gBhEabFnP9z46eaOe+nzww4/BNpiJ6fbQMgWnsRm6dwzxbkbb6cZALUkG5sdIOiwxAbp3LaE3I2zE0BaDiRuI6iF/2Hzb6CWdMPZ6R+I1CKR2AayJUFeOodYW248bLPOPZNguEE6p+BAggERfmHvT398O3dHgrz87PTNHz5U2MkR1AIHBmCVBsQqBwH5BlJUj4JRMApGwYgCAGqyR0DIFeUqAAAAAElFTkSuQmCC","orcid":"","institution":"University of Brasília","correspondingAuthor":true,"prefix":"","firstName":"Taia","middleName":"Maria Berto","lastName":"Rezende","suffix":""}],"badges":[],"createdAt":"2024-06-17 19:23:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4595835/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4595835/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":60598764,"identity":"efc3b76d-2b21-4d8d-a563-b9b24160009c","added_by":"auto","created_at":"2024-07-18 15:55:27","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":185771,"visible":true,"origin":"","legend":"\u003cp\u003eSynoeca-MP and/or chlorhexidine stability in human saliva of different patients with periodontal diagnosis. Kaplan-Meier survival analysis between 0 and 240 min, demonstrating the stability of synoeca-MP (A), chlorhexidine (B) and association of synoeca-MP with chlorhexidine (C) in human saliva, from patients with different periodontal conditions [healthy (green), gingivitis (yellow), periodontitis (red)]. The Log-Rank (Mantel-Cox) showed statistically significant difference in the degradation of synoeca-MP (p \u0026lt; 0.0001) (A), chlorhexidine (p = 0.4529) (B) and association of synoeca-MP with chlorhexidine (p \u0026lt; 0.0001) (C) between all different periodontal conditions.\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-4595835/v1/817b6f2e2f949ea241bd9da3.png"},{"id":60600933,"identity":"320e8852-6608-40cb-a82b-ca072745f2dd","added_by":"auto","created_at":"2024-07-18 16:03:26","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1315111,"visible":true,"origin":"","legend":"\u003cp\u003eCell migration and proliferation of periodontal ligament cells in the presence of FBS 2% (positive control), synoeca-MP peptide and/or chlorhexidine. Yellow spots represent marked migrated cells. Cell proliferation and migration of periodontal ligament cells in the presence of FBS 2% (positive control), synoeca-MP peptide and/or chlorhexidine, at 24h (A, C) and 48h (B, D). Experiments were carried out in technical and biological triplicate. Results were expressed in standard error of the mean. * p \u0026lt; 0.0001, by one-way ANOVA test and Bonferroni post-hoc. Representative images of cell migration of periodontal ligament cells (E, J, O) in the presence of FBS 2% (positive control – F, K, P), synoeca-MP peptide (G, L, Q), chlorhexidine (H, M, R) and association of synoeca-MP peptide with chlorhexidine (I, N, S), at 0h (E, F, G, H, I), 24h (J, K, L, M, N) and 48h (O, P, Q, R, S).\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-4595835/v1/c036424c5d94114b5f68271e.png"},{"id":60598761,"identity":"95e13e20-d676-4355-963a-c1d997e09e43","added_by":"auto","created_at":"2024-07-18 15:55:26","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1380010,"visible":true,"origin":"","legend":"\u003cp\u003eSynoeca-MP peptide and/or chlorhexidine effects on osteoclast differentiation. (A) Cell viability of PBMCs submitted to stimuli of 25 ng.mL\u003csup\u003e-1\u003c/sup\u003e of M-CSF associated with synoeca-MP peptide and/or chlorhexidine, with and without 10 ng.mL\u003csup\u003e-1\u003c/sup\u003e stimulus of sRANKL. (B) Number of TRAP-positive cells differentiated and (C) number of nuclei/osteoclast in the presence of peptide synoeca-MP and/or chlorhexidine, submitted to stimuli of 25 ng.mL\u003csup\u003e-1\u003c/sup\u003e of M-CSF with a stimuli of 10 ng.mL\u003csup\u003e-1\u003c/sup\u003e of sRANKL. (D) PBMCs cells (negative control), (E) PBMCs stimulated with 10 ng.mL\u003csup\u003e-1\u003c/sup\u003e of sRANKL (positive control), (F) PBMCs stimulated with 0.25 μg.mL\u003csup\u003e-1\u003c/sup\u003e of synoeca-MP and 10 ng.mL\u003csup\u003e-1\u003c/sup\u003e of sRANKL, (G) PBMCs stimulated with 0.125 μg.mL\u003csup\u003e-1\u003c/sup\u003e of chlorhexidine and 10 ng.mL\u003csup\u003e-1\u003c/sup\u003e of sRANKL and (H) PBMCs stimulated with 0.25 μg.mL\u003csup\u003e-1\u003c/sup\u003e of synoeca-MP, 0.125 μg.mL\u003csup\u003e-1\u003c/sup\u003e of chlorhexidine and 10 ng.mL\u003csup\u003e-1\u003c/sup\u003e of sRANKL. Experiments were carried out in technical and biological triplicate. * p \u0026lt; 0.0001, comparing to control, by one-way ANOVA test and Bonferroni post-hoc. # p\u0026gt; 0.0001, comparing to chlorhexidine, by one-way ANOVA test and Bonferroni post-hoc.\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-4595835/v1/6f952f5afeeffe4f38bffc46.png"},{"id":60598763,"identity":"4c58a1c4-12c3-40e7-bc21-cba706df22d6","added_by":"auto","created_at":"2024-07-18 15:55:27","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":584017,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of the peptide synoeca-MP and/or chlorhexidine on mineral matrix formation in periodontal ligament and SaOs-2 cells. Cell viability of periodontal ligament (A) and SaOs-2 (B) in the presence of synoeca-MP peptide and/or chlorhexidine, at 24h. Quantitative analysis of mineralized matrix formation by alizarin red stain in periodontal ligament (C) and Saos-2 (D), after 21 days. Representative images of alizarin red stain in cultures of periodontal ligament (C1-C4) and SaOs-2 (D1-D4) with osteogenic conditions. Periodontal ligament cells or SaOs-2 in the presence of osteogenic conditions were considered positive control. Positive control of periodontal ligament (C1) and SaOs-2 (D1), stimulated with synoeca-MP on periodontal ligament (C2) and on SaOs-2 (D2), stimulated with chlorhexidine on periodontal ligament (C3) and on SaOs-2 (D3) or stimulated with synoeca-MP and chlorhexidine association on periodontal ligament (C4) and on SaOs-2 (D4). Alkaline phosphatase production of periodontal ligament cells (E), after 21 days, in the presence of osteogenic conditions were also stimulated with synoeca-MP peptide and/or chlorhexidine. Experiments were carried out in technical and biological triplicate. Results were expressed in standard error of the mean. * p \u0026lt; 0.0001, by one-way ANOVA test and Bonferroni post-hoc.\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-4595835/v1/610dbc54078021e8bc99edbf.png"},{"id":60602848,"identity":"7873f02e-ef60-454b-a856-9ce1a0a159cd","added_by":"auto","created_at":"2024-07-18 16:19:31","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4567993,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4595835/v1/d8a25cfd-3b8c-4404-b66e-4706976dfebd.pdf"},{"id":60598758,"identity":"8b5d23c3-e0bc-425c-9a17-e786fe6d5f98","added_by":"auto","created_at":"2024-07-18 15:55:26","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1547780,"visible":true,"origin":"","legend":"","description":"","filename":"GraphicalAbstract.png","url":"https://assets-eu.researchsquare.com/files/rs-4595835/v1/7521d2f8ed0d4d41e9fc0b1f.png"},{"id":60598760,"identity":"ec3b2f16-b4a1-465f-8b68-0e469a86fa0b","added_by":"auto","created_at":"2024-07-18 15:55:26","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":886244,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryInformationSIBarinetal.docx","url":"https://assets-eu.researchsquare.com/files/rs-4595835/v1/faa61d68744998ed81c2f860.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Antibiofilm, regenerative and bone homeostasis potential of the synergistic association of synoeca-MP peptide with chlorhexidine in oral cavity opportunistic infections","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe control of bacterial biofilm from the oral cavity and respiratory support equipment can avoid nosocomial pneumonia infections in hospital settings [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. It is known that patients admitted to intensive care units (ICU), subject to low immunity, may have respiratory infections, most often caused by nosocomial pneumonia and responsible for up to 50% of deaths in the ICU environment, making them a worldwide public health problem [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Oral immunoinflammatory imbalance can alter oral microbiota, leading to the emergence of opportunistic microbial species [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e], such as \u003cem\u003ePseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus. aureus and Candida albicans\u003c/em\u003e. In this context, the ecological diversity of the oral and periodontal microenvironment may provide suitable conditions for colonization by microorganisms that are not usually considered members of the oral microbiota in subgingival areas [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Such microorganisms have the ability to organize themselves in biofilm and to adhere to dental calculus [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe standard treatment for opportunistic oral infections consists of mechanical microorganisms and biofilm removal [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. It is important to evaluate anatomic involvement and the management of any underlying comorbidities and, if necessary, antibiotic, antifungal, antiviral, or antiparasitic adjuvant drugs can be used [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. However, most conventional antibiotics are uneffective at clearing biofilm-associated infections [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. In a clinical and hospital context, where patients maintain relatively poor oral hygiene conditions, the use of 0.12% chlorhexidine is recommended for oral biofilm control [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. This form of control especially occurs in the presence of ventilation equipment, motor difficulties normally in cases in which patients are in an unconscious or sedated state [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Among the most common adverse effects, the following stand out tooth pigmentation, altered taste perception, increased formation of dental calculus, dry mouth, soft tissue burns, and edema of the lips and glands [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFor this reason, there is a need to develop new agents with antimicrobial, immunomodulatory, and antibiofilm action. Host defense peptides (HDPs) are a class of biomolecules that can have both antimicrobial acivity against bacteria, viruses and fungi, as well as performing important immunomodulatory functions in angiogenesis, wound healing, and chemotaxis [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. In addition, it has been shown that some peptides increase antibiotic action to prevent biofilm formation and eradicate mature biofilms [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. One example is the synergistic combination of peptide IDR 1018 with low levels of the antibiotics ceftazidime, ciprofloxacin, imipenem, or tobramycin, which was able to reduce the concentration of antibiotic required to eradicate biofilms [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn this condition, the synoeca-MP peptide, derived from the venom of \u003cem\u003eSynoeca surinama\u003c/em\u003e, a species of South American wasp, has demonstrated important antimicrobial results against Gram-positive and -negative strains. Synoeca-MP is composed of 14 amino acid residues and is effective mainly against antibiotic-resistant bacteria, such as \u003cem\u003eS. aureus, Escherichia coli\u003c/em\u003e and \u003cem\u003eE. faecalis\u003c/em\u003e [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Likewise, previous studies have also shown this peptide\u0026rsquo;s synergistic antimicrobial activity against \u003cem\u003eP. aeruginosa\u003c/em\u003e and its antibiofilm effect, as well as immunomodulatory activity when associated with chlorhexidine, at low concentrations of both. This association can favor the reduction of undesirable effects of chlorhexidine used alone [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eHowever, there is little information regarding this association [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Therefore, the present study aimed to analyze the association between synoeca-MP peptide and chlorhexidine, regarding its antimicrobial, antibiofilm activity, and its stability in saliva. In addition, its effect on tissue repair, in bone resorption processes, and mineralized matrix formation \u003cem\u003ein vitro\u003c/em\u003e were also analyzed. The hypothesis was that the association between synoeca-MP peptide and chlorhexidine [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] is capable of modulating bone homeostasis, inhibiting osteoclastogenesis, and increasing the formation of the mineral matrix \u003cem\u003ein vitro.\u003c/em\u003e Moreover, it may have repair potential, given the conditions of tissue destruction caused by opportunistic microorganisms, as well as periodontopathogens of the oral cavity and the periodontal microenvironment [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e"},{"header":"Material and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eSynoeca-MP peptide and chlorhexidine digluconate preparation\u003c/h2\u003e \u003cp\u003eSynoeca-MP peptide (INWLKLGKKIIASL-NH\u003csub\u003e2\u003c/sub\u003e) was synthesized in solid-phase using F-moc methodology, purified (purity\u0026thinsp;\u0026gt;\u0026thinsp;95%), lyophilized and stored by AminoTech (S\u0026atilde;o Paulo, Brazil) [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. To confirm molecular mass and purity, Matrix Assisted Laser Desorption Ionization - Time of Flight (MALDI-ToF) mass spectrometry was used (Supplementary Fig.\u0026nbsp;1). The peptide was dissolved in ultrapure water and stored at -20\u0026deg;C until use. Chlorhexidine was manipulated for stock solution (20% concentrated) (Via Magistral, Bras\u0026iacute;lia, Distrito Federal, Brazil) and dissolved in ultrapure water [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eDetermination of minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC) and antibiofilm concentration\u003c/h2\u003e \u003cp\u003eBacteriostatic and bactericidal concentrations were determined according to Clinical \u0026amp; Laboratory Standards Institute guidelines, with some adaptations. Analyses were performed to determine their effectiveness against \u003cem\u003eP. aeruginosa\u003c/em\u003e (ATCC27853), \u003cem\u003eE. faecalis\u003c/em\u003e (ATCC 19433) and \u003cem\u003eS. aureus\u003c/em\u003e (ATCC 25923). Growth curves from each microorganism were previously determined to use its log phase during assays. Antibacterial assays were performed with Luria-Bertani media (Invitrogen, Waltham, Massachusetts, USA) with 5x10\u003csup\u003e5\u003c/sup\u003e CFU.mL\u003csup\u003e-1\u003c/sup\u003e in a 96-well plate for 18h at 37 \u0026ordm;C incubated in a microplate reader with readings at 600 nm absorbance under medium agitation (BioTek PowerWave HT, Winooski, Vermont, USA). Ampicillin (50 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e \u0026ndash; Sigma-Aldrich, San Luis, Missouri, USA) was used as a negative control for antibacterial assays. Chlorhexidine (Via Magistral) was tested up to 512 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (1013 \u0026micro;M) and MIC was determined by no bacteria growth. After MIC determination, each sample was incubated in an agar plate to assess the presence of viable microorganisms for the definition of MBC. Then, antibiofilm properties of synoeca-MP and chlorhexidine were assessed against preformed biofilms of \u003cem\u003eP. aeruginosa\u003c/em\u003e and \u003cem\u003eS. aureus.\u003c/em\u003e Bacterium inoculum was diluted in a BM2 minimum medium, consisting of 62 mM potassium phosphate buffer (pH 7.0 - VETEC, Duque de Caxias, Rio de Janeiro, Brazil), 7 mM (NH\u003csub\u003e4\u003c/sub\u003e)\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e (VETEC), 2 mM MgSO\u003csub\u003e4\u003c/sub\u003e mM (Sigma-Aldrich), 10 \u0026micro;M FeSO\u003csub\u003e4\u003c/sub\u003e (VETEC), and 0.5% glucose (Sigma-Aldrich), so that the final concentration reached was 1/100 v/v per well [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Bacteria were plated and kept for 24h at 37\u0026deg;C. Controls were represented by \u003cem\u003eP. aeruginosa and S. aureus\u003c/em\u003e bacteria (1/100 v/v) as negative control, ciprofloxacin (Sigma-Aldrich) and the BM2 medium [62 mM potassium phosphate buffer (pH 7.0 - VETEC), 7 mM (NH\u003csub\u003e4\u003c/sub\u003e)\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e (VETEC), 2 mM MgSO\u003csub\u003e4\u003c/sub\u003e mM (Sigma-Aldrich), 10 \u0026micro;M FeSO\u003csub\u003e4\u003c/sub\u003e (VETEC), and 0.5% glucose (Sigma-Aldrich)], as positive control. After 24h, planktonic bacteria were discarded and plates were washed twice with 1x PBS. Serial sample dilutions were performed and added to the preformed biofilm microplates, for 24h. Subsequently, to assess the biofilm\u0026rsquo;s viability, the MTT assay (Sigma-Aldrich) was performed, according to the manufacturer's standards [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eDetermination of minimal inhibitory concentration (MIC), minimal fungicidal concentration (MFC) and antibiofilm concentration of\u003c/b\u003e \u003cb\u003eCandida albicans\u003c/b\u003e\u003c/p\u003e \u003cp\u003eAntifungal analyses were performed against \u003cem\u003eC. albicans\u003c/em\u003e (ATCC 10231). Experimental assays were performed with RPMI-1640 media (Sigma-Aldrich) with 0.165 mol.L\u003csup\u003e-1\u003c/sup\u003e MOPS with 2.5x10\u003csup\u003e3\u003c/sup\u003e CFU.mL\u003csup\u003e-1\u003c/sup\u003e in a 96-well plate for 48h at 37 \u0026ordm;C in a microplate reader under medium agitation (BioTek PowerWave HT). Amphotericin B (10 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e \u0026ndash; Sigma-Aldrich) was used as a negative control for antifungal assays. Chlorhexidine (Via Magistral) was tested up to 512 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (1013 \u0026micro;M) and MIC was determined by no fungal growth. After MIC determination, the sample was incubated in an agar medium for determination of MFC. Fungal antibiofilm properties of synoeca-MP and chlorhexidine were assessed against preformed biofilms of \u003cem\u003eC. albicans\u003c/em\u003e. Then, cells were centrifuged, after 48 h growth, at 3000 g, 5 min, at 4 \u0026ordm;C, washed twice with sterile PBS and then resuspended in BM2 media [62 mM potassium phosphate buffer (pH 7.0 - VETEC), 7 mM (NH\u003csub\u003e4\u003c/sub\u003e)\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e (VETEC), 2 mM MgSO\u003csub\u003e4\u003c/sub\u003e mM (Sigma-Aldrich), 10 \u0026micro;M FeSO\u003csub\u003e4\u003c/sub\u003e (VETEC), and 0.5% glucose (Sigma-Aldrich)]. Cells were seeded at 1x10\u003csup\u003e7\u003c/sup\u003e CFU per well in 96-well plates and incubated for 48h at 37 \u0026ordm;C. Antimicrobial agents were added to plates and incubated for 24h more. All agents were tested according to the following concentrations: chlorhexidine up to 128 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (253 \u0026micro;M) and synoeca-MP up to 128 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (80 \u0026micro;M). Amphotericin B (Sigma-Aldrich) at 16 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e was used as a positive control. The effect on biofilms was evaluated by XTT cell viability kit assays (Biotium, Fremont, California, USA) and plates were read at 470 nm absorbance [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eSynergism assay\u003c/h2\u003e \u003cp\u003eAfter the determination of minimum inhibitory concentrations, the synergism assay was assessed for the combination of chlorhexidine and synoeca-MP using the microdilution growth inhibition assay protocol, according to the Clinical and Laboratory Standards Institute (CLSI) [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Thus, concentrations below its MICs were tested according to the checkerboard method with adaptations [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Assays were performed against \u003cem\u003eP. aeruginosa\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eC. albicans\u003c/em\u003e as previously described to determine MIC. The fractional inhibitory concentration (FIC) index determined synergistic interactions according to the following calculation: [(MIC of 1 combined to 2) / (MIC of 1)] + [(MIC of 2 combined to 1) / (MIC of 2)]. Values\u0026thinsp;\u0026lt;\u0026thinsp;0.5 were considered as a synergic interaction.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eDegradation/integrity of synoeca-MP and chlorhexidine in human saliva\u003c/h2\u003e \u003cp\u003eThe protocol of this study was approved by the Human Research Ethics Committee (number 90666218.2.0000.0029). Saliva was collected from control patients (without gingival and/or periodontal diseases; n\u0026thinsp;=\u0026thinsp;7), patients diagnosed with gingivitis (n\u0026thinsp;=\u0026thinsp;6), and periodontitis (n\u0026thinsp;=\u0026thinsp;17), prior to clinical dental care, at the graduate dentistry clinic of Universidade Cat\u0026oacute;lica de Bras\u0026iacute;lia. Patient data for those with periodontal disease were obtained from medical records, and saliva samples were taken between 8 and 11 am. Unstimulated saliva was collected for 5 min [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Subsequently, 80.26 \u0026micro;M (40.565 \u0026micro;g/mL) of chlorhexidine, 80.26 \u0026micro;M (128 \u0026micro;g/mL) of synoeca-MP, or the same concentration of chlorhexidine and synoeca-MP together were exposed to saliva; the control group was represented by no treatment exposed to saliva control group. To assess the integrity of synoeca-MP and chlorhexidine in saliva, molecular mass, and purity were evaluated using the autoflex speed spectrometer mass spectrometry technique (Bruker Daltonics, Billerica, Massachusetts, USA), using the reflected and positive operating method, adjusted for 400 to 3500 Da, with external calibration [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Synoeca-MP and chlorhexidine were diluted in ultrapure water and deposited on the plate (AnchorchipVar-384 \u0026ndash; Bruker Daltonics) together with the matrix in triplicate, awaiting complete crystallization at room temperature after 0, 30, 60, 90, 120, 150, 180, 210 and 240 min. Between the experimental periods, samples were kept at 37\u0026deg;C, to mimic saliva conditions in the body environment. Then, the correlation between the time of degradation of the peptide, chlorhexidine, and their association in the saliva of patients with the different periodontal diagnoses was analyzed.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003ePeriodontal ligament cell culture\u003c/h2\u003e \u003cp\u003eHealthy third molars extracted from patients aged 18 to 30 years were used to obtain periodontal ligament cells. Ethics registration and approval had been obtained from the Human Research Ethics Committee of the Catholic University of Bras\u0026iacute;lia (CAAE: 90666218.2.0000.0029) and all donors signed the understanding and written consent. Right after extraction, extracted teeth were placed in a sterile tube containing Dulbecco Modified Eagle Medium (DMEM; Gibco, Grand Island, New York, USA) without FBS, associated with collagenase type 1 (3 mg.mL\u003csup\u003e-1\u003c/sup\u003e) and dispase (4 mg.mL\u003csup\u003e-1\u003c/sup\u003e), for 1 h at 37 \u0026ordm;C [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. After this period, tooth roots were shaved, and tissue structures of the periodontal ligament adhered to the roots were collected in 6-well cell culture plates (Costar Corp., Cambridge, Massachusetts, USA). Cells were cultured in supplemented DMEM culture medium with 10% FBS (Gibco), 1% MEM amino acid solution (Gibco), 0.05% gentamicin (Gibco), 1% L-glutamine (Gibco), 1% penicillin / streptomycin (1000 U.mL\u003csup\u003e-1\u003c/sup\u003e) (Gibco) and kept at 5% CO\u003csub\u003e2\u003c/sub\u003e, 37 \u0026ordm;C and at 95% humidity [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Every three days, the culture medium was changed, until reaching the number of cells required for the experiment.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eHemolytic assay\u003c/h2\u003e \u003cp\u003eThe hemolytic activity of synoeca-MP and chlorhexidine was based on the methodology described by Bignami, with adaptations [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Thus, a suspension with 1% cells in PBS was prepared and centrifuged twice (1000 x g, for 2 min). Subsequently, 350 \u0026micro;L of the 1% erythrocyte solution was used to test different concentrations of synoeca-MP and chlorhexidine. Controls were represented by saline solution (0% hemolysis, negative control) and triton X-100 (Sigma-Aldrich) at 0.1% (100% hemolysis, positive control). After 1h, tubes were centrifuged and 100 \u0026micro;L of the supernatant was collected to determine the optical density in a microplate reader at 406 nm.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eMTT assay\u003c/h2\u003e \u003cp\u003eHuman peripheral blood mononuclear cells (PBMCs), periodontal ligament cells, and SaOs-2 viability were evaluated according to the experimental groups and incubation period of each proposed assay. The positive control was represented by cells in culture medium (100% of cell viability) and the negative control was represented by cells in lysis solution \u0026minus;\u0026thinsp;10 mM Tris (Sigma-Aldrich), pH 7.4, 1 mM EDTA (Sigma-Aldrich) and 0.1% triton X-100 (Sigma-Aldrich), representing 0% cell viability. At the end of the incubation period, the MTT colorimetric assay (Sigma-Aldrich) was used [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eSynoeca-MP and chlorhexidine on cell proliferation and migration assay\u003c/h2\u003e \u003cp\u003eCells were plated in 96-well plates (Kasvi, S\u0026atilde;o Jos\u0026eacute; dos Pinhais, Paran\u0026aacute;, Brazil) and after 24h, synoeca-MP and chlorhexidine stimuli were added. Then, after 24 h, 48 h and 72 h, cell viability was determined by MTT assay. Cell proliferative potential in the presence of tested substances was performed using the Tripan Blue exclusion technique, after 0h, 24h, and 48h of cell incubation [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Confluent periodontal ligament cell cultures were obtained, and their trypsinization and cell resuspension assays were performed in DMEM medium (Gibco) without FBS (same conditions as those used in cell migration assay). Periodontal ligament cells (1x10\u003csup\u003e5\u003c/sup\u003e cells) were added in 1 mL DMEM (Gibco) without FBS to 24-well plates. Experimental groups contained the lowest antimicrobial synergistic concentration of synoeca-MP and chlorhexidine, associated or alone. Then, after completing the experimental incubation, cells were resuspended and the solution was added to 0.4% trypan blue dye (Sigma-Aldrich), for 1 minute. Cells were counted immediately using a Neubauer chamber (Brand GmbH, Wertheim, Baden-W\u0026uuml;rttemberg, Germany). To assess the effect of synoeca-MP, chlorhexidine and their association on cellular proliferation, a scratch assay was performed [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Cells from the periodontal ligament were kept confluent, and through the accomplishment of 3 markings, cell adhesion in the plate was disrupted. After that, the plate was washed with PBS, followed by the addition of supplemented DMEM culture medium, with the addition of all tested substances. Then, after 0 h, 24 h and 48 h, photographs were taken in an inverted electron microscope, with a 10x increase, in order to verify the migratory activity in these periods [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. The measurement of cell migration was performed through the analysis of photographs by counting cells using Image J software [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eEvaluation of synoeca-MP and chlorhexidine on osteoclastogenesis process\u003c/h2\u003e \u003cp\u003eWhole blood (about 4 mL) was collected from healthy volunteers. PBMCs were isolated by density gradient centrifugation using Ficoll-Paque (Sigma-Aldrich), following the manufacturer's recommendations. Then, the cell pellet was resuspended in 10 mL of supplemented DMEM (Gibco) and 25 ng.mL\u003csup\u003e-1\u003c/sup\u003e of macrophage colony-stimulating factor recombinant (M-CSF; Peprotech, Rocky Hill, Connecticut, USA) [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. After 72h, non-adherent cells were removed, and the adherent cells were plated in 96-well plates (Kasvi). After 24h, synoeca-MP and chlorhexidine stimuli were added. Cellular viability was assessed after 14 days. PBMCs (M-CSF-dependent macrophages) were incubated at 1.6x10\u003csup\u003e4\u003c/sup\u003e cells per well, in 96-well culture plates (Kasvi) with supplemented DMEM (Gibco). Cultures were subjected to stimuli, such as 10 ng.mL\u003csup\u003e-1\u003c/sup\u003e of the soluble receptor activator of nuclear factor-κB ligand (sRANKL; Peprotech) and synoeca-MP at its synergistic concentrations with chlorhexidine, associated or alone, for 14 days [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. After the fourteenth day, the PBMCs were subjected to tartrate-resistant acid phosphatase (TRAP) staining for later counting of the number of differentiated osteoclasts and the number of nuclei per osteoclast in an inverted microscope. For TRAP staining, the acid phosphatase kit, leukocyte (Sigma-Aldrich) was used, following the manufacturer's recommendations. Osteoclasts were considered TRAP-positive cells (red/orange color), with more than three nuclei inside them [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eEvaluation of synoeca-MP and chlorhexidine on mineral matrix formation\u003c/h2\u003e \u003cp\u003eTo evaluate mineral matrix deposition effects, periodontal ligament cells, and SaOs-2 (derived from human osteosarcoma) cells were used [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. Periodontal ligament cells and SaOs-2 cells were cultured at a concentration of 1.6x10\u003csup\u003e4\u003c/sup\u003e cells per well, in 6-well culture plates (Kasvi) for 21 days, in supplemented DMEM medium (Gibco), with osteogenic conditions (50 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e ascorbic acid, 100 nM dexamethasone, 1 mM β-glycerophosphate; Sigma-Aldrich), for 21 days. Medium and all stimuli were changed every 3 days. Cultures were subjected to synoeca-MP and chlorhexidine stimuli in synergistic concentrations, after 24h [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. After the incubation period, the concentration of alkaline phosphatase (ALP) and the formation of the mineral matrix were assessed.\u003c/p\u003e \u003cp\u003e \u003cb\u003eAlkaline phosphatase.\u003c/b\u003e Determination of alkaline phosphatase (ALP) concentration in periodontal ligament and SaOs-2 cell cultures was assessed after 21 days under osteogenic conditions. ALP was measured by the colorimetric method of paranitrophenol, using the Sigmafast p-Nitrophenyl phosphate kit (Sigma-Aldrich). For the ALP assay, the cells were washed with PBS and incubated in 0.05% Triton X-100 for 20 minutes at room temperature, with constant agitation. Cells were transferred, vortexed for 20 seconds, centrifuged for 15 min at 4\u0026deg;C at 2500 RPM, and kept on ice for 20 minutes. Aliquots of cell lysate were incubated with p-Nitrophenol phosphate (p-NF) substrate at 37 \u0026ordm;C for 30 minutes. The reaction was stopped by adding 5 \u0026micro;L 1N NaOH and the absorbance was measured at 405 nm using the microplate reader (Bio-Tek PowerWave HT). A standard p-NF curve was established to determine enzyme activity. The samples were normalized, and protein quantification was determined by Qubit\u0026reg;ฏ [41].\u003c/p\u003e \u003cp\u003e \u003cb\u003eMineral matrix formation assay.\u003c/b\u003e After 21 days of culture, the formation of the mineral matrix of periodontal ligament cells and SaOs-2 cells was determined by alizarin red staining. For this, plates were fixed with 10% (v / v) formaldehyde (VETEC), at room temperature for 15 min. The cell layer was washed twice with distilled water and 1 mL of alizarin red S dye (40 mM; pH 4.1; Sigma-Aldrich) was added per well. The plate was incubated at room temperature for 20 minutes, under constant agitation. After discarding the unincorporated dye, the cell layer was washed 4 times, with 4 mL of distilled water, with a 5-minute stirring between washes. Then, the distilled water was discarded, and the stained cell layer was evaluated under an inverted microscope (Zeiss, Oberkochen, Baden-W\u0026uuml;rttemberg, Germany). To quantify the stained mineral matrix, 800 \u0026micro;L of acetic acid 10% (v/v) (Din\u0026acirc;mica, Indaiatuba, S\u0026atilde;o Paulo, Brazil) was added to each well, followed by incubation for 30 minutes at room temperature, under constant agitation. The cell layer, together with 10% (v/v) acetic acid (Din\u0026acirc;mica), was then scraped from the bottom of the well, vortexing for 30 seconds, covered with 500 \u0026micro;L of mineral oil (VETEC), heated to 85 \u0026ordm;C for 10 minutes and cooled on ice, for 5 minutes. After complete cooling, the suspension was centrifuged at 20,000 x g for 15 minutes and 500 \u0026micro;L of the supernatant transferred to a new tube. For acid neutralization, 200 \u0026micro;L of 10% (v / v) ammonium hydroxide (VETEC) was added. Aliquots (150 \u0026micro;L) of the supernatant were arranged in triplicate in a 96-well plate (Kasvi) for reading in a microplate reader (Bio-Tek PowerWave HT), at 405 nm [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eAll experiments were performed in technical and biological triplicates. Data normality was tested by the Shapiro Wilk test, and parametric variables are described as mean and standard deviation or mean and standard error of the mean. Statistically significant differences were considered when p\u0026thinsp;\u0026lt;\u0026thinsp;0.05. To measure the statistical differences in the continuous variables, the two-way ANOVA was used followed by the Bonferroni post hoc test. The Kaplan-Meier curves were used to show molecule degradation over time (synoeca-MP, chlorhexidine and synoeca-MP plus chlorhexidine), and statistical differences were assessed by Log-Rank (Mantel-Cox) test. Statistical analysis was performed using the GraphPad Prism 5 software (California, USA).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eAntifungal, antibacterial and antibiofilm activity determination\u003c/h2\u003e \u003cp\u003eAntimicrobial activity of synoeca-MP and chlorhexidine was tested against \u003cem\u003eP. aeruginosa, S. aureus, E. faecalis\u003c/em\u003e and \u003cem\u003eC. albicans\u003c/em\u003e. Minimal inhibitory concentrations were determined, and all agents were capable of inhibiting yeast and bacteria growth, at synoeca-MP or chlorhexidine concentrations of up to 32 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e, under the assay conditions. However, the minimum bactericidal or fungicidal concentration was up to 64 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e, except when synoeca-MP was evaluated against \u003cem\u003eP. aeruginosa\u003c/em\u003e, which did not present MBC until values up to 128 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eFurther analysis involving antimicrobial properties might increase the potential use of synoeca-MP and chlorhexidine in the oral environment. To this end, synoeca-MP and chlorhexidine were also evaluated for their inhibitory capacity on biofilms created by \u003cem\u003eP. aeruginosa, S. aureus\u003c/em\u003e and \u003cem\u003eC. albicans.\u003c/em\u003e All tested concentrations of synoeca-MP and chlorhexidine were able to reduce the viability of preformed biofilm of \u003cem\u003eP. aeruginosa\u003c/em\u003e. However, synoeca-MP showed the best results at concentrations between 16 and 128 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (10 to 80 \u0026micro;M), including 50 to 58% of \u003cem\u003eP. aeruginosa\u003c/em\u003e biofilm eradication and 128 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (80 \u0026micro;M) presenting 90% of biofilm eradication of \u003cem\u003eC. albicans\u003c/em\u003e and 40% of \u003cem\u003eS. aureus\u003c/em\u003e biofilm eradication. On the other hand, chlorhexidine showed its greatest result in the concentration of 128 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (253 \u0026micro;M), which corresponded to 49% of biofilm reduction against \u003cem\u003eP. aeruginosa\u003c/em\u003e and approximately 95% of that of \u003cem\u003eS. aureus\u003c/em\u003e. Besides that, 32, 64 and 128 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (63 to 253 \u0026micro;M) of chlorhexidine can inhibit 80\u0026ndash;95% of \u003cem\u003eC. albicans\u003c/em\u003e biofilm preformed (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Given the results obtained, we observed the antimicrobial activity as well the activity against pre-formed biofilms by \u003cem\u003eP. aeruginosa\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, \u003cem\u003eE. faecalis\u003c/em\u003e, and \u003cem\u003eC. albicans\u003c/em\u003e. In comparative analysis, the peptide synoeca-MP showed an advantage in terms of its microbicidal effects against the colonies of the previously mentioned microorganisms, when compared to chlorhexidine. Thus, an important next point would be to evaluate the possible effects of the interaction of both molecules together.\u003c/p\u003e\u003cp\u003eTable 1 Minimal Inhibitory Concentration (MIC), Minimal Bactericidal/Fungicidal Concentration (MBC/MFC) and percentage of Antibiofilm activity of synoeca-MP and chlorhexidine against \u003cem\u003eP. aeruginosa, S. aureus, E. faecalis\u003c/em\u003e and \u003cem\u003eC. albicans in vitro\u003c/em\u003e. NT* Not tested.\u003c/p\u003e\u003cp\u003e\u003cimg 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\" width=\"673\" height=\"151\"\u003e\u003cbr\u003e\u003c/p\u003e\u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eAntimicrobial synergistic analysis of synoeca-MP and chlorhexidine\u003c/h2\u003e \u003cp\u003eOnce synoeca-MP and chlorhexidine had demonstrated promising results for antimicrobial activity, antimicrobial synergistic analyses were performed against \u003cem\u003eP. aeruginosa, S. aureus, E. faecalis\u003c/em\u003e and \u003cem\u003eC. albicans\u003c/em\u003e. Chlorhexidine and synoeca-MP interaction exhibited synergistic antimicrobial activity in different concentrations against \u003cem\u003eP. aeruginosa, S. aureus\u003c/em\u003e, and \u003cem\u003eC. albicans\u003c/em\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). However, interactions involving synoeca-MP and chlorhexidine against \u003cem\u003eE. faecalis\u003c/em\u003e did not demonstrate any synergistic effect. In addition, chlorhexidine association with synoeca-MP reduced 50\u0026ndash;99% of MIC concentration when used against \u003cem\u003eP. aeruginosa, S. aureus\u003c/em\u003e, and \u003cem\u003eC. albicans\u003c/em\u003e. Thus, the synergistic interaction between synoeca-MP and chlorhexidine that demonstrated the best inhibition, at lower concentrations, was 0.125 and 0.25 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e of chlorhexidine and synoeca-MP, respectively, against \u003cem\u003eP. aeruginosa in vitro\u003c/em\u003e. The addition of synoeca-MP to chlorhexidine might promote several different effects on the oral environment in inflammatory and infectious conditions. Local and general use of chlorhexidine also encourages the use of this molecule combined with synoeca-MP.\u003c/p\u003e\u003cp\u003eTable 2 Synergistic concentrations of synoeca-MP (SYN) and chlorhexidine (CHX). Table with detailed analysis of concentrations was demonstrated according to its combination, fractional inhibitory concentration index (FICI) value and percentage of reduction related to previous inhibitory concentration.\u003c/p\u003e\u003cp\u003e\u003cimg 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\" width=\"672\" height=\"179\"\u003e\u003cbr\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eSynoeca-MP and chlorhexidine stability in human saliva\u003c/h2\u003e \u003cp\u003eOnce the antimicrobial potential of using chlorhexidine with or without synoeca-MP had been verified, it was necessary to evaluate the stability of its molecules in human saliva. The monoisotopic mass of synoeca-MP, chlorhexidine, matrix control and saliva of all patients was checked to certify their integrity prior to analysis (Supplementary material - Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). For the molecule\u0026rsquo;s stability in saliva, the last time point at which the ions of the monoisotopic mass of the molecules appeared intact in the spectrum was considered (Supplementary material - Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Saliva from patients with different periodontal diagnoses were used for these analyses.\u003c/p\u003e \u003cp\u003eAll data correlating patients' periodontal diagnoses and the time of degradation of all molecules were found in Supplementary table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. It was found that the time of degradation of synoeca-MP (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA) and the association of synoeca-MP and chlorhexidine (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eC) changed according to the periodontal diagnosis. The degradation time of both molecules was inversely proportional to the severity of periodontal disease; that is, the more severe the periodontal disease, the less time the compounds take to be degraded. On the other hand, chlorhexidine remained stable throughout all evaluated time points (until 240 min) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). These results are favorable for a proposal to use these molecules as mouthwashes in the oral cavity.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eEffect of synoeca-MP, chlorhexidine or both on periodontal ligament cells\u003c/h2\u003e \u003cp\u003eOnce the molecules had shown good integrity in saliva, the next step was to evaluate their effect on periodontal ligament cells. Cell viability of periodontal ligament with stimuli of 0.25 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (0.16 \u0026micro;M) synoeca-MP and/or 0.125 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (0.25 \u0026micro;M) chlorhexidine was performed on periodontal ligament cells. No cytotoxicity was observed after 24 h, 48 h, and 72 h, demonstrating security for human periodontal ligament cells.\u003c/p\u003e \u003cp\u003eThen, to evaluate the molecules\u0026rsquo; possible effects on tissue healing and regeneration, cell cultures of periodontal ligament were subjected to cell proliferation and migration assays. Cultures were stimulated with 0.25 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (0.16 \u0026micro;M) synoeca-MP and/or 0.125 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (0.25 \u0026micro;M) chlorhexidine for 24h (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA and C) and 48h (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB and D). The proliferative potential was increased in the presence of synoeca-MP and the synergistic combination of synoeca-MP and chlorhexidine in both experimental periods (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001 \u0026ndash; Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA and B). These results were also similar to the positive control group (periodontal ligament cell in 2% FBS). However, the presence of chlorhexidine led to a similar proliferation to periodontal ligament cells alone, after 24h and 48h.\u003c/p\u003e \u003cp\u003eCell migration is one of the main events during tissue repair and the wound healing process. Thus, periodontal ligament cultures were stimulated with 0.25 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (0.16 \u0026micro;M) synoeca-MP and/or 0.125 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (0.25 \u0026micro;M) chlorhexidine for 0h (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eE, F, G, H and I), 24 h (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eJ, K, L, M and N) and 48 h (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eO, P, Q, R, and S). It was observed that all tested conditions presented an increase in periodontal ligament cell migration after 24 h, compared to periodontal ligament cells alone (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001 - Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC). However, after 4h there was an increase in periodontal ligament cell migration in cultures exposed to the synergistic association of chlorhexidine and synoeca-MP (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001 - Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eD and S). It can be inferred that, when comparing synoeca-MP and chlorhexidine, both showed no cytotoxicity. Both synoeca-MP and the synergistic combination of synoeca-MP and chlorhexidine demonstrated increased cell proliferation and migration. These findings are highly valuable for use in the tissue regeneration process.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003eEffect of synoeca-MP, chlorhexidine or both on the osteoclastogenesis process\u003c/h2\u003e \u003cp\u003eFrist, it was found that none of tested groups showed hemolytic activity. Low concentrations of synoeca-MP and chlorhexidine (0.25 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (0.16 \u0026micro;M) synoeca-MP and/or 0.125 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (0.25 \u0026micro;M) chlorhexidine), separated or associated, did not present cytotoxicity to human erythrocytes (Supplementary material - Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). This fact was encouraging for the analysis of the effect of these molecules during the osteoclastogenesis processes.\u003c/p\u003e \u003cp\u003eSynoeca-MP and chlorhexidine cytotoxicity was evaluated at synergistic concentrations in peripheral blood mononuclear cells (PBMCs), as well as for its potential for inhibiting osteoclastogenesis. It was shown that cell viability after 14 days remained stable in all tested conditions (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA), demonstrating no cytotoxicity to human PBMCs. It was found that the association between synoeca-MP peptide and chlorhexidine showed the greatest inhibition of the osteoclastogenesis process, reducing it by 86.65% compared to the control group. However, separately, synoeca-MP and chlorhexidine were also able to inhibit osteoclastogenesis by 76.12% and 63.7% respectively, when compared to the sRANKL-stimulated cells (p\u0026thinsp;\u0026gt;\u0026thinsp;0.0001 - Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). Analyzing the ratio, nuclei/osteoclast, an average of 9.6 nuclei per osteoclast were observed in the sRANKL-stimulated cells, while synoeca-MP, chlorhexidine, and their association showed an average of 3.63, 5.3, and 3.58 nuclei per osteoclast, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC), presenting a lower osteoclast size with a diminished number of nuclei.\u003c/p\u003e \u003cp\u003eIn qualitative analysis, it was possible to notice that the multinucleated osteoclasts in sRANKL-stimulated cells were relatively larger compared to other tested groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eE). The chlorhexidine-stimulated group (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eG), synodical-MP-stimulated group (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eF) and synoeca-MP with chlorhexidine-stimulated group (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eH) presented osteoclasts smaller in size and with similar characteristics to PBMCs (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD), corroborating the results of osteoclast count and number of nuclei per osteoclast. Therefore, these findings are encouraging for the application of these molecules in bone reabsorption process, since the absence of cytotoxicity of both chlorhexidine and Synoeca-MP, both combined and individually, for human cells was demonstrated, something essential for the safety of drugs applied to health. Furthermore, it was found that these molecules, when combined, demonstrated greater efficacy in the osteoclastogenesis downregulation process, including a reduction in the number of nuclei and the size of osteoclasts. Thus, the use of these molecules may be promising for the bone remodeling process.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eEffect of synoeca-MP, chlorhexidine or both on the mineralized matrix formation\u003c/h2\u003e \u003cp\u003eGiven the promising results of the effect of these molecules during osteoclastogenesis, we began to evaluate the effects of these molecules on the formation of the mineral matrix. To promote the mineralization process, periodontal ligament cells and SaOs-2 cells, derived from human osteosarcoma, were cultured under osteogenic conditions, allowing for osteoblast differentiation (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The cell viability of periodontal ligament and SaOs-2 cell cultures was analyzed under osteogenic conditions and stimulated with 0.25 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (0.16 \u0026micro;M) synoeca-MP and/or 0.125 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e (0.25 \u0026micro;M) chlorhexidine for 24h, and the other analyses then went on to determine the potential of mineralized matrix formation (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA and B). There was no reduction in cell viability with any tested sample when periodontal ligament (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA) and SaOs-2 (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB) cells were stimulated, under osteogenic conditions.\u003c/p\u003e \u003cp\u003eTo evaluate the effect of synoeca-MP and chlorhexidine on the mineralization process, periodontal ligament, and SaOs-2 cells were cultured under osteogenic conditions for 21 days, and the mineral matrix formation assay was performed by Alizarin red staining. The mineralization potential in periodontal ligament cells (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC) and SaOs-2 cells (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD) stimulated with synoeca-MP and / or chlorhexidine was observed (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). However, periodontal ligament cells stimulated with chlorhexidine showed the greatest potential for matrix mineralization, compared to other tested groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001 - Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC and 4C3). Qualitative analysis of mineral matrix formation in cultures of periodontal ligament cells showed the highest intensity of alizarin red staining in the presence of synoeca-MP (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC2), chlorhexidine (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC3) and synoeca-MP and chlorhexidine-associated (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC4), compared to cells in osteogenic medium alone (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC1). Similarly, a qualitative analysis of mineral matrix formation based on SaOs-2 cells showed greater intensity of alizarin red staining in cultures stimulated with synoeca-MP (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD2), chlorhexidine (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD3) and synoeca-MP and chlorhexidine associated (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD4), compared to cells in osteogenic medium alone (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD1). The alkaline phosphatase production in cultures of periodontal ligament cells, after 21 days, showed no statistically significant differences between any of the tested groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eE).\u003c/p\u003e \u003cp\u003eAdditionally, the results showed that synoeca-MP and chlorhexidine did not reduce cell viability, and they promoted the formation of mineral matrix in periodontal ligament cells and SaOs-2 cells. As previously seen, the combination of Synoeca-MP and chlorhexidine also demonstrated efficacy in inhibiting the osteoclastogenesis process. Although synoeca-MP and chlorhexidine showed mineralization potential, periodontal ligament cells stimulated with chlorhexidine exhibited the greatest potential for mineralizing the matrix. Qualitative analyses confirmed these results, showing intense red alizarin staining in the stimulated cultures. Alkaline phosphatase production, an indicator of osteoblastic differentiation, did not significantly differ between the groups. These findings suggest that synoeca-MP and chlorhexidine may play an important role in tissue regeneration, especially in mineralization processes.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe maintenance of oral health in hospitalized patients, who may be immunosuppressed and in precarious oral hygiene conditions, is a major challenge within dentistry [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. In this sense, the oral cavity is an environment prone to the colonization of opportunistic pathogenic microorganisms such as \u003cem\u003eP. aeruginosa, S. aureus, E. faecalis\u003c/em\u003e, and \u003cem\u003eC. albicans\u003c/em\u003e, which can increase the susceptibility to infections and more serious problems [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Therefore, methods for oral microbial control of hospitalized patients are necessary. Chlorhexidine 0.12% is widely used in the hospital setting to control oral biofilm [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. However, prolonged use of this product can result in adverse effects [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn recent decades, the search for new antimicrobial molecules has sparked researchers' interest in antimicrobial peptides [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Consequently, previous studies have isolated several peptides from wasps, which were then classified by their chemical structures and biological activities. Among them, synoeca-MP, derived from the venom of the wasp \u003cem\u003eSynoeca surinama\u003c/em\u003e, has shown significant antimicrobial and antibiofilm results against \u003cem\u003eP. aeruginosa\u003c/em\u003e and other Gram-positive and Gram-negative strains [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Thus, it may be a promising alternative for controlling pathogenic microorganisms present in the oral microbiota of hospitalized patients.\u003c/p\u003e \u003cp\u003eFurthermore, synoeca-MP exhibits synergistic antimicrobial activity when combined with chlorhexidine, allowing for the reduction of the concentration of both molecules to achieve the desired antimicrobial activity. This fact may favor the reduction of undesirable effects of isolated chlorhexidine [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. In this scenario, the present study brought new knowledge about the association between the synoeca-MP peptide and chlorhexidine. Herein, its antimicrobial and antibiofilm effect, its stability in human saliva, cytotoxicity, regenerative action, and bone homeostasis \u003cem\u003ein vitro\u003c/em\u003e were analyzed.\u003c/p\u003e \u003cp\u003eInitially, the antifungal, antibacterial, and antibiofilm activity of synoeca-MP and chlorhexidine against the opportunistic microorganisms involved in oral pathology was determined. The results obtained demonstrate the effectiveness of synoeca-MP and chlorhexidine in inhibiting the growth of \u003cem\u003eP. aeruginosa, S. aureus, E. faecalis\u003c/em\u003e, and \u003cem\u003eC. albicans\u003c/em\u003e, with minimum inhibitory concentrations of up to 32 \u0026micro;g.mL\u003csup\u003e-1\u003c/sup\u003e. Both substances reduced the viability of biofilms formed by these microorganisms, with synoeca-MP demonstrating a more effective eradication of \u003cem\u003eP. aeruginosa\u003c/em\u003e and \u003cem\u003eC. albicans\u003c/em\u003e biofilms compared to chlorhexidine. The comparative analysis of both molecules suggests that the peptide synoeca-MP may be an encouraging alternative to chlorhexidine in the treatment of oral infections, highlighting the need to investigate a possible synergistic activity between these compounds to develop effective antimicrobial therapeutic strategies.\u003c/p\u003e \u003cp\u003eTherefore, this study also verified the potential synergistic antimicrobial activity of these molecules against \u003cem\u003eP. aeruginosa, S. aureus, C. albicans\u003c/em\u003e, and \u003cem\u003eE. faecalis\u003c/em\u003e [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. In this regard, we confirmed the synergism between chlorhexidine and the peptide synoeca-MP against pre-formed biofilms of \u003cem\u003eP. aeruginosa, S. aureus\u003c/em\u003e, and \u003cem\u003eC. albicans\u003c/em\u003e, but they did not demonstrate any synergistic effect against \u003cem\u003eE. faecalis\u003c/em\u003e. The interaction assays exhibited different concentrations of synergistic antimicrobial activity, reducing from 50\u0026ndash;99% of the MIC of each molecule in this combination.\u003c/p\u003e \u003cp\u003e Additionally, an evaluation of the stability of both chlorhexidine and synoeca-MP molecules in human saliva was necessary, considering the perspective of a possible future mouthwash indicated for patients with different oral conditions. Therefore, our work also investigated the integrity in human saliva after the isolated or combined use of these substances, which revealed fundamental data for their clinical applicability. In this regard, the monoisotopic analysis of these compounds in the saliva of patients with different periodontal diagnoses indicated a relationship between the stability of the molecules and the severity of periodontal disease. It was observed that the greater the periodontal involvement of the individuals, the more rapidly the molecules were degraded. However, chlorhexidine alone maintained the highest degree of stability in the saliva of patients with gingivitis and periodontitis. Furthermore, the combination of synoeca-MP and chlorhexidine was able to increase the stability time of synoeca-MP alone. Such results suggest that chlorhexidine, when combined with synoeca-MP, may be a promising option regarding the development of a new mouthwash. This finding significantly contributes to understanding the stability dynamics of these molecules in human saliva and may guide future therapeutic strategies for the treatment of inflammatory and infectious oral conditions.\u003c/p\u003e \u003cp\u003eAfter infections stemming from oral pathologies, damage and other problems related to tissue integrity can occur. Thus, cell migration is one of the main events during the tissue repair and wound healing process [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. In this regard, periodontal ligament cells were exposed to the combination of synoeca-MP and chlorhexidine for the evaluation of cell proliferation and migration. The results obtained demonstrated that the combination of synoeca-MP and chlorhexidine increased cell migration and proliferation at 24h and 48h, with better wound closure after 48h. In addition to synoeca-MP, other host defense peptides have been reported in the literature with regenerative potential. In this context, IDR-1018 has shown promising regenerative properties using 2D and 3D human skin equivalents of fibroblasts, primary keratinocytes, and MeWo melanocyte cell lines [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. Furthermore, the antimicrobial peptide LL-37 can promote odonto/osteogenic migration and differentiation of stem cells from the apical papilla by activating the Akt/Wnt/β-catenin signaling pathway [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. Therefore, like the other peptides mentioned, synoeca-MP has great potential regarding future clinical applicability, aiming at tissue repair in patients with compromised oral conditions.\u003c/p\u003e \u003cp\u003eMoreover, to assess the potential of the combination of synoeca-MP and chlorhexidine, osteoclastogenesis and mineralized matrix formation processes were analyzed. Data obtained demonstrated that the association between synoeca-MP and chlorhexidine showed greater inhibition of the osteoclastogenesis process \u003cem\u003ein vitro\u003c/em\u003e. Moreover, the association between both molecules was able to reduce the size and number of nuclei per cell in the few differentiated osteoclasts [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. The size and number of nuclei per osteoclast can affect the metabolic activity of osteoclasts. Based on these results, we hypothesize that the increase in metabolic activity may be related to the increase in energy and cell mobility, which can increase areas of bone resorption. Therefore, chlorhexidine and synoeca-MP, when combined, may have promising potential for the treatment of bone resorption.\u003c/p\u003e \u003cp\u003eConsidering that bone homeostasis results from the balance between bone resorption and deposition, mineral matrix formation is a fundamental process [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. Thus, our study evaluated periodontal ligament cells and SaOs-2 cells stimulated with synoeca-MP and/or chlorhexidine, which showed great potential for mineralized matrix formation in the presence of these molecules. However, isolated stimulation with chlorhexidine favored greater formation of mineralized matrix in periodontal ligament cells. Although no damage to osteoblasts and mineralization was observed, probably due to the low concentration of the tested molecules, some studies suggest that chlorhexidine may be harmful to osteoblasts [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. Furthermore, another study concluded that although chlorhexidine has a strong antibacterial capacity against \u003cem\u003eE. faecalis\u003c/em\u003e, this molecule is ineffective in promoting mineralization of tissues around the root apex. These findings suggest that synoeca-MP and chlorhexidine may play an important role in tissue regeneration, especially in mineralization processes [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e].\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eBased on the results obtained, it can be concluded that the association of synoeca-MP and chlorhexidine can be applicable to oral diseases. To assess this, both were evaluated for their antimicrobial activity against microorganisms in planktonic and biofilm states, regenerative potential, stability in saliva, and bone homeostasis activity. Overall, the combination of synoeca-MP and chlorhexidine might be effective primarily in cases of opportunistic infections in the oral cavity and could also be associated with mechanical therapy. These molecules with different actions could potentially optimize dental treatments, as they demonstrated effective activity at low concentrations. However, it is important to emphasize that this is a preliminary \u003cem\u003ein vitro\u003c/em\u003e study, and further \u003cem\u003ein vivo\u003c/em\u003e and clinical analyses must be conducted to confirm these findings.\u003c/p\u003e "},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eThe present contribution of the study author: All authors contributed to the conception and design of the study. Material preparation, data collection and analysis were carried out by Ingrid A. R. Barin, Johnny C. Silva, Raquel F. Ramos, Stella M. F. Lima, Ana P C. Cantu\u0026aacute;ria, Poliana A. O. Silva, Elaine M. G. L. Dantas, Danilo C. M. Martins, Nelson G. O. J\u0026uacute;nior, Osmel F. Martinez, Jesser A. Almeida, Marcelo H. S. Ramada, Oct\u0026aacute;vio L. Franco and Taia M. B. Rezende. The first version of the manuscript was written by Ingrid A. R. Barin, Johnny C. Silva and Raquel F. Ramos, and all authors commented on previous versions of the manuscript. All figures and tables were created by Ingrid A. R. Barin, Johnny C. Silva and Raquel F. Ramos. Bibliographic analysis and corrections were carried out by Stella M. F. Limas and data analysis with the critical resistance of the work was carried out by Taia M. B. Rezende. All authors read and approved the final manuscript.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eDeclarations Section\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e- The present contribution of the study author:\u003c/strong\u003e The present contribution of the study author: All authors contributed to the conception and design of the study.\u0026nbsp;Material preparation, data collection and analysis were carried out by Ingrid A. R. Barin, Johnny C. Silva, Raquel F. Ramos, Stella M. F. Lima, Ana P C. Cantuária, Poliana A. O. Silva, Elaine M. G. L. Dantas, Danilo C. M. Martins, Nelson G. O. Júnior, Osmel F. Martinez, Jesser A. Almeida, Marcelo H. S. Ramada, Octávio L. Franco and Taia M. B. Rezende.\u0026nbsp;The first version of the manuscript was written by Ingrid A. R. Barin, Johnny C. Silva and Raquel F. Ramos, and all authors commented on previous versions of the manuscript. All figures and tables were created by Ingrid A. R. Barin, Johnny C. Silva and Raquel F. Ramos. Bibliographic analysis and corrections were carried out by Stella M. F. Limas and data analysis with the critical resistance of the work was carried out by Taia M. B. Rezende. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e- Conflict of interest:\u0026nbsp;\u003c/strong\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e- Funding:\u0026nbsp;\u003c/strong\u003eThis study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (305242/2022-9), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) (88887.910215/2023-00 and 88887724415/2022-00) and Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF) (00193–00000782/2021-63; 0009-0004-4942-2105 and 00193-00001118/2021-31).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e- Ethical approval:\u003c/strong\u003e 90666218.2.0000.0029\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e- Consent statements:\u0026nbsp;\u003c/strong\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (305242/2022-9), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) (88887.910215/2023-00 and 88887724415/2022-00) and Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF) (00193–00000782/2021-63; 0009-0004-4942-2105 and 00193-00001118/2021-31).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eGershonovitch R, Yarom N, Findler M. Preventing Ventilator-Associated Pneumonia in Intensive Care Unit by improved Oral Care: a Review of Randomized Control Trials. SN Compr Clin Med. 2020;2(6):727-733. doi: 10.1007/s42399-020-00319-8. Epub 2020 May 30. PMID: 32838136; PMCID: PMC7260467. \u003c/li\u003e\n\u003cli\u003eAlvares, F. A., de Oliveira, C. S., Alves, D. C. I., \u0026amp; Braun, G. (2022). Pneumonia associada \u0026agrave; ventila\u0026ccedil;\u0026atilde;o mec\u0026acirc;ni-ca: incid\u0026ecirc;ncia, etiologia microbiana e perfil de resist\u0026ecirc;ncia aos antimicrobianos. Revista De Epidemiologia E Controle De Infec\u0026ccedil;\u0026atilde;o, 11(4). https://doi.org/10.17058/reci.v11i4.16781.\u003c/li\u003e\n\u003cli\u003eHaas, A. N., Prado, R., Rios, F. S., Costa, R. dos S. A., Angst, P. D. M., Moura, M. dos S., Maltz, M., \u0026amp; Jardim, J. J. (2019). Occurrence and predictors of gingivitis and supragingival calculus in a population of Brazilian adults. 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Jun 2006;14(2):67-72.\u003cstrong\u003e\u003c/strong\u003e\u003c/li\u003e\n\u003c/ol\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":"Synergism, Host defense peptide, Periodontal disease, Opportunistic infections","lastPublishedDoi":"10.21203/rs.3.rs-4595835/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4595835/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eObjective\u003c/h2\u003e \u003cp\u003eTo evaluate \u003cem\u003ein vitro\u003c/em\u003e the association between synoeca-MP peptide and chlorhexidine, regarding their antimicrobial and antibiofilm activities, saliva stability, effect on tissue repair, bone resorption processes, and mineralized matrix formation.\u003c/p\u003e\u003ch2\u003eMethodology\u003c/h2\u003e \u003cp\u003eInitially, the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and antibiofilm concentration were determined. The synergism and degradation of synoeca-MP and chlorhexidine in human saliva were assessed. Furthermore, biocompatibility was evaluated using MTT assays, hemolytic assays, and proliferation and migration assays of periodontal ligament cells. Finally, bone homeostasis was evaluated through osteoclastogenesis assays, alkaline phosphatase determination, and mineralized matrix formation assay with SaOs-2 and ligament cells.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eThe antimicrobial and antibiofilm activity against the tested microorganisms was confirmed. Low synergistic concentrations of the synoeca-MP and chlorhexidine combination inhibited tested microorganisms. The association of these molecules remained stable in healthy saliva. Nevertheless, it degraded as the severity of periodontal disease increased. Additionally, lower synergistic concentrations of the combination were not cytotoxic to human cells, promoted the proliferation and migration of ligament cells, inhibited osteoclastogenesis, and increased mineral matrix formation of ligament cells and SaOs-2.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eSynoeca-MP and chlorhexidine combination shows potential for oral diseases treatment, as evidenced by its antimicrobial activity, regenerative potential, saliva stability, and bone homeostasis. It may be particularly effective for opportunistic oral infections and in conjunction with mechanical therapy.\u003c/p\u003e\u003ch2\u003eClinical relevance\u003c/h2\u003e \u003cp\u003eThe results obtained allow us to confirm the safety and expected effects of this combination, making it a promising alternative for hospitalized patients.\u003c/p\u003e","manuscriptTitle":"Antibiofilm, regenerative and bone homeostasis potential of the synergistic association of synoeca-MP peptide with chlorhexidine in oral cavity opportunistic infections","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-18 15:55:21","doi":"10.21203/rs.3.rs-4595835/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":"abcd5db3-3fb7-444f-b414-2ef7c47b2ed5","owner":[],"postedDate":"July 18th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-07-18T15:55:24+00:00","versionOfRecord":[],"versionCreatedAt":"2024-07-18 15:55:21","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4595835","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4595835","identity":"rs-4595835","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}