Objective To explore the mechanism of the new double-layer chitosan membrane combined with platelet-rich fibrin (PRF) on the repair of bone defects around implants in rats. Methods Forty clean-grade male SD rats were selected to establish skull defect models and were randomly divided into a control group, a platelet-rich fibrin group (PRF group), a new double-layer chitosan membrane group (N-CTS group) and a combined group, 10 rats in each group. The defect area of control group was not filled, PRF group was filled with platelet-rich fibrin (PRF), N-CTS group was filled with a new double-layer chitosan membrane (N-CTS), the bone defect area of combined group was filled with a new double-layer chitosan membrane combined with PRF. After 8 weeks, peripheral blood T lymphocyte subsets CD4+, CD8+, and CD4+ /CD8+ were measured by flow cytometry. Micro-CT was used to analyze the bone volume fraction (BV/TV), trabecular number (Tb. N), trabecular thickness (Tb. Th), and trabecular space (Tb. Sp) of new bone tissue. The multi-functional grid test system and micrometer were used to measure the width of the combined bone plate, the thickness of coated bone wall and the coating rate under the microscope. The pathological changes were observed by HE staining. Western blot was used to detect osteopontin (OPN), Runt-related transcription factor 2 (Runx2) and osteocalcin (OCN). Results Compared with control group, CD4+, CD4+/CD8+, Tb.TH, Tb.N, BV/TV, width of bone plate, thickness of coated bone wall, coating rate, OPN, Runx2, and OCN increased, CD8+, Tb.Sp decreased in PRF group and N-CTS group (P<0.05). Compared with PRF group and N-CTS group, CD4+, CD4+/ CD8+, Tb.TH, Tb.N, BV/TV, width of bone plate, thickness of coated bone wall, coating rate, OPN, Runx2 and OCN increased, CD8+ and Tb.Sp decreased in combined group (P<0.05). HE staining showed that the degradation of tissue fibers in control group was slow and the integration with bone defects was poor. The materials in each treatment group were well degraded, and osteoblasts and neovascularization were observed with new bone formation, among which combined group had the most significant effect. Conclusions The new double-layer chitosan membrane combined with PRF can significantly promote the repair of bone defects around rats and accelerate the formation of new bone in defect area.
Key words
Bone defect /
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New double-layer chitosan membrane /
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Platelet rich fibrin /
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Repair
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References
[1] Hasegawa T, Sasaki A, Saito I, et al. Success of dental implants in patients with large bone defect and analysis of risk factors for implant failure: a non-randomized retrospective cohort study[J]. Clin Oral Investig, 2022, 26(3): 2743-2750. DOI:10.1007/s00784-021-04249-0.
[2] Malushi ES, Isufi R, Kadaifciu D, et al. Guided Bone Regeneration Effects on Bone Quantity and Outcomes of Dental Implants in Patients With Insufficient Bone Support: A Single-Center Observational Study[J]. Cureus, 2023, 15(5): e38988. DOI:10.7759/cureus.38988.
[3] Javaid S, Rashid HB, Safdar A, et al. Comparative biochemical analysis of platelet-rich plasma-chitosan and platelet-rich fibrin-chitosan for treating tibial bone defects in rabbits[J]. Vet World, 2024, 17(9): 2036-2043. DOI:10.14202/vetworld.2024.2036-2043.
[4] Albahrawy M, El-Henawey MI, Elmezayyen AS, et al. Ameliorative role of silver nanoparticles incorporated with chitosan solution and leukocyte platelet-rich fibrin scaffold during colon anastomosis in rabbits[J]. J Mater Sci Mater Med, 2025, 36(1): 58. DOI:10.1007/s10856-025-06908-0.
[5] Wijayanti DA, Wirajaya GNKA, Pratiwi NH, et al. Combination of Collagen-Chitosan Hydrogel and Injectable Platelet-Rich Fibrin as a Biomaterial for Bone Regeneration: Characterization and Growth Factor Release Pattern[J]. Eur J Dent, 2025. DOI:10.1055/s-0045-1809144.
[6] Hilmy F, Dilogo I H, Reksodiputro M H, et al. Evaluation of adipose-derived stem cells (ASCS) exosome implantation and platelet-rich fibrin (PRF) on critical long bone defects in Sprague-Dawley rats[J]. Eur J Orthop Surg Traumatol, 2024, 34(5): 2805-2810. DOI:10.1007/s00590-024-03964-0.
[7] Sato R, Matsuura T, Akizuki T, et al. Influence of the bone graft materials used for guided bone regeneration on subsequent peri-implant inflammation: an experimental ligature-induced peri-implantitis model in Beagle dogs[J]. Int J Implant Dent. 2022, 8(1): 3. DOI:0.1186/s40729-022-00403-9.
[8] Razi MA, Mahajan A, Zarrin R, et al. Impact of Platelet-Rich Fibrin (PRF) Versus Freeze-Dried Bone Allograft (FDBA) on Peri-Implant Soft and Hard Tissue in Alveolar Ridge Preservation[J]. J Pharm Bioallied Sci, 2024, 16(Suppl 4): S3550-S3552. DOI:10.4103/jpbs.jpbs_1036_24.
[9] Aldaghir OM, Naje AR, Ghadhban AT, et al. Effectiveness of maxillary cortical bone graft chips harvested by bone scraper, covered with platelet-rich fibrin (PRF), in reconstruction of alveolar clefts: comparative study[J]. Oral Maxillofac Surg, 2024, 28(1): 205-216. DOI:10.1007/s10006-022-01128-0.
[10]Li Y, Tang S, Luo. Chitin whisker/chitosan liquid crystal hydrogel assisted scaffolds with bone-like ECM microenvironment for bone regeneration[J]. Carbohydr Polym, 2024, 15(332): 121927. DOI: 10.1016/j.carbpol.2024.121927.
[11] 孙琛, 朱青青, 王佳琼, 等. BMP2/Smads通路介导富血小板纤维蛋白对人牙髓干细胞成骨分化的作用研究[J]. 口腔颌面修复学杂志, 2021, 22(5): 333-337. DOI:10.19748/j.cn.kqxf.1009-3761.2021.09.003.
Sun C, Zhu QQ, Wang JQ, et al. BMP2/Smads pathway mediates the effect of platelet-rich fibrin on the osteogenic differentiation of human dental pulp stem cells [J]. Journal of Oral and Maxillofacial Repair, 2021, 22 (5): 333-337. DOI:10.19748/j.cn.kqxf.1009-3761.2021.09.003.
[12]Sane VD, Sunil Nair V, Jadhav R, et al. Comparative Evaluation of Efficacy of Platelet Rich Plasma (PRP) and Platelet Rich Fibrin (PRF) in Bone Regeneration after Surgical Removal of Impacted Bilateral Mandibular Third Molars-A Comparative Study[J]. Indian J Otolaryngol Head Neck Surg, 2024, 76(1): 811-818. DOI:10.1007/s12070-023-04285-0.
[13]Dousti M, Golmohamadpour A, Hami Z, et al. Ca-AlN MOFs-loaded chitosan/gelatin scaffolds; a dual-delivery system for bone tissue engineering applications[J]. Nanotechnology, 2024, 35(14): 145101. DOI:10.1088/1361-6528/ad0ef4.
[14]Lv L, Cheng W, Wang S, et al. Poly (β-amino ester) Dual-Drug-Loaded Hydrogels with Antibacterial and Osteogenic Properties for Bone Repair[J]. ACS Biomater Sci Eng, 2023, 9(4):1976-1990. DOI:10.1021/acsbiomaterials.2c01524.
[15]Anaya-Sampayo LM, García-Robayo DA, Roa NS, et al. Platelet-rich fibrin (PRF) modified nano-hydroxyapatite/chitosan/gelatin/alginate scaffolds increase adhesion and viability of human dental pulp stem cells (DPSC) and osteoblasts derived from DPSC[J]. Int J Biol Macromol, 2024, 273(Pt 1): 133064. DOI:10.1016/j.ijbiomac.2024. 133064.
[16]Mittal N, Baranwal HC, Aggarwal H, et al. Comparison of double antibiotic chitosan hydrogel scaffold with platelet-rich fibrin in regeneration in immature necrotic permanent teeth:Randomized controlled trial [J]. J Conserv Dent Endod, 2024, 27(12): 1251-1260. DOI:10.4103/JCDE.JCDE_609_24.
[17]Guo T, Lin S, Zou L, et al. Graphene Oxide Functionalized GelMA Platform Loaded With BFP-1 for Osteogenic Differentiation of BMSCs[J]. J Biomed Mater Res A, 2025, 113(1): e37829. DOI:10.1002/jbm.a.37829.
[18]Rasool N, Negi D, Singh Y. Thiol-Functionalized, Antioxidant, and Osteogenic Mesoporous Silica Nanoparticles for Osteoporosis[J]. ACS Biomater Sci Eng, 2023,9(6):3535-3545. DOI:10.1021/acsbiomaterials. 3c00479.
[19]Gan S, Zheng Z, Zhang M, et al. Lyophilized Platelet-Rich Fibrin Exudate-Loaded Carboxymethyl Chitosan/GelMA Hydrogel for Efficient Bone Defect Repair[J]. ACS Appl Mater Interfaces, 2023, 15(22): 26349-26362. DOI:10.1021/acsami.3c02528.
[20] 吴智恒, 吴睿哲, 荣尚, 等. 基于BMP/Smads/Runx2信号通路探讨桃红四物汤对创伤性股骨头坏死模型大鼠成骨化表达和病理形态学的影响[J]. 中医药导报, 2022, 28(9): 7-13. DOI:10.13862/j.cn43-1446/r.2022.09.002.
Wu ZH, Wu RZ, Rong S, et al. To explore the effects of Taohong Siwu Decoction on ossification expression and pathological morphology in a rat model of traumatic femoral head necrosis based on the BMP/Smads/Runx2 signaling pathway [J]. Chinese Medicine Review, 2022, 28(9): 7-13. DOI:10.13862/j.cn43-1446/r.2022.09.002.
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