Novel collagen membrane in repairing skull bone defects in rats

doi:10.12307/2026.208

Abstract

Abstract: BACKGROUND: The commonly used absorbable barrier membrane in guided bone regeneration technology is Bio-Gide collagen membrane (source of pig skin), which has the disadvantages of poor mechanical strength and fast degradation rate, thus limiting its clinical application. Therefore, it is of great significance to develop collagen membranes with better physicochemical properties.
OBJECTIVE: To prepare a novel collagen membrane derived from porcine bladder, characterize its physicochemical properties and its effect on repairing rat skull bone defects.
METHODS: (1) The serosal layer and partial muscle layer of porcine bladders were removed to prepare a novel collagen membrane. The surface morphology, water absorption, porosity, degradation rate, tensile modulus, and ultimate load of the novel and Bio-Gide collagen membranes were characterized. (2) Rat bone marrow mesenchymal stem cells were co-cultured with the novel and Bio-Gide collagen membranes, respectively. Cells cultured alone were used as controls. Cell proliferation was assessed by CCK-8 assay. Osteogenic differentiation was assessed by alkaline phosphatase staining 7 days after osteogenic induction. (3) A 5-mm-diameter circular, full-thickness bone defect was created on each side of the sagittal suture of the skull in 18 SD rats. Thirty-six defect sites were randomly divided into six intervention groups: the control group (n=6) received no implantation at the defect site; the Bio-Oss group (n=6) had the defect filled with Bio-Oss bone powder; the Bio-Gide group (n=6) had the defect covered with a Bio-Gide membrane; the novel collagen membrane group (n=6) had the defect covered with a novel collagen membrane; the Bio-Oss+Bio-Gide group (n=6) had the defect filled with Bio-Oss bone powder and then covered with a Bio-Gide membrane; and the Bio-Oss+novel collagen membrane group (n=6) had the defect filled with Bio-Oss bone powder and then covered with a novel collagen membrane. Twelve weeks after surgery, the tissue samples were harvested for micro-computed tomography and histological observation.
RESULTS AND CONCLUSION: (1) Scanning electron microscopy revealed that the fibers in the compact layers of the Bio-Gide and novel collagen membranes were denser, with the fibers in the Bio-Gide membrane interlaced. The fibers in the novel collagen membrane were denser and connected into sheets. The fibers in the porous layers of both membranes were looser, with the Bio-Gide membrane having more pores. The porosity and water absorption of the novel collagen membrane were lower than those of the Bio-Gide membrane (P < 0.05), while the tensile elastic modulus and ultimate load were higher than those of the Bio-Gide membrane (P < 0.05). (2) Compared with the Bio-Gide membrane, the novel collagen membrane was more stable to degradation. CCK-8 assay showed that both the Bio-Gide and novel collagen membranes promoted the proliferation of rat bone marrow mesenchymal stem cells. Alkaline phosphatase staining revealed that neither the Bio-Gide nor the novel collagen membranes affected the osteogenic differentiation of rat bone marrow mesenchymal stem cells. (3) Micro-computed tomography revealed a small amount of new bone formation at the bone defect site in the control group, while substantial new bone formation was observed in the other five groups. The Bio-Oss+novel collagen membrane group demonstrated the greatest amount of new bone formation and structural maturity. Hematoxylin-eosin and Masson staining revealed less new bone formation at the defect site in the control group, while more was observed in the other five groups. The Bio-Oss+novel collagen membrane group demonstrated higher new bone density and more mature structure. (4) The results demonstrated that the novel collagen membrane exhibited superior physical and chemical properties to the Bio-Gide membrane. When combined with Bio-Oss bone powder, the novel collagen membrane demonstrated superior bone repair efficacy compared with the Bio-Gide membrane.

Key words: guided bone regeneration, novel collagen membrane, Bio-Gide membrane, rat, skull bone defect, bone marrow mesenchymal stem cell, efficacy, micro-computed tomography

CLC Number:

Yang Ping, Qi Xiaoyang, Lei Zhijie, Chen Yixin, Qiu Xusheng. Novel collagen membrane in repairing skull bone defects in rats[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(32): 8364-8371.

Figures/Tables 9

References

[1] BAI Y, WANG Z, HE X, et al. Application of Bioactive Materials for Osteogenic Function in Bone Tissue Engineering. Small Methods. 2024;8:e2301283.
[2] WANG J, LIU M, YANG C, et al. Biomaterials for bone defect repair: Types, mechanisms and effects. Int J Artif Organs. 2024;47:75-84.
[3] KIM K, SU Y, KUCINE AJ, et al. Guided Bone Regeneration Using Barrier Membrane in Dental Applications. ACS Biomater Sci Eng. 2023;9(10): 5457-5478.
[4] ELGALI I, OMAR O, DAHLIN C, et al. Guided bone regeneration: materials and biological mechanisms revisited. Eur J Oral Sci. 2017;125(5):315-337.
[5] ALQAHTANI AM. Guided Tissue and Bone Regeneration Membranes: A Review of Biomaterials and Techniques for Periodontal Treatments. Polymers (Basel). 2023;15(16):3355.
[6] YANG Z, WU C, SHI H, et al. Advances in Barrier Membranes for Guided Bone Regeneration Techniques. Front Bioeng Biotechnol. 2022;10:921576.
[7] ALI M, MOHD NOOR SNF, MOHAMAD H, et al. Advances in guided bone regeneration membranes: a comprehensive review of materials and techniques. Biomed Phys Eng Express. 2024;10(3):32003.
[8] BEE SL, HAMID ZAA. Asymmetric resorbable-based dental barrier membrane for periodontal guided tissue regeneration and guided bone regeneration: A review. J Biomed Mater Res B Appl Biomater. 2022;110(9):2157-2182.
[9] MATEO-SIDRON ANTON MC, PEREZ-GONZALEZ F, MENIZ-GARCIA C. Titanium mesh for guided bone regeneration: a systematic review. Br J Oral Maxillofac Surg. 2024;62:433-440.
[10] VROOM MG, GRUNDEMANN LJ, GALLO P. Clinical Classification of Healing Complications and Management in Guided Bone Regeneration Procedures with a Nonresorbable d-PTFE Membrane. Int J Periodontics Restorative Dent. 2022;42:419-427.
[11] REN Y, FAN L, ALKILDANI S, et al. Barrier Membranes for Guided Bone Regeneration (GBR): A Focus on Recent Advances in Collagen Membranes. Int J Mol Sci. 2022;23(23):14987.
[12] ABTAHI S, CHEN X, SHAHABI S, et al. Resorbable Membranes for Guided Bone Regeneration: Critical Features, Potentials, and Limitations. ACS Mater Au. 2023;3(5):394-417.
[13] ZHOU Z, YUN J, LI J, et al. Comparison of the efficacy of different biodegradable membranes in guided bone/tissue regeneration: a systematic review and network meta-analysis. Biomed Mater. 2023;18:032003.
[14] MATHEW A, VAQUETTE C, HASHIMI S, et al. Antimicrobial and Immunomodulatory Surface-Functionalized Electrospun Membranes for Bone Regeneration. Adv Healthc Mater. 2017;6(10):1601345.
[15] XUE J, HE M, LIU H, et al. Drug loaded homogeneous electrospun PCL/gelatin hybrid nanofiber structures for anti-infective tissue regeneration membranes. Biomaterials. 2014;35:9395-9405.
[16] HAN F, ZHANG P, SUN Y, et al. Hydroxyapatite-doped polycaprolactone nanofiber membrane improves tendon-bone interface healing for anterior cruciate ligament reconstruction. Int J Nanomedicine. 2015; 10:7333-7343.
[17] KARFELD-SULZER LS, GHAYOR C, SIEGENTHALER B, et al. Comparative study of NMP-preloaded and dip-loaded membranes for guided bone regeneration of rabbit cranial defects. J Tissue Eng Regen Med. 2017;11:425-433.
[18] BENITO-GARZON L, GUADILLA Y, DIAZ-GUEMES I, et al. Nanostructured Zn-Substituted Monetite Based Material Induces Higher Bone Regeneration Than Anorganic Bovine Bone and beta-Tricalcium Phosphate in Vertical Augmentation Model in Rabbit Calvaria. Nanomaterials (Basel). 2021; 12:143.
[19] PARK WB, CRASTO GJ, HAN JY, et al. Bone Regenerative Potential of Cross-Linked Collagen Membrane in Peri-Implant Osseous Defect: Case Series with Histologic/Micro-Computed Tomographic Findings. Medicina (Kaunas). 2023;59:176.
[20] ZHANG C, DU T, MU G, et al. Evaluation and ultrastructural changes of amniotic membrane fragility after UVA/riboflavin cross-linking and its effects on biodegradation. Medicine (Baltimore). 2020;99:e20091.
[21] CIARDELLI G, GENTILE P, CHIONO V, et al. Enzymatically crosslinked porous composite matrices for bone tissue regeneration. J Biomed Mater Res A. 2010;92(1):137-151.
[22] TERZI A, STORELLI E, BETTINI S, et al. Effects of processing on structural, mechanical and biological properties of collagen-based substrates for regenerative medicine. Sci Rep. 2018;8:1429.
[23] ZHANG JY, LIU K, LIU RX, et al. Safety and Efficacy of Midface Augmentation Using Bio-Oss Bone Powder and Bio-Gide Collagen Membrane in Asians. J Clin Med. 2023;12(3):959.
[24] APAZA ALCCAYHUAMAN KA, HEIMEL P, TANGL S, et al. Active and Passive Mineralization of Bio-Gide((R)) Membranes in Rat Calvaria Defects. J Funct Biomater. 2024;15(3):54.
[25] 孙晓嘉,马宁.不同口腔修复膜材料在牙种植中的引导骨再生效果分析[J].中国现代药物应用,2024,18(15):51-54.
[26] 尤鹏越,刘玉华,王新知,等.脱细胞猪心包膜生物相容性及成骨性能的体内外评价[J].北京大学学报(医学版),2021,53(4):776-784.
[27] 林季萩,莫安春.牛心包膜在美学区水平骨增量手术中的应用效果分析[J].口腔疾病防治,2025,33(3):203-210.
[28] 高丹妮,郭星,谢言,等.GTR联合根管治疗左上前牙牙周牙髓联合病变1例[J].牙体牙髓牙周病学杂志,2024,29(12):719-722.
[29] MEYER M. Processing of collagen based biomaterials and the resulting materials properties. Biomed Eng Online. 2019;18(1):24.
[30] 王莉莉,佳严,李东升,等.两种新型胶原膜引导骨组织再生的体内外性能研究[J].口腔医学,2019,39(6):481.
[31] BARBECK M, LORENZ J, KUBESCH A, et al. Porcine Dermis-Derived Collagen Membranes Induce Implantation Bed Vascularization Via Multinucleated Giant Cells: A Physiological Reaction? J Oral Implantol. 2015;41:e238-251.
[32] NOBLE C, MORSE D, LERMAN A, et al. Evaluation of Pericardial Tissues from Assorted Species as a Tissue-Engineered Heart Valve Material. Med Biol Eng Comput. 2022;60:393-406.