EGCG交联仿生矿化马面鱼鱼皮脱细胞基质用作引导骨组织再生膜的实验研究

doi:10.13701/j.cnki.kqyxyj.2024.05.013

摘要/Abstract

摘要： 目的:构建马面鱼鱼皮脱细胞基质,并通过交联和构建仿生矿化模板对其进行改性,为马面鱼鱼皮作为骨再生(guided bone regeneration,GBR)膜开发提供理论依据。方法:选取了海洋来源的马面鱼鱼皮,采用物理化学联合法制备马面鱼鱼皮脱细胞基质(filefish skin decellularized matrix,FS-ECM),通过苏木精-伊红染色法(hematoxylin-eosin staining,HE)、扫描电子显微镜(scanning electron microscope,SEM)、X射线衍射(X-ray diffraction,XRD)和水接触角初步探讨了FS-ECM的结构特征;同时,一方面利用表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)交联修饰EGCG交联改性马面鱼鱼皮脱细胞基质(EGCG crosslinked filefish skin decellularized matrix,E-FS-ECM)以提高材料的抗酶解和机械性能,另一方面通过EGCG的修饰构建EGCG-胶原体外仿生矿化模板EGCG交联仿生矿化改性马面鱼鱼皮脱细胞基质(EGCG crosslinked biomimetic mineralized filefish skin decellularized matrix,EB-FS-ECM),利用胶原仿生矿化策略进一步提高各项性能,采用SEM、mapping、水接触角、弹性模量、热重分析和体外降解评价改性前后材料的理化性能的差异。结果:不同部位FS-ECM的表面结构特性之间无明显差异(P>0.05),且FS-ECM外表面含有一定量的羟基磷灰石晶体;经过EGCG交联和构建仿生矿化模版的改性,E-FS-ECM、EB-FS-ECM的亲水性、机械强度、热稳定性和抗酶解能力均明显高于FS-ECM(P<0.05)。结论:EGCG交联和仿生矿化能够明显地提高FS-ECM的亲水性、热稳定性、机械强度并减慢了降解速率,通过改性后的马面鱼鱼皮脱细胞基质有望成为理想的引导骨再生技术(guided bone regeneration,GBR)膜材料。

关键词: 马面鱼鱼皮, 脱细胞, 表没食子儿茶素没食子酸酯, 交联, 仿生矿化

Abstract: Objective: To construct decellularized matrix of filefish skin and crosslink it to be biomimetic mineralization template as a guided bone regeneration membrane. Methods: Filefish skin of marine origin was selected, and the decellularized matrix of filefish skin (filefish skin decellularized matrix,FS-ECM) was prepared by a combined physicochemical method. The structural features of FS-ECM were preliminarily explored by HE staining, scanning electron microscope (SEM), X-ray diffraction (XRD), and water contact angle. Then, the cross-linking modification of epigallocatechin gallate (EGCG) was utilized (EGCG crosslinked filefish skin decellularized matrix,E-FS-ECM) to improve the material's anti-enzymatic and mechanical properties. The EGCG-collagen in vitro biomimetic mineralization template (EGCG crosslinked biomimetic mineralized filefish skin decellularized matrix, EB-FS-ECM) was constructed through the modification of EGCG, and the collagen biomimetic mineralization strategy was used to further improve the various properties. The differences in the physicochemical properties of the materials before and after modification were evaluated using SEM, mapping, water contact angle, elastic modulus, thermogravimetric analysis, and in vitro degradation. Results: There was no significant difference between the surface structural properties of FS-ECM at different sites (P>0.05), and FS-ECM contained a certain amount of hydroxyapatite crystals on the outer surface. After the modification by EGCG cross-linking and construction of biomimetic mineralization templates, the hydrophilicity, mechanical strength, thermal stability, and resistance to enzymatic degradation of E-FS-ECM and EB-FS-ECM were significantly higher than those of FS-ECM (P<0.05). Conclusion: EGCG cross-linking and biomimetic mineralization significantly improved the hydrophilicity, thermal stability, and mechanical strength, and slowed down the degradation rate of FS-ECM. Decellularized matrix of filefish skin through modification is expected to be an suitable material for GBR membranes.

Key words: filefish skin, decellularization, EGCG, cross-linking, biomimetic mineralized

沈圣洁, 孙宁, 肖停, 李全利. EGCG交联仿生矿化马面鱼鱼皮脱细胞基质用作引导骨组织再生膜的实验研究[J]. 口腔医学研究, 2024, 40(5): 448-455.

SHEN Shengjie, SUN Ning, XIAO Ting, LI Quanli. EGCG Crosslinked Biomimetic Mineralized Decellularized Matrix of Filefish Skin As A Membrane for Guided Bone Tissue Regeneration[J]. Journal of Oral Science Research, 2024, 40(5): 448-455.

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