缓释双生长因子三层支架体内长期促骨软骨缺损再生的研究  

Promotion of Sustained-Release Double Growth Factors from Three-Layer Scaffolds to Osteochondral Defect Regeneration:A Long-Term Study in Vivo

作　　者：董云生 黄佳星 肖慧 王雅菁 王茜霖 王淑芳[1,2] Dong Yunsheng;Huang Jiaxing;Xiao Hui;Wang Yajing;Wang Qianlin;Wang Shufang(College of Life Sciences,Nankai University,Tianjin 300071,China;Key Laboratory of Bioactive Materials for Ministry of Education,Nankai University,Tianjin 300071,China)

机构地区：[1]南开大学生命科学学院,天津300071 [2]南开大学生物活性材料教育部重点实验室,天津300071

出　　处：《天津大学学报（自然科学与工程技术版）》2023年第6期588-594,共7页Journal of Tianjin University：Science and Technology

基　　金：天津市科技支撑计划资助项目(20YFZCSY01020);国家自然科学基金资助项目(31870966);国家重点研发计划资助项目(2020YFA0803701).

摘　　要：外伤或关节使用过度或重度关节炎都会导致关节表面的软骨以及软骨下骨形成难以恢复的损伤.尽管目前临床上的治疗技术在某些方面取得成功,但仍存在治疗的局限性,如自体移植存在供区并发症、移植的组织不完全整合和易降解、骨髓刺激会产生纤维软骨、质量不佳等.为提高骨软骨缺损的治疗效果,前期研究制备了局部缓释生长因子rhBMP-2和rhTGF-β3的丝素蛋白/脱软骨细胞外基质/纳米羟基磷灰石复合三层支架,并在兔子体内短期植入后获得良好的骨软骨修复效果,但并未进行长期体内研究.本研究在兔子双膝滑车沟处建立直径5mm、深度3mm的缺损,并将具有局部缓释生长因子rhBMP-2和rhTGF-β3的复合三层支架进行缺损区植入,且以天然膝关节滑车沟处骨软骨为对照组,修复34周后通过大体观察、组织学染色、免疫组织化学染色、Micro-CT扫描以及力学测定,评价支架对骨软骨缺损长期的修复情况.大体观察、组织学染色、免疫组织学染色扫描结果表明,支架组再生软骨具有与天然组相似的表面平整度和完整度、细胞排列形态以及黏多糖含量;但Ⅱ型胶原染色和软骨纳米压痕结果表明,支架组再生软骨Ⅱ型胶原含量以及力学性能明显低于天然软骨.Micro-CT扫描以及力学结果表明,支架组再生软骨下骨具有与天然软骨下骨相似的结构以及力学强度((48.98±12.7091)MPa).因此,具有局部缓释生长因子rhBMP-2和rhTGF-β3的丝素蛋白复合三层支架在体内长期促进骨软骨再生方面具有显著的优势.Trauma,overuse,or severe knee osteoarthritis of the joint can irreversibly damage the cartilage on the joint and the subchondral bone.Although primary clinical treatments have been found to be effective in a few cases,they are associated with certain limitations,such as complications,incomplete integration,easy degradation,fibrocartilage development,and poor quality.For the treatment of osteochondral defects,a previous study developed a three-layer scaffold composite of silk fibroin/decellularized articular cartilage extracellular matrix/nanohydroxyapatite that could cause local and sustained-release of the growth factors rhBMP-2 and rhTGF-β3.Short-term implantation of this scaffold in rabbits resulted in a good osteochondral repair effect.However,no long-term in vivo studies have been performed on such three-layer scaffolds.In this study,topical sustained-release three-layer scaffolds comprising growth factors rhBMP-2 and rhTGF-β3 were implanted on the osteochondral defects(diameter:5mm,depth:3mm)of the bilateral trochlear groove of rabbit knee joints.The osteocartilage at the trochlear groove of the native knee joint was used as the control.The implanted scaffolds were evaluated for osteochondral defect repair by gross observation,histological staining,immunohistochemical staining,microcomputed tomography(Micro-CT),and mechanical assessment after 34 weeks.The results of gross observation,histological staining,and immunohistologi-cal staining showed that the regenerated cartilage in the scaffold group had similar surface planarity and integrity,cell alignment morphology,and mucopolysaccharide content as the cartilage in the native group.However,the results of typeⅡcollagen staining and cartilage nanoindentation revealed that typeⅡcollagen content and the mechanical properties of regenerated cartilage in the scaffold group were significantly lower than those in the native group.Micro-CT and mechanical testing indicated that both the scaffold and native groups had similar structures of the regenerated subcho

关 键 词：缓释支架 生长因子 骨软骨修复 体内研究 组织工程 

分 类 号：Q819[生物学—生物工程]