摘要
目的构建含和不含天然钙化层的修复材料,比较两者在猪膝关节软骨缺损修复中的效果,探讨钙化层在骨软骨组织工程中的重要性。方法选取普通实验猪膝关节骨,软骨通过Ⅱ型胶原水凝胶冻干技术和天然骨软骨脱细胞技术构建含有钙化层和无钙化层的骨软骨支架,以贵州小香猪自体骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)为种子细胞,构建移植修复材料。30只小香猪按随机数字表法分为空白对照组、无钙化层组、含钙化层组(n=10)。双膝关节滑车骨软骨缺损造模,直径8 mm,深至软骨下骨,植入对应的修复材料。分别于12、24周取材,采用大体、体式显微镜观察,以及MRI检测缺损填充情况,HE染色、番红O-固绿染色,以及Ⅰ、Ⅱ型胶原蛋白免疫组化染色观察新生修复物的组织类别,O’Driscoll组织学评分评价修复效果。结果各组术后24周修复效果较12周均有改善。大体和体式显微镜观察,24周空白对照组新生组织部分填充缺损,为纤维组织,软骨表面凹陷深大;无钙化层组移植物填充缺损,软骨表面凹陷;含钙化层组缺损填充较好,表面微凹陷,三层结构可见,与宿主组织整合佳。组织学观察提示空白对照组为纤维组织填充修复;无钙化层组为骨和纤维软骨修复,三层结构不明显;含钙化层组钙化层结构清晰,移植物与宿主整合,软骨表面微凹陷,透明软骨修复。O’Driscoll组织学评分,12、24周空白对照组分别为(3.80±0.83)、(5.50±0.52)分,无钙化层组分别为(10.30±0.63)、(14.20±0.68)分,含钙化层组分别为(15.10±0.58)、(18.80±0.87)分,各组评分差异均有统计学意义(P<0.01),含钙化层组评分最高。结论含有天然钙化层的骨软骨支架,在猪骨软骨缺损修复中取得了很好的修复效果,明显优于无钙化层支架和空白对照,是今后软骨组织工程支架设计的重要参考。
Objective To explore whether the calcified cartilage zone （CCZ） is essential for tissue- engineered bone scaffold for repairing osteochondral defect in mini-pigs. Methods The bone scaffold materials were prepared by frozen-drying type II collagen cross-linked with trephined on decellularized pig bone with or without natural CCZ. The autologous bone marrow stem cells （BMSCs） of mini-pigs were injected into the cartilage layer of the scaffold to prepare the tissue-engineered implants. Thirty mini-pigs were randomized equally into blank control group, non-CCZ group, and CCZ group, and in each pig, full-thickness osteochondral defects of 8 mm in diameter were created on both knees and repaired using the corresponding implants. At 12 and 24 weeks after the operation, the defects were assessed by general observation, magneticresonance imaging （MRI）, HE staining, safranin O-fast green staining, type-I, type-II collagen immunohistochemical staining and O＇ Driscoll histological scores. Results In each group, obvious improvement in cartilage defect repair was observed at 24 weeks after the operation as compared with that at 12 weeks. In the blank control group, the defect was partly repaired by fibrous tissue, and deep defects remained at 24 weeks; in non-CCZ group, the defect was moderately repaired by the implants with small defects in the cartilage; in CCZ group, the defect was nearly fully repaired by the implant, which was well integrated with the host tissue and clearly showed the three-layered structure. Histological observations showed that the defects was filled with fibrous tissue in the blank control group, by bone and fibrocartilage without an identifiable three-layered structure in non-CCZ group, and with bone and hyaline cartilage without distinct boundaries with the host tissue in CCZ group. The O＇ Driscoll scores in the blank control, non-CCZ, and CCZ groups were 3.8 ±0.83, 10.3 ±0.63, and 15.1 ±0.58 at 12 weeks, and were 5.5 ±0.52, 14.2 ±0.68, and 18.8 ± 0. 87 at 24 weeks, respectivel
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2018年第4期307-315,共9页
Journal of Third Military Medical University
基金
国家自然科学基金重点项目(31130021)
国家重点研发计划(2017YFC1104103)~~
关键词
组织工程
钙化层
软骨缺损
骨软骨支架
自体
贵州小香猪
tissue engineering
calcified cartilage zone
osteochondral defects
osteochondral scaffold
autologous
mini-pigs
