论文部分内容阅读
目的探讨软骨细胞-动物源性骨软骨支架复合体修复兔膝关节骨软骨复合缺损的可行性和影响因素。方法将改良贴壁离心法获取的骨髓间充质干细胞(bone marrowm esenchymal stem cells,BMSCs)/诱导分化的软骨细胞共培养后与经深低温冷冻、脱脂、脱钙、真空冷冻干燥和辐照消毒的动物源性骨软骨支架复合,构建共培养细胞+软骨-骨一体化复合支架。27只新西兰大白兔随机分为实验组(A组)、对照组(B组)和空白组(C组),每组9只。于兔股骨髁间窝处钻一深6mm的骨软骨复合缺损,A组植入共培养细胞+骨软骨复合支架,B组植入骨软骨复合支架,C组不植入任何支架材料和细胞,分别于术后4周、8周和12周取材,行大体观察、苏木精—伊红染色和甲苯胺蓝染色,并对各标本的软骨切片进行组织学评分。结果随着时间的延长,A组大体观察见复合缺损区已完全修复,局部无凹陷,新生组织和周围组织融合;B组新生组织仍不能完全填充缺损;C组缺损区仍明显。苏木精—伊红染色和甲苯胺蓝染色见A组软骨缺损区由新生的透明软骨样组织修复,细胞呈柱状排列,极性好,软骨陷窝明显,骨缺损区由骨样组织修复,新生软骨和软骨下骨以及宿主骨界面耦合良好;B组新生软骨细胞无软骨陷窝,排列混乱,各界面藕合欠理想;C组可见陈旧性肉芽组织生长并突出于缺损区表面。甲苯胺蓝染色阳性率和组织学评分结果表明,A组与B、C两组之间的差异具有统计学意义(P<0.05)。结论 BMSCs/诱导分化的软骨细胞共培养细胞复合动物源性骨软骨支架对兔膝关节软骨和软骨下骨的复合缺损具有修复作用。
Objective To investigate the feasibility and influencing factors of chondrocyte-animal osteochondral scaffold complex in repair of osteochondral composite defects of rabbit knee. Methods BMSCs / induced chondrocytes obtained by improved adherence centrifugation were co-cultured with cryopreservation, degreasing, decalcification, vacuum freeze-drying and irradiation sterilization Animal-derived osteochondral scaffold complex to construct coculture cells + cartilage-bone composite scaffold. Twenty-seven New Zealand white rabbits were randomly divided into experimental group (A group), control group (B group) and blank group (C group). A deep 6mm osteochondral composite defect was drilled in the femoral intercondylar fossa. A group was implanted with cocultured cells + osteochondral composite scaffolds, B group was implanted with osteochondral composite scaffolds, C group was not implanted with any scaffolds and cells, The specimens were harvested at 4, 8 and 12 weeks postoperatively. Gross observation, hematoxylin-eosin staining and toluidine blue staining were performed. The cartilage sections of each specimen were histologically scored. Results With the prolongation of time, in group A, the composite defect area was completely repaired and no local depression was found. The neogenesis and surrounding tissues were fused. The defects in group B still could not be completely filled with defects. The defect in group C was still obvious. Hematoxylin-eosin staining and toluidine blue staining showed cartilage defect in group A was repaired by new hyaline cartilage-like tissue. The cells were arranged in columnar shape with good polarity and obvious cartilage lacuna. The bone defect was repaired by osteoid tissue, The newborn cartilage and subchondral bone and host bone interface were well coupled. In group B, there were no cartilaginous lacunar disorganized cartilage, confused arrangement and less ideal coupling at each interface. In group C, the old granulation tissue grew and protruded on the surface of defect area. Toluidine blue staining positive rate and histological score results showed that the difference between A group and B, C two groups was statistically significant (P <0.05). Conclusion BMSCs / differentiated chondrocytes co-cultured with cell-derived osteochondral scaffolds can repair composite defects of rabbit knee articular cartilage and subchondral bone.