TGF-β3转染脂肪干细胞复合OGP-HA-ChS支架对兔髁突受损软骨的修复作用

目的: 探讨TGF-β3转染脂肪干细胞ADSCs复合OGP（成骨多肽）-HA（透明质酸）-ChS（硫酸软骨素）支架修复兔髁突受损软骨的可行性。方法: 分离培养兔ADSCs,利用重组腺病毒构建携带TGF-β3基因表达载体,并转染至兔ADSCs。14 d后,观察细胞荧光表达情况并计算病毒转染效率,Western 印迹检测TGF-β3蛋白表达。50只大白兔随机分为5组,A组为空白组,B组为TGF-β3转染ADSCs组,C组为OGP-HA-ChS支架组,D组为ADSCs复合OGP-HA-ChS组。E组为TGF-β3转染ADSCs复合OGP-HA-ChS组,兔颞下颌关节骨关节病模型制备完成后,按照实验分组进行材料移植。各组动物分别于材料移植第3、9周后取材,制备标本并行髁突软骨扫描电镜、组织学观察及实时荧光定量PCR检测。采用SPSS17.0 软件包对数据进行统计学分析。结果: 扫描电镜与组织学观察可见,D、E组软骨病变修复改善情况优于B、C组。与模型组比较,实时定量 PCR结果显示,E组MMP-3 表达与A组相对持平但低于C、D组,差异有显著性（P<0.05);E组 TIMP-1 表达与A组相对持平但高于C、D组,差异有显著性（P<0.05)。结论: TGF-β3转染ADSCs复合OGP-HA-ChS支架对兔髁突受损软骨具有修复作用。

Effect of TGF-β3 transfected with adipose derived stem cells and OGP-HA-chondroitin sulfate scaffold on repair of condylar cartilage in rabbits

PURPOSE: To investigate the feasibility of repairing damaged cartilage of rabbit condylar with TGF-β3-transfected adipose stem cells combined with OGP (osteogenesis peptide)-HA (hyaluronic acid)-ChS (chondroitin sulfate) scaffold. METHODS: Rabbit ADSCs were isolated and cultured. The expression vector carrying TGF-β3 gene was constructed using recombinant adenovirus and transfected into rabbit ADSCs. The fluorescence expression of the cells was observed after 14 days and the efficiency of virus transfection was calculated. Western blot was used to detect the expression of TGF-β3 protein. Fifty rabbits were randomly divided into 5 groups: group A as blank group, group B as TGF-β3 transfected ADSCs group, group C as OGP-HA-ChS scaffold group, group D as ADSCs complex OGP-HA-ChS group, E group was TGF-β3 transfected ADSCs with OGP-HA-ChS. After the rabbit temporomandibular joint osteoarthrosis model was established, the animals were transplanted according to the experimental design. The animals in each group were sacrificed after the 3rd and 9th weeks of transplantation. Scanning electron microscopy, histological observation and real-time fluorescent quantitative PCR were performed accordingly. SPSS17.0 software package was used for analysis of variance and t test. RESULTS: Scanning electron microscopy and histological observation showed that the repair of cartilage lesions in group D and E was better than those in group B and C. Compared with group B, the results of real-time quantitative PCR showed that the expression of MMP-3 in group E was similar to that in group A, but significantly lower than that in group C and D (P<0.05). The expression of TIMP-1 in group E was similar to that of group A, but significantly higher than group C and D (P<0.05). CONCLUSIONS: TGF-β3 transfected ADSCs composite OGP-HA-ChS scaffold has repairing effect on rabbit condylar damaged cartilage.