软骨形态发生蛋白1诱导的瘢痕成纤维细胞体内成软骨能力的实验研究

目的探讨软骨形态发生蛋白1(CDMP1)诱导的瘢痕成纤维细胞在体内环境下的软骨构建能力。方法取瘢痕切除术后丢弃的增生性瘢痕组织,提取瘢痕成纤维细胞。将瘢痕成纤维细胞与PGA/PLA支架复合,CDMP1软骨诱导液(CDMP1终浓度为100 ng/m L)进行诱导培养2周,设为诱导组(n=10);将常规培养液培养的瘢痕成纤维细胞-材料复合物植入裸鼠体内作为阴性对照,设为非诱导组(n=4);将软骨细胞-材料复合物植入裸鼠体内作为阳性对照,设为软骨组(n=4)。分别于4周和8周后取材,进行各组湿重、糖胺聚糖(GAG)含量测定,HE染色、Safranine-O染色和Ⅱ型胶原免疫组化染色。结果体内培养4周、8周后各组湿重、GAG含量测定显示,诱导组均高于非诱导组(P<0.05)。体内培养4周后,诱导组HE染色结果显示,瘢痕成纤维细胞诱导后出现软骨细胞陷窝结构;Safranine-O染色结果示,GAG均匀分布于基质;免疫组织化学染色示部分瘢痕成纤维细胞基质中COLⅡ阳性表达。8周时,诱导组的类软骨结果相对4周时更加成熟,更加符合软骨结构分布。结论在CDMP1诱导下,瘢痕成纤维细胞与PGA/PLA材料复合,在体内可以形成类软骨组织,具备一定的成软骨能力。

Chondrogenic Differentiation of Hypertrophic Scar -Derived Fibroblasts Initiated by Cartilage -derived Morphogenetic Protein 1 in Vivo

Objective To explore the chondrogenesis potential of hypertrophic scar fibroblasts （HSFBs） induced by cartilage-derived morphogenetic protein 1 （CDMP1） in vivo. Methods Hypertrophic scar fibroblasts （HSFBs） were isolated from discarded hypertrophic scar. HSFBs combining with PGA/PLA scaffold as HSFBs- PGA/PLA complex were induced by CDMP1 （100 ng/mL） for 2 weeks. Then the complex were transplanted into nude mice as induced group （n=10）. The complex that not induced were treated as negative control, named as un-induced group （ n=4）, while the chondrocytes-PGA/PLA complex were treated as positive control, named as chondrocyte group （n=4）. After 4 and 8 weeks, the complex in each group was harvested for wet weight test, proteoglycan （GAG） content test, HE staining, Safranine-O staining and immunochemistry staining. Results After 4 and 8 weeks in vivo, the wet weight and GAG content in induced group were both higher than in un-induced group （P〈0.05）. In induced group, after cultured for 4 weeks in vivo, chondrocyte-like lacuna structure was observed by HE staining, uniform distribution of GAG was observed by Safranine-O staining, and positive expression of collagen type Ⅱ was revealed by immunohistochemical staining. After 8 weeks of culturing in vivo, chondrocyte-like structure in induced group was more mature and more in line with the structure distribution of cartilage. Conclusion HSFBs combining with PGA/PLA had the potential to develop into polygonal chondrocyte-like tissue by the induction of CDMP 1.