CAJ | 학술논문

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/mL）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。
목적：탐토연골형태발생단백1（CDMP1）유도적반흔성섬유세포재체내배경하적연골구건능력。방법취반흔절제술후주기적증생성반흔조직，제취반흔성섬유세포。장반흔성섬유세포여PGA/PLA지가복합，CDMP1연골유도액（CDMP1종농도위100 ng/mL）진행유도배양2주，설위유도조（n=10）；장상규배양액배양적반흔성섬유세포－재료복합물식입라서체내작위음성대조，설위비유도조（n=4）；장연골세포－재료복합물식입라서체내작위양성대조，설위연골조（n=4）。분별우4주화8주후취재，진행각조습중、당알취당（GAG）함량측정，HE염색、Safranine-O염색화Ⅱ형효원면역조화염색。결과체내배양4주、8주후각조습중、GAG함량측정현시，유도조균고우비유도조（P<0.05）。체내배양4주후，유도조HE염색결과현시，반흔성섬유세포유도후출현연골세포함와결구；Safranine-O염색결과시，GAG균균분포우기질；면역조직화학염색시부분반흔성섬유세포기질중COLⅡ양성표체。8주시，유도조적류연골결과상대4주시경가성숙，경가부합연골결구분포。결론재CDMP1유도하，반흔성섬유세포여PGA/PLA재료복합，재체내가이형성류연골조직，구비일정적성연골능력。
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.