PLLA/PDLA三维支架与滑膜间充质干细胞/软骨细胞构建颞下颌关节盘的研究

Up to now, there is no good treatment for the perforated temporomandibular joint (TMJ) disc, and there is also no article that reported the application of co-cultured synovial mesenchymal stem cells (SMSCs)/articular chondrocytes (ACs) for the tissue engineering of TMJ disc. In this study, SMTCs and ACs were applied as seeding cells, and collagen type I (COL-I) and poly-L-lactic acid (PLLA) as well as poly-D-lactic acid (PDLA) as scaffold. Through the method of electronic spinning, the spongy-like P(L/D)LA 96/4 and membrane-like P(L/D)LA 70/30 were fabricated and sutured together, and then COL-I were added into the P(L/D)LA scaffold. The SMSCs/ACs were cultured onto the P(L/D)LA scaffold and induced by cytokines, and the cells/scaffold constructs were transplanted into the perforated TMJ disc rabbits. Then the constructs were examined to explore its physical and the molecular characterstics. The novelty of this study might be that the SMSCs and ACs were co-cultured as feeding cells in the TMJ disc tissue engineering.

目前，对于颞下颌关节盘破裂与穿孔尚无理想的治疗方法，亦无利用滑膜间充质干细胞与软骨细胞共同构建颞下颌关节盘的报道。所以，本研究选用这两种细胞作为种子细胞。以I型胶原（collagen type I, COL-I）、左旋聚乳酸（poly-L-lactic acid, PLLA）、消旋聚乳酸（poly-D-lactic acid, PDLA）为原料，通过电纺丝法制备海绵状P(L/D)LA 96/4和膜状P(L/D)LA 70/30两种材料，将二者合为上下两层P(L/D)LA材料，COL-I与双层P(L/D)LA复合成COL-I/P(L/D)LA支架。滑膜间充质干细胞/软骨细胞三维培养于支架中，特定的细胞因子诱导细胞支架复合物，然后植入颞下颌关节盘穿孔的动物体内，了解其在颞下颌关节盘组织工程中的作用。本研究创新之处在于探讨这两种细胞混合培养后向纤维软骨分化，选用的支架在结构上更模拟颞下颌关节盘。

目前，对于颞下颌关节盘破裂与穿孔尚无理想的治疗方法，亦无利用滑膜间充质干细胞与软骨细胞共培养构建颞下颌关节盘的报道。为探索共培养负载COL-I/P(L/D)LA复合支架修复关节盘及软骨缺损可行性，一定程度的模拟软骨缺损。我们获取成年雄性大鼠关节软骨及滑膜组织，对其干细胞的生物学特性及其干性进行鉴定并通过不同比例的细胞接触式混合培养，建立、完善接触式混合培养的基本操作流程以及最适合比例，其中以1:2的比例能更好的形成软骨及其外基质。最后鉴定在体外形成了软骨组织。进而负载于支架后回植体内软骨缺损，在体内进行1w、4w、8w的培养，通过形态学、免疫组化等鉴定结果鉴定其合成产物。最后实验结果与预期结果一致，通过本研究，明确SMSCs/软骨细胞混合培养向软骨分化的能力与条件，为组织工程化颞下颌关节盘的构建提供依据。