藻酸盐凝胶三维培养体系在TMJ骨关节病髁突软骨细胞体外培养中的应用

Application of alginate three-dimensional culture system for in vitro culture of mandibular condylar chondrocytes from human osteoarthritic temporomandibular joint

目的 将藻酸盐凝胶三维培养体系应用于颞下颌关节骨关节病 (osteoarthritisoftemporomandibularjoint,TMJOA)髁突软骨细胞的体外培养。方法 利用机械与酶消化的方法从TMJOA患者手术切除患疾髁突软骨中获得髁突软骨细胞 ,部分细胞在二维贴壁培养条件下体外培养 1周 ;部分细胞在高密度条件下转入藻酸盐凝胶培养介质 ,进行三维培养 4周 ;分别对贴壁条件下培养细胞和藻酸盐细胞凝胶微球石蜡包埋切片进行Ⅱ型胶原和软骨特异性蛋白多糖的免疫组化鉴定。结果 从骨关节病髁突软骨组织中收获并行体外培养成活的细胞 ,鉴定为软骨细胞 ;体外培养的藻酸盐凝胶三维培养体系中的髁突软骨细胞生长状态良好 ;培养 4周后细胞保持了良好的分化表型。结论 成功地体外培养成活了人TMJOA髁突软骨细胞 ;成功地将藻酸盐凝胶三维培养体系应用于人TMJOA髁突软骨细胞体外培养 ,在该体系中 ,软骨细胞生长状态与功能蛋白分泌能力良好。

Objective To evaluate the use of alginate three dimensional culture system for mandibular condylar chondrocytes culture in vitro from human osteoarthritic temporomandibular joint. Methods The cultured mandibular condylar chondrocytes from the operatively removed cartilage from a patient with osteoarthritic temporomandibular joint were harvested by mechanical dissection and enzyme digestion. Partial chondrocytes were suspended in the aqueous sodium alginate beads with high seeding density and cultured for 4 weeks, while the others were cultured in monolayer culture condition for 1 week. Thereafter, the alginate beads were embedded in paraffin and sectioned, then studied immunologically with type Ⅱ collagen antibody and aggrecan antibody, same studies were adopted for the monolayer cultures. Results The monolayer cultures were confirmed as chondrocytes. The chondrocytes cultured in the alginate medium showed well. These cells exhibited the excellent differentiated phenotype after 4 weeks culture in alginate gel. Conclusions The condylar chondrocytes from human osteoarthritic temporomandibular joint were successfully cultured in vitro. The alginate three dimensional culture system was successfully adopted for in vitro culture of condylar chondrocytes from human osteoarthritic temporomandibular joint, in which the chondrocytes exhibited the excellent differentiated phenotype.