CAJ | 학술논문

目的观察软骨细胞在同种异体松质骨骨基质明胶(BMG)上的生长、增殖和分化,探讨用松质骨BMG与软骨细胞体外构建组织工程软骨的效果.方法分离1月龄兔关节软骨细胞,扩增后种植在松质骨BMG上体外构建组织工程软骨.于1、2、4、6周取材进行组织学、Ⅱ型胶原免疫组织化学及透射电镜观察.结果软骨细胞在BMG上生长良好,分泌蛋白聚糖和胶原.随培养时间延长,细胞增殖,松质骨BMG空隙内细胞数量增多,软骨陷窝形成,基质分泌增强.培养6周时软骨细胞在BMG表面及孔隙内形成软骨样组织,免疫组织化学染色显示细胞周围基质富含Ⅱ型胶原,分布均匀:透射电镜显示软骨细胞超微结构正常,细胞周围有大量基质产生.结论同种异体松质骨BMG可以促进软骨细胞的生长增殖,维持软骨细胞的分化表型;在体外成功构建了组织工程软骨,它是一种较好的软骨组织工程支架材料.
목적 관찰연골세포재동충이체송질골골기질명효(BMG)상적생장、증식화분화,탐토용송질골BMG여연골세포체외구건조직공정연골적효과.방법 분리1월령토관절연골세포,확증후충식재송질골BMG상체외구건조직공정연골.우1、2、4、6주취재진행조직학、Ⅱ형효원면역조직화학급투사전경관찰.결과 연골세포재BMG상생장량호,분비단백취당화효원.수배양시간연장,세포증식,송질골BMG공극내세포수량증다,연골함와형성,기질분비증강.배양6주시연골세포재BMG표면급공극내형성연골양조직,면역조직화학염색현시세포주위기질부함Ⅱ형효원,분포균균:투사전경현시연골세포초미결구정상,세포주위유대량기질산생.결론 동충이체송질골BMG가이촉진연골세포적생장증식,유지연골세포적분화표형;재체외성공구건료조직공정연골,타시일충교호적연골조직공정지가재료.
Objective To evaluate the use of cancellous bone matrix gelatin (BMG) combined with chondrocytes in constructing tissue-engineered cartilage by observing the growth, proliferation and differentiation of chondrocytes on allogeneic cancellous BMG. Methods The articular chondrocytes isolated from a 1-month-old rabbit were multiplied to a monolayer and seeded onto cancellous BMG to construct tissue-engineered cartilage in vitro during a period of 6 weeks. Samples were taken from the construct after 1, 2,4, and 6 weeks of culture and evaluated by histology, immunohistochemistry and transmission electron microscopy (TEM). Results The chondrocytes excreted matrix proteoglycan and collagen on cancellous BMG. With the prolongation of the culture time, the cells proliferated in the construct and the cells in the lacunae increased. Numerous chondrocytes were present the central region of the cancellous BMG and surrounded by extracellular matrix. By 6 weeks of culture, the BMG was covered with 15-20 layers of chondrocytes and cartilaginous tissue occurred in the pores throughout the cancellous BMG. Immunohistochemical staining showed rich and evenly distributed type Ⅱ collagen around the chondrocytes, and TEM revealed an ultrastructure of the chondrocyte similar to that of native chondroctyes, with abundant extracellular matrix produced around the cells. Conclusion Tissue-engineered cartilage can be constructed in vitro using allogeneic cancellous BMG combined with chondrocytes. Allogeneic cancellous BMG serves as a good scaffold material for tissue-engineered cartilage to promote the growth and proliferation of the seeded chondrocytes and allows maintenance of the differentiation phenotype of the cells.