摘要
目的探讨异体关节软骨细胞作为软骨种子细胞修复关节软骨缺损的可行性。方法无菌取成年新西兰大白兔四肢关节软骨,酶消化法分离细胞,条件培养基体外培养扩增,藻酸钙凝胶包埋培养1周,植入兔膝关节股骨髁间软骨缺损,对照组缺损旷置。术后3、6个月分别取材,观察缺损修复情况;切片特染和免疫组化观察修复组织病理,并W ak itan i评分评估修复质量。结果7/8缺损为成熟透明软骨组织修复,1/8为纤维软骨修复;W ak itan i平均评1.75分;空白对照组评7.65分。实验组3个月组标本未见淋巴细胞或白细胞浸润,6个月组标本未见明显退变;所有标本均未见藻酸残留。结论用藻酸包埋异体关节软骨细胞修复兔膝关节软骨缺损的方法是可行的,异体关节软骨细胞在适当的处理后可成为关节软骨组织工程种子细胞。
Objective To explore the feasibility of repairing the articular cartilage with alloarticular chondrecytes embedded in alginate gel. Methods Allo-articular chondrecytes were isolated from three adult New zealand rabbits. The cells were cultured in DMEM/F-12 supplemented with 20% fetal bovine serum （FBS）. Chondrocytes of 2^nd - 3^rd passage were harvested and were diluted to 5.0 - 10^7cells/ ml with 1. 2% alginate. Then alginate gel was formed by 102 mM CaC12. The gels were cultured subsequently for 1 week and then transfered to the full-thickness defects in the femoral condyles of adult rabbits. In control group the defects were left untreated. The animals were sacrificed in the 3^rd and 6^th month after operation respectively. The specimens were decalcified with 50% formic acid. The histologic sections were stained with safranin O-fast green, hematoxylin-eosin （H＆E） and picric acid-Sirius red and immunohisto- stained for type Ⅱ collagen and aggrecan, the repairing efficiency was evaluated according to Wakaniti scoring. Result In the experiment group all 8 defects acquired repair, 7/8 were repaired with mature hyaline cartilage tissue, and 1/8 was with fibrocartilage tissue for less cell-gel inputted. The thick of repaired tissues were closed to the normal and the tissue integrated smoothly with cartilage around the defects. Safranin O staining of the matrix acted in accordance with the normal and immunostaining for type Ⅱ collagen and aggrecan showed positive. Picric acid-Sirius red staining showed that the chondrocytes lined in lines and the collagen aligned like Gothic architecture structure by polarization microscopy. There was no evidence of residue of alginate and inflammation in 3^rd month specimens and no obvious deterioration at 6^th month. But in control group, only a small amount of fibrous, fibrocartilage, or hyaline-like tissue was seen on the surface of the defects. Wakaniti scoring showed 1.75 points for the experiment group and 7.65 for the control group. Conclusion It is a premising way to repair the articular cartilage with homogeneous articular cbondrocytes embedded in alginate gel.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2006年第13期886-890,共5页
National Medical Journal of China
基金
国家"863"高技术研究发展计划基金资助项目(2002AA205021)
关键词
软骨细胞
藻酸盐
软骨关节
Chondrocytes
Alginates
Cartilage,articular joints
