Yu Fang-yuan, Lu Shi-bi, Zhao Bin, Xu Wen-jing, Huang Li-hong, Yuan Mei, Sun Ming-xue, Zhang Wen-tao
Orthopedics Institute, General Hospital of PLA, Beijing 100853, China.
Zhonghua Yi Xue Za Zhi. 2006 Apr 4;86(13):886-90.
To explore the feasibility of repairing the articular cartilage with allo-articular chondrocytes embedded in alginate gel.
Allo-articular chondrocytes 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 2nd - 3rd passage were harvested and were diluted to 5.0 x 10(7) cells/ml with 1.2% alginate. Then alginate gel was formed by 102 mM CaCl(2). The gels were cultured subsequently for 1 week and then transferred 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 3rd and 6th 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 Wakitani scoring.
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 3rd month specimens and no obvious deterioration at 6th month. But in control group, only a small amount of fibrous, fibrocartilage, or hyaline-like tissue was seen on the surface of the defects. Wakitani scoring showed 1.75 points for the experiment group and 7.65 for the control group.
It is a promising way to repair the articular cartilage with homogeneous articular chondrocytes embedded in alginate gel.
探讨用包埋于藻酸盐凝胶中的同种异体关节软骨细胞修复关节软骨的可行性。
从3只成年新西兰兔分离出同种异体关节软骨细胞。细胞在补充有20%胎牛血清(FBS)的DMEM/F-12培养基中培养。收集第2 - 3代软骨细胞,并用1.2%藻酸盐将其稀释至5.0×10⁷细胞/ml。然后用102 mM氯化钙形成藻酸盐凝胶。随后将凝胶培养1周,然后转移至成年兔股骨髁的全层缺损处。对照组缺损不做处理。分别在术后第3个月和第6个月处死动物。标本用50%甲酸脱钙。组织学切片用番红O - 固绿、苏木精 - 伊红(H&E)和苦味酸 - 天狼星红染色,并对Ⅱ型胶原蛋白和聚集蛋白聚糖进行免疫组织化学染色。根据Wakitani评分评估修复效率。
实验组8个缺损全部获得修复,7/8用成熟透明软骨组织修复,1/8因细胞 - 凝胶输入量较少用纤维软骨组织修复。修复组织厚度接近正常,且与缺损周围软骨组织平滑整合。基质的番红O染色与正常情况一致,Ⅱ型胶原蛋白和聚集蛋白聚糖的免疫染色呈阳性。苦味酸 - 天狼星红染色显示软骨细胞呈线状排列,偏振显微镜下胶原排列呈哥特式建筑结构。在第3个月的标本中未发现藻酸盐残留和炎症迹象,第6个月也无明显退变。但对照组缺损表面仅见少量纤维、纤维软骨或类透明组织。Wakitani评分显示实验组为1.75分,对照组为7.65分。
用包埋于藻酸盐凝胶中的同种异体关节软骨细胞修复关节软骨是一种有前景的方法。