Zimber M P, Tong B, Dunkelman N, Pavelec R, Grande D, New L, Purchio A F
Advanced Tissue Sciences, 10933 North Torrey Pines Road, La Jolla, California 92037-1005.
Tissue Eng. 1995 Fall;1(3):289-300. doi: 10.1089/ten.1995.1.289.
We have investigated the ability of transforming growth factor beta (TGF-beta) to promote the growth and differentiation of chondrocytes in monolayer and on three-dimensional scaffolds. Treatment of chondrocytes with TGF-beta and ascorbate individually stimulated the proliferation of bovine articular chondrocytes about 2-fold when cells were grown in monolayer culture: the combination of TGF-beta and ascorbate resulted in a 3-fold increase in cell number over a 72-h period. Peak stimulation with TGF-beta occurred at about 1.0 ng/ml: bFGF was slightly inhibitory in these assays. TGF-beta led to an increase in glycosaminoglycan synthesis as detected by Western blotting using anti-chondroitin sulfate antibodies. No significant change in collagen type II mRNA or protein was observed. When cells were grown on grown on three-dimensional scaffolds composed of polyglyocolic acid, TGF-beta treatment led to an increase in the size of the cartilage-like constructs produced. This was accompanied by increases in collagen and glycosaminoglycan deposition; immunohistochemical staining showed that the predominant collagen was type II. These results indicate that TGF-beta is capable of increasing the proliferation rate of chondrocytes in monolayer as well as increasing cartilage production on three-dimensional scaffolds and may find utility in the in vitro engineering of cartilage tissue.
我们研究了转化生长因子β（TGF-β）促进单层培养以及在三维支架上培养的软骨细胞生长和分化的能力。当软骨细胞在单层培养中生长时，单独用TGF-β和抗坏血酸处理可刺激牛关节软骨细胞增殖约2倍：TGF-β和抗坏血酸联合使用可使细胞数量在72小时内增加3倍。TGF-β的峰值刺激浓度约为1.0 ng/ml：在这些试验中，碱性成纤维细胞生长因子（bFGF）有轻微抑制作用。通过使用抗硫酸软骨素抗体的蛋白质印迹法检测，TGF-β可导致糖胺聚糖合成增加。未观察到II型胶原蛋白mRNA或蛋白质有显著变化。当细胞在由聚乙醇酸组成的三维支架上生长时，TGF-β处理可使产生的类软骨构建体尺寸增加。这伴随着胶原蛋白和糖胺聚糖沉积的增加；免疫组织化学染色显示，主要的胶原蛋白是II型。这些结果表明，TGF-β能够提高单层培养中软骨细胞的增殖率，同时增加三维支架上软骨的生成，可能在软骨组织的体外工程中具有应用价值。
