The effect of fibroblast growth factor and transforming growth factor-β on porcine chondrocytes and tissue-engineered autologous elastic cartilage
The effect of fibroblast growth factor and transforming growth factor-β on porcine chondrocytes and tissue-engineered autologous elastic cartilage
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DOI：
10.1089/107632700300003314
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发表时间：
2001-02-01
影响因子：
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通讯作者：
Eavey, RD
中科院分区：
文献类型：
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作者：
Arévalo-Silva, CA;Cao, YL;Eavey, RD
弹性软骨在体外对转化生长因子-β（TGF-β）和碱性成纤维细胞生长因子（碱性FGF）有促有丝分裂反应。我们研究了这些生长因子单独或组合对体外培养的猪耳廓软骨细胞和产生的自体弹性软骨的影响。从16- 18-kg约克郡猪的弹性耳软骨中收获细胞。测定细胞的活力和定量。将细胞以相等的浓度铺板，并在四种相同培养基环境之一中进行体外研究，除了生长因子：第I组含有Ham’s F-12和补充剂，但不含生长因子，第II组还含有碱性FGF，第III组还含有TGF-β，第IV组还含有两种生长因子的组合。体外培养3周后，用0.25%胰蛋白酶化学解离细胞。皮下注射由30% Pluronic F-127/Ham's F-12中的3 x 10（7）个细胞/cc组成的细胞悬液。在体内培养6、8、10和12周时收获种植体，然后用组织学染色进行检查。体外培养3周后，细胞总数如下：组I，1.8 x 10（8）;组II，3.5 x 10（8）;组III，1.3 x 10（8）;组IV，2.5 x 10（8）。体内自体植入8周后，各组的平均重量（g）和体积（cm（3））如下：组I，0.7 g/0.15 cm（3）;组II，1.5 g/0.8 cm（3）;组III，0.6 g/0.1 cm（3）;组IV，1.2 g/0.3 cm（3）。在组织学上，第I、II和IV组产生的软骨与天然弹性软骨相似，但第III组标本显示纤维组织向内生长。碱性成纤维细胞生长因子产生的数量和质量的自体组织工程弹性软骨产生在这个猪模型在体外和体内最积极的增强。
Elastic cartilage responds mitogenically in vitro to transforming growth factor-beta (TGF-beta) and basic fibroblast growth factor (basic FGF). We studied the effects of these growth factors separately or in a combination on porcine auricular chondrocytes in vitro and on the autologous elastic cartilage produced. Cells were harvested from the elastic auricular cartilage of 16- to 18-kg Yorkshire swine. Viability and quantification of the cells was determined. Cells were plated at equal concentration and studied in vitro in one of four identical media environments except for the growth factors: Group I contained Ham's F-12 with supplements but no growth factors, Group II also contained basic-FGF, Group III also contained TGF-beta, and Group IV also contained a combination of both growth factors. After 3 weeks in vitro, the cells were chemically dissociated with 0.25% trypsin. Cell suspensions composed of 3 x 10(7) cells/cc in 30% Pluronic F-127/Ham's F-12 were injected subcutaneously. Implants were harvested at 6, 8, 10, and 12 weeks of in vivo culture and then were examined with histologic stains. After 3 weeks of in vitro culture the total number of cells was as follows: Group I, 1.8 x 10(8); Group II, 3.5 x 10(8); Group III, 1.3 x 10(8); Group IV, 2.5 x 10(8). After 8 weeks of in vivo autologous implantation, the average weight (g) and volume (cm(3)) of each group was as follows: Group I, 0.7 g/0.15 cm(3); Group II, 1.5 g/0.8 cm(3); Group III, 0.6 g/0.1 cm(3); Group IV, 1.2 g/0.3 cm(3). Histologically, Groups I, II, and IV generated cartilage similar to native elastic cartilage, but Group III specimens demonstrated fibrous tissue ingrowth. Basic FGF produced the most positive enhancement on the quantity and quality of autologous tissue engineered elastic cartilage produced in this porcine model both in vitro and in vivo.