Zhang W-H, Li X-L, Guo Y, Zhang Y
Tianjin Hospital, Tianjin, China.
Braz J Med Biol Res. 2017 Aug 17;50(10):e6272. doi: 10.1590/1414-431X20176272.
The aim of this study was to determine the proliferation and osteogenic activity of fibroblasts induced with fibronectin and their possible dose-dependent relationship. The fibroblasts obtained by tissue explants adherent method were induced with fibronectin at different concentrations of 0, 10, 20, 40, 60, and 80 μg/mL for 14 days. The 3H-thymidine and 3H-proline incorporation test was used to evaluate the synthesis of DNA and collagen by fibroblasts, respectively. The mineralized nodules and osteocalcin secretion, as vital osteogenic indicators, were detected with tetracycline labeling and 125I-labeled competitive immunoassay, respectively. Fibronectin significantly increased the synthesis of DNA and collagen by fibroblasts, especially at the concentration of 40 μg/mL (P<0.05). The increased secretion of osteocalcin in the supernatant was also statistically significant at the concentration of 40 μg/mL (P<0.05). The mineralized nodules with trabecula-like structure derived from induced fibroblasts were positive for tetracycline labeling. The granulation tissue-derived fibroblasts induced with fibronectin exhibited increased proliferative, functional and osteogenic potential. Fibroblasts are considered a possible in situ stem cell in tissue engineering.
本研究的目的是确定纤连蛋白诱导的成纤维细胞的增殖和成骨活性及其可能的剂量依赖关系。采用组织块贴壁法获取的成纤维细胞,用0、10、20、40、60和80μg/mL不同浓度的纤连蛋白诱导14天。分别采用3H-胸腺嘧啶核苷和3H-脯氨酸掺入试验评估成纤维细胞DNA和胶原蛋白的合成。分别用四环素标记和125I标记的竞争性免疫测定法检测矿化结节和骨钙素分泌这两个重要的成骨指标。纤连蛋白显著增加了成纤维细胞DNA和胶原蛋白的合成,尤其是在40μg/mL浓度时(P<0.05)。在40μg/mL浓度时,上清液中骨钙素分泌增加也具有统计学意义(P<0.05)。诱导成纤维细胞形成的具有小梁样结构的矿化结节对四环素标记呈阳性。用纤连蛋白诱导的肉芽组织来源的成纤维细胞表现出增殖、功能和成骨潜能增加。成纤维细胞被认为是组织工程中一种可能的原位干细胞。