Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, 109 Xue yuan xi Road, Wenzhou, 325000, China.
Int Orthop. 2012 Mar;36(3):647-53. doi: 10.1007/s00264-011-1303-x. Epub 2011 Jun 29.
Many in vitro studies of the analysis of the lactoferrin (LF) effect on cells have been reported. However, no study has yet investigated the effect of LF on osteogenic differentiation of human adipose-derived stem cells (hADSCs). The aim of this study was to evaluate the effect of LF on osteogenic differentiation of human adipose stem cells.
The hADSCs were cultured in an osteogenic medium with 0, 10, 50 and 100 μg/ml LF, respectively. hADSC proliferation was analysed by Cell Counting Kit-8 (CCK-8) assay, and cell osteogenic differentiation was evaluated by alkaline phosphatase (ALP) activity assay, von Kossa staining and real-time polymerase chain reaction (RT-PCR).
Cell proliferation was significantly increased by LF in a dose-dependent manner from days 4 to 14. Cells cultured with 100 μg/ml LF presented a higher activity compared with the control. The deposition of calcium was increased after the addition of LF. The mRNA expression of type I collagen (COL-I), ALP, osteocalcin (OCN) and RUNX2 increased markedly as a result of LF treatment.
We have shown for the first time that LF could promote the proliferation and osteogenic differentiation of hADSCs, which could be a promising approach for enhancing osteogenic capacity of cell-based construction in bone tissue engineering.
已经有许多关于乳铁蛋白（LF）对细胞分析影响的体外研究报告。然而，目前尚无研究调查 LF 对人脂肪来源干细胞（hADSCs）成骨分化的影响。本研究旨在评估 LF 对人脂肪干细胞成骨分化的影响。
分别用含有 0、10、50 和 100μg/ml LF 的成骨培养基培养 hADSCs。通过细胞计数试剂盒-8（CCK-8）检测 hADSC 增殖，通过碱性磷酸酶（ALP）活性检测、茜素红染色和实时聚合酶链反应（RT-PCR）评估细胞成骨分化。
LF 以剂量依赖性方式从第 4 天到第 14 天显著增加细胞增殖。与对照组相比，培养 100μg/ml LF 的细胞具有更高的活性。添加 LF 后钙沉积增加。LF 处理后，I 型胶原（COL-I）、ALP、骨钙素（OCN）和 RUNX2 的 mRNA 表达显著增加。
我们首次表明 LF 可促进 hADSCs 的增殖和成骨分化，这可能是增强骨组织工程中基于细胞构建体成骨能力的有前途的方法。
