聚乳酸-羟基乙酸共聚物支架负载不同比例羟基磷灰石对骨髓间充质干细胞成骨分化的影响。
Osteogenic Differentiation of Bone Marrow Stem Cell in Poly(Lactic-co-Glycolic Acid) Scaffold Loaded Various Ratio of Hydroxyapatite.
机构信息
Departments of BIN Fusion Technology ; Polymer-Nano Science & Technology, Chonbuk National University, Jeonju, Korea.
出版信息
Int J Stem Cells. 2013 May;6(1):67-74. doi: 10.15283/ijsc.2013.6.1.67.
BACKGROUND AND OBJECTIVES
Hydroxyapatite has biocompatibility and bioactivity and similar to bone of in human body. The purpose of this study is to evaluate osteogenic differentiation of bone marrow stem cell (BMSC) in PLGA Scaffold added various ratio of hydroxyapatite (HAp).
METHODS AND RESULTS
PLGA and PLGA/HAp scaffold were prepared using solvent casting/salt-leaching method. BMSC was seeded on the PLGA and PLGA/HAp scaffold and the samples were cultured in 37℃ incubator with 5% CO2 for 28 days. Alkaline phosphatase (ALP) was carried out to evaluate alkaline phosphatase activity at 1, 3, 7, 10 and 14 days. Alizarin Red S stating was performed to identify calcium in scaffold at 1, 7, 14, 21 and 28 days. Compressive strength was measured to evaluate mechanical property of scaffold. To confirm cell viability, MTT was carried out at 1, 3, 7, 14 and 28 days. RT-PCR was performed to verify specific marker expression of osteoblast and stem cell at 7, 14, 21 and 28 days.
CONCLUSIONS
Osteogenic differentiation of BMSC was confirmed through ALP, RT-PCR, and alizarin red S staining in this study. These results suggest that HAp helps osteogenic differentiation of BMSC.
背景与目的
羟基磷灰石具有生物相容性和生物活性，与人体骨骼相似。本研究旨在评估不同比例羟基磷灰石（HAp）添加到聚乳酸-羟基乙酸共聚物（PLGA）支架中对骨髓间充质干细胞（BMSC）成骨分化的影响。
方法和结果
采用溶剂浇铸/盐析法制备 PLGA 和 PLGA/HAp 支架。将 BMSC 接种到 PLGA 和 PLGA/HAp 支架上，在 37℃、5%CO2 的孵育箱中培养 28 天。在第 1、3、7、10 和 14 天进行碱性磷酸酶（ALP）实验，以评估碱性磷酸酶活性。在第 1、7、14、21 和 28 天进行茜素红 S 染色，以鉴定支架中的钙。通过压缩强度测试来评估支架的力学性能。在第 1、3、7、14 和 28 天进行 MTT 实验，以确认细胞活力。在第 7、14、21 和 28 天进行 RT-PCR 实验，以验证成骨细胞和干细胞的特异性标记物表达。
结论
本研究通过 ALP、RT-PCR 和茜素红 S 染色证实了 BMSC 的成骨分化。这些结果表明 HAp 有助于 BMSC 的成骨分化。
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