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通过静电纺丝制备的棉絮状纳米复合生物材料:人间充质干细胞的体外生物活性和成骨分化

Cotton wool-like nanocomposite biomaterials prepared by electrospinning: in vitro bioactivity and osteogenic differentiation of human mesenchymal stem cells.

作者信息

Schneider Oliver D, Loher Stefan, Brunner Tobias J, Uebersax Lorenz, Simonet Marc, Grass Robert N, Merkle Hans P, Stark Wendelin J

机构信息

Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH Zurich, CH-8093 Zurich, Switzerland.

出版信息

J Biomed Mater Res B Appl Biomater. 2008 Feb;84(2):350-62. doi: 10.1002/jbm.b.30878.

DOI:10.1002/jbm.b.30878
PMID:17618506
Abstract

The present study evaluates the in vitro biomedical performance of an electrospun, flexible, and cotton wool-like poly(lactide-co-glycolide) (PLGA)/amorphous tricalcium phosphate (ATCP) nanocomposite. Experiments on in vitro biomineralization, applicability in model defects and a cell culture study with human mesenchymal stem cells (hMSC) allowed assessing the application of the material for potential use as a bone graft. Scaffolds with different flame made ATCP nanoparticle loadings were prepared by electrospinning of a PLGA-based composite. Immersion in simulated body fluid showed significant deposition of a hydroxyapatite layer only on the surface of ATCP doped PLGA (up to 175% mass gain within 15 days for PLGA/ATCP 60:40). Proliferation and osteogenic differentiation of hMSC on different nanocomposites were assessed by incubating cells in osteogenic medium for 4 weeks. Proper adhesion and an unaffected morphology of the cells were observed by confocal laser scanning microscopy for all samples. Fluorometric quantification of dsDNA and analysis of ALP activity revealed no significant difference between the tested scaffolds and excluded any acute cytotoxic effects of the nanoparticles. The osteocalcin content for all scaffolds was 0.12-0.19 ng/ng DNA confirming osteogenic differentiation of human mesenchymal stem cells on these flexible bone implants.

摘要

本研究评估了一种电纺、柔性且类似棉絮的聚(丙交酯-乙交酯)(PLGA)/无定形磷酸三钙(ATCP)纳米复合材料的体外生物医学性能。体外生物矿化实验、在模型缺损中的适用性以及用人骨髓间充质干细胞(hMSC)进行的细胞培养研究,使得能够评估该材料作为骨移植潜在用途的应用情况。通过对基于PLGA的复合材料进行电纺,制备了具有不同火焰法制备的ATCP纳米颗粒负载量的支架。浸入模拟体液中显示,仅在掺杂ATCP的PLGA表面有显著的羟基磷灰石层沉积(对于PLGA/ATCP 60:40,在15天内质量增加高达175%)。通过在成骨培养基中培养细胞4周,评估了hMSC在不同纳米复合材料上的增殖和成骨分化情况。通过共聚焦激光扫描显微镜观察,所有样品的细胞均有良好的黏附且形态未受影响。对双链DNA的荧光定量分析和碱性磷酸酶活性分析表明,测试的支架之间没有显著差异,并且排除了纳米颗粒的任何急性细胞毒性作用。所有支架的骨钙素含量为0.12 - 0.19 ng/ng DNA,证实了人骨髓间充质干细胞在这些柔性骨植入物上的成骨分化。

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