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在涂有脐带华通氏胶的聚乳酸支架上培养的间充质干细胞的成骨分化

Osteogenic differentiation of mesenchymal stem cells cultured on PLLA scaffold coated with Wharton's Jelly.

作者信息

Ahmadi Marziehsadat, Seyedjafari Ehsan, Zargar Seyed Jalal, Birhanu Gebremariam, Zandi-Karimi Ali, Beiki Bahareh, Tuzlakoglu Kadriye

机构信息

Department of Cell and Molecular Biology, School of Biology, College of Science, University of Tehran, Iran.

Department of Biotechnology, College of Science, University of Tehran, Iran.

出版信息

EXCLI J. 2017 May 23;16:785-794. doi: 10.17179/excli2016-741. eCollection 2017.

Abstract

Poly-L-lactic acid (PLLA) electrospun nanofiber scaffold is one of the most commonly used synthetic polymer scaffolds for bone tissue engineering application. However, PLLA is hydrophobic in nature, hence does not maintain proper cell adhesion and tissue formation, moreover, it cannot provide the osteo-inductive environment due to inappropriate surface characteristic and the lack of surface motives participating in the first cellular events. To modify these shortcomings different approaches have been used, among those the most commonly used one is coating of the surface of the electrospun nanofiber with natural materials. In this work Wharton's jelly (WJ), a tissue which surrounds the umbilical cord vessels, reaches in high amounts of extracellular matrix (ECM) components mainly; collagen, hyaluronic acid and several sulphated glycosaminoglycans (GAGs) were used to cover the surface of electrospun PLLA nanofiber scaffolds. The surface morphology of the nanofiber scaffold was evaluated via scanning electron microscope, and the osteogenic differentiation potential was determined by MTT assay and common osteogenic marker tests such as alkaline phosphatase (ALP) activity and calcium deposition tests. Coating of WJ could not change the surface morphology and diameter of the nanofibers. However, WJ-PLLA scaffolds showed higher proliferation of human mesenchymal stem cells (MSC) than tissue culture plate (TCP) and pristine PLLA scaffolds, moreover, WJ-PPLA scaffold demonstrated significant alkaline phosphatase activity and calcium mineralization than either TCP or PLLA nanofiber scaffolds.

摘要

聚左旋乳酸(PLLA)电纺纳米纤维支架是骨组织工程应用中最常用的合成聚合物支架之一。然而,PLLA本质上是疏水性的,因此不能维持适当的细胞粘附和组织形成,此外,由于其不合适的表面特性以及缺乏参与早期细胞事件的表面基序,它无法提供骨诱导环境。为了改善这些缺点,人们采用了不同的方法,其中最常用的是用天然材料对电纺纳米纤维表面进行涂层处理。在这项工作中,华通氏胶(WJ),一种包裹脐带血管的组织,富含大量细胞外基质(ECM)成分,主要是胶原蛋白、透明质酸和几种硫酸化糖胺聚糖(GAGs),被用于覆盖电纺PLLA纳米纤维支架的表面。通过扫描电子显微镜评估纳米纤维支架的表面形态,并通过MTT法以及常见的成骨标志物测试,如碱性磷酸酶(ALP)活性和钙沉积测试,来确定其成骨分化潜力。WJ涂层不会改变纳米纤维的表面形态和直径。然而,WJ-PLLA支架显示出比组织培养板(TCP)和原始PLLA支架更高的人间充质干细胞(MSC)增殖能力,此外,WJ-PPLA支架比TCP或PLLA纳米纤维支架表现出显著的碱性磷酸酶活性和钙矿化能力。

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