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电纺纳米纤维增强脐带来源的无限制体干细胞的成骨分化。

Enhanced osteogenic differentiation of cord blood-derived unrestricted somatic stem cells on electrospun nanofibers.

机构信息

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

出版信息

J Mater Sci Mater Med. 2011 Jan;22(1):165-74. doi: 10.1007/s10856-010-4174-6. Epub 2010 Nov 11.

Abstract

A new stem cell-scaffold construct based on poly-L-lactide (PLLA) nanofibers grafted with collagen (PLLA-COL) and cord blood-derived unrestricted somatic stem cells (USSC) were proposed to hold promising characteristics for bone tissue engineering. Fabricated nanofibers were characterized using SEM, ATR-FTIR, tensile and contact angle measurements. The capacity of PLLA, plasma-treated PLLA (PLLA-pl) and PLLA-COL scaffolds to support proliferation and osteogenic differentiation of USSC was evaluated using MTT assay and common osteogenic markers such as alkaline phosphatase (ALP) activity, calcium mineral deposition and bone-related genes. All three scaffolds showed nanofibrous and porous structure with suitable physical characteristics. Higher proliferation and viability of USSC was observed on PLLA-COL nanofibers compared to control surfaces. In osteogenic medium, ALP activity and calcium deposition exhibited the highest values on PLLA-COL scaffolds on days 7 and 14. These markers were also greater on PLLA and PLLA-pl compared to TCPS. Higher levels of collagen I, osteonectin and bone morphogenetic protein-2 were detected on PLLA-COL compared to PLLA and PLLA-pl. Runx2 and osteocalcin were also expressed continuously on all scaffolds during induction. These observations suggested the enhanced proliferation and osteogenic differentiation of USSC on PLLA-COL nanofiber scaffolds and introduced a new combination of stem cell-scaffold constructs with desired characteristics for application in bone tissue engineering.

摘要

基于聚-L-乳酸(PLLA)纳米纤维接枝胶原蛋白(PLLA-COL)和脐带血来源的无限制体干细胞(USSC)的新型干细胞-支架构建体被提出,具有用于骨组织工程的有前途的特性。使用 SEM、ATR-FTIR、拉伸和接触角测量对制备的纳米纤维进行了表征。使用 MTT 测定法和碱性磷酸酶(ALP)活性、钙矿化沉积和与骨相关的基因等常见成骨标志物评估 PLLA、等离子体处理的 PLLA(PLLA-pl)和 PLLA-COL 支架对 USSC 增殖和成骨分化的支持能力。所有三种支架均表现出纳米纤维状和多孔结构,具有合适的物理特性。与对照表面相比,USSC 在 PLLA-COL 纳米纤维上显示出更高的增殖和活力。在成骨培养基中,ALP 活性和钙沉积在第 7 天和第 14 天在 PLLA-COL 支架上表现出最高值。与 TCPS 相比,这些标志物在 PLLA 和 PLLA-pl 上也更高。与 PLLA 和 PLLA-pl 相比,PLLA-COL 上检测到更高水平的胶原蛋白 I、骨粘连蛋白和骨形态发生蛋白-2。Runx2 和骨钙素在诱导过程中也在所有支架上持续表达。这些观察结果表明,USSC 在 PLLA-COL 纳米纤维支架上的增殖和成骨分化增强,并引入了一种具有应用于骨组织工程所需特性的新型干细胞-支架构建体组合。

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