Department of Tissue Regeneration, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, 7500 AE Enschede, PO BOX 217, The Netherlands.
Macromol Biosci. 2010 Nov 10;10(11):1365-73. doi: 10.1002/mabi.201000145.
Electrospun scaffolds are widely used for various biomedical applications. In this study, we prepared electrospun bioactive composite scaffolds combining hydroxyapatite, collagen (Col) and a synthetic polymer-PolyActive™-to mimic naturally occurring extracellular matrix for in situ bone regeneration. Human mesenchymal stem cells (hMSCs) adhered and proliferated on these scaffolds. Cells on all scaffold types showed an increased metabolic activity with time. On day 4, the metabolic activity of cells cultured on PolyActive™ (PA)-hydroxyapatite (HA)-Col in 1,1,1,3,3,3-hexafluoro-2-propanolhexafluoro-2-propanol (HFIP) was significantly higher than that of cells grown on PA-Col samples. Furthermore, on day 6, cells on PA-HA-Col in HFIP showed significantly higher metabolic activity than those on PA and PA-Col scaffolds. Quantitative PCR analysis for a panel of osteogenic genes showed statistically significant differences between scaffolds. Cells cultured on PA-HA scaffolds had a significantly higher osteonectin and RunX2 expression compared to those on PA-HA-Col scaffolds. Cells on PA-HA-Col in HFIP scaffolds had significantly higher expression of alkaline phosphatase (ALP) and Col 1 compared to PA and PA-Col scaffolds respectively. The bone morphogenetic protein-2 and S100A4 expression of PA-Col and PA-HA-Col constructs was significantly lower than the basal level expression of cells on PA scaffolds. Although not statistically significant in all cases, cells cultured on PA-HA-Col in HFIP and PA-HA scaffolds had the highest expression for most of the genes analysed. The results of the study demonstrate that bioactive composite scaffolds prepared by electrospinning could find potential use in bone regeneration applications.
静电纺丝支架广泛用于各种生物医学应用。在这项研究中,我们制备了静电纺丝生物活性复合支架,将羟基磷灰石、胶原(Col)和合成聚合物-PolyActiveTM-结合在一起,以模拟天然细胞外基质,用于原位骨再生。人骨髓间充质干细胞(hMSCs)在这些支架上黏附和增殖。随着时间的推移,所有支架类型上的细胞代谢活性均增加。在第 4 天,在 1,1,1,3,3,3-六氟-2-丙醇(HFIP)中培养的细胞在 PolyActiveTM(PA)-羟基磷灰石(HA)-Col 上的代谢活性明显高于在 PA-Col 样品上培养的细胞。此外,在第 6 天,在 HFIP 中的 PA-HA-Col 上的细胞表现出比在 PA 和 PA-Col 支架上更高的代谢活性。一组成骨基因的定量 PCR 分析显示支架之间存在统计学上的显著差异。与 PA-HA-Col 支架上的细胞相比,在 PA-HA 支架上培养的细胞骨桥蛋白和 RunX2 的表达明显更高。在 HFIP 中的 PA-HA-Col 支架上的细胞碱性磷酸酶(ALP)和 Col 1 的表达明显高于 PA 和 PA-Col 支架,而 PA-Col 和 PA-HA-Col 支架的骨形态发生蛋白 2 和 S100A4 表达明显低于 PA 支架上细胞的基础表达水平。虽然并非在所有情况下均具有统计学意义,但在 HFIP 中的 PA-HA-Col 和 PA-HA 支架上培养的细胞对大多数分析基因的表达最高。该研究结果表明,通过静电纺丝制备的生物活性复合支架可能在骨再生应用中具有潜在用途。