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利用重组人源核心蛋白聚糖对静电纺丝支架进行表面功能化,可吸引循环内皮祖细胞。

Surface functionalization of electrospun scaffolds using recombinant human decorin attracts circulating endothelial progenitor cells.

机构信息

Department of Cell and Tissue Engineering, Fraunhofer-Institute for Interfacial Engineering and Biotechnology (IGB), 70569, Stuttgart, Germany.

Department of Women´s Health, Research Institute for Women's Health, Eberhard-Karls-University Tübingen, 72076, Tübingen, Germany.

出版信息

Sci Rep. 2018 Jan 8;8(1):110. doi: 10.1038/s41598-017-18382-y.

Abstract

Decorin (DCN) is an important small leucine-rich proteoglycan present in the extracellular matrix (ECM) of many organs and tissues. Endothelial progenitor cells (EPCs) are able to interact with the surrounding ECM and bind to molecules such as DCN. Here, we recombinantly produced full-length human DCN under good laboratory practice (GLP) conditions, and after detailed immunological characterization, we investigated its potential to attract murine and human EPCs (mEPCs and hECFCs). Electrospun polymeric scaffolds were coated with DCN or stromal cell-derived factor-1 (SDF-1α) and were then dynamically cultured with both cell types. Cell viability was assessed via imaging flow cytometry. The number of captured cells was counted and compared with the non-coated controls. To characterize cell-scaffold interactions, immunofluorescence staining and scanning electron microscopy analyses were performed. We identified that DCN reduced T cell responses and attracted innate immune cells, which are responsible for ECM remodeling. A significantly higher number of EPCs attached on DCN- and SDF-1α-coated scaffolds, when compared with the uncoated controls. Interestingly, DCN showed a higher attractant effect on hECFCs than SDF-1α. Here, we successfully demonstrated DCN as promising EPC-attracting coating, which is particularily interesting when aiming to generate off-the-shelf biomaterials with the potential of in vivo cell seeding.

摘要

核心蛋白聚糖(DCN)是一种重要的富含亮氨酸的小蛋白聚糖,存在于许多器官和组织的细胞外基质(ECM)中。内皮祖细胞(EPCs)能够与周围的 ECM 相互作用,并与 DCN 等分子结合。在这里,我们在良好实验室规范(GLP)条件下重组产生全长人 DCN,在经过详细的免疫学表征后,我们研究了其吸引小鼠和人 EPC(mEPC 和 hECFC)的潜力。电纺聚合物支架涂有 DCN 或基质细胞衍生因子-1(SDF-1α),然后与两种细胞类型进行动态培养。通过成像流式细胞术评估细胞活力。计数捕获的细胞数量,并与未涂层的对照进行比较。为了表征细胞-支架相互作用,进行了免疫荧光染色和扫描电子显微镜分析。我们发现 DCN 可减少 T 细胞反应并吸引固有免疫细胞,固有免疫细胞负责 ECM 重塑。与未涂层的对照相比,DCN 和 SDF-1α 涂层支架上附着的 EPC 数量明显更多。有趣的是,DCN 对 hECFC 的吸引力高于 SDF-1α。在这里,我们成功地证明了 DCN 是一种有前途的 EPC 吸引涂层,当目标是生成具有体内细胞播种潜力的现成生物材料时,这尤其有趣。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09cd/5758628/61cc36ededde/41598_2017_18382_Fig1_HTML.jpg

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