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海藻酸钙凝胶作为干细胞基质可产生旁分泌干细胞活性,有助于术后加速愈合。

Calcium alginate gels as stem cell matrix-making paracrine stem cell activity available for enhanced healing after surgery.

作者信息

Schmitt Andreas, Rödel Philipp, Anamur Cihad, Seeliger Claudine, Imhoff Andreas B, Herbst Elmar, Vogt Stephan, van Griensven Martijn, Winter Gerhard, Engert Julia

机构信息

Department of Sports Orthopedics, Technical University Munich, Ismaninger Str. 22, D-81675 Munich, Germany.

Department of Pharmacy, Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-University Munich, Butenandtstr. 5, Haus B, D-81377 Munich, Germany.

出版信息

PLoS One. 2015 Mar 20;10(3):e0118937. doi: 10.1371/journal.pone.0118937. eCollection 2015.

Abstract

Regeneration after surgery can be improved by the administration of anabolic growth factors. However, to locally maintain these factors at the site of regeneration is problematic. The aim of this study was to develop a matrix system containing human mesenchymal stem cells (MSCs) which can be applied to the surgical site and allows the secretion of endogenous healing factors from the cells. Calcium alginate gels were prepared by a combination of internal and external gelation. The gelling behaviour, mechanical stability, surface adhesive properties and injectability of the gels were investigated. The permeability of the gels for growth factors was analysed using bovine serum albumin and lysozyme as model proteins. Human MSCs were isolated, cultivated and seeded into the alginate gels. Cell viability was determined by AlamarBlue assay and fluorescence microscopy. The release of human VEGF and bFGF from the cells was determined using an enzyme-linked immunoassay. Gels with sufficient mechanical properties were prepared which remained injectable through a syringe and solidified in a sufficient time frame after application. Surface adhesion was improved by the addition of polyethylene glycol 300,000 and hyaluronic acid. Humans MSCs remained viable for the duration of 6 weeks within the gels. Human VEGF and bFGF was found in quantifiable concentrations in cell culture supernatants of gels loaded with MSCs and incubated for a period of 6 weeks. This work shows that calcium alginate gels can function as immobilization matrices for human MSCs.

摘要

通过给予合成代谢生长因子可改善术后的再生情况。然而,在再生部位局部维持这些因子存在问题。本研究的目的是开发一种包含人间充质干细胞(MSC)的基质系统,该系统可应用于手术部位,并能使细胞分泌内源性愈合因子。海藻酸钙凝胶通过内部和外部凝胶化相结合的方法制备。研究了凝胶的凝胶化行为、机械稳定性、表面粘附特性和可注射性。以牛血清白蛋白和溶菌酶作为模型蛋白分析了凝胶对生长因子的渗透性。分离、培养人间充质干细胞并将其接种到海藻酸钙凝胶中。通过AlamarBlue检测和荧光显微镜确定细胞活力。使用酶联免疫吸附测定法测定细胞中人血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF)的释放量。制备了具有足够机械性能的凝胶,这些凝胶可通过注射器保持可注射状态,并在应用后在足够的时间内固化。通过添加聚乙二醇300,000和透明质酸改善了表面粘附性。人间充质干细胞在凝胶中6周内保持存活。在接种了人间充质干细胞并孵育6周的凝胶的细胞培养上清液中发现了可定量浓度的人VEGF和bFGF。这项工作表明海藻酸钙凝胶可作为人间充质干细胞的固定基质。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6cd4/4368733/04c2e46f4f94/pone.0118937.g001.jpg

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