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用于间充质干细胞载体的仿生坚固水凝胶

Biomimicking Robust Hydrogel for the Mesenchymal Stem Cell Carrier.

作者信息

Oh Byeongtaek, Melchert Russell B, Lee Chi H

机构信息

Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri, 64108, USA.

Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri, 64108, USA.

出版信息

Pharm Res. 2015 Oct;32(10):3213-27. doi: 10.1007/s11095-015-1698-y. Epub 2015 Apr 25.

Abstract

PURPOSE

This study was aimed to develop a hydrogel-nanofiber as an advanced carrier for adipose derived human mesenchymal stem cells (AD-MSCs) and evaluate its potential for immunomodulatory therapies applicable to surface coating of drug eluting stent (DES) against coronary artery diseases (CAD).

METHODS

A mixture of dispersing-nanofibers (dNFs) and poly (ethylene glycol)-diacrylate (PEGDA) were blended with sodium alginate to achieve robust mechanical strength. The effects of stem cell niche on cell viability and proliferation rates were evaluated using LDH assay and alamar blue assay, respectively. The amount of Nile-red microparticles (NR-MPs) remained in the hydrogel scaffolds was examined as an index for the physical strength of hydrogels. To evaluate the immunomodulatory activity of AD-MSCs as well as their influence by ROS, the level of L-Kynurenine was determined as tryptophan replacement compounds in parallel with IDO secreted from AD-MSCs using a colorimetric assay of L-amino acid.

RESULTS

Both SA-cys-PEG and SA-cys-dNF-PEG upon being coated on stents using electrophoretic deposition technique displayed superior mechanical properties against the perfused flow. d-NFs had a significant impact on the stability of SA-cys-dNF-PEG, as evidenced by the substantial amount of NR-MPs remained in them. An enhanced subcellular level of ROS by spheroidal cluster yielded the high concentrations of L-Kynurenine (1.67 ± 0.6 μM without H2O2, 5.2 ± 1.14 μM with 50 μM of H2O2 and 8.8 ± 0.51 μM with 100 μM of H2O2), supporting the IDO-mediated tryptophan replacement process.

CONCLUSION

The "mud-and-straw" hydrogels are robust in mechanical property and can serve as an ideal niche for AD-MSCs with immunomodulatory effects.

摘要

目的

本研究旨在开发一种水凝胶-纳米纤维作为脂肪来源的人间充质干细胞(AD-MSCs)的先进载体,并评估其在适用于药物洗脱支架(DES)表面涂层以对抗冠状动脉疾病(CAD)的免疫调节治疗中的潜力。

方法

将分散纳米纤维(dNFs)和聚(乙二醇)-二丙烯酸酯(PEGDA)的混合物与海藻酸钠混合,以获得强大的机械强度。分别使用乳酸脱氢酶(LDH)测定法和alamar蓝测定法评估干细胞微环境对细胞活力和增殖率的影响。以水凝胶支架中残留的尼罗红微粒(NR-MPs)数量作为水凝胶物理强度的指标进行检测。为了评估AD-MSCs的免疫调节活性及其受活性氧(ROS)的影响,使用L-氨基酸比色测定法,与AD-MSCs分泌的吲哚胺2,3-双加氧酶(IDO)平行测定L-犬尿氨酸水平,作为色氨酸替代化合物。

结果

使用电泳沉积技术将SA-cys-PEG和SA-cys-dNF-PEG涂覆在支架上后,它们在灌注流作用下均表现出优异的机械性能。d-NFs对SA-cys-dNF-PEG的稳定性有显著影响,这从其中残留的大量NR-MPs可以证明。球状簇导致细胞内亚细胞水平的ROS增强,产生了高浓度的L-犬尿氨酸(无H2O2时为1.67±0.6μM,50μM H2O2时为5.2±1.14μM,100μM H2O2时为8.8±0.51μM),支持了IDO介导的色氨酸替代过程。

结论

“泥草”水凝胶机械性能强劲,可作为具有免疫调节作用的AD-MSCs的理想微环境。

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