Yang Michael C, O'Connor Andrea J, Kalionis Bill, Heath Daniel E
Department of Biomedical Engineering, University of Melbourne, Parkville, VIC, Australia.
Department of Maternal-Fetal Medicine, Pregnancy Research Centre, Royal Women's Hospital, Parkville, VIC, Australia.
Front Med Technol. 2022 Mar 17;4:834123. doi: 10.3389/fmedt.2022.834123. eCollection 2022.
Decellularized extracellular matrix (dECM) deposited by mesenchymal stromal cells (MSCs) has emerged as a promising substrate for improved expansion of MSCs. To date, essentially all studies that have produced dECM for MSC expansion have done so on tissue culture plastic or glass. However, substrate surface chemistry has a profound impact on the adsorption of proteins that mediate cell-material interactions, and different surface chemistries can cause changes in cell behavior, ECM deposition, and the response to a material. This study tested the hypothesis that substrate surface chemistry impacts the deposition of ECM and its subsequent bioactivity. This hypothesis was tested by producing glass surfaces with various surface chemistries (amine, carboxylic acid, propyl, and octyl groups) using silane chemistry. ECM was deposited by an immortalized MSC line, decellularized, and characterized through SDS-PAGE and immunofluorescence microscopy. No significant difference was observed in dECM composition or microarchitecture on the different surfaces. The decellularized surfaces were seeded with primary MSCs and their proliferation and differentiation were assessed. The presence of dECM improved the proliferation of primary MSCs by ~100% in comparison to surface chemistry controls. Additionally, the adipogenesis increased by 50-90% on all dECM surfaces in comparison to surface chemistry controls, and the osteogenesis increased by ~50% on the octyl-modified surfaces when dECM was present. However, no statistically significant differences were observed within the set of dECM surfaces or control surfaces. These results support the null hypothesis, meaning surface chemistry (over the range tested in this work) is not a key regulator of the composition or bioactivity of MSC-derived dECM. These results are significant because they provide an important insight into regenerative engineering technologies. Specifically, the utilization of dECM in stem cell manufacturing and tissue engineering applications would require the dECM to be produced on a wide variety of substrates. This work indicates that it can be produced on materials with a range of surface chemistries without undesired changes in the bioactivity of the dECM.
间充质基质细胞（MSC）沉积的脱细胞细胞外基质（dECM）已成为一种有望用于改善MSC扩增的基质。迄今为止，基本上所有为MSC扩增制备dECM的研究都是在组织培养塑料或玻璃上进行的。然而，基质表面化学对介导细胞 - 材料相互作用的蛋白质吸附有深远影响，不同的表面化学性质可导致细胞行为、ECM沉积以及对材料的反应发生变化。本研究检验了基质表面化学影响ECM沉积及其后续生物活性这一假设。通过使用硅烷化学方法制备具有各种表面化学性质（胺基、羧基、丙基和辛基）的玻璃表面来检验这一假设。通过永生化的MSC系沉积ECM，进行脱细胞处理，并通过SDS - PAGE和免疫荧光显微镜进行表征。在不同表面上的dECM组成或微观结构未观察到显著差异。将原代MSC接种到脱细胞表面，并评估其增殖和分化情况。与表面化学对照相比，dECM的存在使原代MSC的增殖提高了约100%。此外，与表面化学对照相比，所有dECM表面上的脂肪生成增加了50 - 90%，当存在dECM时，辛基修饰表面上的成骨增加了约50%。然而，在dECM表面组或对照表面组内未观察到统计学上的显著差异。这些结果支持了原假设，即表面化学（在本研究测试的范围内）不是MSC来源的dECM组成或生物活性的关键调节因子。这些结果具有重要意义在于它们为再生工程技术提供了重要见解。具体而言，在干细胞制造和组织工程应用中使用dECM将需要在多种基质上生产dECM。这项工作表明可以在具有一系列表面化学性质的材料上生产dECM，而不会使dECM的生物活性发生不良变化。
