Journal
BIOTECHNOLOGY JOURNAL
Volume 18, Issue 4, Pages -Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/biot.202200413
Keywords
mesenchymal stromal cells; nanofibrous scaffolds; PCL-nHA; three-dimensional cell culture
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Human Adipose-Derived Mesenchymal Stem/Stromal Cells (hAD-MSCs) have great potential for tissue regeneration. Culturing these cells under hypoxic conditions improves their post-transplantation survival and regenerative performance. The combination of hAD-MSCs and PCL-nHA nanofibers enhances their osteoblast differentiation. This study reveals cellular adaptation to a three-dimensional environment, highlighting the suitability of PCL-nHA nanofibers for pre-conditioning hAD-MSCs.
Human Adipose-Derived Mesenchymal Stem/Stromal Cells (hAD-MSCs) have great potential for tissue regeneration. Since transplanted hAD-MSCs are likely to be placed in a hypoxic environment, culturing the cells under hypoxic conditions might improve their post-transplantation survival and regenerative performance. The combination of hAD-MSCs and PCL-nHA nanofibers synergically improves the contribution of both components for osteoblast differentiation. In this work, we hypothesized that this biomaterial constitutes a hypoxic environment for hAD-MSCs. We studied the cellular re-arrangement and the subcellular ultrastructure by Transmission Electron Microscopy (TEM) of hAD-MSCs grown into PCL-nHA nanofibers, and we compared them with the same cells grown in two-dimensional cultures, over tissue culture-treated plastic, or glass coverslips. Among the most evident changes, PCL-nHA grown cells showed enlarged mitochondria, and accumulation of glycogen granules, consistent with a hypoxic environment. We observed a 3.5 upregulation (p = 0.0379) of Hypoxia Inducible Factor (HIF)-1A gene expression in PCL-nHA grown cells. This work evidences for the first time intra-cellular changes in three-dimensional compared to two-dimensional cultures, which are adaptive responses of the cells to an environment more closely resembling that of the in vivo niche after transplantation, thus PCL-nHA nanofibers are adequate for hAD-MSCs pre-conditioning.
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