Abstract

Gelatin–dextran hydrogel scaffolds (G-PEG-Dx) were evaluated for their ability to activate the bone marrow human mesenchymal stromal cells (BM-hMSCs) towards mineralization. G-PEG-Dx1 and G-PEG-Dx2, with identical composition but different architecture, were seeded with BM-hMSCs in presence of fetal bovine serum or human platelet lysate (hPL) with or without osteogenic medium. G-PEG-Dx1, characterized by a lower degree of crosslinking and larger pores, was able to induce a better cell colonization than G-PEG-Dx2. At day 28, G-PEG-Dx2, with hPL and osteogenic factors, was more efficient than G-PEG-Dx1 in inducing mineralization. Scanning electron microscopy (SEM) and Raman spectroscopy showed that extracellular matrix produced by BM-hMSCs and calcium-positive mineralization were present along the backbone of the G-PEG-Dx2, even though it was colonized to a lesser degree by hMSCs than G-PEG-Dx1. These findings were confirmed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI), detecting distinct lipidomic signatures that were associated with the different degree of scaffold mineralization. Our data show that the architecture and morphology of G-PEG-Dx2 is determinant and better than that of G-PEG-Dx1 in promoting a faster mineralization, suggesting a more favorable and active role for improving bone repair.

Highlights

  • Regenerative medicine aims to induce self-repair of tissues and organs by unconventional and advanced approaches such as bioengineering of three-dimensional (3D) scaffolds with human mesenchymal stromal cells [1]

  • We investigated the capability of novel three-dimensional gelatin–dextran hybrid hydrogel scaffolds (G-PEG-Dx) having different architecture and morphology to support mineralization in BM-human mesenchymal stromal cells (hMSCs) cultured with growing medium (GM) or osteogenic medium (OM) combined with human platelet lysate (hPL)

  • D-NH2 could provide the possibility of obtaining an interpenetrating polymer network hydrogel that is an entangled combination of two cross-linked polymers, Gelatin-PEG and Dextran-PEG, and they cannot be separated unless chemical bonds are broken

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Summary

Introduction

Regenerative medicine aims to induce self-repair (regeneration) of tissues and organs by unconventional and advanced approaches such as bioengineering of three-dimensional (3D) scaffolds with human mesenchymal stromal cells (hMSCs) [1]. Bone substitute materials are required in critical-sized bone defect repair; they do not meet the clinical requirements of biodegradability, structural support and osteoinductive property [2]. Various strategies have been explored to overcome these critical aspects via bone tissue engineering approaches that incorporate biomimetic scaffolds as a novel platform for phenotypically stable tissue formation and stem cell differentiation. For bone regeneration, they should mimic the 3D bone structure in terms of physical mechanical properties as well as osteoinductive (bone inducing), osteoconductive (bone supporting) and osteogenic (bone forming) features [3]. The hybrid hydrogels, combining the benefits of natural and synthetic polymers, have attracted consideration as support of hMSCs which are able to differentiate into cells of the mesodermal lineages and other embryonic lineages, including osteocytes and chondrocytes, among others [4,5,6]

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