Decellularized Human Umbilical Tissue-Derived Hydrogels Promote Proliferation and Chondrogenic Differentiation of Mesenchymal Stem Cells

Abstract

Tissue engineering is a promising approach for the repair and regeneration of cartilaginous tissue. Appropriate three-dimensional scaffolding materials that mimic cartilage are ideal for the repair of chondral defects. The emerging decellularized tissue-based scaffolds have the potential to provide essential biochemical signals and structural integrity, which mimics the natural tissue environment and directs cellular fate. Umbilical cord-derived hydrogels function as 3D scaffolding material, which support adherence, proliferation, migration, and differentiation of cells due to their similar biochemical composition to cartilage. Therefore, the present study aimed to establish a protocol for the formulation of a hydrogel from decellularized human umbilical cord (DUC) tissue, and assess its application in the proliferation and differentiation of UC-MSCs along chondrogenic lineage. The results showed that the umbilical cord was efficiently decellularized. Subsequently, DUC hydrogel was prepared, and in vitro chondral differentiation of MSCs seeded on the scaffold was determined. The developed protocol efficiently removed the cellular and nuclear content while retaining the extracellular matrix (ECM). DUC tissue, pre-gel, and hydrogels were evaluated by FTIR spectroscopy, which confirmed the gelation from pre-gel to hydrogel. SEM analysis revealed the fibril morphology and porosity of the DUC hydrogel. Calcein AM and Alamar blue assays confirmed the MSC survival, attachment, and proliferation in the DUC hydrogels. Following seeding of UC-MSCs in the hydrogels, they were cultured in stromal or chondrogenic media for 28 days, and the expression of chondrogenic marker genes including TGF-β1, BMP2, SOX-9, SIX-1, GDF-5, and AGGRECAN was significantly increased (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001). Moreover, the hydrogel concentration was found to significantly affect the expression of chondrogenic marker genes. The overall results indicate that the DUC-hydrogel is compatible with MSCs and supports their chondrogenic differentiation in vitro.

Extracellular Matrix Based 3D Scaffold for Directing the Fate of Adult Stem Cells into Osteogenic Lineage

“…According to the literature, the generation of the hydrogel network depends signi cantly on the β-sheet structure [14,41,42]. Recent ndings showed the hydrogel was found to be supportive in terms of both mechanical and biochemical properties, which is consistent with the ndings of the prior reports [22,43,44]. After being identi ed and planted into the scaffold, MSCs, in accordance with earlier scaffold research, were observed to interact in a biocompatible and migratory pattern.…”

“…The smooth bril structures and associated open space pores were visible in the hydrogel architecture. The porous hydrogel scaffolds contained the connected open space pores and smooth lament architecture as mentioned in our previous article [22]. The results showed that the hydrogel topography was wavy, shear, tension, and indentation, with varying magnitudes and heights.…”

“…The MSCs seeded on DUC hydrogel survived, multiplied, and were able to advance toward bone formation in vitro. The e ciency, quality, and composition of the decellularized tissue were optimized in our previous work by mild decellularization, which resulted in a loss of DNA content, sGAG, and collagen brils in the tissue [22] An earlier study showed that ECM was preserved following decellularization. Sulfated GAGs, which are known to increase cell matrix interaction and enhance skeletal bone formation, were present in the DUC matrix [36,37].…”

“…According to our previous nding [22], the ability of hydrogels to absorb water is one of their most important property since it can alter the distribution of nutrients and drugs as well as the proliferation and differentiation of the cells that are trapped within [14,40]. Moreover, the biochemical build and level of crosslinking from lyophilized tissue to hydrogel production were identi ed by FTIR analysis.…”

“…These characteristics in a gel might facilitate bone healing and enhance bone regeneration. Previously we have tested this hydrogel for cartilage formation and it came with positive results, [22] due to this we hypothesized that the UC ECM-based scaffolds may also support osteogenic differentiation. Therefore, they were tested for their ability to support the differentiation potential of MSCs along the osteogenic lineage in basal and osteogenic induction conditions in vitro.…”