Keratose Hydrogels Promote Vascular Smooth Muscle Differentiation from C-kit-Positive Human Cardiac Stem Cells

Abstract

Stem cell-based therapies have demonstrated great potential for the treatment of cardiac diseases, for example, myocardial infarction; however, low cell viability, low retention/engraftment, and uncontrollable in vivo differentiation after transplantation are still major limitations, which lead to low therapeutic efficiency. Biomaterials provide a promising solution to overcome these issues due to their biocompatibility, biodegradability, low/nonimmunogenicity, and low/noncytotoxicity. The present study aimed to investigate the impacts of keratose (KOS) hydrogel biomaterial on cellular viability, proliferation, and differentiation of c-kit human cardiac stem cells (hCSCs). Briefly, hCSCs were cultured on both KOS hydrogel-coated dishes and regular tissue culture dishes (Blank control). Cell viability, stemness, proliferation, cellular morphology, and cardiac lineage differentiation were compared between KOS hydrogel and the Blank control at different time points. We found that KOS hydrogel is effective in maintaining hCSCs without any observable toxic effects, although cell size and proliferation rate appeared smaller on the KOS hydrogel compared to the Blank control. To our surprise, KOS hydrogel significantly promoted vascular smooth muscle cell (VSMC) differentiation (∼72%), while on the Blank control dishes, most of the hCSCs (∼78%) became cardiomyocytes. Furthermore, we also observed "endothelial cell tube-like" microstructures formed by differentiated VSMCs only on KOS hydrogel, suggesting a potential capability of the hCSC-derived VSMCs for in vitro angiogenesis. To the best of our knowledge, this is the first report to discover the preferred differentiation of hCSCs toward VSMCs on KOS hydrogel. The underlying mechanism remains unknown. This innovative methodology may offer a new approach to generate a robust number of VSMCs simply by culturing hCSCs on KOS hydrogel, and the resulting VSMCs may be used in animal studies and clinical trials in combination with an injectable KOS hydrogel to treat cardiovascular diseases.

A keratin-based microparticle for cell delivery

“…Keratin proteins were extracted as previously described. 11 Briefly, human hair fibers were oxidized using a 2% peracetic acid (PAA) solution (pH 2) for 18 hrs using a 20:1 ratio of PAA to hair mass (20 mL PAA: 1 g hair). Oxidized fibers were rinsed with deionized (DI) water to remove residual oxidant and Tris base (100 mM) was used to solubilize keratin proteins from the fiber cortex.…”

“…Prior research suggests keratin and keratin derivatives from human hair exhibit several mechanical and biological properties that support their use as scaffolds for cells that can incorporate various payloads including drugs and growth factors. 11–13 Keratin hydrogels mechanically perform as a flexible, viscoelastic material. However, most reports of keratin biomaterials in tissue engineering utilize cell-free constructs, despite keratin’s intrinsic cell and tissue compatibility and apparent support of cell health and function.…”

Design of Polymeric Culture Substrates to Promote Proangiogenic Potential of Stem Cells

“…StarPEG‐heparin hydrogel enabled multiple modulations of the capillary EC network by delivering multiple elements ( Figure 3 ). Ledford et al demonstrated the effect of keratose (KOS) hydrogel on cardiac differentiation of human cardiac stem cells (hCSCs) cultured in media containing bFGF . Although the underlying mechanism is not known, vascular smooth muscle cells (VSMCs) differentiated from hCSCs exhibited a tubular structure indicating initiation of angiogenesis process.…”

TGFβ1 Improves the Proliferation of Decellularized and Aligned Skeletal Muscle in Extracellular Matrix Hydrogels by m6A Modification-Mediated Integrin Through Inducing ERK Phosphorylation

“…Decellularized extracellular matrix (ECM) is a common material adopted for the rehabilitation and reconstruction of damaged tissues, and it can be obtained by various techniques involving physical methods, chemical reagents and/or biological reagents [6,8,9]. Various decellularized ECM-derived biomaterials, such as small intestine submucosa and urinary bladder tissue, have been adopted; however, these materials have limitations associated with heterogeneity [10][11][12][13][14]. Hydrogels made by ECM are three-dimensional (3D) scaffolds that have a similar three-dimensional microstructure to that of native ECM [12,15,16].…”