2D and 3D Self-Assembling Nanofiber Hydrogels for Cardiomyocyte Culture

Abstract

Collagen is a widely used biomaterial in cardiac tissue engineering studies. However, as a natural material, it suffers from variability between batches that can complicate the standardization of culture conditions. In contrast, synthetic materials are modifiable, have well-defined structures and more homogeneous batches can be produced. In this study, several collagen-like synthetic self-assembling nanofiber hydrogels were examined for their suitability for cardiomyocyte culture in 2D and 3D. Six different nanofiber coatings were used in the 2D format with neonatal rat cardiomyocytes (NRCs) and human embryonic stem-cell-derived cardiomyocytes (hESC-CMs). The viability, growth, and functionality of the 2D-cultured cardiomyocytes were evaluated. The best-performing nanofiber coatings were selected for 3D experiments. Hydrophilic pH-sensitive nanofiber hydrogel coassembled with hyaluronic acid performed best with both NRCs and hESC-CMs. Hydrophilic non-pH-sensitive nanofiber hydrogels supported the growth of NRCs; however, their ability to promote attachment and growth of hESC-CMs was limited. NRCs also grew on hydrophobic nanofiber hydrogels; however, the cell-supporting capacity of these hydrogels was inferior to that of the hydrophilic hydrogel materials. This is the first study demonstrating that hydrophilic self-assembling nanofiber hydrogels support the culture of both NRCs and hESC-CMs, which suggests that these biomaterials hold promise for cardiac tissue engineering.

25th Anniversary Article: Supramolecular Materials for Regenerative Medicine

“…For instance, the bioactive RGD or RGDS has been successfully attached to supramolecular materials ranging from self-assembled monolayers, 42,43,44,45,46 small molecule hydrogelators, 47,48,49,50 vesicle-forming block copolymers 51,52 and supramolecular polymers 53 . In one example, the RGDS has been utilized to functionalize peptide amphiphile (PA) fibers, which are 1D self-assembled nanostructures structures reminiscent of the ECM.…”

Physical cues of cell culture materials lead the direction of differentiation lineages of pluripotent stem cells

“…Subsequently, the cells are cultured on specific somatic cells or tissues, which guide the differentiation of hPSCs into specific tissue cells. 49,[116][117][118][119][120][121][122][123] Human neural crest stem cells (hNCSCs) offer opportunities to investigate neural crest development and provide a clinical and treatment model of neural crest-related disorders. However, it is not easy to differentiate hPSCs into hNCSCs.…”

“…119 Fig. 20 shows six types of molecular schemes for peptide amphiphile molecules where the nanofiber can be formed via the self-assembly of peptide amphiphile molecules.…”

“…20 shows six types of molecular schemes for peptide amphiphile molecules where the nanofiber can be formed via the self-assembly of peptide amphiphile molecules. 119 The differentiation of hESCs (H7) into cardiomyocytes was performed by coculturing hESCs with END-2 cells using the Type F protocol shown in Fig. 2.…”

“…Therefore, several researchers have investigated the effects of nanofibrous structures on the differentiation of hESCs into neural cells, myocytes, and cardiomyocytes. 48,[85][86][87]97,119 Lam et al investigated the effect of using aligned and random nanofibers for hESC (H9) differentiation into neural cells using the Type B and D protocols shown in Fig. 2.…”