Cytocompatible polyion complex gel of poly(Pro-Hyp-Gly) for simultaneous rat bone marrow stromal cell encapsulation

Abstract

Polyion complex (PIC) gel of poly(Pro-Hyp-Gly) was successfully fabricated by simply mixing polyanion and polycation derivatives of poly(Pro-Hyp-Gly), a collagen-like polypeptide. The polyanion, succinylated poly(Pro-Hyp-Gly), and the polycation, arginylated poly(Pro-Hyp-Gly), contain carboxy (pK = 5.2) and guanidinium (pK = 12.4) groups, respectively. Mixing the polyanion and the polycation at physiological pH (pH = 7.4) resulted in PIC gel. The hydrogel formation was optimum at an equimolar ratio of carboxy to guanidinium groups, suggesting that ionic interaction is the main determinant for the hydrogel formation. The hydrogel was successfully used for simultaneous rat bone marrow stromal cell encapsulation. The encapsulated cells survived and proliferated within the hydrogel. In addition, the cells exhibited different morphology in the hydrogel compared with cells cultured on a tissue culture dish as a two-dimensional (2D) control. At day one, a round morphology and homogeneous single cell distribution were observed in the hydrogel. In contrast, the cells spread and formed a fibroblast-like morphology on the 2D control. After three days, the cells in the hydrogel maintained their morphology and some of them formed multicellular aggregates, which is similar to cell morphology in an in vivo microenvironment. These results suggest that the PIC gel of poly(Pro-Hyp-Gly) can serve as a cytocompatible three-dimensional scaffold for stem cell encapsulation, supporting their viability, proliferation, and in vivo-like behavior.

Methods for producing microstructured hydrogels for targeted applications in biology

“…Collagen like polypeptides (CLPs), based on the natural collagen amino acid motifs, Gly-Pro-X and Gly-X-Hyp, adopt a poly(proline) II helix and then further associate into triple helical structures [9][10][11]. The CLPs have yielded materials in which cells can be easily encapsulated with high viability, proliferation, and desired morphology [12,13], characteristics that can be enhanced in hydrogels made with other polymers such as PEG when crosslinked with CLPs [14]. In addition, combining CLPs with other polypeptides imparts new functions to the CLPs, as it has been shown with self-healing upon crosslinking with silk-like polypeptides.…”

Succinylated Bacterial Cellulose Induce Carbonated Hydroxyapatite Deposition in a Solution Mimicking Body Fluid

“…NS = not significant the succinylated-BC. It has been reported that succinylation of a collagen-like polypeptide similarly did not influence the biocompatibility of the polypeptide for rBMSCs [20][21]. Considering its biocompatibility, bioactivity and the ability of the cells to differentiate into bone cells, the succinylated-BC has potential as a 3D scaffold for reconstructing damaged bone.…”