Combination of IKVAV, LRE, and GPQGIWGQ Bioactive Signaling Peptides Increases Human Induced Pluripotent Stem Cell Derived Neural Stem Cells Extracellular Matrix Remodeling and Neurite Extension.

Extracellular matrix (ECM) remodeling is emerging as a modulator of neural maturation and axon extension. Most studies have used rodent cells to develop matrices capable of manipulating extracellular matrix remodeling for regenerative applications. However, clinically relevant human induced pluripotent stem cell derived neural stem cells (hNSC) do not always behave in a similar manner as rodent cells. In this study, hNSC response to a hyaluronic acid matrix with laminin derived IKVAV and LRE peptide signaling that has previously shown to promote ECM remodeling and neurite extension by mouse embryonic stem cells is examined. The addition of enzymatically degradable cross linker GPQGIWGQ to the IKVAV and LRE containing hyaluronic acid matrix is necessary to promote neurite extension, hyaluronic acid degradation, and gelatinase expression over hyaluronic acid matrices containing GPQGIWGQ, IKVAV and LRE, or no peptides. Changes in peptide content alters a number of matrix properties that can contribute to the cellular response, but increases in mesh size are not observed with cross linker cleavage in this study. Overall, these data imply a complex interaction between IKVAV, LRE, and GPQGIWGQ to modulate hNSC behavior.

Effects of free radical initiators on polyethylene glycol dimethacrylate hydrogel properties and biocompatibility.

Many studies have utilized Irgacure 2959 photopolymerized poly(ethylene glycol) (PEG) hydrogels for tissue engineering application development. Due to the limited penetration of ultraviolet light through tissue, Irgacure 2959 polymerized hydrogels are not suitable for use in tissues where material injection is desirable, such as the spinal cord. To address this, several free radical initiators (thermal initiator VA044, ammonium persulfate (APS)/TEMED reduction-oxidation reaction, and Fenton chemistry) are evaluated for their effects on the material and mechanical properties of PEG hydrogels compared with Irgacure 2959. To emulate the effects of endogenous thiols on in vivo polymerization, the effects of chain transfer agent (CTA) dithiothreitol on gelation rates, material properties, Young's and shear modulus, are examined. Mouse embryonic stem cells and human induced pluripotent stem cell derived neural stem cells were used to investigate the cytocompatibility of each polymerization. VA044 and Fenton chemistry polymerization of PEG hydrogels both had gelation rates and mechanical properties that were highly susceptible to changes in CTA concentration and showed poor cytocompatibility. APS/TEMED polymerized hydrogels maintained consistent gelation rates and mechanical properties at high CTA concentration and had a similar cytocompatibility as Irgacure 2959 when cells were encapsulated within the PEG hydrogels. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3059-3068, 2017.

Human Induced Pluripotent Stem Cell Derived Neural Stem Cell Survival and Neural Differentiation on Polyethylene Glycol Dimethacrylate Hydrogels Containing a Continuous Concentration Gradient of N-Cadherin Derived Peptide His-Ala-Val-Asp-Ile.

Although preclinical models of spinal cord injury have shown that matrix inclusion in stem cell therapy leads to greater neurological improvements than that including cells alone, there has been insufficient matrix optimization for human cells. N-Cadherin influences the development and maintenance of neural tissue, but the effects of N-cadherin derived peptide His-Ala-Val-Asp-Ile (HAVDI) on the survival, neurite extension, and expression of neural differentiation markers in human induced pluripotent stem cell derived neural stems (hNSC) have not been widely examined. Using polyethylene glycol hydrogels containing a continuous gradient of HAVDI, this study identifies concentration dependent effects on hNSC survival and neural differentiation.

Oxyma-based phosphates for racemization-free peptide segment couplings.

Glyceroacetonide-Oxyma [(2,2-dimethyl-1,3-dioxolan-4-yl)methyl 2-cyano-2-(hydroxyimino)acetate (1)] displayed remarkable physico-chemical properties as an additive for peptide-forming reactions. Although racemization-free amide-forming reactions have been established for N-urethane-protected α-amino acids with EDCI, 1, and NaHCO3 in water or DMF-water media, amide-forming reactions of N-acyl-protected α-amino acids and segment couplings of oligopeptides still require further development. Diethylphosphoryl-glyceroacetonide-oxyma (DPGOx 3) exhibits relative stability in aprotic solvents and is an effective coupling reagent for N-acyl-protected α-amino acids and oligo peptide segments. The conditions reported here is also effective in lactam-forming reactions. Unlike most of the reported coupling reagents, simple aqueous work-up procedures can remove the reagents and by-products generated in the reactions.