Free thiol groups on poly(aspartamide) based hydrogels facilitate tooth-derived progenitor cell proliferation and differentiation

Abstract

Cell-based tissue reconstruction is an important field of regenerative medicine. Stem and progenitor cells derived from tooth-associated tissues have strong regeneration potential. However, their in vivo application requires the development of novel scaffolds that will provide a suitable three-dimensional (3D) environment allowing not only the survival of the cells but eliciting their proliferation and differentiation. Our aim was to study the viability and differentiation capacity of periodontal ligament cells (PDLCs) cultured on recently developed biocompatible and biodegradable poly(aspartamide) (PASP)-based hydrogels. Viability and behavior of PDLCs were investigated on PASP-based hydrogels possessing different chemical, physical and mechanical properties. Based on our previous results, the effect of thiol group density in the polymer matrix on cell viability, morphology and differentiation ability is in the focus of our article. The chemical composition and 3D structures of the hydrogels were determined by FT Raman spectroscopy and Scanning Electron Microscopy. Morphology of the cells was examined by phase contrast microscopy. To visualize cell growth and migration patterns through the hydrogels, two-photon microscopy were utilized. Cell viability analysis was performed according to a standardized protocol using WST-1 reagent. PDLCs were able to attach and grow on PASP-based hydrogels. An increase in gel stiffness enhanced adhesion and proliferation of the cells. However, the highest population of viable cells was observed on the PASP gels containing free thiol groups. The presence of thiol groups does not only enhance viability but also facilitates the osteogenic direction of the differentiating cells. These cell-gel structures seem to be highly promising for cell-based tissue reconstruction purposes in the field of regenerative medicine.

Analysis of Three-Dimensional Cell Migration in Dopamine-Modified Poly(aspartic acid)-Based Hydrogels

“…This observation suggests that a great amount of PDLCs could adhere to the DA-free gels, but on this gel type, they were not able to proliferate. Previous studies on PASP hydrogels are also in line with our observations, and it has been described that these gels provide a suitable surface for cell adhesion and proliferation of PDLCs [15] and MG-63 osteosarcoma cells [8]. However, the viability of PDLCs did not change notably on DAcontaining gels during the 14-day-long experiment.…”

“…Currently, there are only a few types of synthetic poly(amino acids) available, such as polylysine or poly(aspartic acid) (PASP) that can be suitable for this purpose [ 10 , 11 ]. In our previous studies, the chemical constitution and the physical properties of PASP-based hydrogels have been comprehensively investigated [ 12 , 13 , 14 ] and successfully tailored for applications as a scaffold [ 8 , 15 ]. We have demonstrated that thiol groups in the PASP hydrogels facilitate cell adhesion and proliferation [ 15 ].…”

Hyaluronic Acid Is an Effective Dermal Filler for Lip Augmentation: A Meta-Analysis

“…Theoretically, there is a wide range of possible reconstructive methods that can be applied to rebuild damaged tissues such as tissue engineering using stem cells (9)(10)(11), gene therapy (12,13) and artificial biocompatible scaffolds (14,15), but their use has been not well-established in routine clinical settings. Among the non-surgical regenerative and reconstructive procedures, hyaluronic acid (HA)-based dermal filling is one of the most frequently used treatments (16,17).…”

Culturing and Scaling up Stem Cells of Dental Pulp Origin Using Microcarriers

“…In parallel research we work on the development of scaffolds that allow not only the survival and proliferation of the ectomesenchyme-derived cells, both in vitro and in vivo, but also elicit their differentiation [64][65][66][67][68]. By chemical modification of biocompatible and biodegradable poly(aspartamide) (PASP)-based hydrogels, and parallel studies on their biocompatibility, biodegradability and regenerative potential, we are attempting to develop novel scaffolds that are clinically applicable [64][65][66][67][68]. We expect that in the near future these separate lines of research, on microcarriers and scaffolds, will meet through the development of biomaterials that can serve both for cell expansion and tissue regeneration.…”