ABSTRACT
Alginate is a biocompatible material suitable for biomedical applications, which can be processed under mild conditions on irradiation. This paper investigates the preparation and the rheological behavior of different pre-polymerized and polymerized alginate methacrylate systems for three-dimensional photopolymerization bioprinting. The effect of the functionalization time on the mechanical, morphological, swelling, and degradation characteristics of cross-linked alginate hydrogel is also discussed. Alginate was chemically-modified with methacrylate groups and different reaction times considered. Photocurable alginate systems were prepared by dissolving functionalized alginate with 0.5- 1.5% w/v photoinitiator solutions and cross-linked by ultraviolet light (8 mW/cm2 for 8 minutes).
Subject(s)
Coated Materials, Biocompatible/chemistry , Nanofibers/chemistry , Regeneration/physiology , Skin/metabolism , Tissue Scaffolds/chemistry , Bandages , Biomechanical Phenomena , Cell Culture Techniques , Cell Proliferation , Cell Survival , Fibroblasts/cytology , Gelatin/chemistry , Humans , Infant, Newborn , Male , Polyesters/chemistry , Tissue Engineering/methods , Wound HealingABSTRACT
Skeletal muscle has good regenerative capacity, but the extent of muscle injury and the developed fibrosis might prevent complete regeneration. The in vivo application of human mesenchymal stem cells (HMSCs) of the umbilical cord and the conditioned media (CM) where the HMSCs were cultured and expanded, associated with different vehicles to induce muscle regeneration, was evaluated in a rat myectomy model. Two commercially available vehicles and a spherical hydrogel developed by our research group were used. The treated groups obtained interesting results in terms of muscle regeneration, both in the histological and in the functional assessments. A less evident scar tissue, demonstrated by collagen type I quantification, was present in the muscles treated with HMSCs or their CM. In terms of the histological evaluation performed by ISO 10993-6 scoring, it was observed that HMSCs apparently have a long-term negative effect, since the groups treated with CM presented better scores. CM could be considered an alternative to the in vivo transplantation of these cells, as it can benefit from the local tissue response to secreted molecules with similar results in terms of muscular regeneration. Searching for an optimal vehicle might be the key point in the future of skeletal muscle tissue engineering.
ABSTRACT
In peripheral nerves MSCs can modulate Wallerian degeneration and the overall regenerative response by acting through paracrine mechanisms directly on regenerating axons or upon the nerve-supporting Schwann cells. In the present study, the effect of human MSCs from Wharton's jelly (HMSCs), differentiated into neuroglial-like cells associated to poly (DL-lactide-ε-caprolactone) membrane, on nerve regeneration, was evaluated in the neurotmesis injury rat sciatic nerve model. Results in vitro showed successful differentiation of HMSCs into neuroglial-like cells, characterized by expression of specific neuroglial markers confirmed by immunocytochemistry and by RT-PCR and qPCR targeting specific genes expressed. In vivo testing evaluated during the healing period of 20 weeks, showed no evident positive effect of HMSCs or neuroglial-like cell enrichment at the sciatic nerve repair site on most of the functional and nerve morphometric predictors of nerve regeneration although the nociception function was almost normal. EPT on the other hand, recovered significantly better after HMSCs enriched membrane employment, to values of residual functional impairment compared to other treated groups. When the neurotmesis injury can be surgically reconstructed with an end-to-end suture or by grafting, the addition of a PLC membrane associated with HMSCs seems to bring significant advantage, especially concerning the motor function recovery.
Subject(s)
Disease Models, Animal , Mesenchymal Stem Cells/cytology , Nerve Regeneration/drug effects , Peripheral Nerve Injuries/physiopathology , Peripheral Nerve Injuries/therapy , Polyesters/pharmacology , Wharton Jelly/cytology , Animals , Biomechanical Phenomena/drug effects , Cell Differentiation/drug effects , Humans , Immunohistochemistry , Karyotyping , Membranes, Artificial , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/drug effects , Neuroglia/cytology , Neuroglia/drug effects , Peripheral Nerve Injuries/pathology , Rats , Reaction Time , Reflex/drug effects , Reproducibility of Results , Sciatic Nerve/drug effects , Sciatic Nerve/pathology , Sciatic Nerve/physiopathologyABSTRACT
Cellular adhesion and proliferation inside three-dimensional synthetic scaffolds represent a major challenge in tissue engineering. Besides the surface chemistry of the polymers, it is well recognized that scaffold internal architecture, namely pore size/shape and interconnectivity, has a strong effect on the biological response of cells. This study reports for the first time how polycaprolactone (PCL) scaffolds with controlled micro-architecture can be effectively produced via bioextrusion and used to enhance the penetration of plasma deposited species. Low-pressure nitrogen-based coatings were employed to augment cell adhesion and proliferation without altering the mechanical properties of the structures. X-ray photoelectron spectroscopy carried out on different sections of the scaffolds indicates a uniform distribution of nitrogen-containing groups throughout the entire porous structure. In vitro biological assays confirm that plasma deposition sensitively promotes the activity of Saos-2 osteoblast cells, leading to a homogeneous colonization of the PCL scaffolds.