In VitroEvaluation of a Biomedical-Grade Bilayer Chitosan Porous Skin Regenerating Template as a Potential Dermal Scaffold in Skin Tissue Engineering

Abstract

Chitosan is a copolymer of N-acetylglucosamine and glucosamine. A bilayer chitosan porous skin regenerating template (CPSRT) has been developed for skin tissue engineering. The pore size of the CPSRT was assessed using a scanning electron microscopy (SEM). The in vitro cytocompatibility of the CPSRT was tested on primary human epidermal keratinocyte (pHEK) cultures by measuring lactate dehydrogenase (LDH) levels and skin irritation by western blot analysis of the interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α) secretions. The ability of the CPSRT to support cell ingrowth was evaluated by seeding primary human dermal fibroblasts (pHDFs) on the scaffold, staining the cells with live/dead stain, and imaging the construct by confocal microscopy (CLSM). The CPSRT with pore sizes ranging from 50 to 150 μm was cytocompatible because it did not provoke the additional production of IL-8 and TNF-α by pHEK cultures. Cultured pHDFs were able to penetrate the CPSRT and had increased in number on day 14. In conclusion, the CPSRT serves as an ideal template for skin tissue engineering.

Proliferation and Differentiation of Human Hair Follicle Stem Cells on Chitosan-Skin Engineered Template in Vitro

“…Porosity on a specific scale accelerates specific tissue integration and vascularization. This can be achieved by using a high density of cells to proliferate throughout the scaffold and to ultimately regenerate the specific tissue [11]. In the area of tissue engineering, the function of a scaffold is actually to provide three-dimensional cultivation conditions and to ultimately enhance cell adhesion and proliferation in vitro [9], [15].…”

“…The viability assay of the HFSCs on the chitosan SETs is presented in Table 1; this assay demonstrated that the growth of the HFSCs increased over time. This promising finding of HFSC viability and growth using a chitosan SETs has increased the interest of researchers in employing this scaffold for the fabrication of epidermal substitutes [6], [9], [11]. Chitosan skin substitutes and chitosan SETs have been demonstrated to accelerate wound healing in irradiated rats.…”

“…Chitosan is a poly-(β-1, 4-D-glucosamine) and deacetylated derivative of chitin. Among the attractive features of chitosan are its biomechanical properties [10], which are ideally suited for skin tissue engineering [6], [9], [11]. The potential of chitosan in supporting HFSCs and their differentiation potential in three-dimensional culture is yet unexplored.…”

A Bilayer Engineered Skin Substitute for Wound Repair in an Irradiation-Impeded Healing Model on Rat

“…At 2 h postculture, another 100 lL of culture medium was added to each chitosan. At 6 h postculture, the chitosan discs were transferred into 24-well plates and medium was added adequately as previously explained 7 followed by incubation under 5% CO 2 at 37°C. The growth medium was changed every day.…”

Microstructure, Mechanical, and Biological Properties of Porous Poly(vinylidene fluoride) Scaffolds Fabricated by Selective Laser Sintering

“…Tridimensional porous scaffolds played a crux role in synthetic bone, because the scaffolds could imitate the bone constructions and role of the extracellular matrix and endorse cell conglutination, multiplication, and differentiation. Besides they could transport nutrient and metabolic waste [4][5][6][7].…”