Effective decellularization of human skin tissue for regenerative medicine by supercritical carbon dioxide technique

Abstract

Allotransplantation, performed using an acellular dermal matrix (ADM), plays a significant role in the cultivation of constituted and damaged organs in clinical.Herein, we fabricated an innovative ADM for allografting derived from decellularized human skin by utilizing the supercritical fluid of carbon dioxide to eliminate immunogenic components. By using histological staining, the ADM product demonstrated the successful removal of cellular constituents without exerting any harmful influence on the extracellular matrix. The results from DNA electrophoresis also supported this phenomenon by showing the complete DNA removal in the product, accompanied by the absence of Major Histocompatibility Complex 1, which suggested the supercritical fluid is an effective method for cellular withdrawal.Moreover, the mechanical property of the ADM products, which showed similarity to that of native skin, displayed great compatibility for using our human-derived ADM as an allograft in clinical treatment. Specifically, the cell viability demonstrated the remarkable biocompatibility of the product to human bio-cellular environment which was noticeably higher than that of other products. Additionally, the significant increase in the level of growth factors such as vascular endothelial growth factor, urokinase-type plasminogen activator receptor, granulocytemacrophage colony-stimulating factor suggested the ability to stimulate cellular processes, proving the products to be innovative in the field of regeneration when applied to clinical in the future. This study provides a thoroughly extensive analysis of the new ADM products, enabling them to be applied in industrial and clinical treatment.

Decellularized dermal matrices: unleashing the potential in tissue engineering and regenerative medicine

“…Even though they did not reach the acellular conditions establish by Crapo and Badylak et al (Crapo et al, 2011), of less than 50 ng of dsDNA per mg of dry sample, they were able to remove the majority of the cells. The resultant matrix demonstrated to have excellent biomechanical properties, showing similarities to the native skin, and a very promising content of growth factors and anti-inflammatory cytokines (Giang et al, 2022). These results confirm the potential of this technology to remove cells without substantially alter the ECM's bioactivity and biofunctionality.…”

“…ScCO 2 has affinity for lipids, but in the cell membrane and in nucleic acid there are polar molecules that can be corrupted by the use of co-solvents (e.g., ethanol) enhancing the fluid's solvating power (Chou et al, 2020;Duarte et al, 2022). This method has important advantages, i.e., low EMC damages, is non-toxic, environmentally friendly, does not leave residues, odors and is efficient in eliminating chemical residues (Chou et al, 2020;Giang et al, 2022). Wang et al (Wang C.-H. et al, 2020), have already produced a porcine acellular dermal matrix using scCO 2 as decellularization method, resulting in a non-toxic and biocompatible material capable of accelerating wound healing in a full-thickness in vivo model.…”

“…Wang et al (Wang C.-H. et al, 2020), have already produced a porcine acellular dermal matrix using scCO 2 as decellularization method, resulting in a non-toxic and biocompatible material capable of accelerating wound healing in a full-thickness in vivo model. Another study conducted by Giang et al (Giang et al, 2022), proposed the decellularization of human dermis using scCO 2 with ethanol as co-solvent. Even though they did not reach the acellular conditions establish by Crapo and Badylak et al (Crapo et al, 2011), of less than 50 ng of dsDNA per mg of dry sample, they were able to remove the majority of the cells.…”

Assessment of inflammatory suppression and fibroblast infiltration in tissue remodelling by supercritical CO2 acellular dermal matrix (scADM) utilizing Sprague Dawley models

“…The methods of ADM preparation were reported in the previous study ( Giang et al, 2022 ). In brief, the tissue was defatted, washed in sterile phosphate-buffered saline (PBS) for 30 min, and treated with 1 M NaCl at 37°C overnight to separate the epidermis from the dermis.…”

Unlocking the Promise of Decellularized Pancreatic Tissue: A Novel Approach to Support Angiogenesis in Engineered Tissue

“…Recent advances in regenerative medicine gave rise to a widespread interest in utilizing decellularized extracellular matrix (ECM) as a biological scaffold to develop novel treatment options [1][2][3][4][5] and suggest the possibility of employing ECM as a scaffold to bioengineer functional organs [4,6,7]. Commonly explored decellularization methods include physical [8,9], biological [10,11] and chemical [12][13][14] techniques. Physical methods such as electroporation and hydrostatic pressure are precise and non-toxic, yet their efficacy varies across tissues and may damage the ECM structure [15].…”