Biochemical and mechanical effects of biopreservation methods on decellularized porcine skin grafts

As the demand for tissue and organ transplantation increases, the field of regenerative medicine is rapidly evolving to meet this critical need through innovative methods, such as employing decellularized tissues and organs, for creating engineered transplantable organ and tissue alternatives. This study aims to investigate the decellularization of porcine skin to create biological scaffolds that preserve essential extracellular matrix (ECM) properties and assesses different biopreservation techniques that best preserve morphological, biochemical and mechanical characteristics of the scaffold. We established a decellularization protocol using 1% sodium dodecyl sulfate (SDS) for efficient cellular removal, followed by an evaluation of two biopreservation methods: slow-cooling and freeze-drying. Our results indicate that slow-cooling preserves ECM integrity and composition more effectively than freeze-drying, confirmed by hematoxylin and eosin (H&E) staining and biochemical assays for key ECM components. Quantitative analyses revealed sufficient retention of collagen, glycosaminoglycans (GAGs), and enhanced mechanical properties in slow-cooled samples, which closely resembled fresh decellularized controls. These findings underscore the critical need for optimized preservation techniques to ensure the viability of tissue grafts in regenerative medicine.