USE OF RECONSTRUCTED SKIN DERIVED FROM MESENCHYMAL STEM CELLS IN A SWINE IN VIVO MODEL OF SKIN HEALING

• Grant number: 23/14620-2
• Support Opportunities: Regular Research Grants
• Start date: April 01, 2025
• End date: March 31, 2027
• Field of knowledge: Health Sciences - Medicine - Surgery
• Principal Investigator: Cristina Pires Camargo
• Grantee: Cristina Pires Camargo
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Luiz Felipe Pinho Moreira ; Viviane Abreu Nunes Cerqueira Dantas

Abstract

Introduction: Some clinical situations require coverage of large areas of skin wounds. Although there are several therapeutic alternatives (allograft, dermal substitutes), in vitro skin transplantation technology can improve the treatment of wounds such as varicose ulcers, diabetic foot ulcers and burns, reducing costs for the health system in the medium and long term. Objective: to analyze the healing process in a porcine model using in vitro reconstructed skin made from decellularized porcine extracellular matrix (dMEC) and human adipose tissue-derived stem cells. Methods: To collect adipose tissue for subsequent isolation and stem cell culture, liposuction products from healthy patients will be collected. Pig skin will also be collected for hydrogel production. Human fibroblasts will be added to the hydrogel to produce the dermal equivalent. In order to develop reconstructed and stratified skin in vitro, the air-liquid interface methodology will be used, where the dermal equivalent will be in contact with the culture medium and the epidermal equivalent will be in contact with the air. The stratified skin produced will be transplanted into male domestic pigs (Sus scrofa domesticus, MS60 strain), weighing between 20 and 30 kg. Two animals will undergo surgery to produce 24 skin lesions up to the subcutaneous tissue with a diameter of 2 cm, under general anesthesia. The lesions will be divided into the following groups: 1) Control: dressing with occlusive sterile gauze (n=8 lesions); 2) skin reconstructed with hydrogel containing fibroblasts (dermal equivalent) and basal keratinocytes (epidermal equivalent) (n=8 lesions); 3) skin reconstructed with hydrogel containing fibroblasts and adipose tissue-derived stem cells (n=8 lesions). Four weeks after the procedure, macroscopy will be performed to analyze the proportion of integration of the reconstructed skin on the pig's back, in addition to the viability of the reconstructed skin and monitoring of the wound healing, histology to evaluate the structure and three-dimensional arrangement of the reconstructed skin, inflammation, angiogenesis, and gene and protein expression of epidermal differentiation and healing markers. (AU)