Production and characterization of acellular matrices of porcine skin using a time and cost effective method

• Grant number: 15/03916-1
• Support Opportunities: Scholarships abroad - Research Internship - Scientific Initiation
• Effective date (Start): June 01, 2015
• Effective date (End): September 27, 2015
• Field of knowledge: Biological Sciences - Morphology - Histology
• Principal Investigator: Sonia Maria Oliani
• Grantee: Andréia Rodrigues de Moraes
• Supervisor: Karin Vicente Greco
• Host Institution: Instituto de Biociências, Letras e Ciências Exatas (IBILCE). Universidade Estadual Paulista (UNESP). Campus de São José do Rio Preto. São José do Rio Preto , SP, Brazil
• Research place: University College London (UCL), England
• Associated to the scholarship: 14/18557-4 - Study of inflammatory and angiogenic processes in heterologous implants of rats using porcine acellular matrices, BP.IC

Abstract

The clinical impact of various lesions such as loss of skin due to non-healing ulcers, abdominal wall defects resulting from trauma, extensive tumor resection, necrotizing infections, or extensive burns associated with limited use of autografts, has led an enormous advance in the development of tissue-engineered skin derivatives or acellular matrices. Designed as skin substitutes, both epidermal and dermal equivalents have been under investigation and used clinically for over a decade. In a new era of regenerative medicine, decellularized biological constructs not only have the ability to retain the original three dimensional morphological architecture of the tissue but also preserve the structure and function of a complex mixture of proteins located on the extracellular matrix (ECM). The ECM is highly conserved among many species and consists of molecules such as collagen, fibronectin, laminin, vitronectin, glycosaminoglycans, and growth factors oriented in a specific three-dimensional structure and composition optimized for each tissue of origin. Collagen and glycosaminoglycans (GAGs) are the main mechanostructural elements for dermis, conferring tensile strength and proteolytic resistance to the tissue. ECM molecules signal cells directly or indirectly to trigger biological events and help orchestrate a set of events in a sequence to promote proliferation and differentiation of the host derived cells. These events ultimately lead to tissue remodelling and regeneration with restauration of function of the tissue/organ. The bioactivity of scaffolds can be altered depending on the combination of decellularization reagents used. In addition, residual chemicals from the decellularization process can also impact on the cell's adherence and proliferation capability. A variety of decellularization methods exist to produce skin scaffolds, ranging from osmotic shock to prolonged exposure to a combination of detergents, enzymatic solutions or acids/bases that can last days or several weeks. Nevertheless, decellularization of tissues with preservation of matrix integrity and bioactivity permitting cells recruitment and remodelling can be optimized to produce an "off-the-shelf" product for clinical application which is both time efficient and cost effective. Hence, the scope of this study is to compare different decellularization protocols, to assess their impact on the ECM with regards to structure and composition in order to produce scaffolds that can be a feasible alternative for skin transplants. (AU)