Fabrication of human Wharton’s jelly extra cellular matrix for tissue engineering

Abstract

The development of tissue engineering is based on the use of the extracellular matrix as a construct to which cells migrate and attach for proliferation, differentiation, and long-term functioning. The preparation of the matrix is one of the most important tasks, since it must be non-immunogenic, have optimal mechanical properties, contain cell adhesion molecules and growth factors and degrade at the predicted time. The search for biomaterial for the manufacture of the matrix is limited by a number of circumstances. Tissue-specific for the matrix intravital biomaterial is limited, cadaveric is not acceptable due to age-related changes or diseases that reduce the regenerative capacity of tissues; synthetic materials lack cell adhesion molecules or are not degraded. The umbilical cord is an accessible homologous biomaterial of non- embryonic origin, preserving the features of the embryonic phenotype. The optimal method of decellularization of the Warton jelly of the human umbilical cord in the manufacture of a full-component cell-free matrix is substantiated. Umbilical cord decellularization was carried out using a detergent method with a 0.05% sodium dodecyl sulfate solution for 24 hours. The quality of the decellularization was evaluated microscopically by staining with fluorescent dye and quantification of nucleic acids. The gentle method used to remove cells from the Warton jelly tissue meets the existing criteria for the effectiveness of decellularization, since only single cells and a small amount of deoxyribonucleic acid remain in the processed biomaterial. The technique does not provide centrifugation at high speeds, in which glycosaminoglycans and proteoglycans are lost from the matrix, the enzymatic action that destroys fibrillar collagen structures, and non-physiological conditions of decellularization. The therapeutic success of tissue-engineering structures based on the extracellular matrix will depend not only on the bioactivity of the umbilical cord, but also on the safety of the composition, structure and mechanical characteristics of the matrix. Due to the availability and non-invasiveness of receiving from healthy young donors, provisional organs are an excellent source of homologous biomaterial for matrix production.