METHOD FOR PRODUCING BIODEGRADABLE, BIOCOMPATIBLE TISSUE-ENGINEERED SHEETS USING CULTURED FIBROBLASTS

摘要

The invention may be applied either independently, or in complex surgery treatment of patients with deep burn wounds, extended post-traumatic wound defects of skin and soft tissues, osteomyelitis of different geneses and concomitant with soft tissue defects, and also long-time, poorly healed trophic ulcers. The technical objective of the invention is development of method for producing tissue-engineered sheets representing two-layer bio-engineered constructs that comprise underlayer of biodegradable matrix consisting of natural polysaccharides, chitosane and pectin, supplemented with an antioxidant, quercetin, and performing mechanical and supporting functions. A second layer is formed by skin cells, fibroblasts, and represents artificial analogue to a dermal skin layer that can lower risk of rough scar tissue formation. Cell material represents suspension of live cultured fibroblasts of density 50×10⁴ cells per 1 cm² in nutrient medium with no added xenogenic serum, and it is applied on underlayer surface as suspension and can adhere itself upon matrix surface using centrifugation at 200G (1500 rpm) during 5 to 8 min, so these cells are not cultured. As allogenic material, use is recommended of either human diploid fibroblast strain FECh 2/09 (Innovation Patent No. 25091 of 15.12.2011), or other characterized skin cell culture, or autologous cells obtained from the same patient. Constructive features of transplant performance provide the most beneficial adherence to defect areas and exclude possibility of damage to under- or near-laying tissues. High sorption ability and optimal terms of resorbtion can prevent formation of inflammatory infiltrates and suppuration of wound. Reliability of obtained results was clinically proved. Efficacy in application of tissue-engineered sheets with adhered fibroblasts is provided through non-specific stimulation of epithelium reparation, mainly due to secretion of growth factors, cytokines, and extracellular matrix molecules, which can promote healing, upon wound surface by these cells, while underlayer renders immune-modulation effect. The disclosed method can increase degree of auto transplant engraftment, accelerate epithelisation of perforation spots, and reduce recovery period in both proper and donor wound surfaces.