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Revision as of 03:24, 24 September 2018
Abstract
Cell lines HaCat and L-929 were used to evaluate a tissue regeneration system incorporating a collagen-based scaffold and encapsulated recombinant growth factors. A collagenous synthetic gene comprising collagen type V heparin binding domain (HepV) was expressed in a bacterial system, constituting the main component of the scaffold. For collagen molecule consists of repetitive G-X-Y triplets, being Y frequently hydroxyproline (Hyp), a method of intracellular accumulation of Hyp is used to co-translationally incorporate this non-canonical amino acid. A chitosan crosslinking procedure is carried out on the expressed collagen to prevent contraction and to finally freeze-dry. Further, a mixture of heparin and tenascin C fibronectin type III domain V (TNCIII5) was added to the scaffold, for the heparin-tenascin C binding has a high affinity with a large number of growth factors. Recombinant human leptin β (LepB) is used as a growth factor and we hypothesize that it may be encapsulated to achieve a controlled drug delivery. Due to its suitability, chitosan nanoencapsulation was employed. To evaluate the efficacy of the system, MTT proliferation assay was performed in 96-well plates. Given the cell lines assessed during this project, this tissue regeneration system proves its ability to be used in the treatment of second degree burns. As a widely new biotechnology procedure for skin regeneration the creation of a new, low cost method, for the silencing of miRNA´s involved in the skin regeneration was created and tested. The project was also tested using a skin chip as a practical and small cell proliferation hardware.
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