CİLT YENİLENMESİ İÇİN YENİ BİR FİBRİN MATRİS OLUŞTURULMASI, KİMYASAL/FİZİKSEL ÖZELLİKLERİ VE DOKU İSKELESİ OLUŞTURMA POTANSİYELİ AÇISINDAN İNCELENMESİ

Abstract

There are a considerable amount of wound dressings, fillers and tissue engineered products for the treatment of skin damages, burns, diabetic wounds or skin aging. These products sometimes lead to adverse reactions like allergy and they have the disadvantages of having high cost and limited curing effect. Therefore, there is a need for affordable biomaterials that may be used in skin regeneration efficiently without any risk. The aim of this thesis is to produce a new, cost efficient and easy to prepare kind of fibrin matrix. Therefore, a porous matrix derived from platelet-rich plasma is produced by simply using either calcium chloride or thrombin as gelling agent. The chemical/physical properties and also the potential of these matrices as a skin regenerating biomaterial is analysed using HS27 human fibroblast cells. As a natural scaffold produced from natural fibrin molecules obtained from platelet-rich plasma as PRFM is known to be a good source for growth factors and citokines due to the high number of platelets that it contains. Here, the matrix is produced by simply using calcium chloride or thrombin instead of common combined use. PRFMs obtained by thrombin had a swelling ratio of 83 ± 0,0019 in 60 minutes and calcium chloride as gelling agent. PRFM had a swelling ratio of 198 ± 0,0038 in 60 minutes. Analyses of both PRFMs by scanning electron microscopy (SEM), have revealed that this difference is due to the difference in pore sizes of calcium chloride PRFM (19,4 ± 4,74 µm) and thrombin PRFM (35 ± 5,8 µm). Thrombin unduced PRFMs degraded more than calcium chloride induced PRFM in PBS and DMEM containing 10% FBS in 28 days. The cross linking of fibrin molecules in the matrices were detected by FTIR (Fourier transformed infrared) spectroscopical analysis and their thermal resistance was measured by thermogravimetrical analysis. In this study, the cells cultured for 21 days on these PRFM scaffolds were analysed for their proliferation by MTT test and their collagen I and collagen III production by immunocytochemistry via hemotoxylin/eosin staining and transmission electron microscopy (TEM). Cells growing in the matrix were monitored by scanning electron microscopy (SEM). It is found that fibroblasts attached and proliferated in both matrices, but only the cells on calcium chloride induced matrices have remained intact and synthesized collagen in a three weeks period. It is concluded that, these three dimentional PRFMs, have cell viability and activity promoting properties and thus they may be used in situations which require skin regeneration.