pGM-CSF İçeren Kitozan/PVP Hidrojellerinin GM-CSF Ekspresyonu ve Fibroblast Proliferasyonuna Etkileri

Abstract

Bu çalışmanın amacı, gen taşınması için pGM-CSF içeren kitozan/PVP hidrojel sistemini geliştirmektir. Yarı-interpenetre polimer ağ (yarı-IPN) yapısındaki hidrojeller, %2 kitozan ve %2, %4, %6 PVP’nin glutaraldehitle çapraz bağlanması ile hazırlandı. Hidrojellerin viskozite, şişme özellikleri ve yüzey morfolojisi incelendi. Bu hidrojeller oldukça poroz yapı gösterdi. PVP konsantrasyonunun artması ile porozite arttı. Viskozite hidrojellerin PVP konsantrasyonun artması ile azaldı. Şişme yüzdesi PVP konsantrasyonunun artması ile artış gösterdi. Hidrojellerin hücre tutunması ve proliferasyonuna etkisini incelemek için, NIH 3T3 fibroblast hücreleri kullanıldı. Hücreler, hidrojellerin yüzeyine tutundu. Bununla birlikte hücrelerin tutunması 1 ve 3 gün de yavaş olarak gerçekleşti. 7. ve 14. günlerde hücreler hidrojellerin yüzeyinde ve aralarında sağlıklı normal görünüşlü olup, proliferasyon gözlendi. ELISA çalışması ile en yüksek GM-CSF ekspresyonu pGM-CSF içeren %2 kitozan+%4PVP hidrojellerinde elde edildi. MTT çalışması ile kitozan/PVP hidrojellerinin önemli bir sitotoksik etkiyi indüklemediği gözlendi. Sonuç olarak, bu hidrojeller doku rejenerasyonunda lokal gen taşınması için aday taşıyıcı sistem olarak önerilebilir.

Keywords

• pGM-CSF,PVP,kitozan,hidrojel

The Effects to GM-CSF Expression and Fibroblast Proliferation of pGMCSF Containing Chitosan/PVP Hydrogels

Abstract

The aim of this study was to develop pGM-CSF containing chitosan/PVP hydrogel system for gene delivery. The hydrogels in semi-interpenetrating polymer network (semi-IPN) structure were synthesized by glutaraldehyde crosslinking between 2% chitosan and 2%, %4, %6 PVP. The viscosity, swelling properties and surface morphology of hydrogels were examined. These hydrogels showed highly porous structure. The porosity was increased with enhanced PVP concentration. Viscosity decreased with increasing PVP concentration of the hydrogels. The swelling percentage increased with increasing PVP concentration. In order to examine the effect of cell attachment and proliferation of hydrogels, NIH-3T3 fibroblast cell was used. The cells were able to attach on the top surface of hydrogels. However, the attachment of cells were slow at days 1 and 3. At days 7 and 14, cells looked healthy and normal and proliferated around and top of the hydrogels. With the ELISA study, the highest GM-CSF expression was obtained with pGM-CSF containing 2% chitosan+4% PVP hydrogels. MTT assay showed that chitosan/PVP hydrogels did not induce significant cytotoxic effect. As a result, these hydrogels could be served candidate for local gene delivery in tissue regeneration.

Keywords

• pGM-CSF,chitosan,PVP,hydrogel,gene delivery

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