Effect of Fluorination Modification on Transfection Efficiency of Non-Viral Gene Carrier Systems Based on Chitosan

Öz

The aim of this study was to examine the transfection efficiency of the fluorination modification without the addition of any cationic charge on the chitosan (Chi) molecule. The fluorination reaction on Chi (ChiF) was carried out with using 1H,1H,2H,2H-Perfluorooctyltriethoxysilane (SiF). The characterization of ChiF was realized by Fourier transform infrared (FTIR) analysis and its molecular weight (Mw) and polydispersity index (PDI_Mw) were determined using GPC-SEC system. The physical properties of nanoparticles (nChiF) obtained by ionic gelation method were determined. The gel electrophoresis analysis was applied to the nanoparticles for determine the gene complexing capacity. The cytotoxicity of ChiF onto Human Embryonic Kidney (HEK293) cells was determined via MTT colorimetric assay. The cell confluency (after transfection) and transfection efficiency of nChiF on HEK293 cells were evaluated. The results showed that the nChiF2:pEGFN1 complex (ratio of 35:1) with a particle size of 98.1±2.2 nm and zeta potential of 34.7 ± 6.5 mV, is more superior agent for transfection efficiency in HEK293 cells due to its high transfection effect and higher cell confluency. As a result, it has been showed that the fluorination reaction onto Chi without any cationic charge modification enhance the transfection efficiency for HEK293 cell lines.

Anahtar Kelimeler

• Chitosan,Fluorination,Transfection efficiency,Gene therapy

Kitosan Esaslı Viral Olmayan Gen Taşıyıcı Sistemlerin Transfeksiyon Verimliliği Üzerine Florlama Modifikasyonunun Etkisi

Öz

Bu çalışmanın amacı, kitosan (Chi) molekülü üzerine herhangi bir katyonik yük eklenmeden florinasyon modifikasyonunun transfeksiyon etkinliğini incelemektir. Chi üzerindeki florlama reaksiyonu (ChiF), 1H, 1H, 2H, 2H-Perflorooktiltrietoksisilan (SiF) kullanılarak gerçekleştirildi. ChiF'in karakterizasyonu, Fourier Transform Infrared Spektroskopisi (FTIR) analizi ile gerçekleştirilmiş ve molekül ağırlığı (Mw) ve polidispersite indeksi (PDI_Mw), GPC-SEC sistemi kullanılarak belirlenmiştir. İyonik jelleşme yöntemiyle elde edilen nanopartiküllerin (nChiF) fiziksel özellikleri belirlenmiştir. Gen kompleksleme kapasitesini belirlemek için nanoparçacıklara jel elektroforez analizi uygulandı. ChiF'nin İnsan Embriyonik Böbrek (HEK293) hücreleri üzerindeki sitotoksisitesi, MTT kolorimetrik deneyi ile belirlendi. HEK293 hücreleri üzerine nChiF'nin transfeksiyon etkinliği ve hücre yayılımı (transfeksiyon sonrası) sonuçları değerlendirildi. Sonuçlar, 98.1 ± 2.2 nm partikül büyüklüğüne ve 34.7 ± 6.5 mV zeta potansiyeline sahip nChiF2: pEGFN1 kompleksinin (35: 1 oranı), yüksek transfeksiyon etkisi ve daha yüksek hücre yayılımı gösterdiğinden dolayı HEK293 hücrelerinde transfeksiyon etkinliği için daha üstün bir ajan olduğunu göstermiştir. Sonuç olarak, Chi üzerine herhangi bir katyonik yük modifikasyonu olmadan florinasyon reaksiyonunun, HEK293 hücre hatları için transfeksiyon verimliliğini arttırdığı görülmektedir.

Anahtar Kelimeler

• Kitosan,Florlama,Transfeksiyon etkinliği,Gen terapi

Kaynakça

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