Evaluation of the HGF-1 Cells on Titanium Abutments Fixed by Various Procedures

Abstract

The aim of this study was to compare the effects of fixation protocols in terms of attachment and proliferation of human gingival fibroblasts (HGF-1) on titanium abutment surfaces. HGF-1 cells were seeded on Titanium alloy (Ti6Al4V) plates with dimensions of 10x10x1 cm3. Cells were evaluated at 48 hours. 8 groups were determined (n=4). The specimens were treated with glutaraldehyde (GA) for 30,45 and 60 minutes (Group GA30, GA45, GA60), with formaldehyde (FA) for 6, 12, and 24 hours (Group FA6,FA12,FA24) and with paraformaldehyde (PFA) for 2 hours and 20 minutes (Group PFA2, PFA20) for fixation. Specimens were evaluated under Scanning Electron Microscope (SEM) (n=2) and under Immonoflorescence (n=2). The contact areas of cells in all groups were measured and differences between hours was evaluated statistically. According to SEM images, HGF-1 cell were similar for FA groups as spindled and homogeneous. Cell contact area was 95.83% for FA12 which exhibited the best cell spread among all groups. In GA and PFA groups the cells were elongated, clustered in certain areas and layered with distinct nucleated. According to immunofluorescence images, the densities of actin filaments were observed at similar levels in cells in all three groups, and it was determined that the cell bodies on the titanium surface in the PFA groups were more prominent, and better spread. The fixation process is one of the most critical step of imaging in cell studies. Researchers are needed to consider the most appropriate fixation method in order to obtain successful image results.

Keywords

• Ti6Al4V
• HGF-1
• Fixation
• Immunoflorescence

Titanyum Abutment Üzerinde Çeşitli Prosedürlerle Fikse Edilmiş HGF-1 Hücrelerinin Değerlendirilmesi

Abstract

Bu çalışmanın amacı, titanyum abutment üzerinde, insan gingival fibroblast hücre hattının (HGF-1), tutunma ve proliferasyon açısından fiksasyon solüsyon ve sürelerinin etkilerinin SEM ve İmmunofloresan görüntüleme ile değerlendirilmesi ve kıyaslanmasıdır. Hazır temin edilen insan gingival hücre hattı (HGF-1) 10x10x1cm3 boyutunda 32 adet titanyum alaşım (Ti6Al4V) plaka üzerine ekildi. 8 grup belirlendi (n=4). 48 saat sonucunda hücreler değerlendirildi. Örnekler Gluteraldehit ile 30, 45, 60 dakika (Grup GA30, GA45, GA60), Formaldehit ile 6, 12, 24 saat (Grup FA6, FA12, FA24) ve Paraformeldehit ile 2 saat ve 20 dakika (Grup PFA2, PFA20) süre ile fikse edildi. Fiksasyon sonrası her gruptan 2 örnek Taramalı elektron mikroskobunda (SEM) ve 2 örnek İmmunofloresan mikroskobunda görüntülenmek için hazırlandı. Tüm gruplardaki hücrelerin temas alanları ölçüldü, saatler arasındaki farklılık istatistiksel olarak değerlendirildi. SEM değerlendirmesinde, en uygun HGF-1 hücre morfolojisi görüntülerinin formaldehit gruplarının iğsi şekilli homojen yayılım gösterdiği tespit edildi. FA12 grubunda hücre temas alanı %95,83 bulunmuş olup, tüm deney grupları içerisinde en iyi hücre yayılımını göstermiştir. Gluteraldehit ve paraformaldehit gruplarında birbirleriyle benzer şekilde hücrelerin uzamış, belirli alanlarda öbekleşmiş ve üst üste katmanlanmış belirgin çekirdekli hücre görüntüleri tespit edildi. İmmunofloresan görüntülerinde her üç (gluteraldehit, formaldehit, paraformaldehit) gruptaki hücrelerde de aktin filamentlerinin yoğunlukları benzer seviyelerde görülmesinin yanı sıra paraformaldehit gruplarında titanyum yüzeydeki hücre gövdelerinin diğer fiksatif gruplarına göre daha belirgin, iyi yayılmış ve daha büyük yüzey alanlarına sahip olduğu gözlendi. Fiksasyon hücre çalışmalarında görüntülemenin en kritik basamaklarından biridir. Araştırmacıların başarılı görüntü sonucu elde edebilmek için en uygun fiksasyon yöntemini göz önünde bulundurmaları gerekmektedir.

Keywords

• Ti6Al4V
• HGF-1
• Fiksasyon
• İmmunofloresan

Supporting Institution

Bu çalışma herhangi bir kurum tarafından desteklenmemiştir.

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