ANTIPROLIFERATIVE EFFECTS OF VITAMIN K2 IN OSTEOSARCOMA CELLS: COMPARISON OF DIFFERENT CYTOTOXICITY ANALYZES

Year 2023, , 1 - 8, 14.03.2023

Büşranur Özalper , Tuba Özdemir Sancı , Habibe Özgüner

https://doi.org/10.17343/sdutfd.1099504

Abstract

Objective
The aim of this study was to compare four commonly
used cytotoxicity assays (XTT, neutral red uptake, crys-
tal violet assay, and propidium iodide staining) which
analyzed the antiproliferative effects of vitamin K2.
Material and Method
Saos-2 cells, an osteosarcoma cell line, were
exposed to vitamin K2 at different concentrations (10
μM, 20 μM, 30 μM, 40 μM, 50 μM, 100 μM and 200
μM) for 48 and 72 hours. Tetrazolium salt test (XTT),
neutral red uptake (NR) assay, crystal violet assay,
and propidium iodide (PI) staining were performed to
determine cytotoxic potential of vitamin K2 in terms
of the cell viability and IC50 values. The results were
evaluated with one-way analysis of variance (ANOVA)
and the Tukey test.
Results
Cytotoxic effects of vitamin K2 on osteosarcoma cells
were analyzed with XTT, neutral red, crystal violet
assay, and propidium iodide, respectively. IC50 values
were determined exposure to 61.93; 40.21; 62.11;
70.57 μM vitamin K2 for 48 and 75.44; 68.22; 41.66;
88.01 μM vitamin K2 for 72 hours.
Conclusion
Statistical analysis revealed that there is a significant
statistical difference between four tests used in this
study. In addition, it was determined that the viability
rates in propidium iodide staining were higher than
other tests for cytotoxicity analyses. It has been
concluded that incubations at different concentrations
were required to prevent misinterpretation of results in
cytotoxicity analyses, and tetrazolium salt-based tests
should be supplemented with other non-metabolic
tests.

Keywords

In vitro cytotoxicity, Cell viability tests, Osteosarcoma, Vitamin K.

Supporting Institution

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Thanks

The Saos-2 cell line was a kind gift of Assoc. Prof. Dr. Mustafa Güngörmüş Ankara Yıldırım Beyazıt University Faculty of Dentistry. This study was carried out using the facilities of Ankara Yıldırım Beyazıt University, Central Research Laboratory Application and Research Center (MERLAB).

K2 VİTAMİNİNİN OSTEOSARKOMA HÜCRELERİNDE ANTİPROLİFERATİF ETKİLERİ: FARKLI SİTOTOKSİSİTE ANALİZLERİNİN KARŞILAŞTIRILMASI

Abstract

Amaç
Bu çalışmanın amacı, K2 vitamininin antiproliferatif
etkilerini saptamak için yaygın olarak kullanılan dört
sitotoksisite analizinin (XTT, nötral kırmızı alım, kristal
viyole ve propidyum iyodür boyama) doğruluk potansiyelini
karşılaştırmaktır.
Gereç ve Yöntem
Osteosarkoma hücre hattı olan Saos-2 hücreleri, 48
ve 72 saat boyunca K2 vitaminine (10 μM, 20 μM, 30
μM, 40 μM, 50 μM, 100 μM ve 200 μM) maruz bırakıldı.
Tetrazolyum tuzu testi (XTT), nötral kırmızı alım
testi (NR), kristal viyole testi ve propidyum iyodür (PI)
boyama yapılarak hücrelerin canlılık oranlarına göre
K2 vitamini sitotoksisitesi belirlendi ve farklı dozların
IC50 değerleri hesaplandı. Analiz sonuçları tek yönlü
varyans analizi (ANOVA) ve Tukey testiyle değerlendirildi.
Bulgular
K2 vitamininin osteosarkoma hücreleri üzerinde sitotoksik
etkileri olduğu sırasıyla XTT, NR, kristal viyole
testi ve PI yöntemleri kullanılarak belirlendi ve IC50 değerleri
48 saatlik maruziyet için sırasıyla 61,93; 40,21;
62,11; 70,57 μM ve 72 saatlik maruziyet için sırasıyla
75,44; 68,22; 41,66; 88,01 μM olarak saptandı.
Sonuç
İstatistiksel analizler çalışmada uygulanan testlerden
elde edilen sonuçların farklılık gösterdiğini ortaya koymuştur.
Ayrıca propidyum iyodür boyama sonucundaki
canlılık oranlarının diğer testlerle elde edilen sonuçlara
kıyasla çarpıcı bir şekilde daha yüksek olduğu
belirlenmiştir. Sitotoksisite analizlerinde sonuçların
yanlış yorumlanmasını önlemek için farklı zamanlarda
ve farklı konsantrasyonlarda inkübasyonlar gerektiği
ve ayrıca tetrazolyum tuzu temelli testlerin metabolik
olmayan diğer testlerle desteklenmesi gerektiği sonucuna
varılmıştır.

Keywords

İn vitro sitotoksisite, Hücre canlılık testleri, Osteosarkoma, K vitamini.

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