Methods: Normal human gingival epithelial cells (HGEPp.05, Cellntec®, Bern, Switzerland) were cultured on low electron slide, Cnt-24 medium at 37 °C, 5% CO2, 95% humidity. Cells were activated by TNFα for 6 hours. Anthocyanin derivative, delphinidin-co-cyanidin (0.1μg/ml) was subsequently applied on the cells for 24 hours in comparison with fluocinolone acetonide (0.01μg/ml). Fourier-transform infrared (FTIR) microspectroscopy was used to measure spectra, which were further analyzed using principle component analysis (PCA). Statistical analysis was performed using the Unscrambler®10.1 software (CAMO, Oslo, Norway).
Results: Approximate 50 spectra of HGEPp.05 from each treatment (TNFα, fluocinolone acetonide and anthocyanins) were analyzed. These spectra include apoptotic IR spectra and their second derivatives from 1770 – 800 cm-1, which represent proteins (amide I band ~ 1650 cm-1, amide II ~ 1545 cm-1 and amide III ~ 1300 cm-1), CO stretching from ester (~1730 cm-1) and nucleic acid (asymmetric and symmetric PO2- ~1240 and 1080 cm-1). The difference in amide III and nucleic acid bands between anthocyanins treated and the other two groups were observed. In contrast, FTIR confirmed the presence of characteristic same peak for normal cells and previously treated anthocyanins cells.
Conclusions: Anthocyanins demostrate an anti-apoptotic effect on inflamed human gingival epithelial cells (HGEPp.05).
(This study was supported by grants from the Faculty of Dentistry and Research Group of Chronic Inflammatory Oral Diseases and Systemic Diseases Associated with Oral Health, Khon Kaen University, Khon Kaen, Thailand.)