Adverse effects of Sudanese toombak vs. Swedish snuff on human oral cells.
BACKGROUND
The high incidence of oral cancer in Sudan has been associated with the use of toombak, the local type of smokeless tobacco. However, its specific effects on human oral cells are not known. We aimed to investigate the effects of toombak on primary normal human oral keratinocytes, fibroblasts, and a dysplastic oral keratinocytic cell line, and to compare them with the effects induced by Swedish snuff.


METHODS
Aqueous extracts were prepared from moist toombak and Swedish snuff and added in serial dilutions on in vitro monolayer cultured cells. Cell viability, morphology and growth, DNA double-strand breaks (gammaH2AX staining), expression of phosphatidylserine (Annexin V staining), and cell cycle were assessed after various exposure time periods.


RESULTS
Significant decrease in cell number, occurrence of DNA double-strain breaks, morphological and biochemical signs of programmed cell death were detected in all oral cell types exposed to clinically relevant dilutions of toombak extract, although to a lesser extent in normal oral fibroblasts and dysplastic keratinocytes. G2/M-block was also detected in normal oral keratinocytes and fibroblasts exposed to clinically relevant dilutions of toombak extract. Swedish snuff extract had less adverse effects on oral cells, mainly at non-clinically relevant dilutions.


CONCLUSIONS
This study indicates a potential for toombak, higher than for Swedish snuff, to damage human oral epithelium. Dysplastic oral keratinocytes were less sensitive than their normal counterparts, suggesting that they might have acquired a partially resistant phenotype to toombak-induced cytotoxic effects while still being prone to DNA damage that could lead to further malignant progression.
Costea
Daniela Elena
DE
Section of Pathology, The Gade Institute, University of Bergen, Bergen, Norway. daniela.costea@gades.uib.no
Lukandu
Ochiba
O
Bui
Linh
L
Ibrahim
Muna Jaffar M
MJ
Lygre
Raymond
R
Neppelberg
Evelyn
E
Ibrahim
Salah Osman
SO
Vintermyr
Olav Karsten
OK
Johannessen
Anne Christine
AC
D000328:Adult; D017304:Annexin A5 / Q000032:analysis; D017209:Apoptosis / Q000187:drug effects; D002452:Cell Count; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004396:Coloring Agents; D053903:DNA Breaks, Double-Stranded; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D016195:G2 Phase / Q000187:drug effects; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects; D046529:Microscopy, Electron, Transmission; D009061:Mouth Mucosa / Q000166:cytology* / Q000187:drug effects; D010718:Phosphatidylserines / Q000032:analysis; D010936:Plant Extracts / Q000009:adverse effects; D013397:Sudan; D013548:Sweden; D014030:Tobacco, Smokeless / Q000009:adverse effects*
D017304:Annexin A5; D004396:Coloring Agents; D010718:Phosphatidylserines; D010936:Plant Extracts