Bedoni, Marzia

Donetti, Elena Bianca

Gagliano, Nicoletta

Gualerzi, Alice

Palazzo, Marco

Sciarabba, Michele

Sforza, Chiarella

Tartaglia, Gianluca

Firenze University Press: E-Journals

IJAE Vol. 115, n. 1/2 (Supplement), 2010
58
Comparative morphological analysis of chronic and 
acute cigarette smoke effects on human keratinized 
oral mucosa: ex vivo and in vitro studies
Elena Bianca Donetti1, Alice Gualerzi1, Marzia Bedoni1,2, Marco Palazzo1, Gianluca Tartaglia1, Michele 
Sciarabba1,3, Chiarella Sforza1, Nicoletta Gagliano1
1 Dipartimento di Morfologia Umana e Scienze Biomediche, Città Studi, Università degli Studi di Milano, Italy
2 Fondazione Don Carlo Gnocchi, Polo Tecnologico, Milano, Italy
3 Dipartimento di Informatica e Comunicazione, Università degli Studi di Milano, Italy
Human oral mucosa is the first target organ of cigarette smoke, a mixture of highly 
reactive compounds, well-known causes of cellular molecular alterations. Up to now, 
morphological studies about smoke effects on this organ have not been carried out. The 
aim of the present work was the morphological evaluation of intercellular junctions and 
homeostasis after acute and chronic cigarette smoke exposure. For the acute smoking ef-
fects, biopsies of keratinized human oral mucosa (KHOM) obtained from young healthy 
non-smoking women (n=7) were cultured in a Transwell system previously character-
ized in our laboratory. A comparative morphological analysis was then performed on 
oral mucosa biopsies from healthy chronic smoking women of the same age (n=5). All 
biopsies were obtained after informed consent during the extraction of the wisdom teeth. 
To simulate the acute smoke exposure, each non-smoking KHOM biopsy was divided 
in two fragments: one was exposed to direct cigarette smoke thanks to an ad-hoc tool, 
while the other represented the internal control. After histological processing and paraf-
fin embedding of all experimental groups, the expression of two desmosomal cadherins, 
desmoglein 1 (Dsg1) and desmoglein 3 (Dsg3), and of a cytoskeletal biomarker for basal 
keratinocytes, i.e. keratin 14 (K14), was evaluated through indirect immunofluorescence 
experiments. Dsg1 was distributed as physiologically expected after both chronic and 
acute smoke exposure and immunoreactivity was detected from the middle spinous 
layer up to the stratum corneum. Dsg3 immunolabelling in controls and acute smokers 
was evident in the keratinocytes of basal/lower spinous layers, gradually diminished 
in upper spinous/granular layers, and disappeared in the stratum corneum. In chronic 
smoking women, Dsg3 significantly decreased in lower/upper spinous layers, remain-
ing restricted to a limited area of the epithelium. K14 distribution in control group and 
in chronic smokers was confined to the basal keratinocyte monolayer, but, after acute 
smoke exposure, it was localized also in correspondence of the lower spinous layer. 
Taken together, these results suggest that the early “response-to-injury” to an exogenous 
stimulus such as cigarette smoke involves an alteration in the keratinocyte without any 
impairment of the epithelial barrier function, as shown by the absence of modifications 
in the molecular composition of desmosomes. Future studies are needed to correlate our 
morphological data with the proliferation rate in acute and chronic smokers.


Comparative morphological analysis of chronic and acute cigarette smoke effects on human keratinized oral mucosa: ex vivo and in vitro studies

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Comparative morphological analysis of chronic and acute cigarette smoke effects on human keratinized oral mucosa: ex vivo and in vitro studies

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