ULOGA HUMANIH PAPILOMA VIRUSA (HPV) U RAZVOJU DOBROĆUDNIH I ZLOĆUDNIH PROMJENA SLUZNICE USNE ŠUPLJINE

Objectives: The aim of this study was to determine the prevalence and type of human papillomavirus (HPV) in various oral lesions in comparison to control healthy oral mucosa, and the dynamic of HPV infection course (persistence, change of HPV type and clearance). Material and Methods: The study comprised 426 consented subjects, of which 343 patients with different oral lesions and 83 controls with apparently healthy oral mucosa, referred for oral examination in the period from 2000 to 2015. The diagnoses of oral lesions were established according to clinical criteria and confirmed by histopathology. Oral cytobrush samples were collected from oral lesions and healthy mucosa from different topographic sites and the HPV types were determined by polymerase chain reaction (PCR). The presence of HPV DNA was evaluated by consensus and type-specific (TS) primer-directed PCR. A subset of samples was analysed for the presence of both alpha and beta genus HPV types. Results: Out of 426 specimens, 144 (41.98%) from oral lesions and 15 (18.07%) controls were positive for HPV DNA; 22.3% of all tested samples contained beta-HPV type and 15.02% alpha-PV type. The highest prevalence of beta-HPV and high risk (HR) HPV (alpha-HPV) types were found in benign and premalignant oral lesions, while HR-HPVs were found in a small number of control samples. In 38 subjects sampling was performed more than twice, in average 2.29 times, with time distance between sampling of 5.8 months. Twenty-six of these 38 patients were continuously HPV negative, five had transient infection, five acquired a new HPV infection, one had persistent HPV infection, and one had initially co-infection with different HPV types followed with new one during which initial oral lesion became malignant. The topographic distribution of the alpha-PV and beta-PV types showed affinity for the anterior part of the oral cavity and particular risk localization. Conclusion: Positive finding of HPV in all age groups and on healthy oral mucosa, high prevalence of HPV types in oral premalignant and proliferative lesions, affinity for risk localization in the oral cavity, unpredictable dynamic course and change of HPV type indicates the need for regular oral assessment and HPV control in patients with initially positive findings on oral mucosa.Cilj istraživanja bio je ispitati prevalenciju i tip humanog papiloma virusa (HPV) u različitim oralnim lezijama u usporedbi s kontrolnom, naizgled zdravom sluznicom i dinamiku tijeka HPV infekcije u ustima (ustrajnost, promjenu tipa HPV i prolaznost infekcije). Ispitanici i postupci: U ispitivanju je sudjelovalo 426 ispitanika, od kojih 343 bolesnika s različitim oralnim lezijama i 83 kontrolnih s naizgled zdravom sluznicom koji su upućeni na pregled usne šupljine u razdoblju od 2000. do 2015.godine. Svi ispitanici dali su svoj informirani pristanak prije uključivanja u ovo istraživanje. Dijagnoze oralnih lezija postavljene su na temelju kliničkih kriterija i potvrđene histopatološkom analizom. Citološki obrisci uzeti su citološkom četkicom s oralnih lezija i zdrave sluznice s topografski različitih mjesta sluznice, a HPV je dokazan lančanom reakcijom polimerazom (PCR). Prisutnost HPV DNA dokazana je PCR-om sa zajedničkim i tip-specifičnim početnicama. Određeni dio uzoraka bio je testiran na tipove HPV-a iz roda alfa i beta. Rezultati: U svih 426 testiranih uzoraka, HPV DNA dokazana je u 144 (41,98%) uzoraka s oralnih lezija i 15 (18,07%) kontrolnih; 22,3% ispitanih uzoraka sadržavalo je tip HPV iz roda beta, a 15,02% tip HPV iz roda alfa. Najveća prevalencija beta-HPV i visokorizičnih (HR) HPV tipova (alfa-HPV) pronađeni su u benignim i premalignim oralnim lezijama, a visokorizični HPV tipovi pronađeni su i u malom broju kontrolnih uzoraka. U 38 ispitanika uzorkovanje je provedeno više od dva puta, u prosjeku 2.29 puta, u vremenskom razmaku od prosječno 5,8 mjeseci. Dvadesetšest od 38 ispitanika bilo je stalno negativno na HPV, u pet je dokazana prolazna infekcija, pet je steklo novu infekciju HPV-om, jedan ispitanik imao je stalnu infekciju HPV-om, a jedan je tijekom praćenja početno dokazanu koinfekciju s različitim tipovima HPV-a i stekao novu s HPV tipom visokog rizika tijekom koje je inicijalna premaligna lezija sluznice postala zloćudna. Analiza topografske distribucije alfa-HPV i beta-HPV tipova na oralnoj sluznici pokazala je afinitet virusa za sluznicu prednjeg dijela usne šupljine i osobito za rizične lokalizacije. Zaključak: Pozitivan nalaz HPV-a u svim dobnim skupinama i na zdravoj sluznici te visoka prevalencija HPV tipova u oralnim premalignim i proliferativnim lezijama kao i afinitet za rizične lokalizacije te nepredvidljiv dinamičan tijek uz promjene tipa HPV-a upućuju na potrebu kontrole na HPV u kliničkoj stomatološkoj praksi kod inicijalno pozitivnog nalaza

ULOGA HUMANIH PAPILOMA VIRUSA (HPV) U RAZVOJU DOBROĆUDNIH I ZLOĆUDNIH PROMJENA SLUZNICE USNE ŠUPLJINE

Objectives: The aim of this study was to determine the prevalence and type of human papillomavirus (HPV) in various oral lesions in comparison to control healthy oral mucosa, and the dynamic of HPV infection course (persistence, change of HPV type and clearance). Material and Methods: The study comprised 426 consented subjects, of which 343 patients with different oral lesions and 83 controls with apparently healthy oral mucosa, referred for oral examination in the period from 2000 to 2015. The diagnoses of oral lesions were established according to clinical criteria and confirmed by histopathology. Oral cytobrush samples were collected from oral lesions and healthy mucosa from different topographic sites and the HPV types were determined by polymerase chain reaction (PCR). The presence of HPV DNA was evaluated by consensus and type-specific (TS) primer-directed PCR. A subset of samples was analysed for the presence of both alpha and beta genus HPV types. Results: Out of 426 specimens, 144 (41.98%) from oral lesions and 15 (18.07%) controls were positive for HPV DNA; 22.3% of all tested samples contained beta-HPV type and 15.02% alpha-PV type. The highest prevalence of beta-HPV and high risk (HR) HPV (alpha-HPV) types were found in benign and premalignant oral lesions, while HR-HPVs were found in a small number of control samples. In 38 subjects sampling was performed more than twice, in average 2.29 times, with time distance between sampling of 5.8 months. Twenty-six of these 38 patients were continuously HPV negative, five had transient infection, five acquired a new HPV infection, one had persistent HPV infection, and one had initially co-infection with different HPV types followed with new one during which initial oral lesion became malignant. The topographic distribution of the alpha-PV and beta-PV types showed affinity for the anterior part of the oral cavity and particular risk localization. Conclusion: Positive finding of HPV in all age groups and on healthy oral mucosa, high prevalence of HPV types in oral premalignant and proliferative lesions, affinity for risk localization in the oral cavity, unpredictable dynamic course and change of HPV type indicates the need for regular oral assessment and HPV control in patients with initially positive findings on oral mucosa.Cilj istraživanja bio je ispitati prevalenciju i tip humanog papiloma virusa (HPV) u različitim oralnim lezijama u usporedbi s kontrolnom, naizgled zdravom sluznicom i dinamiku tijeka HPV infekcije u ustima (ustrajnost, promjenu tipa HPV i prolaznost infekcije). Ispitanici i postupci: U ispitivanju je sudjelovalo 426 ispitanika, od kojih 343 bolesnika s različitim oralnim lezijama i 83 kontrolnih s naizgled zdravom sluznicom koji su upućeni na pregled usne šupljine u razdoblju od 2000. do 2015.godine. Svi ispitanici dali su svoj informirani pristanak prije uključivanja u ovo istraživanje. Dijagnoze oralnih lezija postavljene su na temelju kliničkih kriterija i potvrđene histopatološkom analizom. Citološki obrisci uzeti su citološkom četkicom s oralnih lezija i zdrave sluznice s topografski različitih mjesta sluznice, a HPV je dokazan lančanom reakcijom polimerazom (PCR). Prisutnost HPV DNA dokazana je PCR-om sa zajedničkim i tip-specifičnim početnicama. Određeni dio uzoraka bio je testiran na tipove HPV-a iz roda alfa i beta. Rezultati: U svih 426 testiranih uzoraka, HPV DNA dokazana je u 144 (41,98%) uzoraka s oralnih lezija i 15 (18,07%) kontrolnih; 22,3% ispitanih uzoraka sadržavalo je tip HPV iz roda beta, a 15,02% tip HPV iz roda alfa. Najveća prevalencija beta-HPV i visokorizičnih (HR) HPV tipova (alfa-HPV) pronađeni su u benignim i premalignim oralnim lezijama, a visokorizični HPV tipovi pronađeni su i u malom broju kontrolnih uzoraka. U 38 ispitanika uzorkovanje je provedeno više od dva puta, u prosjeku 2.29 puta, u vremenskom razmaku od prosječno 5,8 mjeseci. Dvadesetšest od 38 ispitanika bilo je stalno negativno na HPV, u pet je dokazana prolazna infekcija, pet je steklo novu infekciju HPV-om, jedan ispitanik imao je stalnu infekciju HPV-om, a jedan je tijekom praćenja početno dokazanu koinfekciju s različitim tipovima HPV-a i stekao novu s HPV tipom visokog rizika tijekom koje je inicijalna premaligna lezija sluznice postala zloćudna. Analiza topografske distribucije alfa-HPV i beta-HPV tipova na oralnoj sluznici pokazala je afinitet virusa za sluznicu prednjeg dijela usne šupljine i osobito za rizične lokalizacije. Zaključak: Pozitivan nalaz HPV-a u svim dobnim skupinama i na zdravoj sluznici te visoka prevalencija HPV tipova u oralnim premalignim i proliferativnim lezijama kao i afinitet za rizične lokalizacije te nepredvidljiv dinamičan tijek uz promjene tipa HPV-a upućuju na potrebu kontrole na HPV u kliničkoj stomatološkoj praksi kod inicijalno pozitivnog nalaza

Human Papillomavirus in the Lesions of the Oral Mucosa According to Topography

Background. The association between human papillomavirus (HPV) types and oral lesions has been shown in many studies. In light of the possibility of early detection of HPV genotypes in oral epithelium, and considering the significance which HPV has in the development of malignant and potentially malignant disorders of oral mucosa, the purpose of this study was to investigate the prevalence of HPV DNA in different oral lesions in the Croatian population. In addition, we wanted to elucidate whether HPV infection is associated predominantly with either the lesion or particular anatomic site of the oral cavity. Methodology/Principal Findings. The study included 246 subjects with different oral lesions, and 73 subjects with apparently healthy oral mucosa (controls). Oral lesions were classified according to their surface morphology and clinical diagnosis. Epithelial cells were collected with the cytobrush from different topographic sites in the oral cavity of oral lesions and controls. The presence of HPV DNA was evaluated by consensus and type-specific primer-directed polymerase chain reaction. The HPV positivity was detected in 17.7% of oral lesions, significantly more than in apparently healthy mucosa (6.8%), with a higher presence in benign proliferative mucosal lesions (18.6%). High-risk HPV types were predominantly found in potentially malignant oral disorders (HPV16 in 4.3% and HPV31 in 3.4%), while benign proliferative lesions as well as healthy oral mucosa contained mainly undetermined HPV type (13.6 and 6.8%, respectively). Conclusions/Significance. Distribution of positive HPV findings on oral mucosa seems to be more associated to particular anatomical site than diagnosis itself. Samples taken from vermilion border, labial commissures, and hard palate were most often HPV positive. Thus, topography plays a role in HPV prevalence findings in oral lesions. Because of the higher prevalence of the high-risk HPV types in potentially malignant oral disorders, these lesions need to be continuously controlled and treated

HPV genotypes distribution according to oral lesion morphology and diagnosis irrespective of topography.

a<p>HPV types 18 and 45 were not determined in any case.</p>b<p>three cases with HPV 6/11 and HPV 16, and one case with HPV 6/11 and HPV 31.</p>c<p>HPV X.</p

The frequency of HPV positivity in different combinations of clinical diagnosis and topography of lesions.

a<p>Data is presented as number of HPV positive samples/total number of samples at the intersection of row and column (percentage within subgroup), empty cells contain no samples i.e. 0/0(0);</p>b<p>There were samples from multiple locations and/or diagnosis and such samples were counted in each respective row or column. However, samples described with multiple topographical codes of the same region (i.e. tongue codes are 39 to 45) were counted only once within that subgroup. Thus the number of diagnosis-location pairs is greater than the number of individual samples in the study.</p

Distribution of oral lesion by morphology and diagnosis according to gender and HPV positivity irrespective of topography.

<p>Distribution of oral lesion by morphology and diagnosis according to gender and HPV positivity irrespective of topography.</p

Frequency of HPV detection at different regions of the oral mucosa.

<p>The topography of the oral mucosa is coded according to the modified WHO oral topography codes by Roed-Petersen and Roenstrup <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0069736#pone.0069736-Kramer1" target="_blank">[12]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0069736#pone.0069736-Mattila1" target="_blank">[22]</a>: vermilion border - upper (13), lower (14), labial commissures - right (15), left (16), labial mucosa - upper (17), lower (18), labial sulci - upper (21), lower (22), cheek (bucal mucosa) - right (19), left (20), buccal sulcus - right upper (23) lower (24), buccal sulcus – left upper (25) lower (26), upper gingiva or edentulous alveolar ridge buccally - right (27), left (28), lower gingiva or edentulous alveolar ridge - right (29), left (30), upper anterior gingiva and edentulous ridge labially (31), lower anterior gingiva or edentulous ridge labially (32), upper posterior gingiva or edentulous alveolar ridge palatally - right (33), left (34), lower posterior gingiva or edentulous alveolar ridge lingually - right (35), left (36), anterior gingiva or edentulous ridge palatally (37), and lingually (38), dorsum of the tongue - right (39), left (40), base of the tongue - right (41), left (42), tip of the tongue (43), margine of the tongue - right (44), left (45), surface of the tongue - right (46), left (47), frontal floor of mouth (48), lateral floor of mouth - right (49), left (50), hard palate - right (51), left (52), soft palate - right (53), left (54), anterior tonsillar pillar - right (55), left (56); color scale indicates frequency of HPV positivity in the respective locations; for regions with less than 3 samples analysed the frequency was not calculated; symetrical regions were counted as one for the analysis.</p