Association of oral pathology, oral microbiology, and oral oncology

dgkh000574 10.3205/dgkh000574 urn:nbn:de:0183-dgkh0005741 Research Article Association of oral pathology, oral microbiology, and oral oncology Assoziation von oraler Pathologie, oraler Mikrobiologie und oraler Onkologie Sri Lokaranjan Sri Lokaranjan Divya D

Department of Biochemistry, Tagore Medical College and Hospital, Chennai, India

author Ravi Ravi Kamalam K

Department of Biochemistry, Sree Balaji Medical College and Hospital, Chromepet, India

author Choudhary Choudhary Susmita S

Department of Orthodontics and Dentofacial Orthopaedics, Narsinhbhai Patel Dental College and Hospital, Visnagar, Gujarat, India

author Talukdar Talukdar Anindita A

Department of Pedodontics and Preventive Dentistry, Regional Dental College, Guwahati, Assam, India

author Dandekeri Dandekeri Shilpa S

Department of Prosthodontics and Crown and Bridge, Nitte, University AB Shetty Memorial Institute of Dental Sciences, Mangalore, Karnataka, India

author Shunmugavelu Shunmugavelu Karthik K

Department of Dentistry, PSP Medical College Hospital and Research Institute Tambaram Kanchipuram, main road Oragadam Panruti Kanchipuram district Tamil Nadu 631604, India. Phone: +91 9789885622/9840023697Department of Dentistry, PSP Medical College Hospital and Research Institute Tambaram Kanchipuram, Tamil Nadu, India

drkarthiks1981@gmail.com author German Medical Science GMS Publishing House

Düsseldorf

610 candida oral squamous cell carcinoma HIV Candida orales Plattenepithelkarzinom HIV 20250819 engl This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 License. Dieser Artikel ist ein Open-Access-Artikel und steht unter den Lizenzbedingungen der Creative Commons Attribution 4.0 License (Namensnennung). 2196-5226 20 GMS Hygiene and Infection Control GMS Hyg Infect Control 45 Die ökologische Gemeinschaft von Kommensalen, symbiotischen und pathogenen Organismen teilt sich unseren Körperraum. Veränderungen in der ökologischen Population der Mikroflora führen zu Dysbiose und sind entscheidende Determinanten für die Gesundheit bzw. für Krankheiten, insbesondere im Fall einer Immunsuppression. Das orale Mikrobiom und chronische Entzündungen können bei der Karzinogenese eine Rolle spielen. An ecological community of commensals, symbiotic and pathogenic o</PlainText></TextGroup>rganisms share our body space. Alterations in the ecologically balanced population of microflora result in dysbiosis and are critical determinants of systemic health and diseases, especially in the context of immunosuppression. The oral microbiome and chronic inflammation may have a role in carcinogenesis.</Pgraph></Abstract> <TextBlock name="Introduction" linked="yes"> <MainHeadline>Introduction</MainHeadline><Pgraph>The oral cavity has the second largest and second-most diverse microbiota, with over 700 species of bacteria. It contains bacteria, fungi, viruses and protozoa. The oral cavity, with its various niches, is a complex habitat, e.g., Microbes colonize the hard surfaces of the teeth as well as the soft tissues of the oral mucosa <TextLink reference="1"></TextLink>. Recent studies indicate that the oral microbiome has essential functions in maintaining oral and systemic health, and the emergence of 16S rRNA gene next-generation sequencing (NGS) has greatly contributed to revealing the complexity of its bacterial components <TextLink reference="2"></TextLink>. </Pgraph><Pgraph>Few studies have characterized aspects of the oral microbiome that may be related to oral squamous cell carcinoma (OSCC). Features of the oral microbiome associated with OSCC have been explored by comparing OSCC patients with healthy controls, or by comparing tumor sites with the surrounding normal tissue <TextLink reference="3"></TextLink>. Pushalkar et al. <TextLink reference="4"></TextLink> studied the saliva microbiome of patients with OSCC and proposed its potential use as a diagnostic tool to predict oral cancer. <Mark2>Candida (C.) auris</Mark2> has emerge<TextGroup><PlainText>d a</PlainText></TextGroup>s a multidrug-resistant ascomycete yeast. The saliva microbiome can also contain <Mark2>C. auris</Mark2>, which has emerged and is easily transmissible and highly persistent on environmental surfaces <TextLink reference="5"></TextLink>. It is associated with high mortalities, persistent candidaemia, inconsistencies in testing results, misidentification and treatment failure. This leads to complications in management and prognosis <TextLink reference="6"></TextLink>. </Pgraph><Pgraph>Thus, the aim of this article is to</Pgraph><Pgraph><UnorderedList><ListItem level="1">analyze the prevalent microbial population in healthy individuals and patients with oral squamous cell carcinoma using 16s rRNA sequencing and qPCR,</ListItem><ListItem level="1">examine <Mark2>C. auris</Mark2> in patients with immune suppression (HIV seropositive), denture wearers (diabetics and non-diabetics) and healthy individuals by using qPCR,</ListItem><ListItem level="1">examine the oral microbiome in HIV-seropositive and HIV-seronegative individuals using 16srRNA sequencing and qPCR.</ListItem></UnorderedList></Pgraph></TextBlock> <TextBlock name="Materials and methods" linked="yes"> <MainHeadline>Materials and methods</MainHeadline><Pgraph>The oral microbiome of the following groups was analysed using 16srRNA sequencing:</Pgraph><Pgraph><UnorderedList><ListItem level="1">healthy individuals (group A n=10)</ListItem><ListItem level="1">patients with OSCC (group B, n=10)</ListItem><ListItem level="1">HIV-seropositive patients (group C, n=11),</ListItem><ListItem level="1">HIV-seronegative patients denture wearers (group D, n=11).</ListItem></UnorderedList></Pgraph><Pgraph>In groups C and D, the copy number of <Mark2>C. auris</Mark2> were additionally determined.</Pgraph></TextBlock> <TextBlock name="Results" linked="yes"> <MainHeadline>Results</MainHeadline><SubHeadline>Distribution of overall microbial phyla across all groups (A–D)</SubHeadline><Pgraph>Proteobacteria was identified as the predominant phylum across all sample groups, contributing up to 39% of the microbial population. This was followed by Firmicutes (22%), Actinobacteria (15%), and Bacteroidetes (12%).</Pgraph><SubHeadline2>Microbial composition in different groups</SubHeadline2><Pgraph><Mark1>Group A (OSCC patients):</Mark1> The microbial diversity was great, with the genera Bacillus, Buchnera, Caulobacter, Clostridium, Corynebacterium, Desulfotomaculatum, Enterococcus, Flavobacterium, Gemmata, Hymenobacter, Lactobacillus, Listeria, Lysinibacillus, Marinifilum, Ruminococcus, Streptococcus, Streptomyces, and Thermoanaerobacter represented.</Pgraph><Pgraph><Mark1>Group B (healthy individuals):</Mark1> These individuals showed the presence of Bacillus, Enterococcus, Lactobacillus, Massilia, Paenibacillus, and Streptococcus.</Pgraph><Pgraph><Mark1>Group C (HIV-seronegative individuals):</Mark1> A total of 102 species were observed, including Legionella.</Pgraph><Pgraph><Mark1>Group D (HIV-seropositive individuals)</Mark1>: A total of 30 species were detected, including Neisseria.</Pgraph><SubHeadline2>Microbial overlaps and taxa</SubHeadline2><Pgraph>The following genera were common to both OSCC and healthy individuals: Bacillus, Enterococcus, Lactobacillus, and Streptococcus.</Pgraph><Pgraph>Aphanizomenon, Betaproteobacterium, and Methylococcus were found in both HIV-seropositive and seronegative individuals.</Pgraph><SubHeadline>Functional classification of microbial groups</SubHeadline><Pgraph><Mark1>Saccharolytic bacteria: </Mark1>Bacillus, Buchnera, Clostridium, Corynebacterium, Desulfotomaculatum, Enterococcus, Flavobacterium, Gemmata, Hymenobacter, Lactobacillus, Listeria, Ruminococcus, Streptococcus, Streptomyces, and Thermoanaerobacter.</Pgraph><Pgraph><Mark1>Aciduric bacteria:</Mark1> Bacillus, Caulobacter, Clostridium, Corynebacterium, Desulfotomaculatum, Enterococcus, Lactobacillus, Listeria, Lysinibacillus, Ruminococcus, and Streptococcus.</Pgraph><Pgraph><Mark1>Aerobic bacteria:</Mark1> Buchnera, Caulobacter, Clostridium, Corynebacterium, Gemmata, Hymenobacter, Lysinibacillus, and Streptomyces.</Pgraph><Pgraph><Mark1>Anaerobic bacteria:</Mark1> Bacillus, Desulfotomaculatum, Enterococcus, Flavobacterium, Lactobacillus, Listeria, Marinifilum, Ruminococcus, and Streptococcus.</Pgraph><SubHeadline>Site-specific microbial distribution in OSCC patients (group A)</SubHeadline><Pgraph>Streptomyces was observed in both alveolus (20%) and tongue (20%). Bacillus and Listeria were exclusively present in alveolar lesions (30%). Streptococcus was the predominant bacterium across all OSCC sites:</Pgraph><Pgraph><UnorderedList><ListItem level="1">Tongue: 10%</ListItem><ListItem level="1">Buccal mucosa: 20%</ListItem><ListItem level="1">Alveolus: 10%</ListItem><ListItem level="1">Palate: 20%</ListItem></UnorderedList></Pgraph><SubHeadline>Distribution of phyla in the control group (group B)</SubHeadline><Pgraph>Proteobacteria was the most prevalent phylum, accounting for 35% of the microbial community in healthy individuals.</Pgraph><SubHeadline>Oral microbiome in HIV-positive and HIV-negative individuals</SubHeadline><Pgraph><TextGroup><Mark1>HIV-seronegative individuals (group C):</Mark1><PlainText> A total of 102 m</PlainText></TextGroup>icrobial species were detected, including Legionella.</Pgraph><Pgraph><Mark1>HIV-seropositive individuals (group D):</Mark1> A total of 30 microbial species were identified, including Neisseria.</Pgraph><Pgraph><Mark1>Common microbes in both HIV-seropositive and -seronegative individuals:</Mark1> Aphanizomenon, Betaproteobacterium, and Methylococcus.</Pgraph><SubHeadline>Candida (C.) auris copy numbers in different groups</SubHeadline><Pgraph>The quantification of <Mark2>C. auris </Mark2>showed significant variation across different study groups:</Pgraph><Pgraph><UnorderedList><ListItem level="1">Denture wearers: Highest average copy number of 548,401.1.</ListItem><ListItem level="1">HIV-seropositive patients: Copy number of 474,966.4.</ListItem><ListItem level="1">Healthy controls: Four out of ten control samples showed no detectable <Mark2>C. auris</Mark2>, while the remaining samples had a low average copy number of 9,792.71.</ListItem></UnorderedList></Pgraph><SubHeadline>Anaerobic vs. facultative anaerobic microbial distribution</SubHeadline><Pgraph><Mark1>In OSCC patients (group A):</Mark1> Obligate anaerobes comprised 22% of the microbiota.</Pgraph><Pgraph><Mark1>In healthy individuals (group B):</Mark1> Only facultative anaerobes were present.</Pgraph></TextBlock> <TextBlock name="Discussion" linked="yes"> <MainHeadline>Discussion</MainHeadline><SubHeadline>Method</SubHeadline><Pgraph>16S rRNA sequencing is a very powerful tool for comparative microbiome analysis. The BLAST Basic Local Alignment Search Tool results derived using 16S rRNA, gene DNA sequences was used to identify the evolutionary relationship by a phylogenetic tree. The limitation was that it identifies only the Shine-Dalgarno domain, a domain common in Bacteri and Archaea with an overlap with mitochondrial and chloroplast RNA.</Pgraph><SubHeadline>Results</SubHeadline><Pgraph>The oral microbiome is complex. Our study showed that there were differences between the microbiome of OSCC subjects and healthy individuals. The data from this study will help us to identify the species which need to be studied further to ascertain whether they play a part in oral carcinogenesis. </Pgraph></TextBlock> <TextBlock name="Conclusion" linked="yes"> <MainHeadline>Conclusion</MainHeadline><Pgraph>Proteobacteria was the most prevalent phylum across all groups, followed by Firmicutes, Actinobacteria, and Bacteroidetes. OSCC patients exhibited a unique microbial profile distinct from healthy individuals, with specific genera present only in OSCC cases. Microbial diversity was significantly lower in HIV-seropositive individuals compared to HIV-seronegative individuals. <Mark2>C. auris</Mark2> had the highest prevalence in denture wearers, followed by HIV patients, was minimal in healthy controls, and absent in OSCC patients. Additionally, obligate anaerobes were more dominant in OSCC patients, whereas only facultative anaerobes were found in healthy individuals. These findings contribute additional data on microbial distribution across different health conditions and provide initial clues as to the role of microbial composition in OSCC and HIV as it affects the oral cavity.</Pgraph></TextBlock> <TextBlock name="Notes" linked="yes"> <MainHeadline>Notes</MainHeadline><SubHeadline>Authors’ ORCIDs </SubHeadline><Pgraph><UnorderedList><ListItem level="1">Sri Lokaranjan D:<Hyperlink href="https://orcid.org/0009-0006-1758-8201">https://orcid.org/0009-0006-1758-8201</Hyperlink></ListItem><ListItem level="1">Ravi K: <Hyperlink href="https://orcid.org/0000-0002-9625-3058">https://orcid.org/0000-0002-9625-3058</Hyperlink></ListItem><ListItem level="1">Dandekeri S: <Hyperlink href="https://orcid.org/0000-0001-7814-6067">https://orcid.org/0000-0001-7814-6067</Hyperlink></ListItem><ListItem level="1">Shunmugavelu K: <Hyperlink href="https://orcid.org/0000-0001-7562-8802">https://orcid.org/0000-0001-7562-8802</Hyperlink></ListItem></UnorderedList></Pgraph><SubHeadline>Ethical approval </SubHeadline><Pgraph>Ethical approval obtained</Pgraph><SubHeadline>Funding</SubHeadline><Pgraph>None. </Pgraph><SubHeadline>Competing interests</SubHeadline><Pgraph>The authors declare that they have no competing interests.</Pgraph></TextBlock> <References linked="yes"> <Reference refNo="1"> <RefAuthor>Deo PN</RefAuthor> <RefAuthor>Deshmukh R</RefAuthor> <RefTitle>Oral microbiome: Unveiling the fundamentals</RefTitle> <RefYear>2019</RefYear> <RefJournal>J Oral Maxillofac Pathol</RefJournal> <RefPage>122-8</RefPage> <RefTotal>Deo PN, Deshmukh R. 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