|Year : 2021 | Volume
| Issue : 6 | Page : 1050-1054
Investigation of candidal species among people who suffer from oral potentially malignant disorders and oral squamous cell carcinoma
Mohammed Abidullah1, Sunitha Bhosle2, Bruhathi Komire3, Priyadarshini Sharma4, K Swathi5, L Karthik6
1 Department of Dental and Biomedical Sciences, Faculty of Dentistry, Al Baha University, Al Bahah, Saudi Arabia
2 Department of Oral Medicine and Radiology, Ame's Dental College and Hospital, Raichur, Karnataka, India
3 Department of Oral Medicine and Radiology, Malla Reddy Institute of Dental Sciences, Hyderabad, Telangana, India
4 Department of Oral Medicine and Radiology, New Janapriya Super Speciality Hospital, Bengaluru, Karnataka, India
5 Department of Public Health Dentistry, Malla Reddy Institute of Dental Sciences, Hyderabad, Telangana, India
6 Department of Conservative Dentistry and Endodontics, Meghna Institute of Dental Sciences, Nizamabad, Telangana, India
|Date of Submission||30-Apr-2021|
|Date of Decision||10-Jun-2021|
|Date of Acceptance||13-Jun-2021|
|Date of Web Publication||10-Nov-2021|
Department of Dental and Biomedical Sciences, Faculty of Dentistry, Al Baha University, Al Bahah
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Aim and Objective: The aim of the present study was to evaluate the candidal species among masses with oral potentially malignant disorders and oral squamous cell carcinoma. Materials and Methods: This prospective and observational study was conducted by the Department of Oral Pathology and Microbiology, S. B. Patil Dental College, Bidar, Karnataka, India, from February 2018 to January 2019. The study composed of total of 150 individuals, of which 50 individuals did not had any visible manifestations, 50 were analyzed with potentially malignant disorders (PMDs) in particular oral leukoplakia, oral lichen planus, and oral submucous fibrosis and last group of 50 individuals were suffering from oral squamous cell carcinoma (OSCC). First, the swab samples were elicited from culture technique after that incisional biopsy of the discernible investigated lesion was done for the purpose of justopathological verification. The swab samples were streak on Sabouraud dextrose agar (SDA) and HiCrome Candida Differential HiVeg agar/CHROMagar medium and incubation at 37°C for 24–48 h. Biopsy was done for all the samples. Results: The proportion of candidates as men and women in control was 45 (90%) and 5 (10%), in PMD was 30 (60%) and 20 (40%), and in OSCC was 45 (90%) and 5 (10%), correspondingly. On evaluation on SDA medium in controls, PMD and OSCC groups, Candida was founded in 14 (28%), 20 (40%), and 42 (84%) and not founded in 36 (72%), 30 (60%), and 8 (16%) folks, subsequently. Intragroup contrast illustrated exceedingly necessary distinction with P = 0.000 between both controls versus OSCC and PMD in comparison to OSCC. Nevertheless, controls versus PMD manifested insignificant, P = 0.119. Investigation on CHROM AGAR media among controls, PMD and OSCC groups, Candida species was seen in 11 (22%), 19 (38%), and 40 (80%) and absent in 39 (78%), 31 (62%), and 10 (20%) individuals, respectively. On statistical inspection, the variations noted were enormous, (P = 0.000). On speciation of Candida in CHROM agar among the controls, PMD and OSCC groups, Candida albicans species was present in 9 (18%), 16 (32%), and 6 (12%), Candida krusei in 3 (6%), 6 (12%), and 13 (26%), Candida glabrata in 0, 0, and 8 (16%), and Candida tropicalis in 0, 0, and 3 (6%) cases, respectively. Nonetheless, only OSCC group reveal amalgamation of species such as C. glabrata and C. krusei was present in 2 (4%) case, C. tropicalis and C. krusei in was present 3 (6%) cases, C. tropicalis and C. glabrata was present in 2 (4%) case, C. albicans and C. tropicalis was present in 2 (4%) cases, and C. krusei, C. glabrata with C. albicans was present in 1 (2%) case, respectively. All other types of fungi were regarded as infectious excluding Candida, on analysis on SDA medium, infestation in the form of fungal molds was seen in 18 (36%) in controls, 12 (24%) in PMD and 8 (16%) in OSCC groups. Conclusion: We interpreted that the chief carrier of candidal species in PMDs and OSCC, yet more light is to be thrown on the topic that Candida has particular establishment in PMDs or in malignancy.
Keywords: Candida albicans, carcinogenesis, CHROM agar, oral squamous cell carcinoma, potentially malignant disorders, Sabouraud dextrose agar
|How to cite this article:|
Abidullah M, Bhosle S, Komire B, Sharma P, Swathi K, Karthik L. Investigation of candidal species among people who suffer from oral potentially malignant disorders and oral squamous cell carcinoma. J Pharm Bioall Sci 2021;13, Suppl S2:1050-4
|How to cite this URL:|
Abidullah M, Bhosle S, Komire B, Sharma P, Swathi K, Karthik L. Investigation of candidal species among people who suffer from oral potentially malignant disorders and oral squamous cell carcinoma. J Pharm Bioall Sci [serial online] 2021 [cited 2022 Jun 25];13, Suppl S2:1050-4. Available from: https://www.jpbsonline.org/text.asp?2021/13/6/1050/330106
| Introduction|| |
Clinically, apparent oral potentially malignant conditions are the most common precursors of oral squamous cell carcinoma (OSCC) potentially malignant disorder., According to some findings, Candida spp. isolated from leukoplakia lesions can contain the carcinogen N-nitrosobenzylmethylamine. and due to its ability to nitrosylate N-benzyl methylamine, candida species have been linked to the etiopathogenesis of OSCC and oral potentially malignant disorders.,, There is a connection between epithelial dysplasia and histologically determined fungal infection of the oral mucosa. It is suggested that Candida albicans tubular hyphal structure is needed because it allows for the entrance of precursors from saliva and the release of the nitrosamine agent into keratinocytes, possibly causing OSCC.,, The ability of specific C. albicans strains to promote neoplastic transformation and the development of carcinogenic nitrosamines from saliva has revealed Candida's crucial role in malignant transformation., As compared to noncandidal leukoplakia, candidal leukoplakia is distinguished histologically by persistent (occasionally acute) intraepithelial inflammation with fungal hyphae entering the superficial layers of epithelium and has a significantly higher rate of malignant transformation.,
| Materials and Methods|| |
This prospective and observational study was done in the Unit Department of Oral Pathology and Microbiology, S. B. Patil Dental College, Bidar, Karnataka, India, from February 2018 to January 2019 after taking the approval of the protocol review committee and institutional ethics committee.
Patients with PMDs and OSCC were also included in the report. Patients on oral or systemic corticosteroid medication, long-term broad-spectrum antibiotic therapy, a history of immune compromised illness, and those who had not received chemotherapy, radiotherapy, or antifungal medication were all excluded from the study. In all, 150 participants were studied, 50 of whom were healthy controls with no visible lesions, 50 of whom had PMDs such as oral leukoplakia, oral submucous fibrosis (OSMF), and oral lichen planus, and 50 of whom were OSCC patients. On a custom case history type, all of the study participants' socioeconomic statistics were registered. Using medical instruments, a thorough oral examination of all applicants is conducted. To begin, swab samples were obtained using the culture technique, and then, an incisional biopsy of the discernible examined lesion was performed for just pathological confirmation. Patients were often instructed to rinse their mouths with purified water to remove food particles, followed by salt water for microbiological culture. To collect samples from the buccal mucosa in disease-free controls and the lesion region in PMD and OS cases, sterile cotton swabs with a wooden stick placed on a blue capped high-density polyethylene tube sized 150 mm 12 mm were used. The swab samples were put back in the same polyethylene tube and held at 37°C until they were streaked on Sabouraud dextrose agar (SDA) medium and incubated for 24–48 h at 37°C. A sterile instrument, such as a cotton swab or an inoculation thread, was used to finish the streaking. To keep microbiological colonies alive and avoid contamination of the growth medium, aseptic techniques were used. After that, the creamy/white pasty smooth colonies were inoculated again on HiCrome Candida Differential HiVeg agar/CHROMagar and incubated at 37°C for 24–48 h. According to HiMedia Laboratories' orders, microbial-colored colonies representing different species of Candida organisms were interpreted for color and unique colony characteristics. After biopsy, the tissue sample was fixed in neutral-buffered formalin, then processed and embedded in paraffin wax as normal. The 4–5 mm thick sections were primed and stained with hematoxylin and eosin for histopathological confirmation using a binocular analysis microscope (Motic BA400) with a 5 MP camera for photomicrography (Moticam 2500, USB2.0).
The whole set of statistics was then moved to a Microsoft Excel sheet. The Chi-square test was performed using the Statistical Package for the Social Sciences (SPSS edition 19.0, Chicago, IL, USA). P ≤ 0.05 was considered statistically significant and was often used in statistical studies.
| Results|| |
There was age range for assortment of the participants in controls, PMD and OSCC of 15–58, 20–60 and 25–70 years, subsequently, and the mean age ± standard deviation was 30.92 ± 13.12, 43.1 ± 5.26 and 50.12 ± 12.96 years, correspondingly. The dispersal of the candidates between men and women in controls was 45 (90%) and 5 (10%), in PMD was 30 (60%) and 20 (40%) and in OSCC was 45 (90%) and 5 (10%), correspondingly. On evaluation on SDA medium in controls, PMD and OSCC groups, Candida was found in 14 (28%), 20 (40%) and 42 (84%) and not seen in 36 (72%), 30 (60%) and 8 (16%) people, respectively. On statistical investigations of intergroup comparison, huge difference (P = 0.000) was noted [Table 1]. As well as in intragroup comparison, there was colossal distinction with (P = 0.000) among both controls versus OSCC and PMD versus OSCC. However, controls versus PMD showed a nonsignificant, (P = 0.119) [Table 2]. On evaluation on CHROM agar media among controls, PMD and OSCC groups, Candida species was present in 11 (22%), 19 (38%), and 40 (80%) and absent in 39 (78%), 31 (62%) and 10 (20%) individuals distributively. On statistical analysis, an extremely significant difference was noted (P = 0.000) [Table 3]. On speciation of Candida in CHROM agar among the controls, PMD and OSCC groups, C. albicans species was present in 9 (18%), 16 (32%), and 6 (12%), Candida krusei in 3 (6%), 6 (12%), and 13 (26%), Candida glabrata in 0, 0, and 8 (16%) and Candida tropicalis in 0, 0, and 3 (6%) cases, respectively. Nonetheless, OSCC group solely showed union of species such as C. glabrata and C. krusei was present in 2 (4%) case, C. tropicalis and C. krusei in was present 3 (6%) cases, C. tropicalis and C. glabrata was present in 2 (4%) case, C. albicans and C. tropicalis was present in 2 (4%) cases, and C. krusei, C. glabrata with C. albicans was present in 1 (2%) case, respectively [Table 4]. All the types of the fungi excluding Candida were regarded as infection. On evaluation on SDA medium, contamination in the form of fungal molds was present in 18 (36%) in control, 12 (24%) in PMD, and 8 (16%) in OSCC groups [Table 5].
|Table 1: Estimation of Candida in Sabouraud dextrose agar media among the study batches|
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|Table 2: Intergroup contrast of Candida in Sabouraud dextrose agar media|
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|Table 4: Recognition of different candidal species in chrom agar in the study groups|
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|Table 5: Infestation in control, potentially malignant disorders, and oral squamous cell carcinoma groups|
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| Discussion|| |
In oral Candida-based experiments, one of the key investment strategies is to approximate the occurrence of Candida and the species specificity for certain societal classes.,, OSCC has been shown to grown from PMDs such as leukoplakia, OSMF, and lichen planus in many ways., Any researchers believe that candidal inflammation increases the risk of premalignant lesions transforming into cancer. 22 Yeasts are usually found commensal organisms in masses, accounting for around 40% of the total, with C. albicans being the most common species isolated and its contribution to development of OSCC. C. albicans, C. tropicalis, and C. glabrata are the most commonly isolated Candida species, while Candida parapsilosis, Candida stellatoidea, Candida guilliermondii, Candida krusei, and Candida pseudotropicalis are the least often isolated Candida species from medical specimens.,,,,
The samples in this analysis were taken using a sterile swab. In contrast to Saigal et al., who used saliva instead of swab for their culture technique. The swab technique was simple to use and resulted in the isolation of viable cells from a specific location, while the saliva technique did not. For speciation, the collected swab sample was first inoculated on SDA medium, then on CHROM agar (a differential medium). Nadeem et al. and Odds and Bernaerts. used the same mixture of culture media, while Saigal et al. used corn meal agar as the main culture media, preceded by the germ tube examination, which only detects C. albicans and Candida dubliniensis and is unable to distinguish other Candida species. Candida was found in 14 (28%) of the samples, 20 (40%) of the PMD patients, and 42 (84%) of the OSCC patients, respectively, and was absent in 36 (72%) of the controls, 30 (60%) of the PMD patients, and 8 (16%) of the OSCC patients. An extraordinarily large gap (P = 0.000) was found in the statistical study of intergroup comparison. Similarly, different researchers, such as Hongal et al., have shown that the findings of Vuckovic et al., Saigal et al., Anila et al., and Saigal et al. in the present study, the most prevalent Candida in CHROM agar among the controls, PMD and OSCC groups, C. albicans species was present in 9 (18%), 16 (32%) and 6 (12%), C. krusei in 3 (6%), 6 (12%) and 13 (26%), C. glabrata in 0, 0, and 8 (16%) and Candida tropical is in 0, 0 and 3 (6%) cases, respectively. However, only OSCC group showed a combination of species such as C. glabrata and C. krusei was present in 2 (4%) case, C. tropicalis and C. krusei in was present 3 (6%) cases, C. tropicalis and C. glabrata was present in 2 (4%) case, C. albicans and C. tropicalis was present in 2 (4%) cases and C. krusei, C. glabrata with C. albicans was present in 1 (2%) case, respectively. This was in consistent to the study by Gall et al., He was the one who demonstrated that was shift in the dispersal of Candida species with PMD going through malignant transformation from C. albicans to nonalbicans species such as C. glabrata, C. tropicalis, C. krusei and C. parapsilosis.
| Conclusion|| |
We summarized that the prime carriage of Candidal species in PMDs and OSCC, yet conclusion and more evidences are required to prove whether Candida has specific establishment in PMDs or in malignancy.
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Conflicts of interest
There are no conflicts of interest.
| References|| |
Warnakulasuriya S. Global epidemiology of oral and oropharyngeal cancer. Oral Oncol 2009;45:309-16.
Khan Z. An overview of oral cancer in indian subcontinent and recommendations to decrease its incidence. Webmed Central Cancer 2012;3:WMC003626.
Mohd Bakri M, Mohd Hussaini H, Rachel Holmes A, David Cannon R, Mary Rich A. Revisiting the association between candidal infection and carcinoma, particularly oral squamous cell carcinoma. J Oral Microbiol 2010;2:5780-86.
Patil S. Analyzing the association between Candida prevalence, species specifcity, and oral squamous cell carcinoma: A systematic review and meta-analysis. Appl Sci 2020;10:1099.
Barrett AW, Kingsmill VJ, Speight PM. The frequency of fungal infection in biopsies of oral mucosal lesions. Oral Dis 1998;4:26-31.
Kalantar E, Marashi SM, Pormazaheri H, Mahmoudi E, Hatami S, Barari MA, et al.
First experience of Candida non-albicans isolates with high antibiotic resistance pattern caused oropharyngeal candidiasis among cancer patients. J Cancer Res Ther 2015;11:388-90.
Glažar I, Prpić J, Muhvić Urek M, Pezelj-Ribarić S. Identification of Candida
spp. In the oral cavity in patients with malignant diseases. Vojnosanit Pregl 2017;74:1066-70.
Rautemaa R, Hietanen J, Niissalo S, Pirinen S, Perheentupa J. Oral and oesophageal squamous cell carcinoma – A complication or component of autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED, APS-I). Oral Oncol 2007;43:607-13.
Shield KD, Ferlay J, Jemal A, Sankaranarayanan R, Chaturvedi AK, Bray F, et al.
The global incidence of lip, oral cavity, and pharyngeal cancers by subsite in 2012. CA Cancer J Clin 2017;67:51-64.
Bartie KL, Williams DW, Wilson MJ, Potts AJ, Lewis MA. PCR fingerprinting of Candida albicans associated with chronic hyperplastic candidosis and other oral conditions. J Clin Microbiol 2001;39:4066-75.
O'Grady JF, Reade PC. Candida albicans as a promoter of oral mucosal neoplasia. Carcinogenesis 1992;13:783-6.
Krogh P, Hald B, Holmstrup P. Possible mycological etiology of oral mucosal cancer: Catalytic potential of infecting Candida albicans
and other yeasts in production of N-nitroso benzyl methylamine. Carcinogenesis 1987;8:1543-8.
Gall F, Colella G, Di Onofrio V, Rossiello R, Angelillo IF, Liguori G. Candida spp. in oral cancer and oral precancerous lesions. New Microbiol 2013;36:283-8.
Jahanshahi G, Shirani S. Detection of Candida albicans in oral squamous cell carcinoma by fluorescence staining technique. Dent Res J (Isfahan) 2015;12:115-20.
McManus BA, Coleman DC. Molecular epidemiology, phylogeny and evolution of Candida albicans
. Infect Genet Evol 2014; 21:166-78.
Hulimane S, Maluvadi-Krishnappa R, Mulki S, Rai H, Dayakar A, Kabbinahalli M. Speciation of Candida using CHROM agar in cases with oral epithelial dysplasia and squamous cell carcinoma. J Clin Exp Dent 2018;10:e657-60.
Katiraee F, Teifoori F, Soltani M. Emergence of azole-resistant Candida
species in AIDS patients with oropharyngeal candidiasis in Iran. Curr. Med. Mycol 2015;1:11-6.
Ur. Rahaman SM, Ahmed Mujib B. Histopathological correlation of oral squamous cell carcinoma among younger and older patients. J Oral Maxillo fac Pathol 2014;18:183-8.
Hongal BP, Kulkarni VV, Deshmukh RS, Joshi PS, Karande PP, Shroff AS. Prevalence of fungal hyphae in potentially malignant lesions and conditionsdoes its occurrence play a role in epithelial dysplasia? J Oral Maxillo fac Pathol 2015;19:10-7.
Williams DW, Kuriyama T, Silva S, Malic S, Lewis MA. Candida biofilms and oral candidosis: Treatment and prevention. Periodontology 2000 2011;55:250-65.
Cawson RA. Chronic oral candidiasis and leukoplakia. Oral Surg Oral Med Oral Pathol 1966;22:582-91.
McCullough M, Jaber M, Barrett AW, Bain L, Speight PM, Porter SR. Oraly east carriage correlates with presence of oral epithelial dysplasia. Oral Oncol 2002;38:391-3.
Deorukhkar SC, Saini S. Non-albicans Candida
species: Its isolation pattern, species distribution, virulence factors and antifungal susceptibility profile. Int J Med Sci Public Health 2013;2:533-8.
Ramirez-Garcia A, Rementeria A, Aguirre-Urizar JM, Moragues MD, Antoran A, Pellon A, et al. Candida albicans
and cancer: Can this yeast induce cancer development or progression? Crit Rev Microbiol 2016;42:181-93.
Saigal S, Bhargava A, Mehra SK, Dakwala F. Identification of Candida albicans
by using different culture media sand its association in potentially malignant and malignant lesions. Contemp Clin Dent 2011;2:188-93.
] [Full text]
Singh A, Verma R, Murari A, Agrawal A. Oral candidiasis: An overview. J Oral Maxillofac Pathol 2014;18:S81-5.
Nadeem SG, Hakim ST, Kazmi SU. Use of CHROMagar Candida for the presumptive identification of Candida species directly from clinical specimens in resource-limited settings. Libyan J Med 2010;5:1-6.
Odds FC, Bernaerts R. CHROMagar Candida
, differential isolation medium for presumptive identification of clinically important Candida
species. J Clin Microbiol 1994;32:1923-9.
Anila K, Hallikeri K, Shubhada C, Naikmasur V, Kulkarni R. Comparative study of Candida
in oral submucous fibrosis and healthy individuals. Rev Odonto Ciência 2011;26:71-6.
Vučković N, BokorBratić M, Vučković D, Pićurić I. Presence of Candida albicans
in potentially malignant oral mucosal lesions. Arch Oncol 2004;12:51-4.
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5]