|Year : 2018 | Volume
| Issue : 2 | Page : 8-13
Cytomorphometric changes of oral mucosa during normal hormonal turnovers in healthy young menstruating women
Usha Balan1, Master Luqman1, Abdelnasser Mohamed M. Soliman2, Hussain Almubarak1
1 Assistant Professor in Oral Pathology, Department of Diagnostic sciences and Oral Biology (DDS), King, King Khalid University College of Dentistry, Abha, Kingdom of Saudi Arabia
2 Associate Professor in Oral Biology, Department of Diagnostic sciences and Oral Biology (DDS), King Khalid University College of Dentistry, Abha, Kingdom of Saudi Arabia
|Date of Web Publication||13-Aug-2020|
MDS Master Luqman
College of Dentistry, King Khalid University Grainger, Abha
Kingdom of Saudi Arabia
Source of Support: None, Conflict of Interest: None
Oral mucous membrane is an excellent indicator of the constitutional state of a patient. The oral mucosa is under tropic influence of various hormones and thus reflects the systemic status of an individual. Very few cytomorphometric studies are done to evaluate the role of sex hormones on oral mucosa. This study was done to determine the morphometric changes in a cell and nuclear diameter of exfoliated cells from the oral mucosa during the course of normal menstrual cycle in healthy women. The purpose of this study was to determine whether oral smears reflect hormonal state of menstrual cycle. In this study consisting of 40 study subjects and 20 control subjects, smears were collected from buccal mucosa twice per week for a period of three months. Morphometric analysis of the cell and nuclear diameter was done and results were statistically analysed. Cell diameter showed significant changes during various phases of the menstrual cycle of healthy young women. In Comparison of both study and control groups, no significant changes were seen in cell and nuclear diameter of the exfoliated cells. The present study showed changes in cell diameter of the study group which could be related to the role of female sex hormones.
Keywords: Oral mucosa, Estrogen, Progesterone, Hormones, Cytomorphometry
|How to cite this article:|
Usha Balan, Luqman M, M. Soliman AM, Almubarak H. Cytomorphometric changes of oral mucosa during normal hormonal turnovers in healthy young menstruating women. King Khalid Univ J Health Sci 2018;3:8-13
|How to cite this URL:|
Usha Balan, Luqman M, M. Soliman AM, Almubarak H. Cytomorphometric changes of oral mucosa during normal hormonal turnovers in healthy young menstruating women. King Khalid Univ J Health Sci [serial online] 2018 [cited 2021 Jun 12];3:8-13. Available from: https://www.kkujhs.org/text.asp?2018/3/2/8/291953
| Introduction|| |
Oral mucous membrane is an excellent indicator of the constitution of a patient. The oral mucosa possesses hormone receptors and is under tropic influence of various hormones, thus it is a true reflector of the systemic status of an individual. Menstrual cycle is the result of the sequence of hormonal influences that in a normal woman follow each other with great regularity from puberty to menopause except during pregnancy.
Menstrual cycle commences with menstruation and has four phases -Bleeding, Proliferative, Ovulation and Secretory. Hormone levels fluctuate during various phases of the cycle. During first half of the menstrual cycle there is an increase in estrogen level followed by its decline. Second half of the cycle shows increased progesterone. During menstrual period both the hormone levels decline.
Vaginal and buccal epithelia are microscopically similar having stratified squamous epithelium and a desquamative growth pattern. Both epithelia are predominantly non-keratinised. Vaginal mucosa has been shown to respond to changes in hormonal levels and thus cytohormonal evaluation has been used for accurate assessment of hormonal effect on vaginal mucosa. Whether oral mucosa exhibits properties similar to vaginal mucosa in relation to sensitivity to ovarian steroids and whether oral mucosa is an index of hormonal milieu is still a topic for debate. Estrogen receptors have been identified in oral tissues including oral mucosal keratinocytes, thus variation in this hormone can affect oral mucosal cells. With the advancement in the field of cytology, parameters like cytoplasmic and nuclear area have been shown significant in diagnosis of oral lesions. Oral cytomorphometric studies are mainly done for screening of oral malignant and premalignant lesions. The present study uses cytology to assess the effect of hormonal changes during various phases of the menstrual cycle by estimating the cell and nuclear diameter of normal exfoliated buccal mucosal cells.
| Materials And Methods|| |
This study was initiated after obtaining an ethical clearance from the institution with an order number KMCTDC/IEC/2014/25. The experimental group was divided into study group and control group. Study group consisted of forty healthy young women volunteers with normal menstrual cycle of 28-30 days without any local or systemic diseases, abnormal endocrinal or immunological status. Control group consisted of twenty healthy young age matched men volunteers. With an informed written consent a thorough oral examination was done for both these groups of volunteers to rule out clinically evident and habit related lesions. All subjects were apparently healthy without any habits and were non anemic. None of them had any fixed or removable appliances.
Hemoglobin estimation was done to rule out anemia. A proper menstrual history was recorded from the study subjects and the entire cycle was divided into four phases consisting of bleeding phase, proliferative phase, ovulation phase and secretory phase Oral smears were collected from both the study and control groups at regular intervals twice a week for a period of three months. Smears were taken using cyto brush from the buccal mucosa anterior to the stenson’s duct, which according to Timonen is histologically similar to vaginal epithelium. Scrapings were smeared on a plain glass slide, fixed in absolute alcohol and stained using Rapid-papTMPapinicolau stain kit.
Pap stained smears were observed under a binocular light microscope LABOMED. Cell and nuclear diameter of the exfoliated cells were measured using a calibrated 5x eyepiece and 40x objective for both groups. Eyepiece graticule was calibrated for 40x objective by reference to a stage micrometer. The stage micrometer had gradations as small as 10 μm (0.01mm). By superimposition of eyepiece graticule on cytological smears, a direct measurement of individual epithelial cells was done.
Only clearly defined cells were measured avoiding clumped or folded cells. Fifty cells were measured for cell diameter (CD) and nuclear diameter (ND), from each slide which included the superficial, parabasal and intermediate cells. Diameters were obtained along the longest axis and shortest axis of cells and nuclei [Figure 1].
|Figure 1: PAP stained cytologic smear of an intermediate cell-Linear measurements of cell|
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For the study group, the mean value was obtained for each case during various phases of the menstrual cycle. Mean value of cell and nuclear diameter in all the three cycles of the study group were then compared with that of the control group.
Statistical analysis included estimation of value of significance using student’s unpaired’t’ test and analysis of variance was performed using Fischers test and Tukey’s Honestly Significant difference test.
| Results|| |
In our study significant changes were observed in the cell diameter and nuclear diameter of the exfoliated buccal cells in the study group during various phases of the menstrual cycle [Table 1].
|Table 1: Showing cell and nuclear diameter during various phases of a menstrual cycle in study Group|
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Comparing cell diameter and nuclear diameter of the exfoliated cells of one phase with other phases of the menstrual cycle, cell diameter showed statistically significant changes in all the phases of menstrual cycle whereas nuclear diameter showed significant changes in a few phases of all the three cycles studied in study group [Table 2].
|Table 2: Showing comparison between the cell diameter and nuclear diameter of one phase with the other phases in the menstrual cycle|
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Comparison of morphometry of cells between the study and control groups during the three month study showed no significant changes in the cell diameter and nuclear diameter statistically [Table 3].
|Table 3: Showing comparison of cell diameter and nuclear diameter between the study group and control group|
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| Discussion|| |
In our study significant changes were observed in the cell diameter and nuclear diameter during various phases of the menstrual cycle in all the three cycles studied. Comparing cell diameter and nuclear diameter of one phase with other phases of the menstrual cycle, a significant change in the cell diameter is seen from bleeding to secretory phase. Nuclear diameter shows a change during a few phases in the three cycles studied. Increasing cell diameter seen in the second half of the cycle could be attributed to the role of progesterone, which is seen in excess during this period. Age matched men were taken as controls to correlate the morphometric changes to be age- related. On comparing cytomorphometry of cells between study and control groups, no significant changes were seen in the cell diameter and nuclear diameter statistically. Thus in our study we could not establish a distinct difference in oral cytology with regard to morphometry between men and women.
Oral cellular changes associated with phases of menstrual cycle remains a controversial issue. Size of individual epithelial cells undergoes changes in the female reproductive tract as a result of cyclic changes in the menstrual cycle. Very few studies,, have been done with regard to the cellular parameters like cell diameter and nuclear diameter of exfoliated oral cells in the course of normal menstrual cycle in women and most of them showed that buccal squames are unaffected by menstrual cycle. Some authors’ have proposed nucleus: cytoplasmic ratio as a powerful parameter in reflecting the effect of hormones.
According to Driesch’s Law of volume invariance maximal cell size is genetically regulated and hormonal over stimulation has no effect on cell size. Wingrove FA (1979) on cytomorphometric analysis of gingival epithelium during various phases of menstrual cycle found no change in cell size and nuclear surface area. Cowpe JG et al (1985) found no change in the cell size and nuclear size in buccal squames during various phases of menstrual cycle and concluded that oral smears unlike vaginal smears do not clearly demonstrate stage of menstrual cycle or time of ovulation.
Gerasimov JG et al (1996) found changes in buccal cell size, cell areas of cytoplasm, nucleus during menstrual cycle occurring throughout menstrual bleeding indicating a tensed state of female body during this period shown as smallest sizes in buccal epithelial cells. Chretion (1998) on cytomorphometric analysis of superficial vaginal cells in women of various groups found increased nucleus: cytoplasmic ratio in women with oral contraceptives compared to women on in-vitro fertilization stimulation protocol.
Significant variations were seen in nuclear and cytoplasmic area between different sites and with the advancing age (Cowpe JG et al1985).
Studies have shown that various factors like alcohol, smoking,,, age,,‘ radiotherapy, nutritionaldeficiencies, anemia, field change effect etc influence the cytomorphology of cells. Cytomorphometric analysis of exfoliated cells in premalignant and malignant lesions show variation from normal values.
In the present study in which cytomorphometric measurements were done with the help of an eyepiece graticule and stage micrometer, only linear measurements of cell and nuclear diameter were possible. A study of longer duration including more number of subjects would be more appropriate in determining the actual role of female sex hormones on oral mucosa. Cytomorphometric studies comparing cell and nuclear diameter changes in young females during various phases of menstruation with those of age-matched men are not much reported in the literature. Statistically no differences were found in the cell and nuclear diameter between the study and control groups. Thus in our study we could not establish a distinct difference in oral cytology with regard to morphometry between men and women.
| Conclusion|| |
The present study showed significant cytomorphometric changes in the exfoliated oral cells in the study group during various phases of the three cycles studied. An increase in cell diameter was seen during secretory phase of menstrual cycle which could be attributed to the role of progesterone. Comparison between study and control groups showed no changes in the cell and nuclear diameter statistically. This may be due to the influence of sex hormones on the oral mucosa both in men and women.
A study of longer duration including more number of subjects would be more appropriate in determining the actual role of female sex hormones on oral mucosa.
Source of Support: Nil
Conflict of Interest: None declared
| References|| |
Balan U, Gonsalves N, Jose M, Girish KL. Symptomatic changes of oral mucosa during normal hormonal turnover in healthy young menstruating women. J Contemp Dent Pract. 2012 Mar 1;13(2):178- 81.
Dayal J, Pandya D, Dayal PK, Bhat A. Oral health amongst females during hormonal turnover, A clinical and cytological study. JIAOMR 2000; 11:5-22.
PM Donald, George R, SriramG, Kavitha B, Shivapathasundharam.B. Hormonal changes in exfoliated normal buccal mucosal cells. Cytol. 2013 Oct-Dec; 30(4): 252-256.
Laufer N, Navot D, Schenker JG. The pattern of luteal phase plasma progesterone and estradiol in fertile cycle. Am J ObstetGynecol 1982; 143:808-813.
Vittek J, Hernandez MR, Wenk EJ, Rappaport SC, Southern AL. Specific estrogen receptors in human gingiva. J ClinEndocrinolMetab 1982; 54:608-12.
Forabosco A, CriscuoloM, Coukos G, Uccelli E, Weinstein R, Spinato S et al. Efficacy of hormone replacement therapy in postmenopausal women with oral discomfort. Oral Surg Oral Med Oral Pathol 1992; 73:570-4.
Virtanen LR, Pennanen R, Syrjanen K, Syrjanen S. Estrogen response in buccal mucosa-A cytological and immunohistological assay. Maturitas 1997; 27(1): 41-5.
Bercovici B, Gron S, Pisanty S. Vaginal and oral cytology of the menopause, a comparative study. Actacytol 1985; 29: 805-9.
Wingrove FA, Rubright WC, Kerber PE. Influence of ovarian hormone stimulation on atropy, hypertrophy and/or desquamation of human gingiva in premenopausal and postmenopausal women. J Periodontol 1979; 50:445-49.
Cowpe JG and Semmens E. Assessment of the effects of menstrual cycle on the nuclear and cell size of buccal squames. J of Dent Res 1985; 78:995-1004.
Gerasimov IG, Kalutskaia OA. The dynamics of the cellular parameters of the buccal epithelium in the course of menstrual cycle in women. Tsitologia 1996; 38(11): 1152-7.
Chretien MF, Lebouvier B, Denis A, Chappard D. Cytomorphometric analysis of vaginal cells during normal cycle, under oral contraceptive therapy or in- vitro fertilization stimulation protocol. Hum Reprod 1998; 13(10): 2767-71.
Cowpe JG, Longmore RB, Green MW. Quantitative exfoliative cytology of normal oral squames an age, site and sex related survey. JR Soc Med 1985; 78:995-1004.
Ogden GR, Wight AJ, Rice P. Effect of alcohol on the oral mucosa assessed by quantitative cytomotphometry. J Oral Pathol Med 1999; 28:216-20.
Einstein TB, Sivapathasundharam B. Cytomorphometric analysis of the buccal mucosa of tobacco users. Indian J Dent Res. 2005;16:42-6.
Hande AH, Chaudhary MS. Cytomorphometric analysis of buccal mucosa of tobacco chewers Rom J MorpholEmbryol. 2010;51:527-32.
Hegde V. Cytomorphometric analysis of squames from oral premalignant and malignant lesions. J ClinExp Dent. 2011;3:441-4.
Nayar AK, Sundharam BS. Cytomorphometric analysis of exfoliated normal buccal mucosa cells. Indian J Dent Res. 2003;14:87-93.
Jacobs A. Cornification in buccal smears. Brit Dent J 1959; 110:249-50.
Swarnameenakshi.S, Nithyajagannathan. Cytomorphometric analysis of oral epithelial cells in Menstrual cycle.Int J Pharma Res Health Sci. 2017;5(3),1695- 1697.
Ogden GR, Cowpe JG and Green MW: Effect of radiotherapy on oral mucosa assessed by quantitative exfoliative cytology. J ClinPathol 1989; 42:940-43.
Krejei CB, Bissada NF. Womens health issues and their relationship to Periodontitis. JADA 2002; 133:323-28.
Monto RW, Rizek RA, Fine G. Observations on the exfoliative cytology and histology of the oral mucous membranes in Iron deficiency. Oral Surg Oral Med and Oral Pathol 1961; 14:965-73.
[Table 1], [Table 2], [Table 3]