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Indian Journal of Otolaryngology and Head & Neck Surgery

04-01-2025 | Original Article

Detection of Oral Submucous Fibrosis at the Crossroads of Altering Nucleogeometry, Nuclear Spatial Distribution and Nuclear Molecular Abundance

Authors: Sourangshu Chakraborti, Ranjan Rashmi Paul, Mousumi Pal, Jyotirmoy Chatterjee, Raunak Kumar Das

Published in: Indian Journal of Otolaryngology and Head & Neck Surgery

Abstract

The manuscript concentrates on spatial distribution and various nucleomorphometric parameters of epithelium to diagnose Oral Submucous Fibrosis (OSF). Histologically confirmed OSF and normal submucosa tissue samples were procured and stained with diamidino phenylindole (DAPI) to visualize nuclei. E-cadherins and p63 were also immunohistochemically stained. Microphotographs were analyzed to quantify the spatial distance among the nuclei of the stained tissue samples. Intensity of the DAPI stained nuclei and p63 was quantified. In addition, morphometric analysis of nuclei was done with the help of ImageJ software to quantify the geometric alterations in OSF tissue. Spatial distances among the nuclei of OSF tissue samples were found to be significantly higher than that of normal tissue. We also observed a significant decrease in the mean intensity of DAPI and p63 in OSF tissue samples. In addition, we have found statistically significant alterations of various morphometric quantifications in OSF tissue nuclei. There was a considerable change in the spatial distribution of nuclei, as well as some distinct changes in the nucleogeometry of OSF tissue, which corresponds to histological abnormalities. Decreased intensity of DAPI and p63 advocate disease progression. The biomarkers in this study are the accountable role-players for early detection of oral carcinoma.

Rao NR, Villa A, More CB, Jayasinghe RD, Kerr AR, Johnson NW (2020) Oral submucous fibrosis: a contemporary narrative review with a proposed inter-professional approach for an early diagnosis and clinical management. J Otolaryngol Head Neck Surg 49(1):3. https://doi.org/10.1186/s40463-020-0399-7 PMID: 31915073; PMCID: PMC6951010CrossRefPubMedPubMedCentral

Shieh DH, Chiang LC, Lee CH, Yang YH, Shieh TY (2004) Effects of arecoline, safrole, and nicotine on collagen phagocytosis by human buccal mucosal fibroblasts as a possible mechanism for oral submucous fibrosis in Taiwan. J Oral Pathol Med.;33(9):581-7. https://doi.org/10.1111/j.1600-0714.2004.00229.x. PMID: 15357680

Tilakaratne WM, Klinikowski MF, Saku T, Peters TJ, Warnakulasuriya S (2006) Oral submucous fibrosis: review on aetiology and pathogenesis. Oral Oncol 42(6):561–568. https://doi.org/10.1016/j.oraloncology.2005.08.005 Epub 2005 Nov 28. PMID: 16311067CrossRefPubMed

Trivedy CR, Warnakulasuriya KA, Peters TJ, Senkus R, Hazarey VK, Johnson NW (2000) Raised tissue copper levels in oral submucous fibrosis. J Oral Pathol Med.;29(6):241-8. https://doi.org/10.1034/j.1600-0714.2000.290601.x. PMID: 10890553

Haque MF, Meghji S, Khitab U, Harris M (2000) Oral submucous fibrosis patients have altered levels of cytokine production. J Oral Pathol Med.;29(3):123-8. https://doi.org/10.1034/j.1600-0714.2000.290304.x. PMID: 10738939

Trivedy C, Warnakulasuriya KA, Tavassoli M, Steingrimsdottir H, Penhallow J, Maher R et al (1998) p53 aberrations in oral submucous fibrosis and oral squamous cell carcinoma detected by immunocytochemistry and PCR-SSCP. J Oral Pathol Med.;27(2):72– 7. https://doi.org/10.1111/j.1600-0714.1998.tb02097.x. PMID: 9526733

Karthik H, Nair P, Gharote HP, Agarwal K, Ramamurthy Bhat G, Kalyanpur Rajaram D (2012) Role of hemoglobin and serum iron in oral submucous fibrosis: a clinical study. ScientificWorldJournal 2012:254013. https://doi.org/10.1100/2012/254013CrossRefPubMedPubMedCentral

Angadi PV, Rao S (2010) Management of oral submucous fibrosis: an overview. Oral Maxillofac Surg.;14(3):133– 42. https://doi.org/10.1007/s10006-010-0209-x. PMID: 20177949

Biradar SB, Munde AD, Biradar BC, Shaik SS, Mishra S (2018) Apr-Jun;14(3):597–603 Oral submucous fibrosis: A clinico-histopathological correlational study. J Cancer Res Ther. https://doi.org/10.4103/0973-1482.176177. PMID: 29893325

10.

Pindborg JJ, Sirast SM (1966) Oral submucous fibrosis. Oral Surg Oral Med Oral Pathol 22:764–779CrossRefPubMed

11.

Yang PY, Chen YT, Wang YH, Su NY, Yu HC, Chang YC (2017) Malignant transformation of oral submucous fibrosis in Taiwan: a nationwide population-based retrospective cohort study. J Oral Pathol Med 46(10):1040–1045. https://doi.org/10.1111/jop.12570 Epub 2017 Apr 6. PMID: 28295637CrossRefPubMed

12.

Chaturvedi P, Malik A, Nair D, Nair S, Mishra A, Garg A et al (2017) Oral squamous cell carcinoma associated with oral submucous fibrosis have better oncologic outcome than those without. Oral Surg Oral Med Oral Pathol Oral Radiol.;124(3):225–230. doi: 10.1016/j.oooo.2017.04.014. Epub 2017 May 4. PMID: 28606826

13.

Neville B, Damm DD, Allen C, Chi A (2015) May. Oral and Maxillofacial Pathology. 4th Edition. Elsevier. 4th Page no. 366–368

14.

Rajendran R, Sivapathasundharam B (2012) Shafer’s Textbook of Oral Pathology. 7th Edition. Elsevier India. Page no. 97–103

15.

Qin X, Ning Y, Zhou L, Zhu Y (2023) Oral Submucous Fibrosis: etiological mechanism, Malignant Transformation, therapeutic approaches and targets. Int J Mol Sci 24(5):4992CrossRefPubMedPubMedCentral

16.

Nag R, Kumar Das R (2018) Analysis of images for detection of oral epithelial dysplasia: A review. Oral Oncol.;78:8–15. https://doi.org/10.1016/j.oraloncology.2018.01.003. Epub 2018 Jan 10. PMID: 29496062

17.

Moore MJ, Sebastian JA, Kolios MC (2019) Determination of cell nucleus-to-cytoplasmic ratio using imaging flow cytometry and a combined ultrasound and photoacoustic technique: a comparison study. J Biomed Opt 24(10):1–10. https://doi.org/10.1117/1.JBO.24.10.106502CrossRefPubMed

18.

Hansen JP, McDonald IR Theory of Simple of Liquids: Academic Press. Published Date: 8th February 2006

19.

Conway JH, Sloane NJA (1998) Sphere packings. Springer, Lattices and Groups

20.

Peebles PJE (1993) Principles of Physical Cosmology: Princeton University Press. Published Date: May 9, 1993

21.

Gevertz JL, Torquato SA, Novel Three-phase Model of Brain Tissue Microstructure (2008). PLoS Comput Biol.;4(8):e1000152. https://doi.org/10.1371/journal.pcbi.1000152. Erratum in: PLoS Comput Biol. 2009;5(1). https://doi.org/10.1371/annotation/c9faa83b-3c7b-4f38-8d74-1a4309403688. PMID: 18704170; PMCID: PMC2495040

22.

Farhadifar R, Roper JC, Aigouy B, Eaton S, Frank Julicher (2007) The influence of cell mechanics, cell-cell interactions, and proliferation on epithelial packing. Curr Biol 17:2095–2104

23.

Staple DB, Farhadifar R, Roper JC, Aigouy B, Eaton S et al (2010) Mechanics and remodelling of cell packings in epithelia. Eur Phys J E 33:117–C127CrossRefPubMed

24.

Feichtenbeiner A, Haas M, Buttner M et al (2014) Critical role of spatial interaction between CD8(þ) and Foxp3(þ) cells in human gastric cancer: the distance matters. Cancer Immunol Immunother 63:111–119CrossRefPubMed

25.

Balsat C, Signolle N, Goffin F et al (2014) Improved computer-assisted analysis of the global lymphatic network in human cervical tissues. Mod Pathol 27:887–898CrossRefPubMed

26.

Syverud K, Chinga G, Johnssen PO, Leirset I, Wiik K (2007) Analysis of lint particles from full-scale printing trials. Appita J 60(4):286–290

27.

Clark PJ, Evans FC (1954) Distance to Nearest Neighbor as a measure of spatial relationships in populations. Ecology, vol 35. JSTOR, pp 445–453. 4

28.

Lobo J, See EY, Biggs M, Pandit A (2016) An insight into morphometric descriptors of cell shape that pertain to regenerative medicine. J Tissue Eng Regen Med 10(7):539–553 Epub 2015 Mar 11. PMID: 25757807CrossRefPubMed

29.

Andrei P, Sommer D, Zhu T, Scharnweber H-J, Fecht (2010) On the Social Behaviour of Cells. J Bionic Eng Volume 7(1):1672–6529

30.

Gobaa S, Gayet RV, Lutolf MP (2018) Artificial niche microarrays for identifying extrinsic cell-fate determinants. Methods Cell Biol 148:51–69. https://doi.org/10.1016/bs.mcb.2018.06.012 Epub 2018 Jul 20. PMID: 30473074CrossRefPubMed

31.

Arakeri G, Brennan PA (2018) TFM classification and staging of oral submucous fibrosis: a new proposal. J Oral Pathol Med 47(5):539. https://doi.org/10.1111/jop.12700 Epub 2018 Mar 24. PMID: 29473974CrossRefPubMed

32.

Guberman E, Sherief H, Regan ER (2020) Boolean model of anchorage dependence and contact inhibition points to coordinated inhibition but semi-independent induction of proliferation and migration. Comput Struct Biotechnol J 18:2145–2165 PMID: 32913583; PMCID: PMC7451872CrossRefPubMedPubMedCentral

33.

Lodish H, Berk A, Zipursky SL et al (2000) Molecular Cell Biology. 4th edition. New York: W. H. Freeman; Sect. 15.7, Transport across Epithelia

34.

West J, Schenck RO, Gatenbee C et al (2060) (2021) Normal tissue architecture determines the evolutionary course of cancer. Nat Commun 12. https://doi.org/10.1038/s41467-021-22123-1

35.

Xu R, Boudreau A, Bissell MJ (2009) Tissue architecture and function: dynamic reciprocity via extra- and intra-cellular matrices. Cancer Metastasis Rev 28(1–2):167–176. https://doi.org/10.1007/s10555-008-9178-zCrossRefPubMedPubMedCentral

36.

Alberts B, Johnson A, Lewis J et al (2002) Molecular Biology of the Cell. 4th edition. New York: Garland Science; Cell Junctions. Available from: https://www.ncbi.nlm.nih.gov/books/NBK26857

37.

Melino G (2011) p63 is a suppressor of tumorigenesis and metastasis interacting with mutant p53. Cell Death Differ 18:1487–1499. https://doi.org/10.1038/cdd.2011.81CrossRefPubMedPubMedCentral

38.

Urist MJ, Di Como CJ, Lu ML, Charytonowicz E, Verbel D, Crum CP, Ince TA, McKeon FD, Cordon-Cardo C (2002) Loss of p63 expression is associated with tumor progression in bladder cancer. Am J Pathol 161(4):1199–1206. https://doi.org/10.1016/S0002-9440(10)64396-9 PMID: 12368193; PMCID: PMC1867279CrossRefPubMedPubMedCentral

39.

Koga F, Kawakami S, Fujii Y, Saito K, Ohtsuka Y, Iwai A, Ando N, Takizawa T, Kageyama Y, Kihara K (2003) Impaired p63 expression associates with poor prognosis and uroplakin III expression in invasive urothelial carcinoma of the bladder. Clin Cancer Res 9(15):5501–5507 PMID: 14654529PubMed

40.

Barbieri CE, Tang LJ, Brown KA, Pietenpol JA (2006) Loss of p63 leads to increased cell migration and up-regulation of genes involved in invasion and metastasis. Cancer Res.;66(15):7589-97. https://doi.org/10.1158/0008-5472.CAN-06-2020. PMID: 16885358

41.

Bergholz J, Xiao ZX (2012) Role of p63 in Development, Tumorigenesis and Cancer Progression. Cancer Microenviron 5(3):311–322. https://doi.org/10.1007/s12307-012-0116-9 Epub 2012 Jul 31. PMID: 22847008; PMCID: PMC3460051CrossRefPubMedPubMedCentral

42.

Tan EH, Morton JP, Timpson P, Tucci P, Melino G, Flores ER, Sansom OJ, Vousden KH, Muller PA (2014) Functions of TAp63 and p53 in restraining the development of metastatic cancer. Oncogene 33(25):3325–3333. https://doi.org/10.1038/onc.2013.287 Epub 2013 Jul 22. PMID: 23873029; PMCID: PMC4181588CrossRefPubMed

43.

Chazotte B (2011) Labeling nuclear DNA using DAPI. Cold Spring Harb Protoc. 2011(1). https://doi.org/10.1101/pdb.prot5556 PMID: 21205856

44.

Illeperuma RP, Kim DK, Park YJ, Son HK, Kim JY, Kim J, Lee DY, Kim KY, Jung DW, Tilakaratne WM, Kim J (2015) Areca nut exposure increases secretion of tumor-promoting cytokines in gingival fibroblasts that trigger DNA damage in oral keratinocytes. Int J Cancer 137(11):2545–2557. https://doi.org/10.1002/ijc.29636 Epub 2015 Jun 24. PMID: 26076896; PMCID: PMC4744697CrossRefPubMedPubMedCentral

45.

Chen CL, Chi CW, Chang KW, Liu TY (1999) Safrole-like DNA adducts in oral tissue from oral cancer patients with a betel quid chewing history. Carcinogenesis.;20(12):2331-4. https://doi.org/10.1093/carcin/20.12.2331. PMID: 10590228

46.

Chung FL, Krzeminski J, Wang M, Chen HJ, Prokopczyk B (1994) Jan-Feb;7(1):62– 7 Formation of the acrolein-derived 1,N2-propanodeoxyguanosine adducts in DNA upon reaction with 3-(N-carbethoxy-N-nitrosamino)propionaldehyde. Chem Res Toxicol. https://doi.org/10.1021/tx00037a009. PMID: 8155826

47.

Sundqvist K, Liu Y, Nair J, Bartsch H, Arvidson K, Grafström RC (1989) Cytotoxic and genotoxic effects of areca nut-related compounds in cultured human buccal epithelial cells. Cancer Res 49(19):5294–5298 PMID: 2766297PubMed

48.

Kulasekaran C, Devi M, Dhivya K, Vijayalakshmi D, Sathishkumar M, Madhanmohan A (2020 Sep-Dec) Immunohistochemical detection of 8-hydroxydeoxyguanosine: a biomarker of oxidative DNA damage in oral submucous fibrosis. J Oral Maxillofac Pathol 24(3):536–541. https://doi.org/10.4103/jomfp.JOMFP_42_20 Epub 2021 Jan 9. PMID: 33967493; PMCID: PMC8083425

49.

Lepekhin EA, Walmod PS, Berezin A et al (2001) Evaluation of cell morphology. Methods Mol Biol 161:85–100PubMed

50.

Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9:671–675CrossRefPubMedPubMedCentral

51.

Takashimizu Y, Iiyoshi M (2016) New parameter of roundness R: circularity corrected by aspect ratio. Prog Earth Planet Sci 3:2. https://doi.org/10.1186/s40645-015-0078-xCrossRef

52.

Zdilla MJ, Hatfield SA, McLean KA, Cyrus LM, Laslo JM, Lambert HW, Circularity (2016) Solidity, axes of a best fit ellipse, aspect ratio, and roundness of the Foramen Ovale: a morphometric analysis with neurosurgical considerations. J Craniofac Surg 27(1):222–228. https://doi.org/10.1097/SCS.0000000000002285 PMID: 26703050; PMCID: PMC4706773

53.

Soltys Z, Ziaja M, Pawlínski R et al (2001) Morphology of reactive microglia in the injured cerebral cortex. Fractal analysis and complementary quantitative methods. J Neurosci Res 63:90–97CrossRefPubMed

54.

Payne CM, Bjore CG, Cromey DW, Roland F (1989) A comparative mathematical evaluation of contour irregularity using form factor and PERBAS, a new analytical shape factor. Anal Quant Cytol Histol 11:341–352PubMed

55.

Mitruț R, Pirici D, Stepan AE, Mărgăritescu C, Simionescu CE, Kesse AM, Militaru C Histopathological and morphometric study of Fibrosis and Nuclear Pleomorphism in Dilated Cardiomyopathy. Curr Health Sci J 2019 Jan-Mar;45(1):73–78. https://doi.org/10.12865/CHSJ.45.01.10. Epub 2019 Mar 31. PMID: 31297266; PMCID: PMC6592661.

56.

Gadiwan M, Madhushankari G, Mandana D, Praveen S, Selvamani M, Pradeep D (2014) Nuclear features in different grades of epithelial dysplasia in leukoplakia: a computer assisted microscopic study. J Oral Maxillofac Pathol 18(2):194–200 PMID: 25328298; PMCID: PMC4196286CrossRefPubMedPubMedCentral

57.

Fischer EG (2020) Nuclear morphology and the Biology of Cancer cells. Acta Cytol 2 November 64(6):511–519. https://doi.org/10.1159/000508780CrossRef

58.

Fischer AH, Taysavang P, Jhiang SM (2003) Nuclear envelope irregularity is induced by RET/PTC during interphase. Am J Pathol 163(3):1091–1100CrossRefPubMedPubMedCentral

Title: Detection of Oral Submucous Fibrosis at the Crossroads of Altering Nucleogeometry, Nuclear Spatial Distribution and Nuclear Molecular Abundance
Authors: Sourangshu Chakraborti
Ranjan Rashmi Paul
Mousumi Pal
Jyotirmoy Chatterjee
Raunak Kumar Das
Publication date: 04-01-2025
Publisher: Springer India
Published in: Indian Journal of Otolaryngology and Head & Neck Surgery
Print ISSN: 2231-3796
Electronic ISSN: 0973-7707
DOI: https://doi.org/10.1007/s12070-024-05305-3

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Springer Medicine

Detection of Oral Submucous Fibrosis at the Crossroads of Altering Nucleogeometry, Nuclear Spatial Distribution and Nuclear Molecular Abundance

Abstract

Keynote webinar | Spotlight on adolescent vaping

Springer Medicine

Abstract

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