Exfoliative cytologic studies on normal oral cavity and on the postexodontic wound healing of infants were carried out and the following results were obtained: 1) The keratinization of cells was found marked in such regions as
buccal mucosa, mucobuccal fold, gingiva and palate in that order. 2) As for changes in the distribution of cells and leucocytes, the cell distribution in the period of 15-21 postexodontic days proved to be identical with that of normal exfoliated cells. Namely, the wound healing from exfoliative cytologic aspects takes place during the period of 15-21 postexodontic days.</p

Kiguchi, Kenichiro

Nishijima, Katsumi

Okayama University Scientific Achievement Repository

Acta Medica OkayamaVolume 24, Issue 2 1970 Article 8APRIL 1970Exfoliative cytologic study on normalchildren’s oral cavity and their postexodonticwound healingKatsumi Nishijima∗ Kenichiro Kiguchi†∗Okayama University,†Okayama University,Copyright c©1999 OKAYAMA UNIVERSITY MEDICAL SCHOOL. All rights reserved.Exfoliative cytologic study on normalchildren’s oral cavity and their postexodonticwound healing∗Katsumi Nishijima and Kenichiro KiguchiAbstractExfoliative cytologic studies on normal oral cavity and on the postexodontic wound healingof infants were carried out and the following results were obtained: 1) The keratinization of cellswas found marked in such regions as buccal mucosa, mucobuccal fold, gingiva and palate in thatorder. 2) As for changes in the distribution of cells and leucocytes, the cell distribution in theperiod of 15-21 postexodontic days proved to be identical with that of normal exfoliated cells.Namely, the wound healing from exfoliative cytologic aspects takes place during the period of15-21 postexodontic days.∗PMID: 4247894 [PubMed - indexed for MEDLINE] Copyright c©OKAYAMA UNIVERSITYMEDICAL SCHOOLActa Med. Okayama 24, 227-235 (1970)EXFOLIATIVE CYTOLOGIC STUDY ON NORMAL CHILD-REN'S ORAL CAVITY AND THEIR POSTEX-ODONTIC WOUND HEALINGKatsumi NISHI]IMA* and Kenichiro KIGUCHI**Department of Oral Surgery, Okayama University Medical School, Okayama, Japan(Director: Pro]. Y. Watanabe)Received for publication, October 3, 1969Cytologic study of the oral cavity dates back to MILLER'S paper (1890)on the epithelial cells and leucocytes in the human saliva (10), and thereare studies on the relationship between keratinization of the epithelialcells and cellular components by WEINMANN (1935, 1936, 1940), ORBANand WEINMANN (1939), MULLER (1940), KUME (1941, 1944), and BRADLEY(1948)(12,13,15). Keratinization of the gingiva was investigated by AURER.KOZEL] (1967) (1) recently. WATANABE (1951) (11) made a study on theroentgen effect upon the cellular elements in the human saliva by meansof sialogram.MILLER (1951) (5), MONTGOMERY (1951) (6) and HIDAKA (1960) (4)made comparative studies on the difference of the percentage of keratinizedcells at different regions of the normal oral mucosa statistically by Papani-colaou'S staining technic. In addition, there are studies on histologicchanges in normal oral mucosa according to the age by SUZUKI et aZ. (8),and those on the changes due to mouth washing and gurgling in newbornand infants by SHIRAISHI (1951) (9) of our Department, and those byMATSUI (1958) (7) on oral exfoliated and keratinized cells by scraping.Though there have been many descriptions on the keratinization ofthe oral mucosa, only a few studies on the exfoliative cytologic study inpostexodontic healing were made by HAHN (1958) (3), FUKUI and MAKI(1958) (2), WATANABE and TOMOZAWA (1963) (14). And no study can befound on oral cytology after extraction of deciduous teeth. In order tomake this point clear, exfoliative cytologic observations were made innormal oral cavity and in wound healing after extraction of deciduousteeth of infants.Presented at the Fifth Japanese Society of Fedodontics, Tokyo, Japan, 1967 and theSixth Autumn Meeting of the Japanese ~ociety of Clinical Cytology, Morioka, Japan, 1967* Assistant Professor and **Clinical Assistant of Oral Surgery, Oka; ama University MedicalSchool2271Nishijima and Kiguchi: Exfoliative cytologic study on normal children's oral cavity andProduced by The Berkeley Electronic Press, 1970228 K. NISHIJIMA and K. KIGUCHIMATERIALS AND METHODSA. Study on Exfoliated Cells in Normal Oral Cavity of Infants1. Collecting and Staining: Subjects of the study were 20 infants of the ageranging 2-5 years with apparently normal oral mucosa who saw us at theDepartment of Oral Surgery in Okayama University Hospital. Materials werecollected from the following 5 sites: a) Buccal mucosa corresponding to theposition of the crown of the upper first deciduous molar. b) Palatal mucosacorresponding to the apical area of the root of the upper first deciduous molar.c) Mucobuccal fold of the lower first deciduous molar. d) Buccal gingival mucosaof the lower first deciduous molar (about 1-2 mm below the neck of the tooth).e) Lingual gingival mucosa of the lower first deciduous molar (about 1-2 mmbelow the neck of the tooth).The materials collected from these sites by scraping the mucosa softly witha cotton swab about 2 mm in diameter were thinly smeared over a clean slideglass as evenly as possible about 2 cm2 in the area and immersed quickly in asolution of 95 % ethyl alcohol and equal part of ether to avoid drying. Afterfixing over 2 hours the smears were stained by Papanicolaou method.2. Cell Classification: Microscopic observations of smears were made at 200-400 x magnification and all the cells were classified into Type I to V accordingto the morphology and stainability of cytoplasm and nucleus.Type I: Cells with round small cytoplasm and somewhat large nucleusstained deep (Fig. 1). Type II: Cells with ovoid cytoplasm and a nucleusslightly larger than Type I. Spindle-shaped cytoplasm is included in this group(Fig. 2). Type III: Cells with polygonal cytoplasm of medium to large size,and the nucleus showing no degeneration (Figs. 3, 4-a). Type IV: Cells withpolygonal large cytoplasm and the nucleus showing degeneration (Figs. 3, 4-b).Cells with very poorly stained nucleus. Type V: Cells with nucleus mostly des-troyed (Fig. 4-c).B. Study on Exfoliated Cells in Postexodontic Wound Healing1. Collecting and Staining: Subjects of this study were 74 postexodonticwounds in case of infants of the age ranging 2-5 years who saw us at the Depart-ment of Oral Surgery, Okayama University Hospital, and extractions of theirdeciduous teeth were indicated clinically and radiologically.Daily follow-up observations were carried out on these cases and smearswere taken from the center of the postexodontic wounds by the same method asin the normal control, avoiding hemorrhage as much as possible, and they werestained by Papanicolaou technic.2. Cell Classification: Cells were classified in the same way as mentionedabove.3. Classification of Leucocytes: Smears were examined at 200-400 x magnifi-cation and cell counts were made in 5 different fields at random. They weredivided into the following 5 groups according the number of cells: 1) one under4 cells, 2) 5-9 cells, 3) 10-19 cells, 4) 20-49 cells and 5) one with over 50 cells.No red blood cells were counted.2Acta Medica Okayama, Vol. 24 [1970], Iss. 2, Art. 8http://escholarship.lib.okayama-u.ac.jp/amo/vol24/iss2/8Exfoliative Cytology of Oral Cavity 2291. Fig. 1 Type I 2. Fig. 2 Type II3. Fig. 3 Types III & IV 4. Fig. 4 Type Va: Type III a: Type IIIb: Type IV b: Type IVc: Type VRESULTSIn the central portion of the specimen where cells were smearedevenly cell counts were taken under the microscope (200-400 X) andclassified according to the classification mentioned above. In the normaloral cavity of infants cells of Type I and Type II were observed in eachof the fIve sites of the oral cavity, though they were a few in number. Cells3Nishijima and Kiguchi: Exfoliative cytologic study on normal children's oral cavity andProduced by The Berkeley Electronic Press, 1970230 K. NISHIJIMA and K. KWUCHIof Type III were found lowest in the number in the palatal region (25.6 percent) and were highest in the buccal mucosa (40.4 per cent). These cellsof Type III were observed about 36-37 per cent both in the mucobuccalfold and gingival region. They hardly showed any difference in thenumber. Cells of Type IV were highest in the number in the palatalregion showing about 60.5 per cent, followed by the mucobuccal fold(56.8 per cenO, and no differences were observed in other sites. Cellsof Type V were most predominant in the palate showing 13.9 per cent,followed by 11.1 per cent in the buccal gingiva. A minimal number ofleucocytes and no erythrocytes were observed.After tooth ex:traction the distribution pattern of cells appearing inoral changed with lapse of time.No cells other than blood cells could be observed immediately aftertooth extraction. Cells of Types I and II appeared prominently 2-3 daysafter extraction and showed a decrease with lapse of time (as the cellcount of Type I was so small, it was included with that of Type II). Cellsof Type III were numerous, But they reached their peak in about 4-5days later, and gradually decreased in the number thereafter. The cellsof Type IV showed a tendency of increase in number till 2 postexodonticweeks, and during about 10-21 days their number became higher thanthat of Type III cells. Up to the period of 10-21 days, while the cells ofTypes I and II were decreased in nurnbel' and those of Type IV wereincreased more than the number of Type III cells (Fig. 5).,-- ••~'''' It'./Fig. 5During 2-3 postexodontic days the groups of over 50 cells occupiedmore than half of the total cell count. During 4-5 days the groups ofover 50 cells grew less in numb~r, and groups of 5-9, 10-19, and 20-49 cells were seen scatteringly. In more than 10 days the groups of over50 cells could no longer be observed and the groups of less than 4 cellsgrew more numerous. The groups of less than 4 cells became predomi-nantly marked 15 days later. Leucocytes were hardly visible 22 days later.4Acta Medica Okayama, Vol. 24 [1970], Iss. 2, Art. 8http://escholarship.lib.okayama-u.ac.jp/amo/vol24/iss2/8Table I NUMBER PER CENT OF EACH SPECIES, EXFOLIATED CELL FROM NORMAL CAVITY OF 20 INFANTSCase 1'\0. TypeII III IV Va b c d e a b c d e a b c d e a b c d e a b c d eI 2 36 23 35 32 40 54 62 59 58 48 8 15 6 10 122 I 40 30 35 32 34 51 60 60 59 55 9 10 5 8 II3 2 I 41 27 35 32 37 56 67 60 47 54 3 6 5 19 84 I 2 45 17 35 52 42 49 72 60 38 53 6 II 6 8 5 trlX5 42 27 35 19 35 52 61 59 59 54 6 12 6 21 II §:6 I 47 35 39 46 39 48 59 56 51 55 5 6 5 3 5 ~.7 44 14 37 40 35 49 67 60 56 54 7 19 3 4 II <'('1)8 43 25 37 37 38 53 57 54 53 53 4 18 9 10 9 0'<9 I 2 45 32 31 31 38 49 59 62 52 53 4 9 7 16 9 00-10 I 44 31 37 39 27 52 61 59 50 61 4 8 4 10 12aq'<II 36 21 36 45 31 58 59 60 52 58 6 20 4 3 II 0.....12 43 17 36 26 39 52 66 61 52 56 5 17 3 22 5 0...13 47 29 41 33 36 49 59 55 47 55 4 12 4 20 9 e'..47 34 42 45 38 49 58 53 50 56 4 8 5 6014 1 5 II)::.IS I 33 22 38 33 39 56 56 57 54 59 II 22 5 12 2 ~16 31 25 38 33 37 62 53 55 62 56 7 22 7 5 517 37 30 43 35 36 48 58 51 52 57 15 12 6 13 7III 37 29 40 37 42 56 50 51 54 50 7 21 9 9 819 34 32 43 29 35 60 55 53 59 61 6 13 4 12 420 35 12 42 40 37 57 71 50 50 58 8 17 8 10 5average 0.1 0.1 0.2 0.1 0.3 0.1 40.4 25.6 37.7 35.8 36.9 53.0 60.5 56.8 52.8 55.3 6.5 13.9 5.6 I I.1 7.8Note: a: Cheek b: Palate c: M ucobuccal Fold d: Buccal Gingiva e: Lingual Gingiva~UJ.....5Nishijima and Kiguchi: Exfoliative cytologic study on normal children's oral cavity andProduced by The Berkeley Electronic Press, 1970232 K. NISHIJIMA and K. KIGUCHIDuring 2-3 postexodontic days numerous erythrocytes could be observedbut they gradually decreased in number thereafter (Tabl~ 2).Table 2 FLU'::TUATION OF THE NUMBER OF LEUCOCYTE GROUPS DURING THEPOSTEXODONTIC PERIODgroups of leucocytes under over4 5-9 10-19 20-49 50days cells cells cells cells cells2 -- 3 2 2 2 84 -- 5 3 5 4 36- 9 2 3 6 4 310 - 14 6 3 215 - 21 5 I22 - 28 9Note: The leucocyte count in each group showing the average of five fields selected atrandomSigns of degeneration were seen during inflammatory changes, andvacuolization in cytoplasm or nucleus was prominent feature: Cells show·ing vacuolization in cytoplasm and in nucleus (Fig. 6), cells showingvacuolization in cytoplasm (Figs. 7, 8), cells showing large granules incytoplasm (Fig. 9).LI3CUSSIONThere are many studies dealing with problems of postexodontic woundhealing, but the majority of them are histopathologic or roentgenologicstudy, and exfoliative cytologic studies in this field are few in number,such as the papers by HAHN (3), FUKUI and MAKI (2) and WATANABE andTOMOZA WA (13 J. And exfoliative cytologic study on the wound healingafter extraction of human deciduous teeth in normal oral cavity is notavailable so far.According to MATSUI (7) the epithelial cells in various layers of theepithelium were stained blue, red and yellow in the order as the keratini.zation advances. WATANABE and TOMOZAWA (13) reported on stainabilityof the cytoplasm of postexodontic wound healing in dogs and classifiedthe epithelial cells into deep. layer cells, intermediate cells, precornifiedcells and keratinized cells.From the viewpoint of the appearance of parabasal cells which canhardly be observed in normal exfoliated cells, it was considered that thestainability of the cells alone would not serve as a sufficient clue to pursuepostexodontic wound healing. Therefore, further observations were madeon exfoliated epithelial cells and they were classified into Types I, II, III,6Acta Medica Okayama, Vol. 24 [1970], Iss. 2, Art. 8http://escholarship.lib.okayama-u.ac.jp/amo/vol24/iss2/8Exfoliative Cytology of Oral Cavity 2336. Fig. 68. Fig. 87. Fig. 79. Fig. 9IV and V, according to the shape and stainability of the cytoplasm andmorphology of the nucleus by the method of WATANABE and TOMOZAWA.WEINMANN (10) stated that the percentage of keratinized cells increasedin the order of buccal mucosa, tongue root, gingiva and hard palate, andMILLER (5) obtained practically similar results. MONTGOMERY (6) observedthat the order of keratinization was soft palate, buccal mucosa, anterumof oral cavity, the anterior part of tongue and gingiva, while MATSUI (7)in the order of buccal mucosa, tongue root, gingiva and hard palate.It is difficult to make comparison of our results directly with theirsbecause of difference in the age of the subjects, but the fact that Type7Nishijima and Kiguchi: Exfoliative cytologic study on normal children's oral cavity andProduced by The Berkeley Electronic Press, 1970234 K. NISHIJIMA and K. KIGUCHIIV and V cells are more predominant than in other regions, showing thetotal of 74.4 per cent, indicates marked keratinization in this region agree·ing with those findings of MATSUI, WEINMANN and others, both gingivaand mucobuccal fold show Type IV and V cells in total of 62-64 percent with little difference between the two regions and the buccal mucosaleast with 59.5 per cent. The fact that in a few cases Type I and II cellswhich do not generally appear among exfoliated cells of normal oral cavitywere observed, may imply a marked metabolism of mucosa in infant orwandering of the cells out of the periodontal pockets.WATANABE and TOMOZAWA (13) described that the wound becamecovered with a new regenerated epithelium 15--20 days after extractionof canine teeth.In the authors' study, up to 10-14 postexodontic days Type I and IIcells appeared at the wound site relatively many which decreased in num·ber with lapse of time. In the meantime Type III cells usually appearedmore numerous than Type IV cells, and the latter tended to increasegradually, while Type III cells tended to decrease during 4-5 days.In other words, the result in the present observation was reverse of thatobservable in the distribution of normal exfoliated cells. During 10-14days when Type I and II cells started to decrease in number, Type IVcells grew more than Type III cells, showing normal cell distribution.On the other hand, as for leucocytes during 2-3 postexodontic daysmore than half of the cases belonged to the group of over 50 cells andduring 10-14 days more than half of the cases belonged to the groups ofless than 4 cells. And during 15-21 days most of the groups becameunder 4·cell group and all leucocytes during 22-29 days belonged to theunder 4-cell group. As for erythrocytes, although the specimens wereobtained avoiding bleeding as much as possible, red blood cells appearedup to one postexodontic week in every case.From these changes in the exfoliated cells and leucocytes, the woundhealing after deciduous tooth extraction must be completed during theperiod of 15-21 days.SUMMARYExfoliative cytologic studies on normal oral cavity and on the post-exodontic wound healing of infants were carried out and the followingresults were obtained:1) The keratinization of cells was found marked in such regions asbuccal mucosa, mucobuccal fold, gingiva and palate in that order.8Acta Medica Okayama, Vol. 24 [1970], Iss. 2, Art. 8http://escholarship.lib.okayama-u.ac.jp/amo/vol24/iss2/8Exfoliative Cytology of Oral Cavity 2352) As for changes in the distribution of cells and leucocytes, the celldistribution in the period of 15-21 postexodontic days proved to beidentical with that of normal exfoliated cells. Namely, the wound healingfrom exfoliative cytologic aspects takes place during the period of 15-21postexodontic days.ACKNOWLEDGEMENTThe authors express their profound thanks to Dr. YOSHIO WATANABE for his kind gui-dance throughout this study.REFERENCES1. AURER-KoZELJ, J.: Zytologishe Unterschiede im Prozess der Keratinisierung des Ober-fHichenepithels der gesunden und pathologisch vedinderten Gingiva. Dtsch. Zahn-Mund-Kieferhk. 48, 446, 19672. FUKUI, T. and K. MAKI: Experimental study on postexodontic wound healing. j.japan. Stomat. Soc. 7, 430, 1958 (in japanese)3. HAHN, W.: Beobachtungen bei der Heilung von Extraktionswunden. Dtsch. Zahn-Mund-Kieferhk. 28, 114, 19584. HIDAKA, T.: Oral cytology in case of leprosy (I) Normal exfoliative cytology. Okayama1. Z. 72, 45, 1960 (in japanese)5. MILLER, S. C. et al.: A study of the cornification of the oral mucosa of young maleadults. j. Dent. Res. 30, 4, 19516. MONTGOMERY, P. W.: A study of exioliative cytology of normal human oral mucosa.j. Dent. Res. 30, 12, 19517. MATSUI, T.: Normal exfoliative cytology of the oral cavity. Okayama I. Z. 71, 6089,1958 (in japanese)8. SUZUKI, A. et al.: Histological studies on changes in the oral mucosa according to theage. (1) j. japan. Stomat. Soc. 5, 365, 1956 (in japanese)9. SHIRAISHI, Y.: Exfoliative cytology of the mouth washings and gurglings, (I) Exfolia-tive cytology of normal oral cavity. Okayama I. Z. 71, 6059, 1958 (in japanese)10. WEINMANN, j.: The keratinization of the human oral mucosa, j. Dent. Res. 19, 57,194011. WATANABE, Y.: The effect of roentgen irradiation upon the cellular elements in thehuman saliva. Oral Surg., Oral Med. and Oral Path. 4, 89, 195112. WATANABE, Y.: Exfoliative cytology of the oral cavity with special reference to cyto-logical diagnosis of oral cancer recent advance in oral surgery (II ed.). Ishi-Yaku Pub-lish. Co., Tok)'o, pp 131, 1957 (in japanese)13. WATANABE, Y.: Exfoliative cytology of the oral cavity. j. japan. Stomat. Soc. 8, 371,1959 (in japanese)14. WATANABE, Y. and M. TOMOZAWA: Experimental study on exfoliative cytology ofposte.wdontic wound healing. j. japan. Stomat. Soc. 12, 176, 1963 (in japanese)15. WATANABE, Y.: RevieN of the literature of oral cytology. Dent. Outlook. 31, 321, 1968(in japanese)9Nishijima and Kiguchi: Exfoliative cytologic study on normal children's oral cavity andProduced by The Berkeley Electronic Press, 1970

Exfoliative cytologic study on normal children's oral cavity and their postexodontic wound healing

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