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Revista chilena de anatomía

versión impresa ISSN 0716-9868

Rev. chil. anat. v.16 n.1 Temuco  1998

http://dx.doi.org/10.4067/S0716-98681998000100006 

CHARACTERISTICS OF THE PALATINE MUCOSA OF THE Calomys callosus: LIGHT AND SEM STUDY

CARACTERISTICAS DE LA MUCOSA PALATINA DEL Calomys callosus: ESTUDIO A LOS MICROSCOPIOS DE LUZ Y ELECTRONICO DE BARRIDO

* Marino, J. C.
** Watanabe, I.
*** Lopes, R.A.
*** Mizusaki, C.I.
*** Iyomasa, M.M.
*** Semprini, M.

Part of this paper presented as fullfilement of Master of Science Degree* to the School of Medicine, Federal University of São Paulo, Brazil
* Deparment of Anatomy, Faculty of Medicine University of Alfenas, MG - Brazil
** Departmentof Anatomy, Institute of Biomedical Sciences, University of São Paulo - Brazil.
*** Department of Stomatology, School of Dentisty of Ribeirão Preto- USP - São Paulo - Brazil.

SUMMARY: The characteristics of the surface and the interface epithelium-connective tissue of the hard palatine mucosa of Calomys callosus were studied using light and scanning electron microscopy. Six animals were fixed in Bouin and modified Karnovsky solution. The results obtained revealed that the anterior and medium regions contain the transversal plicae and adjacent flat surfaces. The surface shows squamous queratinized epithelium. The lamina propria of palatine mucosa present numerous connective tissue papillae of several forms. In the near region to the teeth, the connective tissue papillae are more numerous. The transversal plicae measured at 150 a 300µm in diameter. The surface of hard palatine mucosa of Calomys callosus revealed the presence of poligonal epithelial cells. Some desquamating cells are clearly noted. The surface of interface of epithelium-connective tissue presented numerous primary and secundary connective tissue papillae. The network of thick bundles of collagen fibers are noted in each papilla, demonstrating in a three-dimensional images.

KEY WORDS: 1. Palatine mucosa; 2. Calomys callosus; 3. Anatomy; 4. SEM.

INTRODUCTION

There are several kind of epithelial cells in the oral mucosa, surch as lingual and gingival mucosa, hard and soft palate, and sublingual mucosae.

KUTUZOV and SICHER (1953) verified the structure of the intermolar gingiva; SOGNNAES and ALBRIGHT (1958), in the queratinized and desquamated epithelial cells; BRODY (1959) the process of queratinization of the epithelial cells.

On the other hand, OSMANSKI & MEYER (1967); SQUIER (1968) studied the fine structure of the queratinized epithelium in the several regions of the oral mucosa. However, MEYER and SCHROEDER (1975); APPLETON & TYLDESLEY (1971); and McMILLAN (1974) revealed the characteristics of the stratum basale, spinosum and granulosum of oral mucosa. DALLE et al. (1977) using transmission electron microscopy studied the aspects of queratinized epithelial cells. BEZERRA (1979) reported the presence of tonofilaments, mitochondrias, keratohialin granules, desmossomes and intercellular junctions of epithelial cells.

The purpose of this paper is to show the characteristics of the interface epithelium-connective tissue of the palatine mucosa of Calomys callosus using light and scanning electron microscopy.

MATERIAL AND METHOD

In this paper were used six palatine mucosa of Calomys callosus, fixed in Bouin solution and modified Karnovsky solution.

For light microscopy, the specimens were decalcified in a solution containing 20% of sodium citrate and 50% of formic acid (MORSE,1945) during 12 days at room temperature. The tissue were rinsed, dehydrated in a increasing series of ethanol and embedded in parafin. The section of 10mm thick were monted and stained with hematoxilin-eosin and Azo-carmim. For scanning electron microscopy, four palatine mucosae of Calomys callosus were fixed in modofied Karnovsky solution, containing 2.5% glutaraldehyde, 2% paraformaldehyde in 0.1M (pH 4) in sodium phosphate buffer, at 4C. Then, the tissue were treated in 5% NaOH solution, during 2-4 days, at room temperature, according to the report described by OHTANI (1987).

The tissues were rinsed and postfixed in 1% osmium tetroxide solution, during 2h at 4C. Rinsed again in distilled water during 5h, imersed in 1% tannic acid and dehydrated in ethanol. The samples were dhied in critical point Balzers, SCD- 030 with CO2 liquid, coated with gold and examined in scanning electron microscopy JEOL, JSM- 6100 and JSM- 5300.

RESULTS

The anterior and medium palatine mucosae of the Calomys callosus presented a stratified queratinized epithelium, containing the pointed and flat areas of connective tissue (Fig.1). The interface epithelium-connective tissue present papillae of different forms. There are numerosus primary and secondary papillae. The lamina propria of this anterior region showed a delimited connective tissue papillae.

The medium region of hard palatine mucosa of Calomys callosus contains a epithelial layer showing stratified queratinized epithelial cells (Fig.2). The different epithelial layers are clearly showed containing the cytoplasm of cells. The lamina propria is flat, and there are few small projections. The projections are concentrated in the primary plica. The small papillae are noted in the connective tissue papilla, fibroblasts and small capillaries (Fig.3).

In the SEM observations the anterior region of palatine mucosa of Calomys callosus revealed the transversal plicae (Fig.4). The adjacent areas are flat and showed poligonal epithelial cells (Fig.5). The each elevation of mucosa measure form 150 to 300µm at the base.



 Fig. 1. Light microscopy image of the anterior portion of the palatine mucosa of Calomys callosus. Shows a pointed fomation of the mucosa (small arrow), horizontal portion (two arrows) of the epithelial layer and connective tissue (large arrow). X75.

The medium portion of hard palatine mucosa present primary plicae and flat adjacents surfaces (Fig.6). There are some irregular elevations in the lateral parts. Numerous desquamated epithelial cells are noted on the surface presenting three-dimensional SEM images (Fig.7).

In the samples treated by NaOH solution may note the original characteristics of the presence of different sizes of papillae. The surface of inter-face epithelium-connective tissue revealed transversal primary projection layerof connective tissue papillae (Figs. 8 and 9) and flat surfaces of adjacents areas. At high magnification, may noted the flat surface revealing the processure of collagen fiber bundles (Fig.10). In the medium portion of palate, the primary papillae and secondary papillae are numerous. The primary papillae are located parallely and small irregular projetions are noted in the three-dimensional charac-teristics. Between the primary papillae revealed a flat surface with the network of collagen fibers.



Fig. 2. Light microscopy image of the medium portion of the palatine mucosa of the Calomys callosus. Shows the epithelial layer, connective tissue papillae (small arrow), periodontal membrane (large arrow), and enamel surface (*). X80.



Fig. 3. Light microscopy image of the medium portion of the palatine mucosa of Calomys callosus. At high magnification, shows the superficial layer (small arrow), epithelial cells (two arrows), capillaries (large arrow), and connective tissue (three arrows). X190.


Fig. 4. SEM image of the anterior portion of the palatine mucosa of Calomys callosus. Shows the transversal plicae (*), round formations (arrows), and a flat surface with epithelial cells. X100.



Fig. 5. SEM image of the anterior portion of the palatine mucosa of Calomys callosus. Shows a transversal plica (*), two pointed projections (large arrows), and numerous epithelial cells in desquamation (small arrows). X220.


Fig. 6. SEM image of the medium portion of the palatine mucosa of Calomys callosus. Shows transversal palatine plica (arrow), and flat surface of the mucosa (*). X 110.



Fig. 7. SEM image of the medium portion of the palatine mucosa of Calomys callosus. Shows the transversal plicae (large arrows), and numerous epithelial cells in desquamation . X220.


Fig. 8. NaOH treated specimen of the interface epithelium-connective tissue surface of the anterior portion. Shows a transversal plicae (arrows) revealing primary and secondary papillae, and a flat surface. X 110.


Fig. 9. NaOH treated specimen. SEM image of the medium portion of the hard palatine mucosa of Calomys callosus. The primary papillae corresponding to the transversal plicae and secundary papillae are noted in three-dimensional images. X100.


Fig. 10. NaOH treated specimen. SEM image, at high magnification, showing a surface with several secundary connective tissue papillae (arrows). X600.

DISCUSSION

Our data demonstrated the structural characteristics of the palatine mucosa of the Calomys callosus, using the method of the light and scanning electron microscopy. The observations showed that the anterior and medium regions are covered with stratified squamous queratinized epithelium. Similar observations were made by COHEN (1967) in the oral mucosa of the Macaca irus; MARTINEZ et al. (1995) in the palatine mucosa of the tatu galinha Dasypus novemcinctus. The queratinized area in the oral mucosa are the palatine, gengival and lingual mucosa and in the mouse, the queratinization extend for almost all areas according to the data reported by WEIMANN (1940).

HAYWARD (1973) reported that the oral mucosa of the rats has a active queratinization. Our data confirm that the hard palatine mucosa of the Calomys callosus present a queratinization which may compared with those observed in the mouse.

Our histological aspects of the lamina propria demonstrated the presence of large connective tissue papillae of several forms and the bundle of collagen fibers.

In the cornefied layer of palatine mucosa revealed numerous poligonal epithelial cells and some in desquamation. The anterior and medium regions of palatine mucosa have poligonal epithelial cells well delimited.

In the treatment of the palatine mucosa of Calomys callossus in NaOH solution, permited to remove the epithelial layer and may visualize lamina propria in original localization. The structures of the lamina propria in our results are noted similar appearance those showed by several authors (KOBAYASHI et al., 1987; OHTANI,1987; WATANABE et al., 1995).

Although, the utilization of the technique of the maceration with NaOH solution, at room temperature, obtained good characterization of the surface of the lamina propria, with three-dimensional image of the primary and secundary papillae.

The characteristics of the connective tissue papillae examined by SEM demonstrated the detail of ultrastructure, identifying the large and small connective tissue papillae.

RESUMEN: Se estudiaron las características de la superficie e interfase del tejido epitelio-conjuntivo de la mucosa palatina del paladar duro del Calomys callosus, a través del microscopio de luz y de la microscopía electrónica de barrido. Seis ejemplares fueron fijados en soluciones de Bouin y de Karnovsky modificada. Los resultados mostraron que las regiones anterior y media presentan pliegues transversales y superficies planas adyacentes. La superficie mostraba un epitelio escamoso queratinizado. La lámina propia de la mucosa palatina presentaba abundante tejido conjuntivo papilar, de variadas formas. Próximo a la región de los dientes, el tejido conjuntivo papilar era más abundante. El pliegue transversal medía entre 150 µm y 300µm de diámetro. La superficie de la mucosa palatina del paladar duro del Calomys callosus presentaba células epiteliales poligonales. Algunas células descamativas se notaban claramente. La superficie de la interfase del tejido epitelio-conjuntivo presentaba abundante tejido conjuntivo papilar primario y secundario. La red de gruesos haces de fibras colágenas se observaban en cada papila, mostrando imágenes tridimensionales.

PALABRAS CLAVE: 1. Mucosa palatina; 2. Calomys callosus; 3. Anatomía; 4. SEM.

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Dirección para correspondencia:
Prof. Dr. José Carlos Marino
Departamento de Anatomia
Faculdade de Ciências Médicas
Universidade de Alfenas
37130-000
Alfenas - MG
BRASIL

Recibido : 18-12-1997
Aceptado : 30-01-1998

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