Accessible sialic acid content of oral epithelial cells from healthy and gingivitis subjects.

The purpose of the present study was to determine if there were differences in the quantity of accessible sialic acid on superficial epithelial cells collected from different areas of the mouth, and from healthy subjects with good oral hygiene, as compared to subjects with gingivitis. Superficial epithelial cells were collected by gently scraping the tongue dorsum, hard palate, free gingiva and buccal epithelium. The cells were washed and treated with clostridial neuraminidase to release accessible sialic acid; this was quantitated using a fluorometric assay. Buccal cells released an average of 62.6 ng sialic acid per 10,000 cells, which was nearly 3-fold more than cells from the hard palate (24.1 ng), free gingiva (21.9 ng), or tongue (15.4 ng). Buccal and free gingival cells collected from 5 healthy subjects had significantly higher levels of accessible sialic acid on their surface than cells collected from 5 subjects with gingivitis. These differences were significant at the p less than 0.001 and p less than 0.01 levels, respectively. The data obtained suggest that the oral hygiene status of an individual can influence the quantity of accessible sialic acid residues on oral epithelium; this would be expected to influence the attachment and colonization of bacteria which bind to sialic acid-containing receptors.

The antiperinuclear factor. I. Clinical and serologic associations.

The antiperinuclear factor (APF) test raises two main problems: the unpredictability of the cells used as substrate and the difficulty in expressing the results. We propose that 10% of the cells have to be stained by a given serum in order for it to be considered positive. APF were found to be present in 76% of rheumatoid arthritis (RA) patients, 3% of healthy controls and occasionally in disease controls. The production of APF was significantly (p less than 0.01) related to the presence of rheumatoid factor in RA, and IgG antibody was predominant in the APF test.

The antiperinuclear factor. II. Variability of the perinuclear antigen.

Since the antiperinuclear factor (APF) test on human buccal cells is rather unpredictable, we have investigated the possible factors determining the expression of appropriate antigens by the cells. We failed to find any relationship of the expression of perinuclear antigens to the donor's smoking habits, the degree of contamination with saprophytic bacteria, the presence or absence of blood group substances in saliva, or the titers of serum antibodies to Epstein-Barr virus. Family studies were also performed to further elucidate a genetic predisposition to the expression of the APF antigen.

Fatty acid composition of clinically healthy and malignant oral epithelium.

The fatty acid composition of epithelium from clinically healthy oral mucosa and or oral squamous cell carcinoma was analyzed by gas/liquid chromatography following extraction using a modified Folch technique and conversion of fatty acids to methyl esters. There were significant reductions in the relative proportions of palmitoleic and oleic acids and elevation of the relative proportions of palmitic, steric and arachidonic acids in samples from squamous cell carcinoma. These differences could reflect changes in cell membranes and/or fatty acid metabolism. Further studies are required to assess their functional, diagnostic and prognostic significance.

RAS oncogene product expression in normal and malignant oral mucosa.

Proto-oncogenes are important in both normal cellular differentiation and in carcinogenesis. The majority of transforming genes belong to the ras family and the ras gene product has been shown to be elevated in some oral carcinomas. RAP-5 monoclonal antibody was used to determine the expression of the p21ras protein in normal and neoplastic oral mucosa in an immunohistological study. The expression of p21ras protein was generally restricted to acanthous cells with strong staining in normal oral mucosa and well-differentiated carcinomas. In contrast, the p21ras protein was not detected in significant amounts in severely dysplastic lesions and poorly differentiated carcinomas. These results suggest that expression of p21ras is a normal feature of more fully differentiated tissues, both normal and neoplastic, and is not useful as an indicator of cell proliferation or 'malignant potential'.

Endogenous levels of beta-carotene in human buccal mucosa cells by reversed-phase high-performance liquid chromatography.

We have developed a reversed-phase high-performance liquid chromatographic assay for the measurement of low nanogram levels of beta-carotene in a single sample of human buccal mucosa cells. The method includes a simple sonification step for cell disruption and release of the compounds into the supernatant. The limits of detection were 0.02, 0.02 and 0.07 ng/mg of protein for beta-carotene, retinol and retinol palmitate, respectively. Two patient populations were analysed. Average endogenous levels for beta-carotene normalized to protein were 0.25 ng/mg of protein (range 0.04-1.9 ng/mg, twelve patients). No evidence of endogenous retinol or retinol palmitate could be detected in the human samples. An oral dosing study of four normal individuals showed a wide variation of beta-carotene uptake. This rapid and sensitive method will enable investigators to use the non-invasive technique of buccal mucosa cell harvesting to determine cellular depot levels of beta-carotene in various patient populations.

Hemoglobin concentration and oxygen saturation of clinically healthy and inflamed gingiva in human subjects.

The hemoglobin concentration (Hb index) and oxygen saturation (apparent SO2) in human gingiva were estimated by tissue reflectance spectrophotometry (TRS). The gingiva had significantly lower Hb index and higher apparent SO2 than those in alveolar mucosa, but there was no difference in either parameter among different gingival areas. The reproducibility in repeated measurements was high for both Hb index and apparent SO2 in gingiva. In inflamed gingiva, Hb index was significantly higher than that in clinically healthy gingiva. A lower apparent SO2 was observed in inflamed gingiva. This suggests that the increase in blood supply is insufficient to meet the oxygen demand in inflamed gingiva. There were significant correlations between either the Hb index or the apparent SO2 and the clinical parameters of gingival inflammation such as gingival index, plaque index, Periotron score and probing depth. Thus, TRS may be clinically available to estimate the blood volume and oxygen saturation in inflamed gingiva.

Buccal mucosa fibroblasts and periodontal ligament cells perturbed by tensile stimuli in vitro.

Human buccal mucosa fibroblasts and periodontal ligament cells grown in tissue culture were subjected to tensile forces approximating those used for orthodontic bodily tooth movement. The cells were synchronized into pre S phase and positively tested for response to nonmechanical physical stimuli. Two-dimensional gel analysis and immunohistochemical analysis of the three cytoskeletal components showed a lack of response. Similar negative results were found when the cells were perturbed in the presence of substance P. We hypothesize that perhaps these cells respond more readily to injury, a secondary effect of the forces of tooth movement, than to tensile forces.

Amalgam associated mercury accumulations in normal oral mucosa, oral mucosal lesions of lichen planus and contact lesions associated with amalgam.

Fourty-three patients with oral mucosal lesions were divided into 3 groups based on the relationship between lesions and amalgam restorations. Group I consisted of patients with contact lesions confined to mucosal areas in contact with amalgam fillings. Group II patients had lichen planus lesions exceeding the area of contact with an amalgam filling and Group III comprised patients with lichen planus lesions without relation to amalgam fillings. Biopsies were embedded in epon and subjected to autometallography in order to demonstrate a possible accumulation of mercury in the affected mucosa. In 20 our of 21 patients in Group I, 4 of 11 patients in Group II and 4 of 11 patients in Group III, mercury was found in the lysosomes of macrophages and fibroblasts. In Group I the number of cells loaded with mercury was much higher than in Group II and in particular Group III. In the latter groups autometallographically demonstrated mercury was found almost exclusively in macrophages. Nineteen biopsies taken from patients with normal mucosa served as controls. Ten had occlusal (Group IV) and seven buccal fillings (Group V). The biopsies from the latter group were taken from areas opposing amalgam restorations. Two patients had no amalgam fillings (Group VI). The histochemical technique showed that three biopsies in Group IV (occlusal fillings only) and two in Group V (opposing buccal fillings) contained traces of mercury in the juxtaepithelial connective tissue. The silver enhanced mercury was found in macrophages. The two controls (Group VI) without amalgam fillings were devoid of precipitates.(ABSTRACT TRUNCATED AT 250 WORDS)

Histochemical localization of hyaluronate in human oral epithelium using a specific hyaluronate-binding probe.

Biochemical data suggest that gingival epithelium contains hyaluronate, but there is little histochemical information about its localization. Hyaluronate was here visualized in gingival and buccal mucosa using a specific probe derived from the hyaluronate binding region of cartilage proteoglycan. Hyaluronate was found both in the gingival and buccal epithelium, but its localization was correlated with the type of keratinization. In the keratinized epithelium of gingiva, whether ortho- or parakeratotic, the intercellular spaces from basal to upper spinous layers displayed strong staining, most intense in the middle spinous cell layer. The uppermost vital cell layers as well as the cornified cell layer remained unstained. In the non-keratinized epithelium of buccal mucosa and the local non-keratinized areas of gingiva, only the basal cells and the lowermost spinous cell layers stained for hyaluronate, whereas the majority of the upper epithelium was negative. Electron microscopic examination of the basal and spinous cell layers displayed hyaluronate, both associated with the cell surface and free in the intercellular space. The subepithelial connective tissue showed positive but diffuse staining in all specimens.

Monoclonal antibody MAb C-50 in buccal mucosa cancer cells.

Staining patterns of cancerous and normal tissues from 20 cases of proven buccal cancer and ten control individuals respectively to monoclonal antibody MAb C-50 were studied. Nine of 20 patients and 4 of 10 control individuals showed positive immunofluorescence respectively. There was no correlation between clinical stage of disease and MAb C-50 staining nor any significant difference between the ulcerative or exophytic growth groups. One section with T4 lesion of buccal cancer with normal salivary tissue showed positive immunofluorescence of normal salivary cells. In our study the presence of receptor for MAb C-50 on buccal mucosa cancer cells was found to be statistically insignificant.

Ha-ras oncogene product in human oral squamous cell carcinoma.

The expression of Ha-ras oncogene product (Ha-ras p21) in the biopsy or operation materials derived from 70 oral squamous cell carcinomas, 10 oral leukoplakias and 10 normal oral mucosae was examined immunohistochemically using anti-Ha-ras p21 antibody. Ha-ras p21 was detected in 43 (61%) of 70 carcinomas, 3 (30%) of 10 oral leukoplaskias and 3 (30%) of 10 normal oral mucosae. Patients with Ha-ras p21-positive carcinomas had a significantly worse prognosis than those with Ha-ras p21-negative carcinomas. These observations strongly suggest that the expression of Ha-ras p21 is a common event in oral squamous cell carcinomas, and that Ha-ras p21 serves as a tissue tumor marker for determining the prognosis of patients with oral squamous cell carcinomas.

Dental and jaw changes in primary hyperoxaluria.

A case of teeth and jaw changes in primary hyperoxaluria is described. The patient, a 25-yr-old man, was treated by kidney transplantation twice, and finally by combined liver and kidney transplantation. The teeth and jaw changes consisted of deposition of oxalate crystals in the gingiva, in the pulp and in the vicinity of bone and dentin. The resorption of bone and dentin was extensive.

Metal release from plates used in jaw fracture treatment. A pilot study.

This study investigated the occurrence of corrosion associated with the use of metallic implants to stabilize jaw fractures. Three different types of plates, Co-Cr and Ni-Cr alloys and Titanium, were connected in vivo to the mandibular bone surface of monkeys (Cercopithecus aethiops). The animals were killed after 3 and 6 months. The mucous membrane and bone tissue were analysed for concentrations of Co, Cr, Ni, Mo, Al, and Ti by atomic absorption spectrophometry and a radiochemical neutron activation technique. With the exception of Ti, higher concentrations of all the above elements were found in the tissue near the implants when compared with contralateral controls. However, no signs of corrosion, macroscopic or microscopic, were observed on the surface of the implants.

Sex determination from human somatic cells.

The method and its reliability for sex determination by examining X-chromatin in the nucleus of hair root cell and buccal mucosa cell are presented: 1) X-chromatin in the nucleus of hair cortex cell was stained by Feulgen reaction after bleaching melanin granules and the frequency of X-chromatin was calculated under a ordinary microscope. An another method, fluorescent Feulgen reaction with acriflavine using a fluorescence microscope, was attempted. The results obtained from both methods indicated significantly higher values in the female samples than those in the male ones. 2) The other hand, the frequency of Y-chromatin was determined after quinacrine staining using a fluorescence microscope, and was also distinguishable between the male and female samples. 3) Using a single specimen, sex determination from the frequency of X- or Y-chromatin was practicable through combined treatment of quinacrine staining and fluorescent Feulgen reaction. 4) These methods were available during 16 to 64 weeks at least regarding samples kept dry. It is concluded that sex determination from frequency of sex chromatin after treatment described above is reliable.

Factor XIIIa-containing cells and fibrosis in oral and maxillofacial lesions: an immunohistochemical study.

The distribution of subunit A of blood coagulation factor XIII (FXIIIa) was investigated by the avidin-biotin-peroxidase complex (ABC) method in various oral and maxillofacial tissues. These tissues were from normal tongue, gingiva, lip, and submandibular gland, and from Dilantin gingival hyperplasia (one case), pyogenic granuloma (three cases), peripheral fibroma (four cases), squamous cell carcinoma (seven cases), chronic sclerosing submandibular adenitis (two cases), and fibrous dysplasia of the mandibular bone (one case). The distribution of collagenous components was examined in the same tissues by means of the Sirius red F3BA method. By means of the ABC method, FXIIIa was detected in the cytoplasm of certain connective tissue cells in each of the tissues examined. These FXIIIa-containing cells were sparse in the normal tissues but evidently abundant in the fibrous connective tissue of inflammatory and neoplastic lesions. In the present study, the close relationship between the distribution of FXIIIa-containing cells and that of collagenous components is demonstrated. The role that FXIIIa-containing cells play in the process of fibrosis is discussed.

Ultrastructure of cultured canine oral keratinocytes.

Keratinocytes from explants of the oral mucosa of dogs were grown in culture for five passages. The ultrastructure of primary cultures and fully developed subcultures passaged 1, 3, and 5 times was examined. At every stage, the cells had the morphologic characteristics of epithelial cells and formed a multilayered squamous epithelium. The basal cells had the characteristics of metabolically active cells, whereas the suprabasal cells and the cells at the media interface expressed many, but not all, of the organelles and cell surface characteristics associated with keratinocyte differentiation. Keratohyalin granules were located in the suprabasal and superficial cells. Cell size and shape and the relationship between cells in the layers also reflected the morphologic characteristics of the parent tissue. Cells maintained this typical structure through all passages and the cultures changed minimally for up to a week after development.

[Immunohistochemical localization of laminin and its relation to the grade of histological malignancy in oral cancer].

The immunohistochemical localization of Laminin has been investigated with a monoclonal antibody in biopsy specimens from 27 patients with an invasive squamous cell carcinoma of the oral cavity. In normal mucosa, laminin was found distributed in a linear pattern in the epithelial-stromal junction, around the blood vessels, the nerve fibres, and the skeletal muscle bundles. In the cancerous tissues a variety in the linear staining around the cancer cell nests was seen. Extreme attenuation or complete loss of the linear staining of laminin was demonstrated in 8/27 specimens, and these tumors histologically showed a high grade of malignancy with H.E. staining.

Automated Feulgen staining with a temperature-controlled staining machine.

A Shandon Varistain 24-3 staining machine was modified in order to run automated DNA Feulgen staining. Initial studies showed a strict dependence of the staining intensity (integrated optical density [IOD]) on the temperature of the DNA hydrolysis in 4 N HCl: a difference of 0.5 degrees C around the optimum hydrolysis temperature of 27.5 degrees C resulted in IOD differences of up to 7.8% in epithelial cells and up to 12.0% in lymphocytes. A temperature-controlled stainless steel cuvette, covered with a 4 N HCl-resistant material, was developed and integrated into the machine. Temperature measurements were performed at different positions in the cuvette and on glass slides with copper-constantan electrodes fixed on them; no temperature gradient could be detected within the cuvette. The adjusted temperature of 27.5 degrees C remained constant over 24 hours. The coefficient of variation (CV) of the staining intensity in lymphocytes between different areas on the same slide and between different slides of the same staining cycle was less than 0.6%. The CV between different staining cycles was 5.9%. This system for automated Feulgen staining thus gives reproducible and reliable results and may be introduced into routine diagnostic procedures.