Expression of estrogen receptor (ER) in oral mucosa and salivary glands.

OBJECTIVES: To assess the expression of estrogen receptor (ER) in oral mucosa and salivary glands, buccal mucosal biopsies from ten postmenopausal women (taken before and during the hormone replacement therapy), as well as, single biopsies from 20 healthy 19-year-old women were analyzed for ER expression. Salivary gland biopsies were taken from the minor labial salivary glands (n=6), submandibular glands (n=5) and parotid gland (n=1) from women at different ages. METHODS: total RNA extracted from the tissue samples was reverse-transcripted (RT) to single-stranded cDNA, and the RT-polymerase chain reaction (RT-PCR) product was subjected to nucleotide sequencing to confirm the match with ER cDNA. Immunohistochemistry (IHC) with a monoclonal ER antibody (ER-ICA, Abbott) and Western blot analysis with monoclonal antibody against ER-related antigen (ER-D5, Amersham) were performed on the biopsies taken from the postmenopausal women. RESULTS: ER mRNA was expressed in 18/20 (90%) and 20/20 (100%) of the mucosal biopsies in the postmenopausal and 19-year-old women, respectively. The expression of mRNA was detected in all the submandibular gland samples, in the single parotid gland, as well as, in 4/6 (67%) of the labial glands. ER expression could not be detected by IHC, indicating either a very low level of expressed protein or difficulties in recognizing the epitopes by IHC. However, Western blot demonstrated 8/8 (100%) of the mucosal biopsies of postmenopausal women positive for ER-related antigen. CONCLUSIONS: the presence of ER mRNA and immunoreactive ER protein suggests that estrogens have a biological role in oral mucosa and salivary glands.

The effect of hormone replacement therapy on salivary calcium concentrations in menopausal women.

Hormone replacement therapy in menopausal women is known to affect the general calcium turnover of the body. No information is available about the effects of hormone therapy on salivary electrolytes and on calcium in particular. A group of 16 healthy peri- and postmenopausal women, all recommended to start hormone replacement therapy, were studied longitudinally for 5 months. Paraffin-stimulated whole saliva was collected at baseline, at 3 and at 5 months after the onset of therapy, and analysed for calcium, sodium and potassium concentrations. In response to hormone replacement therapy, calcium concentration decreased (p = 0.037), that of sodium increased (p = 0.019), while no change was observed in the potassium concentrations during the follow-up period.

Oral ulcers in AIDS patients frequently associated with cytomegalovirus (CMV) and Epstein-Barr virus (EBV) infections.

Oral ulcers are common in AIDS patients, with a wide spectrum of underlying causes, including different viruses. In the present study, the presence of cytomegalovirus (CMV), Epstein-Barr virus (EBV) and human herpesvirus-8 (HHV-8) DNA was analysed in 21 biopsies from oral ulcers of 17 male homosexual AIDS patients. The methods used were in situ hybridization (ISH) and the polymerase chain reaction (PCR) with subsequent non-radioactive Southern blot hybridization to confirm the specificity of PCR products. With ISH, 4 biopsies were CMV DNA-positive and 11 contained EBV-DNA. Using PCR, an additional 4 CMV- and 7 EBV-positive samples were detected, and HHV-8 DNA was present in three oral ulcers. Six of the patients (35%) had oral ulcers co-infected by two or three viruses. The overall figures for patients with the detectable EBV-, CMV-, and HHV-8 DNA were 82% (14/17), 35% (6/17) and 18% (3/17), respectively. This is the first study to show the frequent presence of EBV-DNA in oral ulcers of AIDS patients. Because ISH-positivity signifies active virus replication, these results implicate an etiological role of EBV in AIDS-associated oral ulcers. The causal role of HHV-8 has to be considered as well, because this virus was detected in three such ulcers, which were not associated with Kaposi's sarcoma. To conclude, three common members of the herpesvirus family (CMV, EBV, HHV-8) were detected in all but three ulcers in AIDS patients, warranting the inclusion of these viral analyses in the diagnosis of ulcerative lesions of the oral mucosa in all immunosuppressed individuals.

The effect of repeated sampling on paraffin-stimulated salivary flow rates in menopausal women.

Paraffin-stimulated whole-saliva samples of 12 post- and perimenopausal women were taken five times over a 7-week period. After 1 min of prestimulation, saliva was collected for 5 min under standardized conditions. Saliva was first collected on three consecutive days, and two additional samples were collected 6 and 7 weeks after the first. Salivary flow rates showed significant variation between samplings. The greatest difference appeared between the first and second collections. The increase in flow rates between these collections was significant (p = 0.003). The third successive collection also gave a significantly (p = 0.005) higher flow rate than the first. The 6-week and baseline samples were similar. The 7-week sample showed a significantly higher flow rate (p = 0.018) compared with the baseline value. These results stress the importance of standardization of sampling intervals when salivary flow rates are studied.

Effect of snuff and smoking on tenascin expression in oral mucosa.

We have shown, by using two monoclonal antibodies (143DB7 and 100EB2), that the expression of the extracellular matrix protein tenascin (Tn) is increased in the connective tissue of biopsies taken from snuff users' and tobacco smokers' oral mucosa. In normal oral mucosa Tn was seen to underlie the epithelium as a thin delicate band. The most increase in Tn reaction was observed in snuff users' mucosa while the immunoreaction in smokers' mucosa was less conspicuous. Often the most prominent Tn reaction took place in association with round cell inflammatory infiltration, indicating epithelial irritation. Tn has been shown to take part in epithelial-mesenchymal interactions during embryogenesis, wound healing and tumorigenesis. Here, a superficial epithelial irritant has been shown to cause conspicuous alterations not only in the epithelial cell layers but also in the underlying connective tissue by increasing its Tn content. As a result of our findings we suggest a further link for Tn in a dynamic epithelial-mesenchymal interplay by virtue of this marked connective tissue reaction in snuff users' and smokers' oral mucosa.

Hormone replacement therapy and some salivary antimicrobial factors in post- and perimenopausal women.

OBJECTIVES: The effect of hormone replacement therapy (Cyclabil) on non-immunoglobulin (peroxidase) and immunoglobulin (total IgA, IgG, IgM) antimicrobial factors as well as on total protein and microorganisms in whole saliva was assayed in 19 postmenopausal and 8 perimenopausal women. METHODS: Paraffin-stimulated whole saliva was collected before as well as 3 and 5 months after the onset of the treatment. Time- and group-related differences between post- and perimenopausal women were analyzed. RESULTS: Peroxidase and total protein output per min increased significantly (P = 0.004 and 0.001) during the treatment in both groups. No significant time- or group-related differences in the mean concentrations of the respective variables were found. The mean concentrations of salivary IgA and IgG showed a significant time-related decrease in both groups (P = 0.012 and 0.010). Salivary IgM concentration in perimenopausal women also showed a significant time-related decline (P = 0.017) and the difference in changes of salivary IgM between the two groups was significant (P = 0.033). Total IgA output per min increased in perimenopausal whereas it decreased in postmenopausal women (interaction; P = 0.021). Hormone treatment had no effect on the amount of salivary bacterial floras. CONCLUSIONS: The composition of saliva in post- and perimenopausal women was found to be estrogen-dependent. The second finding was that all women participating in the study reported a sense of enhanced oral well-being including relief of oral dryness.

Estrogen response in buccal mucosa -- a cytological and immunohistological assay.

OBJECTIVES: The aims of the present study were firstly, to evaluate the effect of estrogen on the cytology of buccal mucosa during the menstrual cycle and at menopause, and, secondly, to analyze the presence of estrogen receptors in the epithelial cells of buccal mucosa using immunohistochemical methods. METHODS: The cytological samples of buccal mucosa from 10 healthy young women (mean age 24 years) were taken on every day of the menstrual cycle and stained according to Papanicolaou. A single cytological sample of buccal mucosa was also collected from 20 healthy postmenopausal women (mean age 66 years) using no hormone replacement therapy. For each specimen, a maturation index was calculated on the basis of percentages of the three (parabasal, intermediate and superficial) cell types. Moreover, 50 incisional biopsies were taken from buccal mucosa of 50 young healthy women (19 years old) for analysis of estrogen receptor expression by immunohistochemistry using an indirect immunoperoxidase technique. RESULTS: On cytohormonal evaluation, intermediate cells were dominant in postmenopausal women and also in young women at all stages of the menstrual cycle indicating a nearly complete cell maturation. Estrogen receptor positive cells were not detected in buccal epithelium by immunohistochemistry. CONCLUSIONS: As the cell pattern of buccal mucosa indicated a nearly complete cell maturation both in young and in postmenopausal women, estrogen may not be the only factor causing maturational changes in buccal epithelial cells. Alternatively, the antibody used in immunohistochemistry may not detect the epitope of the estrogen receptor present in buccal mucosa or the level of expression of this protein may be under the detection limit.

Effect of hormone replacement therapy on salivary flow rate, buffer effect and pH on perimenopausal and postmenopausal women.

The effects of hormone replacement therapy (HRT) on salivary gland functions were examined in a longitudinal study. The flow rate, buffer effect and pH of paraffin-stimulated whole saliva were analysed in 8 perimenopausal and 19 postmenopausal women. The saliva samples were collected 1-2 days before as well as 3 and 5 months after the start of HRT. Salivary flow rates were higher (p <0.033) in the perimenopausal than in the postmenopausal group. The flow rates increased significantly (p <0.001) during HRT in both groups. Similarly, buffer effect and pH showed significant (p = 0.004 and p = 0.009, respectively) time-related changes without any differences between the groups. It appears that HRT improves both the quantity and the quality of salivary gland function in peri- and postmenopausal women.

Lack of effect of hormonal treatment and smoking on cytokeratin expression in buccal mucosa.

Differences in cytokeratin expression of clinically normal buccal mucosa were studied in 50 healthy women by indirect immunofluorescence staining with monoclonal antibodies. The subjects were divided into four groups: control group (N = 18), smokers (N = 8), oral contraceptive users (N = 8) and smokers receiving oral contraceptives (N = 16). Our findings indicate that cytokeratin expression in noncornified stratified epithelium is not influenced by smoking or oestradiol/progesterone treatment. Only cytokeratin No. 19 showed variable patterns of expression but the differences could not be ascribed to smoking or contraceptives. Cytokeratin No. 19 gave a positive reaction in the basal and suprabasal layers in 34 subjects (68%). In 9 (18%) specimens, the staining was positive in the basal cells and showed a positive heterogeneous cytoplasmic reaction in the suprabasal cells. Interestingly, cytokeratin No. 7 was recognized in all epithelial cells except the basal cells. Our results suggest that changes in the serum oestradiol levels do not affect the cytokeratin pattern in buccal mucosa.

ITI titanium plasma-sprayed screw implants in the treatment of edentulous mandibles: a follow-up study of 39 patients.

The outcomes of 153 ITI titanium plasma-sprayed screw implants were studied in 39 patients. Four implants were placed in the edentulous mandible between the mental foramina and used as support for a denture prosthesis. Thirteen implants in six patients had been lost during the follow-up time of 3 to 10 years (mean 5.6 years). The mean annual bone resorption was 0.25 mm (SD 0.29 mm; range 0 to 1.37 mm). Bone loss was smaller in patients with prosthetic loading applied within 7 postoperative days than in those with delayed prosthetic treatment. No evidence of bone loss was seen around 37 (26.4%) implants. The cumulative and overall success rates for titanium plasma-sprayed implants were 80.8% and 91.5%, respectively. The corresponding figures for the patients were 86.8% and 94.9%.

Cytomegalovirus (CMV) and Helicobacter pylori (HP) found in oral mucosal ulcers.

The possible involvement of Cytomegalovirus (CMV) and Helicobacter pylori (HP) in oral mucosal ulcers is suggested by their role in the development of ulceration at other mucosal sites of the gastrointestinal tract. A series of 29 incisional biopsies from 29 consecutive and apparently immunocompetent patients attending the clinic for oral ulceration were examined by routine histopathology as well as by in situ hybridisation (ISH) with biotinylated CMV and HP DNA probes. In 14/29 biopsies, Giemsa staining disclosed spiral bacteria. Six (20.7%) of these 14 Giemsa-positive samples showed HP DNA on ISH and 3 ulcers (10.3%) contained CMV DNA. In none of the specimens were CMV and HP detected simultaneously. Two of the ulcers containing CMV DNA were found on the labial mucosa and one on the posterior palatal mucosa, whereas all HP DNA-positive ulcers were located on the buccal mucosa. The results indicate that CMV and HP DNA can be found in separate oral mucosal ulcers in apparently immunocompetent adults.

Comparison of periapical and detailed narrow-beam radiography for diagnosis of periodontal pathology.

The diagnostic performance of the detailed narrow-beam (DNB) technique of the Scanora multimodal X-ray system was compared with periapical radiography for detecting periodontal pathology. In total, 253 sites in 133 patients were examined. Receiver operating characteristics (ROC) of both imaging modalities were analyzed in relation to the ratings of five observers for the whole dentition, three dental regions and five types of periodontal lesion. ROC analysis demonstrated that the overall diagnostic performance of DNB radiography was better (P < 0.05) than that of periapical radiography. DNB radiography was significantly superior (P < 0.01) for detecting marginal widening of periodontal membrane space, but there was no significant difference between the two techniques for crestal erosion, vertical bone loss, furcation involvement or calculus. The regional differences found in overall diagnostic performance of the two imaging modalities were not significant. The sensitivity for periapical radiography was 71% and for DNB radiography 85%, and the specificities 82% and 81% respectively. It is concluded that DNB radiography is a good radiographic examination for periodontal disease, and an acceptable alternative to periapical radiography.