TGF-beta1 enhances the expression of alpha-smooth muscle actin in cultured human pulpal fibroblasts: immunochemical and ultrastructural analyses.

Transforming growth factor-beta 1 (TGF-beta1) has been related to induce the expression of alpha-smooth muscle actin (alpha-SMA) in fibroblasts during repair. Because pulpal fibroblasts seem to be somewhat different from other fibroblasts, the present study investigated in vitro whether TGF-beta1 enhances the expression of alpha-SMA in human pulpal fibroblasts. TGF-beta1 was added in doses between 5-10 ng/mL to cultures of both dental pulp and gingival human fibroblasts. The expression of alpha-SMA was analyzed by immunofluorescence and Western blotting, whereas the ultrastructure was evaluated by electron microscopy. In addition, the expression of tenascin, osteonectin, and vimentin was also investigated. Both cell types were immunoreactive for alpha-SMA even without TGF-beta1. When TGF-beta1 was added to cell cultures, the expression of alpha-SMA increased dramatically in pulpal fibroblasts, independent of the concentration used. It was confirmed by the Western blotting analysis. Ultrastructure revealed myofilaments and indented nuclei in both fibroblasts treated with TGF-beta1. Tenascin and osteonectin were only immunolabeled in pulpal fibroblasts treated or not with TGF-beta1. Both fibroblast types were positive for vimentin. The present findings showed that TGF-beta1 up-regulated the expression of alpha-SMA, thus inducing pulpal fibroblasts to acquire the myofibroblast phenotype.

Simple salting-out method for DNA extraction from formalin-fixed, paraffin-embedded tissues.

The aim of this study was to standardize a method of DNA extraction from formalin-fixed and paraffin-embedded tissues (PETs) using a salt solution to precipitate protein and isopropanol to precipitate DNA. The samples were submitted to a DNA extraction method in which two different concentrations of ammonium acetate (2 and 4M) were compared with a phenol-chloroform extraction method and with a commercial DNA isolation kit. DNA was qualified and quantified by spectrophotometer analysis, electrophoresis, and amplification by PCR. The 167 and 268bp fragments of APC and beta-globin genes, respectively, were amplified equally from DNA extracted by all tested methods and in all cases. However, the 536bp fragment of beta-globin gene was not amplified in all cases. According to our results, the extraction method using ammonium acetate proved to be simple and suitable for obtaining DNA of good quality, which can be easily amplified by PCR.

Oral squamous cell carcinoma: an analysis of 1,564 cases showing advances in early detection.

The present study aimed to detect possible differences in the data of oral squamous cell carcinomas (OSCC) cases diagnosed in a Dental School in São Paulo city over the last 40 years. The records of patients diagnosed as having OSCC between the years 1960 and 2008 were retrieved. The whole period was divided into four time periods. A total of 1,564 cases were reviewed. The variables analyzed were: sex, age, race, anatomical site, lesion duration, and lesion size. The chi-square test was used for statistical analysis. Overall, males were more affected than females (3:1), but when comparing the first and last time periods, the ratio decreased significantly (5.8:1 to 2.8:1). A significant increase in the rate of OSCC in patients over 80 years was observed in the last time periods. The gingiva was the most affected site, but the frequency of lower lip involvement increased in the last time period. Regarding lesion size and duration of symptoms at the time of diagnosis, there was a significant difference between the first and last time periods. Smaller lesions were found and the time of lesion development was shorter in the last few years of the study. These findings support the optimistic view that, in recent years, earlier diagnosis has resulted from early oral cancer detection in São Paulo city.

Intraosseous myofibroma simulating an odontogenic lesion.

Myofibroma is a rare benign nodular tumor of the soft tissues, bones, or internal organs, and may affect both children and adults. It is mostly found in the head and neck region, although uncommon in the jaw bones, where only a few lesions have been described. Radiographically, it may appear as a well-defined unilocular radiolucent entity simulating a cystic or odontogenic lesion. Histopathologic findings alone may be insufficient for a final diagnosis, and immunostaining for markers such as vimentin, alpha-smooth muscle actin, desmin and S-100 may be required. Intraosseous myofibroma is a diagnostic challenge and requires careful histopathologic, immunohistochemical and radiographic correlation. The present article describes a conservatively treated intraosseous myofibroma that occurred in a 7-year-old girl. The clinical, radiographic, histopathologic and immunohistochemical features are discussed.

Altered cytokeratin expression in actinic cheilitis.

BACKGROUND: Actinic cheilitis (AC) is a widely recognized precancerous lesion of the lip. Varying degrees of epithelial dysplasia may be present. However, no studies have correlated epithelial changes with cytokeratin expression that might reflect the disordered maturation that is probably occurring. METHODS: Thirty-four cases diagnosed as AC were classified according to dysplasia degree, and submitted to immunohistochemical staining for the detection of cytokeratins (CKs) 7, 8, 13, 14, 16 and 19. Normal mucosa adjacent to the lesions was also evaluated. RESULTS: The results obtained showed that CK10 immunostained only superficial keratinized epithelial layers in 11 cases, and also intermediate spinous layers in 18 cases. Cytokeratin 14 was expressed in all epithelial layers of 31 cases, in two cases its expression was in the basal and intermediate layers, and one case was negative. Cytokeratin 13 immunostained 26 cases and was negative in eight cases. In these eight cases, CK13 was apparently replaced by CK16. Cytokeratin 16, besides these eight cases, was also expressed in the spinous intermediate layers of a further eight cases. The remaining CKs tested were all negative. No relation between the degree of dysplasia and the CK expression was noted. CONCLUSIONS: Cytokeratin expression in AC is different from that of normal oral mucosa, and is not related to the degree of dysplasia.