Multiplexed blood-brain barrier organ-on-chip.

Organ-on-chip devices are intensively studied in academia and industry due to their high potential in pharmaceutical and biomedical applications. However, most of the existing organ-on-chip models focus on proof of concept of individual functional units without the possibility of testing multiple experimental stimuli in parallel. Here we developed a polydimethylsiloxane (PDMS) multiplexed chip with eight parallel channels branching from a common access port through which all eight channels can be addressed simultaneously without the need for extra pipetting steps thus increasing the reproducibility of the experimental results. At the same time, eight outlets provide individual entry to each channel with the opportunity to create eight different experimental conditions. A multiplexed chip can be assembled as a one-layer device for studying monocultures or as a two-layer device for studying barrier tissue functions. For a two-layer device, a ∼2 µm thick transparent PDMS membrane with 5 µm through-hole pores was fabricated in-house using a soft lithography technique, thereby allowing visual inspection of the cell-culture in real-time. The functionality of the chip was studied by recapitulating the blood-brain barrier. For this, human cerebral microvascular endothelial cells (hCMEC/D3) were cultured in mono- or coculture with human astrocytes. Immunostaining revealed a cellular monolayer with the expression of tight junction ZO-1 and adherence junction VE-cadherin proteins in endothelial cells as well as glial fibrillary acidic protein (GFAP) expression in astrocytes. Furthermore, multiplexed permeability studies of molecule passage through the cellular barrier exhibited expected high permeability coefficients for smaller molecules (4 kDa FITC-dextran) whereas larger molecules (20 kDa) crossed the barrier at a lower rate. With these results, we show that our device can be used as an organ-on-chip model for future multiplexed drug testing.

Immunohistochemical study of GLUT-1 in oral peripheral nerve sheath tumors.

AIM: To investigate the immunoexpression and diagnostic applicability of human erythrocyte-type glucose transporter protein (GLUT-1) in oral peripheral nerve sheath tumors. MATERIAL AND METHODS: Specimens diagnosed as oral peripheral nerve sheath tumors archived in the Oral Pathology Service of Universidade Federal de Minas Gerais from 1966 to 2006 were evaluated. Thirty-four lesions were included: 15 traumatic neuromas, 11 neurofibromas, four neurilemmomas, and four malignant peripheral nerve sheath tumors (MPNST). One case of neurofibroma was associated with neurofibromatosis type I. Immunohistochemistry for S-100 and GLUT-1 was performed. S-100 was immunopositive in all lesions. RESULTS: Benign lesions were immunopositive for GLUT-1 except in two (18.2%) cases of neurofibromas. In the traumatic neuroma, the perineuriums were immunopositive for GLUT-1. In the neurofibroma, the immunoreactivity was heterogeneous. Immunopositivity was observed at levels of 54.5% in the periphery of the lesion, 9.1% in the center, and 18.2% in both. The neurilemmoma demonstrated immunopositivity in the capsule. One case (25%) of MPNST presented GLUT-1 positive stain in occasional cells distributed homogeneously in all the tumor area. CONCLUSION: GLUT-1 is a useful marker for perineurial cells and should be included in the oral peripheral nerve sheath tumors immunophenotyping thus aiding in the correct diagnosis of these lesions.

Oral candidiasis treatment with Brazilian ethanol propolis extract.

The Brazilian commercial ethanol propolis extract, also formulated to ensure physical and chemical stability, was found to inhibit oral candidiasis in 12 denture-bearing patients with prosthesis stomatitis candidiasis association.

Prevalence of oral mucosal alterations in children from 0 to 12 years old.

BACKGROUND: As the exact prevalence of oral alterations in children is still not known and the findings are controversial, other studies about this subject are necessary. METHODS: A cross-sectional survey was carried out on 1211 Brazilian children divided in two age groups: 0-4 and 5-12 years. The patients were economically classified and data of medical history were obtained from medical records. RESULTS: The frequency of children presenting alterations was 27.0%, and it was higher in older children. The most common lesions were geographic tongue, cheek biting, and melanotic macule. Candidiasis was associated with antibiotic therapy and use of pacifiers. Fissured tongue was associated with congenital anomalies, allergy; age from 5 to 12 years. There was a lack of association of patient's economic status and prevalence of oral mucosal alterations. CONCLUSIONS: The frequency of mucosal alterations in children is high and increases with age, and some of them are associated with habits and medical history of the patients.

Argyrophilic nucleolar organizer regions (AgNORs) in odontogenic myxoma (OM) and ameloblastic fibroma (AF).

Ten cases of odontogenic myxoma (OM) and six cases of ameloblastic fibroma (AF) were subjected to comparative analysis by the AgNOR technique, in order to determine a possible difference in cell proliferation index between these lesions. The mean AgNOR number of the mesenchymal component of AF was compared with its epithelial component and the difference was not found to be statistically significant. The mean AgNOR index of the AF group was significantly higher than that of the OM group. Moreover, the mesenchymal component of AF demonstrated increased AgNOR numbers compared with that of OM (P<0.05). These results suggest that the epithelial and mesenchymal components of AF may have similar cell proliferative activity. However, the cell proliferative index of this lesion seems to be higher than that of OM.

Identification of the AgNORs, PCNA and ck16 proteins in oral lichen planus lesions.

The expression of proliferating cell nuclear antigen (PCNA), cytokeratin 16 (ck16) and Ag nucleolar organizer regions (AgNORs) were assessed in 20 cases of lichen planus, 20 cases of keratosis and 20 cases of normal oral mucosa in order to evaluate the rate of keratinocyte proliferation in these tissues. Three hundred cells were counted in each sample: 100 basal cells, 100 suprabasal cells and 100 squamous cells. The mean number of AgNORs and the percentage of PCNA positive cells were calculated. Except from similar staining of suprabasal cells of lichen planus and keratosis, PCNA and AgNORs values were higher in all layers of lichen planus than in both keratosis and normal oral mucosa. The three groups showed similar ck16 immunostaining: all of the cells were positive, except those of the basal layer. The results suggest that the keratinocyte proliferation index is higher in lichen planus than in keratosis and normal mucosa. Besides, ck16 should not be used to differentiate the entities studied.

AgNORs in hyperplasia, papilloma and oral squamous cell carcinoma.

Ten inflammatory fibrous hyperplasias, ten papillomas, and nineteen oral squamous cell carcinomas were analyzed by the AgNOR technique to determine if different disturbances of oral epithelia presented different AgNOR counts. The papilloma group showed higher mean AgNOR counts (3.15 +/- 0.58) than the hyperplasia group (1.98 +/- 0.24) and smaller than the well-differentiated oral squamous cell carcinoma group (6.56 +/- 1.25) and poorly differentiated oral squamous cell carcinoma group (7.07 +/- 1.60). The differences among the groups of lesions were statistically significant (P < 0.05) except between the well differentiated oral squamous cell carcinoma group and the poorly differentiated oral squamous cell carcinoma group. Our findings suggest that the cellular proliferation ratio in papillomas is greater than hyperplasias and smaller than carcinomas.

Regional odontodysplasia: an unusual case with a conservative approach.

A case of a 14-year-old male with regional odontodysplasia is reported. In this presentation many atypical clinical and radiographical features of this condition are present. The chief complaint of the patient was the enlargement of the gingiva and, according to the literature, inflammatory processes are the main reason why patients look for care. Moreover, there was no radiographic evidence of unerupted teeth in this report. The functional and psychological benefits of the conservative approach are emphasised.

Argyrophilic nucleolar organizer regions (AgNORs) in ameloblastomas and adenomatoid odontogenic tumours (AOTs).

Twenty-two cases of ameloblastoma and ten cases of adenomatoid odontogenic tumour (AOT) were analyzed comparatively by the AgNOR technique. Ameloblastomas were distributed into three groups according to their clinical behaviour: primary lesions without recurrences (PLWTR), 5 cases; primary lesions with recurrences (PLWR), 4 cases; and recurrences, 13 cases. The cases were also regrouped according to their histological pattern: follicular (9 cases), plexiform (7 cases), acanthomatous (4 cases) and unicystic (2 cases). Considering histological patterns, there was a significant statistical difference only between follicular and plexiform types. There were no significant differences between the group of ameloblastomas and the group of AOTs or between the three groups of ameloblastomas with different clinical behaviour. Our results strongly suggest that the distinct clinical behaviour of ameloblastomas and AOT is not correlated with their cellular proliferation ratio. Thus, the infiltrative ability of the ameloblastomas is probably not related to the cellular proliferation index of these tumours.