Remodeling of cortical and corticocancellous fresh-frozen allogeneic block bone grafts - a radiographic and histomorphometric comparison to autologous bone grafts

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

RANKL expression is differentially modulated by TLR2 and TLR4 signaling in fibroblasts and osteoblasts

Resident, non-immune cells express various pattern-recognition receptors and produce inflammatory cytokines in response to microbial antigens, during the innate immune response. Alveolar bone resorption is the hallmark of destructive periodontitis and it is caused by the host response to bacteria and their mediators present on the biofilm. The balance between the expression levels of receptor activator of nuclear factorkappa B ligand (RANKL) and osteoprotegerin (OPG) is pivotal for osteoclast differentiation and activity and has been implicated in the progression of bone loss in periodontitis. To assess the contribution of resident cells to the bone resorption mediated by innate immune signaling, we stimulated fibroblasts and osteoblastic cells with LPS from. Escherichia coli (TLR4 agonist), Porphyromonas gingivalis (TLR2 and -4 agonist), and interleukin-1 beta (as a control for cytokine signaling through Toll/IL-1receptor domain) in time-response experiments. Expression of RANKL and OPG mRNA was studied by RT-PCR, whereas the production of RANKL protein and the activation of p38 MAPK and NF-kB signaling pathways were analyzed by western blot. We used biochemical inhibitors to assess the relative contribution of p38 MAPK and NF-kB signaling to the expression of RANKL and OPG induced by TLR2, -4 and IL1β in these cells. Both p38 MAPK and NFkB pathways were activated by these stimuli in fibroblasts and osteoblasts, but the kinetics of this activation varied in each cell type and with the nature of the stimulation. E. coli LPS was a stronger inducer of RANKL mRNA in fibroblasts, whereas LPS from P. gingivalis downregulated RANKL mRNA in periodontal ligament cells but increased its expression in osteoblasts. IL-1β induced RANKL in both cell types and without a marked effect on OPG expression. p38 MAPK was more relevant than NF-kB for the expression of RANKL and OPG in these cell types.

Effect of LPS treatment on the viability and chemokine synthesis by epithelial cells and gingival fibroblasts

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Green tea polyphenol epigallocatechin-3-gallate and cranberry proanthocyanidins act in synergy with cathelicidin (LL-37) to reduce the LPS-induced inflammatory response in a three-dimensional co-culture model of gingival epithelial cells and fibroblasts

Objectives: The human antimicrobial peptide cathelicidin (LL-37) possesses anti-inflammatory properties that may contribute to attenuating the inflammatory process associated with chronic periodontitis. Plant polyphenols, including those from cranberry and green tea, have been reported to reduce inflammatory cytokine secretion by host cells. In the present study, we hypothesized that A-type cranberry proanthocyanidins (AC-PACs) and green tea epigallocatechin-3-gallate (EGCG) act in synergy with LL-37 to reduce the secretion of inflammatory mediators by oral mucosal cells. Methods: A three-dimensional (3D) co-culture model of gingival epithelial cells and fibroblasts treated with non-cytotoxic concentrations of AC-PACs (25 and 50 mg/ml), EGCG (1 and 5 mg/ml), and LL-37 (0.1 and 0.2 mM) individually and in combination (AC-PACs + LL-37 and EGCG + LL-37) were stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS). Multiplex ELISA assays were used to quantify the secretion of 54 host factors, including chemokines, cytokines, growth factors, matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases (TIMPs). Results: LL-37, AC-PACs, and EGCG, individually or in combination, had no effect on the regulation of MMP and TIMP secretion but inhibited the secretion of several cytokines. ACPACs and LL-37 acted in synergy to reduce the secretion of CXC-chemokine ligand 1 (GRO-a), granulocyte colony-stimulating factor (G-CSF), and interleukin-6 (IL-6), and had an additive effect on reducing the secretion of interleukin-8 (IL-8), interferon-g inducible protein 10 (IP-10), and monocyte chemoattractant protein-1 (MCP-1) in response to LPS stimulation. EGCG and LL-37 acted in synergy to reduce the secretion of GRO-a, G-CSF, IL-6, IL-8, and IP-10, and had an additive effect on MCP-1 secretion. Conclusion: The combination of LL-37 and natural polyphenols from cranberry and green tea acted in synergy to reduce the secretion of several cytokines by an LPS-stimulated 3D coculture model of oral mucosal cells. Such combinations show promising results as potential adjunctive therapies for treating inflammatory periodontitis.

Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblasts

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Gene expression of estrogen receptor-alpha in orbital fibroblasts in Graves’ ophthalmopathy

Graves’ ophthalmopathy (GO) is one of the most severe clinical manifestations of Graves’ disease (GD), and its treatment might involve high-dose glucocorticoid therapy. The higher incidence of GO among females, and the reported association between polymorphisms of estrogen receptor (ER) and GD susceptibility have led us to question the role of estrogen and its receptor in GO pathogenesis. We, thus, assessed estrogen receptor-alpha (ERA) gene expression in cultures of orbital fibroblasts from a patient with GO before (controls) and after treatment with 10 nM and 100 nM dexamethasone (DEX). Orbital fibroblasts showed ERA gene expression. In the cells treated with 10 nM and 100 nM DEX, ERA gene expression was, respectively, 85% higher and 74% lower, than in the control group. We concluded that ERA gene expression is found in the orbital fibroblasts of patient with GO, which may be affected by glucocorticoids in a dose-related manner. Arch Endocrinol Metab. 2015;59(3):273-6

Cortical correlates of affective syndrome in dementia due to Alzheimer’s disease

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Expression of Vascular Endothelial Growth Factor (VEGF) in Macrophages, Fibroblasts, and Endothelial Cells in Pterygium Treated with 5-Fluorouracil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Antiageing mechanisms of a standardized supercritical CO2 preparation of Black Jack (Bidens pilosa L.) in human fibroblasts and skin fragments

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytotoxicity of peracetic acid in L929 fibroblasts

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Widespread pH abnormalities in patients with malformations of cortical development and epilepsy: a phosphorus-31 brain MR spectroscopy study

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Purely cortical anaplastic ependymoma

Ependymomas are glial tumors derived from ependymal cells lining the ventricles and the central canal of the spinal cord. It may occur outside the ventricular structures, representing the extraventicular form, or without any relationship of ventricular system, called ectopic ependymona. Less than fifteen cases of ectopic ependymomas were reported and less than five were anaplastic. We report a rare case of pure cortical ectopic anaplastic ependymoma.

In vitro study of cyclosporine A 0.05 % on primary and recurrent pterygium fibroblasts

To compare the cyclosporine 0.05 % exposure effect on fibroblasts from primary and recurrent pterygium. Primary culture of fibroblasts from primary and recurrent pterygium was performed until the third passage, which was exposed to cyclosporine 0.05 % in a group and the other remaining unexposed (control group), in triplicates. After 3, 6, 12, and 17 days of exposure the viable cell counting was performed by hemocytometer. The results were statistically analyzed using the technique of analysis of non-parametric variance model for repeated measures with three factors. There was a significant reduction in both fibroblast proliferation, in primary as in the recurrent pterygium cultures exposed to cyclosporine when compared not exposed cultures, with statistical significance (P < 0.05). Comparing primary and recurrent pterygium that received the drug, there was no significant difference in cell proliferation in relation to primary or recurrent pterygium. Cyclosporine 0.05 % is effective in inhibiting fibroblast proliferation in culture, both in primary and as in recurrent pterygium.

Mandibular cortical bone evaluation on cone beam computed tomography images of patients with bisphosphonate related osteonecrosis of the jaws

Objectives The objective of this study was to develop a technique for detecting cortical bone dimensional changes in patients with bisphosphonate-related osteonecrosis of the jaw (BRONJ). Study Design Subjects with BRONJ who had cone-beam computed tomography imaging were selected, with age- and gender-matched controls. Mandibular cortical bone measurements to detect bisphosphonate-related cortical bone changes were made inferior to mental foramen, in 3 different ways: within a fixed sized rectangle, in a rectangle varying with the cortical height, and a ratio between area and height. Results Twelve BRONJ cases and 66 controls were evaluated. The cortical bone measurements were significantly higher in cases than controls for all 3 techniques. The bone measurements were strongly associated with BRONJ case status (odds ratio 3.36-7.84). The inter-rater reliability coefficients were high for all techniques (0.71-0.90). Conclusions Mandibular cortical bone measurement is a potentially useful tool in the detection of bone dimensional changes caused by bisphosphonates. Long-term administration of bisphosphonates (BPs) affects bone quality and metabolism following accumulation in bone.1 Since the first cases of bisphosphonate-related osteonecrosis of the jaw (BRONJ) were published in 2003,2 there has been a search for factors that can predict the onset of the condition. Oral and intravenous BPs reduce bone resorption, increase mineral content of bone, and alter bony architecture.3, 4, 5 and 6 Previous studies have demonstrated these changes both radiographically and following histologic analysis.1, 3, 7, 8, 9 and 10 The BP-related jaw changes may present radiological features, such as thickening of lamina dura and cortical borders, diffuse sclerosis, and narrowing of the mandibular canal3 and 11; however, oral radiographs of patients taking BPs do not consistently show radiographic changes to the jaws.11 and 12 The challenge is to find imaging tools that could improve the detection of changes in the bone associated with BP use. Various skeletal radiographic features associated with BRONJ in conventional periapical and panoramic radiographs, computed tomography, magnetic resonance imaging, and nuclear bone scanning have been described.3, 8, 9, 10 and 11 There has also been a search for BP-related quantitative methods for the evaluation of radiographic images, to avoid observer subjectivity in interpretation. Factors thought to be important include trabecular and cortical structure, and bone mineralization.4 Consequently, measurable bone data have been reported in subjects taking BPs through many techniques, including bone density, architecture, and cortical bone thickness.1, 4, 7 and 13 Trabecular microarchitecture of postmenopausal women has been evaluated with noninvasive techniques, such as high-resolution magnetic resonance images showing less deterioration of the bone 1 year after initiation of oral BP therapy.4 A decrease in bone turnover and a trend for an increase in the bone wall thickness has been detected by histomorphometry in subjects taking BPs.1 Alterations in the cortical structure of the second metacarpal have been detected in digital x-ray radiogrammetry of postmenopausal women treated with BPs.7 Mandibular cortical width may be measured on dental panoramic radiographs, and it has been suggested as a screening tool for referring patients for bone densitometry for osteoporosis investigation.14 and 15 Inhibition of the intracortical bone remodeling in the mandible of mice taking BPs has been reported.16 Thus, imaging evaluation of the mandibular cortical bone could be a biologically plausible way to detect BP bone alterations. Computed tomography can assess both cortical and trabecular bone characteristics. Cone-beam computed tomography (CBCT) can provide 3-dimensional information, while using lower doses and costing less than conventional CT. The CBCT images have been studied as a tool for the measurement of trabecular bone in patients with BRONJ.13 Therefore, cortical bone measurements on CBCT of the jaws might also help to understand bone changes in patients with BRONJ. There is no standard in quantifying dimensional changes of mandibular cortical bone. We explored several different approaches to take into consideration possible changes in length, area, and volume. These led to the 3 techniques developed in this study. This article reports a matched case-control study in which mandibular cortical bone was measured on CBCT images of subjects with BRONJ and controls. The aim of the study was to explore the usefulness of 3 techniques for detecting mandibular cortical bone dimensional changes caused by BP.

Mercury toxicity in Amazon gold miners: Visual dysfunction assessed by retinal and cortical electrophysiology

Amazonian gold mining activity results in human exposure to mercury vapor. We evaluated the visual system of two Amazonian gold miners (29 and 37 years old) by recording the transient pattern electroretinogram (tPERG) and transient pattern visual evoked potential (tPVEP). We compared these results with those obtained from a regional group of control subjects. For both tPERG and tPVEP, checkerboards with 0.5 or 2 cycles per degree (cpd) of spatial frequency were presented in a 16 degrees squared area, 100% Michelson contrast, 50cd/m(2) mean luminance, and 1 Hz square-wave pattern-reversal presentation. Two averaged waveforms (n = 240 sweeps, Is each) were monocularly obtained for each subject in each condition. Both eyes were monocularly tested only in gold miners. Normative data were calculated using a final pooled waveforin with 480 sweeps. The first gold miner, LCS, had normal tPERG responses. The second one, RNP, showed low tPERG (P50 component) amplitudes at 0.5cpd for both eyes, outside the normative data, and absence of response at 2 cpd for his right eye. Delayed tPVEP responses (P 100 component) were found at 2 cpd for LCS but the implicit times were inside the normative data. Subject RNP also showed delayed tPVEP responses (all components), but only the implicit time obtained with his right eye was outside the normative data at 2cpd. We conclude that mercury exposure levels found in the Amazon gold miners is high enough to damage the visual system and can be assessed by non-invasive electrophysiological techniques. (C) 2007 Elsevier Inc. All rights reserved.