Silva J.A.F.,University of Campinas |
Lopes Ferrucci D.,University of Campinas |
Peroni L.A.,University of Campinas |
Carvalho H.F.,University of Campinas |
And 2 more authors.
Cells Tissues Organs | Year: 2012
Alveolar bone resorption results from the inflammatory response to periodontal pathogens. Systemic diseases that affect the host response, such as type 1 diabetes mellitus (DM1), can potentiate the severity of periodontal disease (PD) and accelerate bone resorption. However, the biological mechanisms by which DM1 modulates PD are not fully understood. The aim of this study was to determine the influence of DM1 on alveolar bone resorption and to evaluate the role of receptor activator of nuclear factor-kappaB ligand (RANKL)/ osteoprotegerin (OPG) in osteoclastogenesis in rats. PD was induced by means of ligature in nondiabetic and in streptozotocyn-induced DM1 rats. Morphological and morphometric analyses, stereology and osteoclast counting were performed. RANKL and OPG mRNA levels, protein content, and location were determined. PD caused alveolar bone resorption, increased the number of osteoclasts in the alveolar bone crest and also promoted changes in RANKL/OPG mRNA expression. DM1 alone showed alveolar bone destruction and an increased number of osteoclasts at the periapical and furcal regions. DM1 exacerbated these characteristics, with a greater impact on bone structure, resulting in a low OPG content and a higher RANKL/OPG ratio, which correlated with prominent osteoclastogenesis. This work demonstrates that the effects of PD and DM1 enhance bone destruction, confirms the importance of the RANKL signaling pathway in bone destruction in DM1 in animal models and suggests the existence of alternative mechanisms potentiating bone degradation in PD. © 2012 S. Karger AG, Basel.
Machado L.C.,University of Sao Paulo |
Pelegati V.B.,Applied Cell Sciences |
Oliveira A.L.,University of Sao Paulo
Journal of Supercritical Fluids | Year: 2016
Supercritical CO2 was studied for impregnation or encapsulation of essential oils in modified starches via Particle from Gas Saturated Solutions or Suspensions (PGSS). Modified starches were choose as function of its low cost. The advantage of this method over conventional encapsulation that use modified starch via spray drier refers to the low temperatures used and absence of water in the process. Modified starch presents hydrophobic elements and this molecules present amphiphilic character. Usually it is employed in the encapsulation of essences as wall material with excellent volatiles retention due to its polar and nonpolar interface. Considering its hydrophobic characteristics, interactions between the modified starch and supercritical CO2 occurred, resulting in two different structural interactions of the limonene and modified starch in the PGSS (50 °C and 60 °C at 100 bar and 120 bar). When hydrous ethanol was used in the suspension, impregnation occurred and, when anhydrous ethanol was used, encapsulation occurred. Analysis of particle morphology via scanning electron and confocal microscopy, thermo-oxidative characterization by differential scanning calorimetry and determination of microencapsulated limonene via gas chromatography coupled to mass spectrometry indicated limonene microencapsulation and impregnation occurred despite the highly solubility of limonene in supercritical CO2. The retention of limonene by Purity Gum Ultra® was 86% when encapsulated and, 53% when impregnated, similar values to those obtained in conventional microencapsulation methods via a spray drier or via PGSS-drying. © 2015 Elsevier B.V. All rights reserved.
Adur J.,National University of Entre Rios |
Adur J.,Applied Cell Sciences |
Carvalho H.F.,Applied Cell Sciences |
Cesar C.L.,Applied Cell Sciences |
Casco V.H.,National University of Entre Rios
Cancer Informatics | Year: 2014
This work reviews the most relevant present-day processing methods used to improve the accuracy of multimodal nonlinear images in the detection of epithelial cancer and the supporting stroma. Special emphasis has been placed on methods of non linear optical (NLO) microscopy image processing such as: second harmonic to autofluorescence ageing index of dermis (SAAID), tumor-associated collagen signatures (TACS), fast Fourier trans-form (FFT) analysis, and gray level co-occurrence matrix (GLCM)-based methods. These strategies are presented as a set of potential valuable diagnostic tools for early cancer detection. It may be proposed that the combination of NLO microscopy and informatics based image analysis approaches described in this review (all carried out on free software) may represent a powerful tool to investigate collagen organization and remodeling of extracellular matrix in carcinogenesis processes. © the authors.
Singh A.,Applied Cell Sciences |
Singh A.,Maastricht University |
Rajasekaran N.,Applied Cell Sciences |
Rajasekaran N.,Stanford University |
And 6 more authors.
Arthritis Research and Therapy | Year: 2013
Introduction: Matrix metalloproteinases (MMPs) are important in tissue remodelling. Here we investigate the role of collagenase-3 (MMP-13) in antibody-induced arthritis.Methods: For this study we employed the K/BxN serum-induced arthritis model. Arthritis was induced in C57BL/6 wild type (WT) and MMP-13-deficient (MMP-13-/-) mice by intraperitoneal injection of 200 μl of K/BxN serum. Arthritis was assessed by measuring the ankle swelling. During the course of the experiments, mice were sacrificed every second day for histological examination of the ankle joints. Ankle sections were evaluated histologically for infiltration of inflammatory cells, pannus tissue formation and bone/cartilage destruction. Semi-quantitative PCR was used to determine MMP-13 expression levels in ankle joints of untreated and K/BxN serum-injected mice.Results: This study shows that MMP-13 is a regulator of inflammation. We observed increased expression of MMP-13 in ankle joints of WT mice during K/BxN serum-induced arthritis and both K/BxN serum-treated WT and MMP-13-/- mice developed progressive arthritis with a similar onset. However, MMP-13-/- mice showed significantly reduced disease over the whole arthritic period. Ankle joints of WT mice showed severe joint destruction with extensive inflammation and erosion of cartilage and bone. In contrast, MMP-13-/- mice displayed significantly decreased severity of arthritis (50% to 60%) as analyzed by clinical and histological scoring methods.Conclusions: MMP-13 deficiency acts to suppress the local inflammatory responses. Therefore, MMP-13 has a role in the pathogenesis of arthritis, suggesting MMP-13 is a potential therapeutic target. © 2013 Singh et al.; licensee BioMed Central Ltd.
Schmidt-Bleek K.,Charité - Medical University of Berlin |
Kwee B.J.,Applied Cell Sciences |
Mooney D.J.,Applied Cell Sciences |
Duda G.N.,Charité - Medical University of Berlin
Tissue Engineering - Part B: Reviews | Year: 2015
Delayed healing or nonhealing of bone is an important clinical concern. Although bone, one of the two tissues with scar-free healing capacity, heals in most cases, healing is delayed in more than 10% of clinical cases. Treatment of such delayed healing condition is often painful, risky, time consuming, and expensive. Tissue healing is a multistage regenerative process involving complex and well-orchestrated steps, which are initiated in response to injury. At best, these steps lead to scar-free tissue formation. At the onset of healing, during the inflammatory phase, stationary and attracted macrophages and other immune cells at the fracture site release cytokines in response to injury. This initial reaction to injury is followed by the recruitment, proliferation, and differentiation of mesenchymal stromal cells, synthesis of extracellular matrix proteins, angiogenesis, and finally tissue remodeling. Failure to heal is often associated with poor revascularization. Since blood vessels mediate the transport of circulating cells, oxygen, nutrients, and waste products, they appear essential for successful healing. The strategy of endogenous regeneration in a tissue such as bone is interesting to analyze since it may represent a blueprint of successful tissue formation. This review highlights the interdependency of the time cascades of inflammation, angiogenesis, and tissue regeneration. A better understanding of these inter-relations is mandatory to early identify patients at risk as well as to overcome critical clinical conditions that limit healing. Instead of purely tolerating the inflammatory phase, modulations of inflammation (immunomodulation) might represent a valid therapeutic strategy to enhance angiogenesis and foster later phases of tissue regeneration. Copyright 2015, Mary Ann Liebert, Inc.
PubMed | Gifu University, High Energy Accelerator Research Organization and Applied Cell Sciences
Type: | Journal: Bioorganic & medicinal chemistry | Year: 2017
Selenium-incorporated fucoses (seleno-fucoses) differing in the position of the seleno-substituent were synthesized and applied to the X-ray structural determination of a carbohydrate-lectin complex using single/multi-wavelength anomalous dispersion (SAD/MAD) phasing. The hydroxyl groups at the C-1, -2, -3 and -4 position of fucose were individually substituted with a methylseleno group via a transacetalization reaction using MeSeCH
Thiel J.,University Hospital Freiburg |
Kimmig L.,University Hospital Freiburg |
Salzer U.,University Hospital Freiburg |
Grudzien M.,University Hospital Freiburg |
And 19 more authors.
Journal of Allergy and Clinical Immunology | Year: 2012
Background: Complement receptor 2 (CR2/CD21) is part of the B-cell coreceptor and expressed by mature B cells and follicular dendritic cells. CD21 is a receptor for C3d-opsonized immune complexes and enhances antigen-specific B-cell responses. Objective: Genetic inactivation of the murine CR2 locus results in impaired humoral immune responses. Here we report the first case of a genetic CD21 deficiency in human subjects. Methods: CD21 protein expression was analyzed by means of flow cytometry and Western blotting. CD21 transcripts were quantified by using real-time PCR. The CD21 gene was sequenced. Wild-type and mutant CD21 cDNA expression was studied after transfection of 293T cells. Binding of EBV-gp350 or C3d-containing immune complexes and induction of calcium flux in CD21-deficient B cells were analyzed by means of flow cytometry. Antibody responses to protein and polysaccharide vaccines were measured. Results: A 28-year-old man presented with recurrent infections, reduced class-switched memory B cells, and hypogammaglobulinemia. CD21 receptor expression was undetectable. Binding of C3d-containing immune complexes and EBV-gp350 to B cells was severely reduced. Sequence analysis revealed a compound heterozygous deleterious mutation in the CD21 gene. Functional studies with anti-immunoglobulin- and C3d-containing immune complexes showed a complete loss of costimulatory activity of C3d in enhancing suboptimal B-cell receptor stimulation. Vaccination responses to protein antigens were normal, but the response to pneumococcal polysaccharide vaccination was moderately impaired. Conclusions: Genetic CD21 deficiency adds to the molecular defects observed in human subjects with hypogammaglobulinemia. © 2011 American Academy of Allergy, Asthma & Immunology.
Silva J.A.,University of Campinas |
Ferrucci D.L.,University of Campinas |
Peroni L.A.,University of Campinas |
Abrahao P.G.,University of Campinas |
And 5 more authors.
Journal of Cellular Physiology | Year: 2012
Molecular mechanisms responsible for periodontal disease (PD) and its worsening in type 1 Diabetes Mellitus (DM1) remain unknown. Cytokine profile and expression levels of collagenases, Mmp14, and tissue inhibitors were determined, as were the numbers of neutrophils and macrophages in combined streptozotocin-induced DM1 and ligature-induced PD models. Increased IL-23 (80-fold) and Mmp8 expression (25-fold) was found in DM1. Ligature resulted in an IL-1β/IL-6 profile, increased expression of Mmp8, Mmp13, and Mmp14 (but not Mmp1), and transient expression of Timp1 and Reck in non-diabetics. PD in DM1 involved IL-1β (but not IL-6) and IL-23/IL-17, reduced IL-6 and IL-10, sustained Mmp8 and Mmp14, increased Mmp13 and reduced Reck expression in association with 20-fold higher counts of neutrophils and macrophages. IL-23 and Mmp8 expression are hallmarks of DM1. In association with the IL-1/IL-6 (Th1) response in PD, one found a secondary IL-17 (Th17) pathway in non-diabetic rats. Low IL-6/TNF-α suggest that the Th1 response was compromised in DM1, while IL-17 indicates a prevalence of the Th17 pathway, resulting in high neutrophil recruitment. Mmp8, Mmp13, and Mmp14 expression seems important in the tissue destruction during PD in DM1. PD-associated IL-1/IL-6 (Th1), IL-10, and Reck expression are associated with the acute-to-chronic inflammation transition, which is lost in DM1. In conclusion, IL-23/IL-17 are associated with the PD progression in DM1. © 2011 Wiley Periodicals, Inc.
Singh A.,Applied Cell Sciences |
Vastert S.J.,University Utrecht |
Prakken B.J.,University Utrecht |
Illges H.,Applied Cell Sciences
Rheumatology International | Year: 2012
A soluble form of CD21 (sCD21) and CD23 (sCD23) is released from the surface of human white blood cells upon shedding of the extracellular domain. sCD21 circulates in a complex with cleavage fragments of C3 and sCD23, which were previously identified as ligands of membrane and soluble CD21. sCD21 seems to be a marker of chronic infiammatory disease. To assess the sCD21 and sCD23 status in patients with subsets of juvenile arthritis (JA), we determined plasma levels sCD21 and sCD23. Plasma sCD21 levels were significantly decreased in all JA subtypes (O-JA P < 0.0068; P- and S-JA P < 0.0001) compared to healthy controls. Plasma sCD23 levels were significantly decreased in P-JA and S-JA (both P < 0.0001), but not in O-JA (P < 0.3843) in comparison with healthy controls, and data statistically analyzed. Our results suggest that pathological mechanisms relevant to autoimmune disorders interfere with the regulation of both CD21 and CD23 shedding. © Springer-Verlag 2011.