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F1-protein fraction (F1) is a natural bioactive compound extracted from the rubber tree, Hevea brasiliensis, and has been recently studied for its therapeutic potential in wound healing. In this study, we investigated the concentration-dependent effects of F1 (0.01%, 0.025%, 0.05%, and 0.1%) incorporated into deproteinized bovine bone (DBB) and porous biphasic calcium phosphate (pBCP), on the repair of rat calvarial critical-size bone defects (CSBD). The defects were analyzed by 3D-microtomography and 2D-histomorphometry at 12 weeks postsurgery. The binding efficiency of F1 to pBCP (96.3 ± 1.4%) was higher than that to DBB (67.7 ± 3.3%). In vivo analysis showed a higher bone volume (BV) gain in all defects treated with DBB (except in 0.1% of F1) and pBCP (except in 0.05% and 0.1% of F1) compared to the CSBD without treatment/control group (9.96 ± 2.8 mm3 ). DBB plus 0.025% F1 promoted the highest BV gain (29.7 ± 2.2 mm3 , p .05). Additionally, a small BV occurred in defects treated with pBCP plus 0.1% F1 (10.4 ± 1.4 mm3, p < .05). In conclusion, F1 showed a higher bone formation potential in combination with DBB than with pBCP, in a concentration-dependent manner. Incorporation of 0.25% F1 into DBB showed the best results with respect to bone formation/repair in CSBD. These results suggest that DBB plus 0.25% F1 can be used as a promising bioactive material for application in bone tissue engineering.

Purpose: To evaluate in-vivo degradation of two bioabsorbable interference screws. Methods: Twenty-two crossbred Santa Inês ewes were used. A poly-DL-lactide (PDLLA) screw (70%/30%) was inserted in the right pelvic limb, and a PDLLA screw (70%) + β-tri-calcium phosphate (β-TCP) (30%) in the left pelvic limb. Animals were euthanized at one, four, seven and a half and 18 months after surgery. Plain radiography, computed tomography (CT), microCT, and histological analysis were accomplished. Results: PDLLA screw was hypodense at all evaluation moments, but with progressive density increase along the central axis, whereas PDLLA/β-TCP was initially hyperdense and progressively lost this characteristic. No adverse reactions were observed on histological evaluation. Conclusions: The inclusion of β-TCP favors screw degradation since the PDLLA/β-TCP screws evidenced a more intense degradation process than the PDLLA screws at the last evaluation. PDLLA screws showed higher bone production, evident around the screw thread, inside the lateral perforations, and in the central canal, whereas the PDLLA/β-TCP screws presented less bone tissue at the implantation site.

Background This study evaluated the long-term effects of platelet-rich plasma (PRP) on bone formation and regeneration when associated with autogenous bone graft (AB), porous biphasic calcium phosphate (pBCP), or deproteinized bovine bone (DBB) in maxillary sinus augmentation (MSA) of rabbit. Methods In 54 rabbits, bilateral MSA procedure was performed and randomly one sinus was filled with 200 mm3 material plus blood clot (AB/clot, DBB/clot, and pBCP/clot) and other with the same graft plus PRP (AB/PRP, DBB/PRP, and pBCP/PRP). After 30, 60, and 180 days, microtomographic were performed to analyze the three-dimensional MSA volume and histomorphometric analyses for the percentage of bone and soft tissues ingrowth. Data were compared by two-way ANOVA and the means were compared by the Tukey test, at p < 0.05. Results The percentage of pBCP and DBB were nearly unchanged throughout the whole period and bone formation occurred in the spaces between particles. The MSA volume filled with DBB and pBCP agglutinated with clot and PRP maintained constant during all experimental periods (147.2 mm3 and 154.9 mm3, respectively, p = 0.7377), and no significant changes in the new formatted bone and soft tissue were observed between treatments. In AB/clot and AB/PRP, the MSA volume was similar at 30 days (140.3 mm3 and 137.9 mm3, respectively), but a higher and gradual reduction was observed until 180 days. In the AB/PRP, this reduction was significantly higher (44.2%) than AB/clot (22.5%) (p = 0.01792). Histologically, the addition of PRP to AB accelerated the new bone formation/remodeling maintaining the percentage of new bone similar to AB/clot during all experimental volume (p = 0.6406), while the AB particles showed a higher resorption in AB/PRP than AB/clot until 60 days (mean of 7.8% and 15.1%, respectively, p = 0.0396). Conclusion The association of PRP with the autogenous graft accelerates the process of bone formation/remodeling in MSA, but not had influence on the pBCP and DBB groups.

Made available in DSpace on 2020-12-10T20:01:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-09-01 Water fluoridation is an important public health measure for the control of dental caries. Recent animal studies have shown that low doses of fluoride (F) in the drinking water, similar to those found in public water supplies, increase insulin sensitivity and reduce blood glucose. In the present study we evaluated the effects of low-level F exposure through the drinking water on glucose homeostasis in female NOD mice. Seventy-two 6-week mice were randomly divided into 2 groups according to the concentration of F in the drinking water (0-control, or 10 mg/L) they received for 14 weeks. After the experimental period the blood was collected for analyses of plasma F, glucose and insulin. Liver and gastrocnemius muscle were collected for proteomic analysis. Plasma F concentrations were significantly higher in the F-treated than in the control group. Despite treatment with fluoridated water reduced plasma levels glucose by 20% compared to control, no significant differences were found between the groups for plasma glucose and insulin. In the muscle, treatment with fluoridated water increased the expression of proteins related to muscle contraction, while in the liver, there was an increase in expression of antioxidant proteins and in proteins related to carboxylic acid metabolic process. Remarkably, phosphoenolpyruvate carboxy-kinase (PEPCK) was found exclusively in the liver of control mice. The reduction in PEPCK, a positive regulator of gluconeogenesis, thus increasing glucose uptake, might be a probable mechanism to explain the anti-diabetic effects of low doses of F, which should be evaluated in further studies. (C) 2020 Elsevier Ltd. All rights reserved. Univ Sao Paulo, Bauru Sch Dent, Dept Biol Sci, Al Octavio Pinheiro Brisolla 9-75, BR-17012901 Bauru, SP, Brazil Sao Paulo State Univ, Inst Biosci, Bauru, SP, Brazil Sao Paulo State Univ, Fac Sci, Dept Phys Educ, Bauru, SP, Brazil Sao Paulo State Univ, Inst Biosci, Bauru, SP, Brazil Sao Paulo State Univ, Fac Sci, Dept Phys Educ, Bauru, SP, Brazil

Objectives This study compares the osseointegration of machined-zirconia implants containing yttria (M-Y-TZP) with machined (M-Ti) and resorbable blast media (RBM-Ti) titanium implants. Material and Methods M-Y-TZP, M-Ti and RBM-Ti implants were randomly placed in rabbit tibiae. Fluorochrome bone labels (tetracycline, alizarin and calcein) were administered at different time periods. After 8 weeks, osseointegration was evaluated in terms of bone-to-implant contact (BIC), new bone area (nBA), remaining cortical bone area (rBA) and temporal quantification of fluorochromes, using micro-CT and histomorphometric analyses. Results RBM-Ti implants showed higher resorption of the remaining cortical bone and bone formation (rBA = 36.9% and nBA = 38.8%) than M-Y-TZP implants (rBA = 48% and nBA = 26.5%). The BIC values showed no differences among the groups in the cortical region (mean = 52.2%) but in the medullary region, they were 0.45-fold higher in the RBM-Ti group (51.2%) than in the M-Y-TZP group (35.2%). In all groups, high incorporation of tetracycline was observed (2nd to 4th weeks), followed by alizarin (4th to 6th weeks) and calcein (6th to 8th weeks). In the cortical region, incorporation of tetracycline was similar between RBM-Ti (49.8%) and M-Y-TZP (35.9%) implants, but higher than M-Ti (28.2%) implants. Subsequently, alizarin and calcein were 1.1-fold higher in RBM-Ti implants than in the other implants. In the medullary region, no significant differences were observed for all fluorochromes. Conclusion All implants favored bone formation and consequently promoted primary stability. Bone formation around the threads was faster in RBM-Ti and M-Y-TZP implants than in M-Ti implants, but limited bone remodeling with M-Y-TZP implants over time can have significant effects on secondary stability, suggesting caution for its use as an alternative substitute for titanium implants.

The bone-induction capacity of a porous biphasic calcium phosphate (pBCP) using heterotopic implantation in mouse (mHI-model) and its efficacy as substitute for autograft in mandibular critical-size defect in rabbit (rabMCSD-model) was investigated. In mHI-model, pBCP was implanted into the thigh muscles and bone formation was histomorphometrically and immunohistochemically evaluated. In rabMCSD-model, 13 mm bone defects were treated with pBCP or autograft and bone repair comparatively evaluated by radiographic and histomorphometric methods. In mHI-model, formed bone and immunolabeling for bone morphogenetic protein-2 and osteopontin were observed in 90% of pBCP implanted samples after 12 weeks. In rabMCSD-model neither statistically significant difference was found in newly formed bone between pBCP and autograft groups at 4 weeks (18.8 ± 5.5% vs 27.1 ± 5.6%), 8 weeks (22.3 ± 2.7% vs 26.2 ± 5.1), and 12 weeks (19.6 ± 4.7% vs 19.6 ± 2.3%). At 12 weeks, the stability and contour of the mandible were restored in both treatments. Near tooth remaining, pBCP particles were covered by small amount of mineralized tissue exhibiting perpendicular attachments of collagen fiber bundles with histological characteristic of acellular cementum. Within the limitations of this study, it was concluded that pBCP is osteoinductive and able to stimulate the new formation of bone and cementum-like tissues in rabMCSD-model, suggesting that it may be an alternative to treatment of large bone defect and in periodontal regenerative therapy. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1546-1557, 2018.

Type 1 diabetes (T1D) is associated with delayed tissue healing and bone loss. Periodontal tissues during tooth movement (OTM) in T1D and under diabetic treatment are poorly understood. We aimed to study the effect of metformin as an add-on to insulin therapy on periodontal structures during OTM in T1D rats.Rats were divided into normoglycemic (NG, n = 20) and streptozotocin-induced diabetic groups that were untreated (T1D, n = 20), treated with insulin (I-T1D, n = 20), or treated with insulin plus metformin (IM-T1D, n = 20). After 7 days of treatment, the first right upper molar (M1) was moved mesially. At days 0, 3, 7 and 14, the pattern of OTM and the periodontal tissues were analyzed by micro-CT, histomorphometry, and immunohistochemistry for TRAP.In T1D, major osteoclastogenic activity and bone loss versus other groups were confirmed by a greater TRAP-positive cell number and reabsorption surface on both the pressure and tension sides for 14 days (p 0.01). Additionally, we observed low bone volume density. Metformin plus insulin resulted in a daily insulin dose reduction and major glycemic control versus I-T1D. Although no significant differences were observed between I-T1D and IM-T1D, the tooth displacement and inclination, periodontal ligament thickness, and alveolar bone density on the pressure side in IM-T1D were similar to that of NG (p 0.05).Antidiabetic treatment reduces severe periodontal damage during applied orthodontic force in T1D untreated rats. Metformin as an add-on to insulin therapy resulted in glycemic control and a periodontal tissue response to orthodontic forces that was similar to that of normoglycemic rats.

Statement of problem Antifungal agents incorporated into interim denture resilient liners have been suggested as an adjunct treatment for denture stomatitis (DS). However, before applying this protocol to humans, biocompatibility analysis of such drugs in animal models is required. Purpose The purpose of this animal study was to evaluate the in vivo biocompatibility of an interim resilient liner modified with minimum inhibitory concentrations (MICs) of antifungal drugs for Candida albicans biofilm. Material and methods Sixty Wistar rats were divided into 6 groups (n=5): PC=positive control/no protocol; IOD (intraoral device)=rats using an acrylic resin palatal device (PD); Tru=rats using a PD relined with Trusoft; and Ny (nystatin), Chx (chlorhexidine diacetate), and Ke (ketoconazole) groups=rats using a PD relined with Trusoft + drug MICs. The rats were sacrificed at 7 or 14 days of trial. Histopathological qualitative analysis was performed by comparing photomicrographs of histological sections of the intermolar region. Morphological changes in the epithelium and keratin were quantitatively analyzed by computerized planimetry, and data were analyzed by using 2-way ANOVA and the Tukey HSD test (α=.05). Results Quantitative analysis showed that only PD containing Ke significantly decreased the thickness and area of the keratin compared with the other groups (P .05). These results agreed with those of qualitative analysis. Conclusions Incorporation of MICs of Ny and Chx in Trusoft did not induce histopathological changes in the rat palatal mucosa, suggesting the in vivo biocompatibility of this DS treatment.

Objectives The aim of this study was to evaluate the putative influence of diabetes without metabolic control in the loss of tooth structure as well as histological changes in dentin and pulp tissue in rats. Design Diabetes was induced in Wistar rats ( n = 25) by intravenous administration of alloxan (42 mg/kg). Diabetic and non-diabetic control rats were evaluated at 1, 3, 6, 9 and 12 months of follow-up. In order to evaluate the presence and progression of dental caries and periapical lesions, hemimandibles were removed and submitted to radiographical, histological, and morphometrical procedures. Results Dental caries were detected after radiographical and histological evaluations in diabetic group from the third month of diabetes onset, increasing gradually in frequency and severity in periods. Diabetic rats dental pulps also presented significant reduction in volume density of collagen fibers and fibroblasts at third month, parallel with a trend towards the increase in inflammatory cells volume density. Diabetic rats presented a generalized pulp tissue necrosis after 6 months of diabetes induction. Moreover, periapical lesions were not detected in control group, while these lesions were observed in all rats after 3, 6, 9, and 12 months of diabetes induction. Conclusions Uncontrolled diabetes seems to trigger the loss of tooth structure, associated to histological dental changes and mediates its evolution to progressive severe pulp and periapical lesions in rats. Therefore, diabetes may be considered a very important risk factor regarding alterations in dental pulp, development of dental caries, and periapical lesions.

In this work, bone formation/remodeling/maturation was correlated with the presence of multinucleated giant cells (MGCs)/osteoclasts (tartrate-resistant acid phosphatase [TRAP]-positive cells) on the surface of beta-tricalcium phosphate (β-TCP), sintered deproteinized bovine bone (sDBB), and carbonated deproteinized bovine bone (cDBB) using a maxillary sinus augmentation (MSA) in a New Zealand rabbit model. Microtomographic, histomorphometric, and immunolabeling for TRAP-cells analyses were made at 15, 30, and 60 days after surgery. In all treatments, a faster bone formation/remodeling/maturation and TRAP-positive cells activity occurred in the osteotomy region of the MSA than in the middle and submucosa regions. In the β-TCP, the granules were rapidly reabsorbed by TRAP-positive cells and replaced by bone tissue. β-TCP enabled quick bone regeneration/remodeling and full bone and marrow restoration until 60 days, but with a significant reduction in MSA volume. In cDBB and sDBB, the quantity of TRAP-positive cells was smaller than in β-TCP, and these cells were associated with granule surface preparation for osteoblast-mediated bone formation. After 30 days, more than 80% of granule surfaces were surrounded and integrated by bone tissue without signs of degradation, preserving the MSA volume. Overall, the materials tested in a standardized preclinical model led to different bone formation/remodeling/maturation within the same repair process influenced by different microenvironments and MGCs/osteoclasts. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 108B:282-297, 2020.

Purpose: The aim was to assess the effect of a relevant regimen of zoledronic acid (ZA) treatment for the study of bisphosphonate-related osteonecrosis of the jaw on alveolar bone microstructure and vasculature. A sub-objective was to use 3-dimensional imaging to describe site-specific changes induced by ZA in the alveolar bone. Materials and Methods: Five Wistar rats received ZA (0.6 mg/kg) and five (controls) received saline solution in the same volume. The compounds were administered intraperitoneally in 5 doses every 28 days. The rats were euthanized 150 days after therapy onset. The mandibles were scanned using high-resolution (14-mu m) micro-computed tomography (micro-CT), decalcified, cut into slices for histologic analysis (5 mu m), and stained with hematoxylin-eosin. Bone quality parameters were calculated using CTAnalyser software (Bruker, Kontich, Belgium) in 2 different volumes of interest (VOIs): the region between the first molar roots (VOI-1) and the periapical region under the first and second molars' apex (VOI-2). Blood vessel density and bone histomorphometric parameters were calculated only for the region between the roots of the first molar using AxioVision Imaging software (version 4.8; Carl Zeiss, Gottingen, Germany). Results: ZA-treated rats showed a significant increase in percentage of bone volume and density (P < .05), with thicker and more connected trabeculae. Furthermore, the ZA group showed a significant decrease in the size of the marrow spaces and nutritive canals and in blood vessel density (P < .05). In the micro-CT evaluation, VOI-2 showed better outcomes in measuring the effect of ZA on alveolar bone. Conclusions: ZA treatment induced bone corticalization and decreased alveolar bone vascularization. VOI-2 should be preferred for micro-CT evaluation of the effect of bisphosphonates on alveolar bone. This analysis allowed the effect of ZA on alveolar bone and its vascularization to be characterized. The results of this analysis may add further knowledge to the understanding of the physiopathology of osteonecrosis of the jaw. (C) 2018 American Association of Oral and Maxillofacial Surgeons International Collaboration Latin America Fund - KU Leuven, Leuven, Belgium; Coordination for the Improvement of Higher Education Personnel (CAPES) Foundation; Ministry of Education of Brazil [99999.007007/2015-09]; Research Foundation-Flanders (FWO) [11.ZU.117N]

AIM The effects of green tea on the modulation of vascularization during the progression of spontaneous periodontitis in long-term hyperglycaemia in streptozotocin-induced type 1 diabetic (T1D) rats were evaluated. MATERIALS AND METHODS Wistar rats normoglycaemic (NG) and T1D were divided into two control groups, which received water (NG-W and T1D-W) and two experimental groups that received green tea (NG-GT and T1D-GT). Periodontal structures were evaluated by microtomographic and histological analyses. Number of immunostained cells for VEGF (NcVEGF+/mm2 ) and CD31 (NcCD31+/mm2 ), as well microvessel density (MVD) in the periodontal ligament (PDL) were evaluated. RESULTS Long-term hyperglycaemia in T1D-W rats induced vascular alterations in PDL with a reduction of 36% in MVD, a decrease of 33% in NcCD31+/mm2 and an increase of 53% in NcVEGF+/mm2 . Concomitantly, a severe degree of periodontitis with higher reduction in bone volume and periodontal bone level was observed. In T1D-GT, green tea maintained the MVD, NcCD31+/mm2 and NcVEGF+/mm2 in the PDL similar to normoglycaemic groups. Clinically, in T1D-GT rats, green tea reduced dental plaque accumulation and the degree of periodontitis when compared to T1D-W. CONCLUSION Daily green tea consumption has a therapeutic effect on the diabetic vascular disorder in PDL and the progression of periodontitis in long-term hyperglycaemia in T1D rats.

PURPOSE To evaluate if an inorganic graft applied before implant insertion interferes with osseointegration. MATERIALS AND METHODS The bilateral mandibular incisors of 12 rabbits were extracted. One of the sockets was randomly filled with an inorganic xenogenic bone graft, whereas the remaining socket was allowed to heal naturally and served as a control. After 60 days, titanium implants were inserted into healing areas. The animals were killed 60 days after. Bone depositions were marked with fluorochrome oxytetracycline, alizarin, and calcein and evaluated using confocal laser scanning microscopy. Bone-to-implant contact (BIC) and bone area (BA) within the limits of the implant threads were analyzed. Data were compared statically by paired t tests, one-way ANOVA, and Bonferroni post hoc tests (α = 0.05). RESULTS No differences between the control and experimental groups in bone deposition for each marker, in either the BIC or BA analysis were observed. The bone deposition marked by alizarin (14-21 days) was the highest, followed by oxytetracycline (0 and 7 days) and calcein (45 and 52 days) in both groups (P < 0.05). CONCLUSION The bone healing or the course of osseointegration was not impaired by the use of an inorganic xenogenic graft before insertion of a titanium implant.

The bone-induction capacity of a porous biphasic calcium phosphate (pBCP) using heterotopic implantation in mouse (mHI-model) and its efficacy as substitute for autograft in mandibular critical-size defect in rabbit (rabMCSD-model) was investigated. In mHI-model, pBCP was implanted into the thigh muscles and bone formation was histomorphometrically and immunohistochemically evaluated. In rabMCSD-model, 13 mm bone defects were treated with pBCP or autograft and bone repair comparatively evaluated by radiographic and histomorphometric methods. In mHI-model, formed bone and immunolabeling for bone morphogenetic protein-2 and osteopontin were observed in 90% of pBCP implanted samples after 12 weeks. In rabMCSD-model neither statistically significant difference was found in newly formed bone between pBCP and autograft groups at 4 weeks (18.8 ± 5.5% vs 27.1 ± 5.6%), 8 weeks (22.3 ± 2.7% vs 26.2 ± 5.1), and 12 weeks (19.6 ± 4.7% vs 19.6 ± 2.3%). At 12 weeks, the stability and contour of the mandible were restored in both treatments. Near tooth remaining, pBCP particles were covered by small amount of mineralized tissue exhibiting perpendicular attachments of collagen fiber bundles with histological characteristic of acellular cementum. Within the limitations of this study, it was concluded that pBCP is osteoinductive and able to stimulate the new formation of bone and cementum-like tissues in rabMCSD-model, suggesting that it may be an alternative to treatment of large bone defect and in periodontal regenerative therapy. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1546-1557, 2018.

Made available in DSpace on 2018-12-11T17:04:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-09-01 Autogenous bone grafts are used to repair bone defects, and the stabilization is needed for bone regeneration. Laser photobiomodulation is a modality of treatment in clinical practice for tissue regeneration, and it has therapeutic effects as an anti-inflammatory, analgesic and modulating cellular activity. The aim of the present study was to evaluate the effects of low-level laser therapy (LLLT) on an autogenous bone graft integration process stabilized with a new heterologous fibrin sealant. Forty rats were divided into two groups: Autogenous Fibrin Graft (AFG, n = 20), in which a 5 mm dome osteotomy was conducted in the right parietal bone and the graft was adhered to the left side using fibrin sealant; and Autogenous Fibrin Graft Laser (AFGL, n = 20), which was subjected to the same procedures as AFG with the addition of LLLT. The treatment was performed immediately following surgery and then three times a week until euthanasia, using an 830 nm laser (30 mW, 6 J/cm2, 0.116 cm2, 258.6 mW/cm2, 2.9 J). Five animals from each group were euthanized at 10, 20, 30 and 40 days postoperative, and the samples were submitted to histomorphological and histomorphometric analysis. Partial bone regeneration occurred, with new bone tissue integrating the graft to the recipient bed and small areas of connective tissue. Comparative analysis of the groups at the same intervals revealed minor interfaces in group AFGL, with statistically significant differences (p

Made available in DSpace on 2013-08-28T14:12:59Z (GMT). No. of bitstreams: 1 WOS000304960800001.pdf: 4021714 bytes, checksum: 4460e658a306b120b22c525c025ab8ea (MD5) Made available in DSpace on 2013-09-30T18:16:23Z (GMT). No. of bitstreams: 2 WOS000304960800001.pdf: 4021714 bytes, checksum: 4460e658a306b120b22c525c025ab8ea (MD5) WOS000304960800001.pdf.txt: 45587 bytes, checksum: 15aa9ac2ba99e56016b010c09c749e6d (MD5) Previous issue date: 2012-01-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:32:43Z No. of bitstreams: 2 WOS000304960800001.pdf: 4021714 bytes, checksum: 4460e658a306b120b22c525c025ab8ea (MD5) WOS000304960800001.pdf.txt: 45587 bytes, checksum: 15aa9ac2ba99e56016b010c09c749e6d (MD5) Made available in DSpace on 2014-05-20T13:32:43Z (GMT). No. of bitstreams: 2 WOS000304960800001.pdf: 4021714 bytes, checksum: 4460e658a306b120b22c525c025ab8ea (MD5) WOS000304960800001.pdf.txt: 45587 bytes, checksum: 15aa9ac2ba99e56016b010c09c749e6d (MD5) Previous issue date: 2012-01-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Diabetes mellitus is a heterogeneous group of disorders, in which hyperglycemia is a main feature. The objective was to evaluate the involvement of RAGE, inflammatory cytokines, and metalloproteinases in spontaneous periodontitis triggered by diabetes induction. Immunohistochemical procedures for MMP-2, MMP-9, TNF-alpha, IL-1 beta, IL-6, RANKL, and RAGE were performed in rats after 1, 3, 6, 9, and 12 months of diabetes induction. Total DNA was extracted from paraffin-embedded tissues and evaluated by Real-TimePCR for 16S total bacterial load and specific periodontopathogens. Our data did not demonstrate differences in microbiological patterns between groups. In diabetic groups, an increase in RAGE-positive cells was detected at 6, 9, and 12 months, while TNF-alpha-stained cells were more prevalent at 6 and 12 months. In experimental groups, IL-beta-positive cells were increased after 12 months, IL-6 stained cells were increased at 9 and 12 months, and RANKL-positive cells at 9 months. Diabetes resulted in widespread expression of RAGE, followed by expression of proinflammatory mediators, without major alterations in oral microbial profile. The pervasive expression of cytokines suggests that spontaneous periodontitis development may be independent of microbial stimulation and may be triggered by diabetes-driven imbalance of homeostasis. São Paulo Univ FOB USP, Sch Dent Bauru, Dept Biol Sci, BR-17012901 Bauru, SP, Brazil São Paulo State Univ UNESP, Sch Med Botucatu, Dept Surg & Orthoped, BR-18618970 Botucatu, SP, Brazil São Paulo State Univ UNESP, Sch Med Botucatu, Dept Surg & Orthoped, BR-18618970 Botucatu, SP, Brazil FAPESP: 07/07681-2

O objetivo do atual trabalho, foi correlacionar os eventos celulares e teciduais com a expressão das proteínas VEGF, BMP-7, RANKL e OPG durante a osteogênese ectópica e ortotópica, induzida pela matriz óssea (MO) e dentinária (MD) alogênica. Matrizes alogênicas desmineralizada em HCl a 0,6N, obtidas de fêmur e incisivo de ratos, fori implantada entre as fáscias musculares da coxa e em defeito trans-ósseo de 8mm de diâmetro nos ossos parietais. As análises radiográfica e histomorfométrica da neoformação óssea e, a imunohistoquímica e o western blotting para as proteínas VEGF, BMP, RANKL e OPG, mostraram que: a) o volume da região do enxerto nos sítios ortotópicos reduziu 19% em 42 dias; b) em ambos tipos de enxerto e locais de implantação, ocorreu formação de tecido cartilaginoso e ósseo; c) nos sítios intramusculares, a reabsorção da matriz alogênica e a remodelação do tecido cartilaginoso, ósseo e medular foi mais acelerado, em relação a implantação ortotópico; d) o aumento na densidade de volume dos vasos sanguíneos e no número de osteoblastos/osteócitos e osteoclastos ocorreu simultaneamente e estava associado à maior reabsorção da matriz alogênica e à formação do tecido medular (hematopoiético); e) as proteínas VEGF, BMP-7, RANKL, OPG foram expressas em condrócitos, osteoblastos ativos, osteócitos recém aprisionados na matriz e em células estromais próximas aos osteoblastos ou às áreas da matriz alogênica reabsorvida; e f) a expressão das proteínas VEGF, BMP-7, RANKL e OPG foi maior no grupo MO. O pico de expressão dessas proteínas ocorreu nos períodos de 14 aos 21 dias no grupo da MO e 21 e 28 dias no grupo da MD. Concluímos que, a capacidade osteoindutora da matriz alogênica desmineralizada está relacionado a origem da matriz e ao sítio de implantação e que, as proteínas VEGF, BMP-7, RANKL e OPG estão associadas a maior reabsorção da matriz implantada, promovendo uma rápida e contínua liberação dos morfógenos contidos em seu interior que, induzem temporal e espacialmente a formação óssea/medular. The aim of the present work was to correlate the cellular and tissue events with the expression of VEGF, BMP-7, RANKL and OPG during ectopic and orthotopic osteogenesis, induced by bone and dentin allogeneic matrix. Allogenic matrices obtained from femur and incisor of rats and demineralized in 0.6 N HCl were implanted into a intramuscular pocket and a 8mm-diameter bone defect in the skull. The radiographic and histomorphometric analysis of new bone formation, and immunohistochemistry and western blotting for VEGF, BMP, RANKL and OPG proteins, showed that: a) the total volume of the graft region in orthotopic site decreased 19% at 42 days b) in both graft types and implantation sites occurred formation of cartilaginous and bone tissues, c) in intramuscular sites, the resorption of allogenic matrix and remodeling of the new formed cartilage and bone were faster, in relation to orthotopic implantation sites; d) the increase in the volume density of blood vessels and in the number of osteoblasts/osteocytes and osteoclasts occurred simultaneously and was associated with greater reabsorption of the allogenic matrix and hematopoietic bone marrow formation; e) VEGF, BMP-7, RANKL, OPG proteins were expressed in chondrocytes, active osteoblasts, newly osteocytes confined and stromal cells located near the osteoblasts or in the surface of the reabsorbed matrix; and f) the VEGF, BMP-7, RANKL and OPG expression was higher in MO grafts than in the MD. The peak of expression of these proteins each occurred at 14 and 21 days in MO and 21 and 28 days in MD. We concluded that, the osteoinductive capacity of allogeneic demineralized matrix is related to matrix origin and implantation site and that the VEGF, BMP-7, RANKL and OPG proteins are associated with greater reabsorption of the implanted matrix, promoting rapid and continuous matrix-release morphogens that induces spatially and temporally the bone and bone marrow formation.

Objective: To histomorphometrically investigate the repair of critical size defects (CSDs) and bone augmentation in cranial walls using block of sintered bovine-derived anorganic bone (sBDAB) graft. Material and methods: Forty guinea-pigs were divided into test (n ¼20) and CSD control (n ¼20) groups. In each animal, a full-thickness bone defect with 9.5mm diameter was made in the frontal bone. The defects were filled with an sBDAB block soaked in blood in the test group and with blood clot in the CSD control group. The skulls were collected at 0h (n ¼2) and 30, 90 and 180 days (n ¼6/group and period) postoperatively. The volume density and total volume of newly formed bone, sBDAB, blood vessels and connective tissue, vertical thickness of removed bone plug, sBDAB block and graft area were evaluated. Results: The vertical thickness of the adapted sBDAB block was 3.8 times higher than that of the removed bone plug and did not show significant difference between periods, filling in average 29.8% of the total graft region. The sBDAB block exhibited complete osseointegration with the borders of the defect at 90 days. At 90 and 180 days, the vertical thickness of the graft was 279% in the average, and the total volume of bone augmentation was, respectively, 78.8% and 148.5% higher compared with the removed bone plug. The defects of the CDS control group showed limited osteogenesis and filling by connective tissue plus tegument. Conclusion: The sBDAB block can be used to promote repair of CSDs and bone augmentation in the craniomaxillofacial region, due to its good osteoconductive and slow resorptive properties.

Background: Vascular endothelial growth factor (VEGF) is a macromolecule of importance in inflammation that has been implicated in periodontitis. The aims of this study were to investigate VEGF expression during the progression of periodontal disease and to evaluate the effect of a preferential cyclooxygenase (COX)-2 inhibitor meloxicam on VEGF expression and alveolar bone loss in experimentally induced periodontitis. Methods: A total of 120 Wistar rats were randomly separated into groups 1 (control) and 2 (meloxicam, 3mg/kg/day,intraperitoneally,for3,7,14,or30days).Silkligatureswereplacedatthegingivalmarginlevel of the lower right first molar of all rats. VEGF expression was assessed by reverse transcription-polymerase chain reaction (RT-PCR), Western blot (WB), and immunohistochemical (IHC) analyses. The hemiarcades wereprocessedforhistopathologicanalysis.RT-PCRandWBresultsweresubmittedtoanalysisofvariance, the Tukey test, and Pearson correlation analysis (P

Diabetes mellitus comprises a heterogeneous group of disorders with the main feature of hyperglycemia. Chronic hyperglycemia increases the severity of periodontal disease via an exacerbated inflammatory response, activated by advanced glycation end products and their receptor, RAGE. Therefore, anti-inflammatory agents represent potential inhibitors of this pathological interaction. In particular, green tea has been shown to possess anti-inflammatory properties mediated by its polyphenol content. Objectives: This study investigated the mechanisms by which green tea attenuates the spontaneous onset of diabetes-induced periodontitis. Methods: Diabetes was induced in rats via a single intraperitoneal injection of streptozotocin (STZ). Diabetic and control animals were divided into water-treated and green tea-treated subgroups and were analyzed at 15, 30, 60 and 90 days after diabetes induction. Immunohistochemistry was performed to quantitatively evaluate tumor necrosis factor-α (TNF-α), receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG), interleukin-10 (IL-10) and runt-related transcription factor 2 (RUNX-2) expression in serial sections of each hemimaxilla. Morphometric measurements of the distance from the cementum-enamel junction (CEJ) of the superior distal root of the first molar to the alveolar bone crest (ABC) were performed to assess bone loss. Results: Diabetes resulted in significant bone loss and alterations in the number of cells that stained positive for inflammatory mediators. In the diabetic rats treated with green tea, we observed a decreased number of cells expressing RANKL and TNF-α compared with that observed in the diabetic rats treated with water. Additionally, green tea increased the numbers of cells that stained positive for OPG, RUNX-2 and IL-10 in the diabetic rats. Conclusion: Green tea intake reduces expression of the pro-inflammatory cytokine TNF-α and the osteoclastogenic mediator RANKL to normal levels while increasing expression of the anti-inflammatory cytokine IL-10, the osteogenesis-related factor RUNX-2 and the anti-osteoclastogenic factor OPG. Therefore, green tea represents a potential therapeutic agent for the treatment of diabetes-related periodontal disease.

Vascular endothelial growth factor (VEGF) plays an important role during angiogenesis and bone repair. This study investigated whether the use of meloxicam alters bone repair via downregulation of VEGF and receptor expression.One hundred twenty male Wistar rats had their maxillary right incisor extracted. Animals were divided into a control group (CG; n = 60) and a meloxicam-treated group (TG; n = 60) that received either a single daily intraperitoneal injection of 0.9% NaCl or meloxicam 3 mg/kg, respectively, for 7 consecutive days. Alveolar bone repair was evaluated histomorphometrically, whereas VEGF and its receptors were analyzed by immunohistochemistry and quantitative polymerase chain reaction (qPCR). Data were submitted to two-way analysis of variance and Tukey post hoc test with P0.05.Bone volume density increased significantly (P = 0.001) in both groups with a strong correlation between treatment and periods (P = 0.003). In the TG, a small amount of bone formation occurred compared with the CG between 3 and 21 days. No significant differences in the number of VEGF-positive cells per square millimeter (P = 0.07) and VEGF messenger RNA (mRNA) expression (P = 0.49) were found between groups. Immunostained cells per square millimeter and mRNA expression for VEGF receptor (VEGFR)-1 (P = 0.04 and P0.001) and VEGFR-2 (P0.001 for both analysis) showed a strong interaction between treatment groups and periods. In the TG, immunostained cells per square millimeter and mRNA expression for VEGFR-1 were, respectively, 89% and 37% lower from 3 to 10 days compared with the CG, whereas for VEGFR-2, these values were 252% and 60%, respectively, from 3 to 7 days.In rat alveolar bone repair, meloxicam did not affect VEGF expression but downregulated VEGFR expression, which may cause a delay in the bone repair/remodeling process.

Poucos estudos demonstram as reações patológicas devidas à inalação de fumaça pelas vias aéreas de ratos. OBJETIVO: Estudar e analisar os possíveis efeitos histopatologicos produzidos pela inalação crônica de fumaça de cigarro nas pregas vocais de ratos. DESENHO DE ESTUDO: Experimental. MATERIAL E MÉTODOS: 36 ratos masculinos (Rattus norvergicus Wistar), de 60 dias de idade, foram mantidos em gaiolas e expostos à inalação da fumaça produzida por 10 cigarros, 3 vezes ao dia, 7 dias por semana, para períodos de 25, 50 e 75 dias, e os controles respectivos. Os animais foram sacrificados e as suas laringes dissecadas e submetidas a análise histológica com coloração de Hematoxilina e Eosina e analisados através de microscopia simples. RESULTADOS: Os ratos expostos ao cigarro exibiram menor (p Few studies have demonstrated the pathologic reactions yielded by smoke inhalation on the airway in rats. AIM: The aim of this study was to analyze the possible histopathological effects produced by chronic cigarette smoke inhalation on the vocal folds of rats. STUDY DESIGN: Experimental. MATERIAL AND METHOD: 36 male rats (Rattus norvergicus Wistar strain), aged 60 days, were kept in cages and exposed to inhalation of the smoke produced by 10 cigarettes lit 3 times a day, 7 days a week, for periods of 25, 50 and 75 days, and their respective controls. Thereafter the animals were killed and their larynxes were dissected and submitted to histological processing for achievement of histological sections, which were stained with Hematoxylin and Eosin and analyzed by light microscopy. RESULTS: The rats exposed to smoke displayed smaller (p< 0,05) body mass than the control group. There was hyperplasia and squamous metaplasia in the free edge of the vocal fold and squamous hyperplasia on the middle portion of the vocal fold in all 3 study periods. Moreover, the 50-day group revealed keratinizing metaplasia in this area. Morphological alterations in other areas of the larynx and inflammatory reaction of the lamina propria were also not observed. CONCLUSION: It was concluded that the passive inhalation of cigarette smoke yields important morphological changes in the vocal fold epithelium, which may progress to neoplasia.

SummaryFew studies have demonstrated the pathologic reactions yielded by smoke inhalation on the airway in rats. Aim: The aim of this study was to analyze the possible histopathological effects produced by chronic cigarette smoke inhalation on the vocal folds of rats. Study design: Experimental. Material and Method: 36 male rats (Rattus norvergicus Wistar strain), aged 60 days, were kept in cages and exposed to inhalation of the smoke produced by 10 cigarettes lit 3 times a day, 7 days a week, for periods of 25, 50 and 75 days, and their respective controls. Thereafter the animals were killed and their larynxes were dissected and submitted to histological processing for achievement of histological sections, which were stained with Hematoxylin and Eosin and analyzed by light microscopy. Results: The rats exposed to smoke displayed smaller (p< 0,05) body mass than the control group. There was hyperplasia and squamous metaplasia in the free edge of the vocal fold and squamous hyperplasia on the middle portion of the vocal fold in all 3 study periods. Moreover, the 50-day group revealed keratinizing metaplasia in this area. Morphological alterations in other areas of the larynx and inflammatory reaction of the lamina propria were also not observed. Conclusion: It was concluded that the passive inhalation of cigarette smoke yields important morphological changes in the vocal fold epithelium, which may progress to neoplasia.

The ability of a pool of bovine bone morphogenetic proteins bound to synthetic microgranular hydroxyapatite (BMPb-HA) to stimulate bone repair was determined in rat critical size defects. An 8-mm diameter defect was created in the calvaria of 25 rats. In 15 rats, the defects were filled with BMPb-HA homogenized with blood (experimental group), and in 10 rats the defects were filled only with blood clots (control). The calvariae of experimental rats were collected 1, 3 and 6 months after surgery and of the control rats at the end of surgery and 6 months thereafter. The morphometric results obtained in the radiographs showed an absence of new bone formation at 1 and 3 months post-surgery and, histologically, the defects were filled with fibrous connective tissue and numerous foci of a foreign body-type granulomatous reaction around hydroxyapatite agglomerates. At the end of 6 months, the number and size of the granulomatous foci decreased and the area of the defects was reduced by 22% compared to the 0-hour control due to the formation of new bone at their borders, although the mean area was similar to the 6-month control. We conclude that the use of BMPb-HA in the treatment of critical size bone defects of the rat skull leads to the formation of a foreign body-type granulomatous reaction that markedly inhibits new bone formation, suggesting that synthetic microgranular hydroxyapatite does not represent a good carrier for BMP-induced bone formation. A capacidade de um pool de proteínas morfogenéticas bovinas ligada a hidroxiapatita sintética absorvível microgranular (BMPb-HA) em estimular a reparação óssea foi avaliada em defeito de tamanho crítico em calvária de rato. Defeitos de 8 mm diâmetro foi produzido na calvária de 25 ratos, sendo 15 preenchidos com BMPb-HA homogeinizado com sangue (grupo experimental) e 10 somente com coágulo sanguíneo (grupo controle). As calvárias dos animais do grupo experimental foram coletadas 1, 3 e 6 meses após as cirurgias e as do grupo controle, ao término da cirurgia e após 6 meses. Os resultados morfométricos obtidos nas imagens radiográficas apresentou ausência de formação óssea nos períodos de 1 e 3 meses e, histologicamente os defeitos estavam preenchidos por tecido conjuntivo fibroso e inúmeros focos de reação granulomatosa, tipo corpo estranho, ao redor de aglomerados de hidroxiapatita. Ao final de 6 meses, os focos de reação granulomatosa diminuiu em número e tamanho e os defeitos diminuíram a sua área em 22% em relação ao controle 0 hora, devido a neoformação óssea nas suas bordas, mas apresentaram área semelhante ao controle 6 meses. Concluímos que a utilização de BMPb-HA no tratamento de defeitos ósseos cranianos em ratos, promove a formação de reação granulomatosa tipo corpo estranho que inibe profundamente a neoformação óssea, sugerindo que a hidroxiapatita sintética microgranular não é um bom carreador para as BMPs expressarem o seu potencial indutor.

Made available in DSpace on 2021-07-14T10:29:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2004-03. Added 1 bitstream(s) on 2021-07-14T11:31:43Z : No. of bitstreams: 1 S1678-77572004000100012.pdf: 352622 bytes, checksum: 2cef959e33abacc47fb471f71f516a4a (MD5) Nos últimos anos, vários materiais de origem bovina para enxerto ósseo têm sido lançados no comércio para serem utilizados em cirurgias bucomaxilofaciais. Na presente pesquisa avaliamos a capacidade de um material de enxerto ósseo orgânico em bloco (Gen-ox, Baumer SA - Brasil) preparado à partir de osso medular bovino, em promover a reparação de lesões ósseas de tamanho crítico em calvária de ratos. Uma lesão trans-óssea de aproximadamente 8mm de diâmetro foi realizada com trefina cirúrgica nos ossos parietais de 25 ratos, sendo que em 15 os defeitos foram preenchidos com bloco de 8mm de diâmetro do material de enxerto, embebida com sangue do próprio animal e em 10 somente com coágulo sanguíneo. As calvárias dos ratos tratados com o material foram coletadas após 1, 3 e 6 meses após as cirurgias e dos tratados com coágulo sanguíneo, imediatamente após as cirurgias e decorridos 6 meses. Durante as cirurgias verificamos que esse material enxertado é de fácil manuseio e se adapta perfeitamente ao leito receptor após sua embebição com sangue. Os resultados obtidos mostraram que na maioria dos casos tratados, o material foi reabsorvido lentamente e serviu como material de preenchimento e mantenedora de espaço, favorecendo a angiogênese, migração e adesão celular e a neoformação óssea à partir das bordas da lesão. No entanto, em 2 casos no subgrupo tratado - 1 mês e 1 caso no subgrupo tratado - 3 meses, ocorreu a presença de reação granulomatosa tipo corpo estranho com a presença de inúmeras células gigantes, que inibiu a neoformação óssea local. Esses casos foram interpretados como decorrentes de falhas na desmineralização e na retirada de potenciais fatores antigênicos durante a produção do biomaterial. Concluímos que com um aprimoramento do controle de qualidade na linha de produção do material, a matriz orgânica medular em bloco poderá ser uma boa alternativa para ser usada no reparo de defeitos ósseos na região buco-maxilofacial, devido a sua alta capacidade osteocondutora. Over the last few years, various bone graft materials of bovine origin to be used in oromaxillofacial surgeries have entered the market. In the present study, we determined the capacity of a block organic bone graft material (Gen-ox, Baumer SA, Brazil) prepared from bovine cancellous bone to promote the repair of critical size bone injuries in rat calvaria. A transosseous defect measuring approximately 8mm in diameter was performed with a surgical trephine in the parietal bone of 25 rats. In 15 animals, the defects were filled with a block of graft material measuring 8mm in diameter and soaked in the animal's own blood, and in the other 10 animals the defects were only filled with blood clots. The calvariae of rats receiving the material were collected 1, 3 and 6 months after surgery, and those of animals receiving the blood clots were collected immediately and 6 months after surgery. During surgery, the graft material was found to be of easy handling and to adapt perfectly to the receptor bed after soaking in blood. The results showed that, in most animals treated, the material was slowly resorbed and served as a space filling and maintenance material, favoring angiogenesis, cell migration and adhesion, and bone neoformation from the borders of the lesion. However, a foreign body-type granulomatous reaction, with the presence of numerous giant cells preventing local bone neoformation, was observed in two animals of the 1-month subgroup and in one animal of the 3-month subgroup. These cases were interpreted as resulting from the absence of demineralization and the lack of removal of potential antigen factors during production of the biomaterial. We conclude that, with improvement in the quality control of the material production, block organic bone matrix will become a good alternative for bone defect repair in the oromaxillofacial region due to its high osteoconductive capacity. Universidade Estadual Paulista, Araçatuba Dental School Universidade de São Paulo, Bauru Dental School Universidade Estadual Paulista, Araçatuba Dental School

The presence of morphological differences in the sublingual gland of male and female adult rats was determined by morphometry. Absolute and relative glandular mass was 21% lower and 31% higher, respectively, in females than in males. The fractions of glandular volume occupied by the mixed acini, intercalated ducts and striated ducts did not differ significantly between genders; however, their absolute volume was respectively 29, 42 and 58% higher in males. Despite the differences in the volume of these morphological compartments, the number of cells did not differ significantly between genders, except for the excretory duct compartment, for which a larger number was observed in males. With respect to cell volume, 13, 33 and 47% higher volumes were observed in males for mucous acinar cells and striated and excretory duct cells, respectively, while a 38% higher volume of serous demilune cells was observed for females. The surface-to-volume ratio of acini and striated ducts was respectively 16 and 35% higher in females. Based on these results, we conclude that the sublingual gland of female rats possesses smaller acini, and shorter ducts whose caliber is narrower, smaller mucous acinar and larger serous cells than the ones found in the male gland, indicating the presence of sexual dimorphism as well as suggesting sexual differences in the quality of the secreted product. A ocorrência de diferenças morfológicas entre sexos na glândula sublingual de ratos adultos foi verificada pela morfometria. As massas glandular absoluta e relativa das fêmeas foi, respectivamente, 21% menor e 31% maior que as dos machos. As frações de volume glandular ocupadas pelos ácinos mistos, ductos intercalares e ductos estriados não mostraram diferenças significantes entre sexos, no entanto, os seus volumes absolutos foram, respectivamente, 29%, 42% e 58% maiores nos machos. Apesar dessas diferenças nos volumes compartimentais, os seus conteúdos em número de células não apresentaram diferenças significantes entre sexos, exceto o compartimento dos ductos excretores, que mostrou maior número nos machos. Quanto ao volume celular, as células acinosas mucosas, as dos ductos estriados e as dos ductos excretores mostraram volumes, respectivamente, 13%, 33% e 47% maiores nos machos, e o das células das semiluas serosas foi 38% maior nas fêmeas. A relação superfície-volume dos ácinos e dos ductos estriados foi, respectivamente, 16% e 35% maior nas fêmeas. Baseados nos resultados obtidos, concluímos que as glândulas sublinguais das fêmeas exibem ácinos menores e ductos mais curtos e menos calibrosos e células acinosas mucosas menores e serosas maiores do que nos machos, indicando a ocorrência de dimorfismo sexual, e sugerindo que possa haver também diferenças na qualidade do produto secretado.

Background: In recent years, important advances have occurred in the determination of diagnostic criteria for the disease diabetes mellitus and in new strategies for its treatment. The purpose of this research was to develop a new method for diabetes diagnosis by microscopic and cytomorphometric analyses of the oral epithelium. Methods: The smears were obtained from three distinct oral sites: buccal mucosa (cheek), tongue dorsum, and floor of the mouth in 10 control individuals and 10 type II diabetic patients. The oral smears were stained with Papanicolaou EA-36 solution. The nuclear (NA) and cytoplasmic (CA) areas were evaluated from 50 integral cells predominant in each oral site by the use of the KS 300™ image analysis system (Carl Zeiss, Germany), by which the cytoplasmic/nuclear ratio (C/N) was calculated. Results: The results showed that: (i) the epithelial cells of the diabetic group exhibited figures of binucleation and occasional karyorrhexis in all layers; (ii) the NA was markedly higher (P 0.05) between these two groups; and (iv) the C/N mean was 37.4% lower in the type II diabetic group. Conclusions: These results associated with clinical observations suggest that diabetes mellitus can produce alterations in oral epithelial cells, detectable by microscopy and cytomorphometry, which can be used in the diagnosis of this disease.

This study aimed to evaluate morphometrically the bone formation and immunohistochemically the expression of vascular endothelial growth factor (VEGF) and metalloproteinase (MMP)-2 and -9 during the healing of critical-size defects treated with sintered anorganic bone (sAB). The 8-mm diameter full-thickness trephine defects created in the parietal bones of rats were filled with sAB (test group) or blood clot (CSD-control group). At 7, 14, 21, 30, 90 and 180 days postoperatively (n = 6/period) the volume of newly formed bone and total number of immunolabeled cells (Ntm) for each protein were determined. Bone formation was smaller and faster in the CSD-control group, stabilizing at 21 days (6.74 mm(3)). The peaks of VEGF, MMP-2 and MMP-9 occurred at 7 and 14 days in fibroblasts and osteoblasts, with mean reduction of 0.80 time at 21 days, keeping constant until 180 days. In the test group, sAB provided continuous bone formation between particles throughout all periods. The peak of MMP-2 was observed at 7-14 days in connective tissue cells and for VEGF and MMP-9 at 30 days in osteoblasts and osteocytes. Ntm for VEGF, MMP-2 and MMP-9 were in average, respectively, 3.70, 2.03 and 5.98 times higher than in the control group. At 180 days, newly formed bone (22.9 mm(3)) was 3.74 times greater in relation to control. The physical and chemical properties of sAB allow increased autocrine expression of VEGF, MMP-2 and MMP-9, favoring bone formation/remodeling with very good healing of cranial defects when compared to natural repair in the CSD-control.

The morphometric dimensions of the various structures of the pancreas of adult Syrian golden hamsters, of both sexes, were evaluated using stereological methods. The average body mass of the animals used was 133.8 +/- 2.45 g and 140.6 +/- 7.98 g for the males and females, respectively, and the pancreatic mass, 389.9 +/- 14.88 and 409.7 +/- 21.42 mg, respectively. The analysis of variance of the obtained data showed that: a) the acini, intercalated ducts and stroma did not present statistically significant differences in any of the dimensions evaluated, with the exception of the nucleus volume of the acinar cells which was 8.5% larger in the female (P < 0.05); b) the excretory ducts exhibited surface density, total external surface, surface-to-volume ratio, and absolute cell number, 18%, 33%, 14%, and 44%, respectively, larger in the females (P < 0.05); and c) the pancreatic islets of the females exhibited volume density, total volume and absolute cell number, 20%, 27% and 27%, respectively, larger than those of the males (P < 0.05).

Fluoride (F) is a potent anti-cariogenic element, but when ingestion is excessive, systemic toxicity may be observed. This can occur as acute or chronic responses, depending on both the amount of F and the time of exposure. The present study identified the profile of protein expression possibly associated with F-induced chronic hepatotoxicity. Weanling male Wistar rats (three-weeks old) were divided into three groups and treated with drinking water containing 0, 5 or 50 mg/L F for 60 days (n=6/group). At this time point, serum and livers were collected for F analysis, which was done using the ion-sensitive electrode, after hexamethyldisiloxane-facilitated diffusion. Livers were also submitted to histological and proteomic analyses (2D-PAGE followed by LC-MS/MS). Western blotting was done for confirmation of the proteomic data A dose-response was observed in serum F levels. In the livers, F levels were significantly increased in the 50 mg/L F group compared to groups treated with 0 and 5 mg/L F. Liver morphometric analysis did not reveal alterations in the cellular structures and lipid droplets were present in all groups. Proteomic quantitative intensity analysis detected 33, 44, and 29 spots differentially expressed in the comparisons between control vs. 5 mg/L F, control vs. 50 mg/L F, and 5 mg/L vs. 50 mg/L F, respectively. From these, 92 proteins were successfully identified. In addition, 18, 1, and 5 protein spots were shown to be exclusive in control, 5, and 50 mg/L F, respectively. Most of proteins were related to metabolic process and pronounced alterations were seen for the high-F level group. In F-treated rats, changes in the apolipoprotein E (ApoE) and GRP-78 expression may account for the F-induced toxicity in the liver. This can contribute to understanding the molecular mechanisms underlying hepatoxicity induced by F, by indicating key-proteins that should be better addressed in future studies.

Due to issues in the typesetting process, there were errors in Figures 3,4, and 6. Correct versions of these Figures are available below. Figure 3: Figure 4: Figure 6

OBJECTIVE: This study evaluated experimentally-induced periapical bone loss sites using digital radiographic and histopathologic parameters. MATERIAL AND METHODS: Twenty-seven Wistar rats were submitted to coronal opening of their mandibular right first molars. They were radiographed at 2, 15 and 30 days after the operative procedure by two digital radiographic storage phosphor plates (Digora®). The images were analyzed by creating a region of interest at the periapical region of each tooth (ImageJ) and registering the corresponding pixel values. After the sacrifice, the specimens were submitted to microscopic analysis in order to confirm the pulpal and periapical status of the tooth. RESULTS: There was significant statistically difference between the control and test sides in all the experimental periods regarding the pixel values (two-way ANOVA; p

Heterotopic osteogenesis induced by allogenic bone matrix graft in the mouse muscle was analyzed by radiography, morphology and morphometry. Diaphyses of the femurs of 18 fifty-day-old mice were demineralized with 0.6 M HCl and implanted into the adductor muscle of the right thigh of 36 mice. Two, 5, 7, 14, 21 and 28 days after implantation, groups of 6 animals/period were sacrificed and radiographied, and specimens were collected and processed histologically. The presence of mineralized material in the graft was detected by radiography only on day 21. Morphologic and morphometric analysis of the histological sections showed that: a) the matrix remained intact up to day 7, followed by resorption of 16% by fibroblast-like mononuclear cells up to day 28; b) cartilage tissue was first detected on day 14 at the highest quantity observed in the experiment, occupying 5% of the graft volume and containing 4 x 102 chondrocytes/mm3 graft, followed by a substantial decline up to day 28 when it reached a volume density of 0.5% and 0.6 x 102 chondrocytes/mm3; c) bone cells and matrix were detected in minute amounts from day 14 on, with a volume density of 0.4% and containing 0.2 x 102 cells/mm3, which increased about 15 and 32 times, respectively, by the end of the 28-day, reaching 6% of the graft volume and 6 x 102 cells/mm3. We conclude that, despite the relatively low percentage of graft matrix resorption (total of 16%), the matrix was able to induce all events of the endochondral ossification cascade, i.e., formation and subsequent disappearance of hyaline cartilage, osteoblast differentiation, new bone formation and, later, the formation of hematogenous bone marrow.

OBJECTIVE: To analyze and to quantify morphological acinar postmortem changes in rat sublingual glands (SLG). MATERIAL AND METHODSs: Fifty rats were divided into two groups of 25 animals each. Group I was used for morphological and morphometric evaluations and group II for the determination of gland density and processed gland volume. Acinar autolytic changes were studied at 0 (control group), 3, 6, 12 and 24 h postmortem periods. The morphometric analysis of the volume density (Vv) and total volume (Vt) of intact (ia) and autolyzed (aa) acini was performed under light microscopy using a Zeiss II integration grid with 100 symmetrically distributed points. RESULTS: Morphologically, temporal progressive nuclear alterations and gradual loss of the structural architecture of acinar cells were found. Regarding quantitative results, both the Vvaa and the Vvia showed statistically significant differences among all postmortem periods (p0.05), respectively. Vtaa increased from 0.18 mm³ at 0 h to 38.17 mm³ at 12 h, while Vtia showed a decrease from 33.47 mm³ to 0 mm³ between 3-24 h postmortem. Data concerning Vtaa were adjusted by two-variable linear regression, obtaining the equation: y=-3.54 + 3.38x (r²=0.90). The Vtaa growth rate calculated by this equation was 3.38 mm³/h between 0-12 h. CONCLUSION: Acinar autolysis on rat SLG demonstrated the most significant signs during the first 6 h postmortem and was widely spread through the gland at 12 h.

The development of the rat sublingual gland during the first month of postnatal life was analyzed by morphometry. The absolute number of cells in the various morphological glandular compartments - mixed acini with serous demilunes, intercalated ducts, striated ducts and stroma - was determined by the Aherne II morphometric method for particle counting. The data of fresh glandular volume and the number of cells were analyzed by linear, exponential and parabolic regression, with the best fits being obtained with the linear equation. Based on these equations, the growth rate of glandular volume and of each cell population was calculated. Glandular volume increased about 12-fold during the period from 2 to 30 days of postnatal life, corresponding to a rate of 1.34 mm3/day. This volumetric growth of the gland was mainly due to significant 16-, 10-, 4-, 7- and 8-fold increases in the absolute number of mucous acinar, serous demilune, intercalated duct, striated duct and stromal cells, corresponding to a growth rate of 301 x 103, 228 x 103, 19 x 103, 54 x 103 and 247 x 103 cells/day, respectively. Based on the present results, we conclude that the mucous and serous cell populations of the mixed acini grew at a rate close to that of stromal cells, but considerably higher than that obtained for the intercalated duct and striated duct populations, with the lowest growth rate being observed for the ntercalated duct cell population.

This study evaluated the biomechanical and microscopic response of previously grafted bone to titanium implants. The lower incisors of 16 rabbits were surgically extracted, and bilateral perforations communicating with the remaining sockets were created distally. A socket/perforation defect on each mandible was chosen at random to be immediately filled with a xenogenic graft, whereas the contralateral perforation was left to heal naturally and served as a paired control. After 60 days, titanium implants were installed in the previously operated areas. After periods of 2 and 6 months, the animals were killed, and the force necessary to retrieve implants as well as the bone-implant contact (BIC) and bone mass (BM) were quantified and statistically compared by 2-way analysis of variance and Tukey's test (α = .05). No significant differences in removal torque were observed, either by time or by treatment condition. Differences in BIC and BM between experimental and control groups were not statistically significant through the intervals studied (P < .05). The presence of a xenogenic graft did not influence the microscopic tissue response to titanium implants or fixation into newly formed or mature bone.