Influence of microcurrent on the modulation of remodelling genes in a wound healing assay.

The literature has shown the beneficial effects of microcurrent (MC) therapy on tissue repair. We investigated if the application of MC at 10 µA/90 s could modulate the expression of remodeling genes transforming growth factor beta (Tgfb), connective tissue growth factor (Ctgf), insulin-like growth factor 1 (Igf1), tenascin C (Tnc), Fibronectin (Fn1), Scleraxis (Scx), Fibromodulin (Fmod) and tenomodulin in NIH/3T3 fibroblasts in a wound healing assay. The cell migration was analyzed between days 0 and 4 in both fibroblasts (F) and fibroblasts + MC (F+MC) groups. On the 4th day, cell viability and gene expression were also analyzed after daily MC application. Higher expression of Ctgf and lower expression of Tnc and Fmod, respectively, were observed in the F+MC group in relation to F group (p < 0.05), and no difference was observed between the groups for the genes Tgfb, Fn1 and Scx. In cell migration, a higher number of cells in the scratch region was observed in group F+MC (p < 0.05) compared to group F on the 4th day, and the cell viability assay showed no difference between the groups. In conclusion, MC therapy at an intensity/time of 10 µA/90 s with 4 daily applications did not affect cell viability, stimulated fibroblasts migration with the involvement of Ctgf, and reduced the Tnc and Fmod expression.

Diabetes mellitus and periodontitis: Inflammatory response in orthodontic tooth movement.

OBJECTIVES: This study evaluated, in experimental model, the inflammatory alterations in gingival tissue and alveolar bone during the orthodontic tooth movement (OTM) in diabetes mellitus (D) and periodontitis (P). SETTING AND SAMPLE POPULATION: Forty male Wistar rats, 90 days old and weighing 300 g. MATERIALS AND METHODS: The sample was divided into four groups (n = 10). OTM: orthodontic movement (10 days, 0.4 N force); P + OTM: periodontitis (ligature-induced periodontitis, 3-0 silk suture thread) and orthodontic movement; D + OTM: diabetes (Alloxan-induced diabetes, 150 mg/kg) and orthodontic movement; and D + P + OTM: diabetes, periodontitis and orthodontic movement. Tooth displacement was measured; fibroblast, inflammatory cells, osteoclast and blood vessels were quantified by histomorphometric analysis. Inflammatory markers, interleukin-6 (IL-6) and tumour necrosis factor (TNF-α) were quantified by ELISA (Enzyme-Linked Immunosorbent Assay) in gingival tissue. The fibroblastic growth factor (bFGF), transforming growth factor (TGF-ß1) and the vascular endothelial growth factor (VEGF) were measured via Western blotting in the alveolar bone. The results were analysed by ANOVA and Tukey's test at a 5% significance level. RESULTS: The quantification of inflammatory cells and the expression of IL-6, TNF-α, TGF-ß1 and bFGF were increased in diabetes and periodontitis. However, the number of fibroblasts and blood vessels and the percentage of birefringent collagen fibres were higher in healthy animals. There was greater tooth displacement in the OTM group. CONCLUSION: Diabetes Mellitus modifies the inflammatory response. The increased expression of inflammatory markers IL-6, TNF-α and TGF-ß1 in diabetic animals impairs neovasculogenesis and tissue reorganization during orthodontic tooth movement, which may be aggravated by periodontitis.

Do electrical current and laser therapies improve bone remodeling during an orthodontic treatment with corticotomy?

OBJECTIVES: Evaluate the bone remodeling during orthodontic movement with corticotomy when submitted to low-intensity electrical stimulation application (microcurrent-MC) and low-level laser therapy (LLLT). MATERIAL AND METHODS: One hundred and fifty Wistar rats were divided into the following 5 groups: (C) submitted to tooth movement; (Cort) tooth movement/corticotomy; (Cort-L) tooth movement/corticotomy/laser AsGaAl 808 nm (4.96J/50s); (Cort-Mc) tooth movement/corticotomy/microcurrent (10 µA/5 min); (Cort-L-Mc) tooth movement/corticotomy and laser/microcurrent alternated. Inflammation, angiogenesis, and osteogenesis were evaluated in the periodontal ligament (PDL) and alveolar bone on the 7th, 14th, and 21st days of orthodontic movement. RESULTS: The quantification of inflammatory infiltrate, angiogenesis and expression of TGF-ß1, VEGF, and collagen type I were favorably modulated by the application of therapies such as low-level laser therapy (LLLT), MC, or both combined. However, electrical stimulation increased fibroblasts, osteoclasts and RANK numbers, birefringent collagen fiber organization, and BMP-7 and IL-6 expression. CONCLUSIONS: Low-level laser therapy (LLLT) and MC application both improved the process of bone remodeling during orthodontic treatment with corticotomy. Still, electrical current therapy promoted a more effective tooth displacement but presented expected root resorption similar to all experimental treatments. CLINICAL RELEVANCE: It is important to know the effects of minimally invasive therapies on cellular and molecular elements involved in the bone remodeling of orthodontic treatment associated with corticotomy surgery, in order to reduce the adverse effects in the use of this technique and to establish a safer clinical routine.

Electrical stimulation enhances tissue reorganization during orthodontic tooth movement in rats.

OBJECTIVE: This study evaluated the effects of a low-intensity electric current on tissue reorganization during experimental orthodontic tooth movement. MATERIALS AND METHODS: Thirty-two animals were divided into two groups evaluated on days 3 and 7: OTM-orthodontic tooth movement and OTM + MC-orthodontic tooth movement and microcurrent application (10 µA/5 min). The samples were processed for histological, morphometric, and Western blotting analysis. RESULTS: Analysis of the periodontal ligament (PL) showed a significantly smaller number of granulocytes in the OTM + MC group on day 7.The number of fibroblasts was significantly higher in the OTM + MC group on days 3 and 7. The area of birefringent collagen fibers was more organized in the OTM + MC group on days 3 and 7. The number of blood vessels was significantly higher in the OTM + MC group on day 7. Microcurrent application significantly increased the number of osteoclasts in the compression region of the PL. In the OTM + MC group on day 7 of tooth movement, the expression of TGF-ß1 and VEGF was significantly reduced whereas the expression of bFGF was increased in PL. CONCLUSIONS: Electrical stimulation enhances tissue responses, reducing the number of granulocytes and increasing the number of fibroblasts, blood vessels, and osteoclasts and modulates the expression of TGF-ß1, VEFG, and bFGF. CLINICAL RELEVANCE: This technique is used in many areas of medicine, but poorly explored in dentistry and orthodontics. This treatment is cheap and non-invasive and can be applied by own orthodontist, and it can improve the treatment with a faster and safe tooth movement, without pain.

InGaP 670-nm laser therapy combined with a hydroalcoholic extract of Solidago chilensis Meyen in burn injuries.

Therapies that accelerate the healing of burn injuries, improving the quality of life of the patient and reducing the cost of treatment are important. This study evaluated the effects of InGaP 670-nm laser therapy combined with a hydroalcoholic extract of Solidago chilensis leaves on burn wound healing in rats. Seventy-two rats were divided randomly into four groups: control untreated (C), treated with InGaP 670-nm laser with power density of 0.41 W/cm(2) and energy density of 4.93 J/cm(2) (L), treated with S. chilensis extract (S) and treated with S. chilensis extract and laser (LS). Second-degree burns were produced on the back of the animals with metal plate. Wound samples were collected on days 7, 14 and 21 of treatment for structural analysis, morphometry and Western blotting to quantify the expression of transforming growth factor beta 1 (TGF-ß1) and vascular endothelial growth factor (VEGF). The results showed that InGaP laser irradiation at 670 nm alone and combined with extract of S. chilensis promoted significant tissue repair responses in this experimental model, increasing the number of fibroblasts, collagen fibres and newly formed blood vessels throughout the experimental period and decreasing the number of granulocytes in burn wounds of second degree in all treated groups. Exclusive treatment of burn wounds with the hydroalcoholic extract of S. chilensis provided similar quantitative results to those seen in the untreated group throughout the experimental period. Therefore, it was observed in the L and LS groups different responses in the expression of TGF-ß1 and VEGF. The application of 670-nm laser alone or combined with the extract of S. chilensis promoted favourable responses in tissue repair of second-degree burns in this experimental model.

Effects of laser irradiation (670-nm InGaP and 830-nm GaAlAs) on burn of second-degree in rats.

This study investigated the effects of 670-nm indium gallium phosphide (InGaP) and 830-nm gallium aluminum arsenide (GaAlAs) laser therapy on second-degree burns induced on the back of Wistar rats. Sixty-three male Wistar rats were anesthetized, and second-degree burns were made on their back. The animals were then divided randomly into three groups: control (C), animals treated with 670-nm InGaP laser (LIn), and animals treated with 830-nm GaAlAs laser (LGa). The wound areas were removed after 2, 6, 10, 14, and 18 days of treatment and submitted to structural and morphometric analysis. The following parameters were studied: total number of granulocytes and fibroblasts, number of newly formed blood vessels, and percentage of birefringent collagen fibers in the repair area. Morphometric analysis showed that different lasers 670-nm InGaP and 830-nm GaAlAs reduced the number of granulocytes and an increase of newly formed vessels in radiated lesions. The 670-nm InGaP laser therapy was more effective in increasing the number of fibroblasts. The different treatments modified the expression of VEGF and TGF-ß1, when compared with lesions not irradiated. The different types of light sources showed similar effects, improved the healing of second-degree burns and can help for treating this type of injury. Despite the large number of studies with LLTI application in second-degree burns, there is still divergence about the best irradiation parameters to be used. Further studies are needed for developing a protocol effective in treating this type of injury.

Electrical stimulation therapy and rotary jet-spinning scaffold to treat bone defects.

The combination of electrical stimulation (ES) and bone tissue engineering (BTE) has been successful in treatments of bone regeneration. This study evaluated the effects of ES combined with PCL + ß-TCP 5% scaffolds obtained by rotary jet spinning (RJS) in the regeneration of bone defects in the calvaria of Wistar rats. We used 120 animals with induced bone defects divided into 4 groups (n = 30): (C) without treatment; (S) with PCL+ ß-TCP 5% scaffold; (ES) treated with ES (10 µA/5 min); (ES + S) with PCL + ß-TCP 5% scaffold. The ES occurred twice a week during the entire experimental period. Cell viability (in vitro: Days 3 and 7) and histomorphometric, histochemical, and immunohistochemical (in vivo; Days 30, 60, and 90) analysis were performed. In vitro, ES + S increased cell viability after Day 7 of incubation. In vivo, it was observed modulation of inflammatory cells in ES therapy, which also promoted blood vessels proliferation, and increase of collagen. Moreover, ES therapy played a role in osteogenesis by decreasing ligand kappa B nuclear factor-TNFSF11 (RANKL), increasing alkaline phosphatase (ALP), and decreasing the tartarate-resistant acid phosphatase. The combination of ES with RJS scaffolds may be a promising strategy for bone defects regeneration, since the therapy controlled inflammation, favored blood vessels proliferation, and osteogenesis, which are important processes in bone remodeling.

Poly (Lactic Acid) membrane and Sedum dendroideum extract favors the repair of burns in rats.

PURPOSE: To evaluate the healing potential of the electrospinning membranes of Poly (Lactic Acid) (PLA) associated with Sedum dendroideum extract in burn injuries in rats. METHODS: Seventy-five rats were submitted to burn injury on their back skin: (C) untreated; (F) with daily topical application of S. dendroideum extract; (M) with electrospinning membranes of PLA; (MF10) with electrospinning membranes of PLA with 10% S. dendroideum extract; (MF25) with electrospinning membranes of PLA with 25% S. dendroideum extract. Tissue samples were taken after 2, 6 and 14 days of the burn injury and were subjected to histomorfometric analysis of quantification of fibroblasts, collagen fibers, blood vessels, and inflammatory infiltrate Results: The histomorphometric analysis showed an increase in the number of fibroblasts, collagen fibers and blood vessels in the burns treated with membranes of PLA, associated or not with the 10% and 25% extract. The extract of S. dendroideum promoted the increase of collagen fibers. CONCLUSION: The electrospinning PLA membrane, isolated or associated with the S. dendrodeum extract, favored the healing of burn injuries in this experimental model, with an increase of fibroblasts, collagen fibers, and blood vessels. S. dendroideum isolated only stimulated the collagenesis.

Acupuncture and moxibustion stimulate fibroblast proliferation and neoangiogenesis during tissue repair of experimental excisional injuries in adult female Wistar rats.

OBJECTIVE: To investigate the effects of acupuncture and moxibustion on the repair of excisional skin injuries on the back of adult female Wistar rats. METHODS: 90 animals were divided into three groups: C, control; A, acupuncture treatment (needled at traditional acupuncture points BL13, BL17 and ST36); M, moxibustion treatment (overlying same traditional acupuncture points). They were euthanased on days 7, 14 and 21 after injury for removal and preparation of tissue for analysis. RESULTS: The treated groups (A and M) showed no changes regarding the structural analysis relative to the control (C) group. The total number of fibroblast cells in the A and M groups were significantly higher than those in the C group on days 14 and 21. The number of granulocytes was significantly less in the A and M groups compared with the C group on days 14 and 21. The total number of newly formed vessels increased on day 21 and was significantly higher in the A and M groups. The amount of birefringent collagen fibre detected on day 21 was significantly higher in the C group. The amount of glycosaminoglycan and hydroxyproline was similar between the groups. The amount of collagen I did not differ between the groups in any period, despite the increased amount detected over time. The amount of type III collagen did not differ between the groups but the detected amount decreased over the course of the experiment. The amount of transforming growth factor ß1 (TGF-ß1) and vascular endothelial growth factor (VEGF) in the A and M rats was similar but inferior to C rats across all experimental periods. CONCLUSIONS: Acupuncture and moxibustion stimulated fibroblast proliferation and neoangiogenesis, and extended the period of collagen fibre reorganisation in the repair of excisional injuries in adult female rats.

Inhibitory effect of red LED irradiation on fibroblasts and co-culture of adipose-derived mesenchymal stem cells.

The objective of this study was to evaluate the effects of red Light Emiting Diode (red LED) irradiation on fibroblasts in adipose-derived mesenchymal stem cells (ASC) co-culture on the scratch assay. We hypothesized that red LED irradiation could stimulate paracrine secretion of ASC, contributing to the activation of genes and molecules involved in cell migration and tissue repair. ASC were co-cultured with NIH/3T3 fibroblasts through direct contact and subjected to red LED irradiation (1.45 J/cm2/5min6s) after the scratch assay, during 4 days. Four groups were established: fibroblasts (F), fibroblasts + LED (FL), fibroblasts + ASC (FC) and fibroblasts + LED + ASC (FLC). The analyzes were based on Ctgf and Reck expression, quantification of collagen types I and III, tenomodulin, VEGF, TGF-ß1, MMP-2 and MMP-9, as well as viability analysis and cell migration. Higher Ctgf expression was observed in FC compared to F. Group FC presented higher amount of tenomodulin and VEGF in relation to the other groups. In the cell migration analysis, a higher number of cells was observed in the scratched area of the FC group on the 4th day. There were no differences between groups considering cell viability, Reck expression, amount of collagen types I and III, MMP-2 and TGF-ß1, whereas TGF-ß1 was not detected in the FC group and the MMP-9 in none of the groups. Our hypothesis was not supported by the results because the red LED irradiation decreased the healing response of ASC. An inhibitory effect of the LED irradiation associated with ASC co-culture was observed with reduction of the amount of TGF-ß1, VEGF and tenomodulin, possibly involved in the reduced cell migration. In turn, the ASC alone seem to have modulated fibroblast behavior by increasing Ctgf, VEGF and tenomodulin, leading to greater cell migration. In conclusion, red LED and ASC therapy can have independent effects on fibroblast wound healing, but the combination of both does not have a synergistic effect. Therefore, future studies with other parameters of red LED associated with ASC should be tested aiming clinical application for tissue repair.

Activators of SIRT1 in wound repair: an animal model study.

Caloric restriction (CR) and resveratrol activate SIRT1 and induce anti-inflammatory and antioxidant properties. We perform excisional lesion on the dorsum of four groups anesthetized animals: ad libitum-AL diet, AL diet with topical application of 2% resveratrol-Rv, 30% calorie restricted, and finally 30% calorie restricted with 2% resveratrol and we examine CR and Rv effects in wound repair. Restricted animals remained with CR for 31 days. The lesion was performed on day 18 of CR, and resveratrol application was started on day 19. Lesion samples were then collected on days 3 and 10 of treatment for structural, morphometric, and protein analyses. Our results showed that CR and Rv group as well as R group had enhanced numbers of blood vessels, VEGF, fibroblast, birefringent collagen fiber areas in the lesion. We conclude that effects in wound repair suggests that both CR and resveratrol may modulate angiogenesis, fibroplasia, and collagenesis, which could be ascribed to the action of SIRT1.

Electrical stimulation: Complementary therapy to improve the performance of grafts in bone defects?

The limitations of bone reconstruction techniques have stimulated the tissue engineering for the repair of large bone defects using osteoconductive materials and osteoinductive agents. This study evaluated the effects of low intensity electric current on the inorganic bovine graft in calvaria defects. Bone defects were performed with piezoelectric system in the calvaria of Wistar rats divided into four groups (n = 24): (C) without grafting and without electrical stimulation; (E) with grafting; (MC) without grafting and submitted to electrical stimulation; (MC + E) with grafting and submitted to electrical stimulation. Inflammatory, angiogenic and osteogenic events during bone repair at the 10th, 30th, 60th, and 90th days were considered. Several inflammatory markers demonstrated the efficacy of grafting in reducing inflammation, particularly when subjected to electrical stimulation. Angiogenesis and collagen organization were more evident by electrical stimulation application on the grafts. Moreover, the osteogenic cell differentiation process indicated that the application of microcurrent on grafting modulated the homeostasis of bone remodeling. It is concluded that microcurrent favored the performance of grafts in calvarial rat model. Low-intensity electrical current might improve the osteoconductive property of grafting in bone defects. Therefore, electrical current becomes an option as complementary therapy in clinical trials involving bone surgeries and injuries. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 924-932, 2019.

Evolution of Periodontal Disease: Immune Response and RANK/RANKL/OPG System.

The aim of this study was to evaluate markers of bone loss and immune response present in evolution of periodontal disease. One hundred and two Wistar rats were divided into three animals groups: PD0, without ligation and PD15 days and PD60 days, submitted to ligation placement with a sterile 3-0 silk cord in the cervical region of the upper first molar on both sides. Samples were obtained from the gingival tissue for histomorphometric analysis, immunohistochemical analysis of RANK, RANKL, OPG, characterization of the inflammatory infiltrate, quantification of nitric oxide, MCP-1, RANTES, IP10 chemokines, and expression of the TGF-b1, VEG, and bFGF. The number of inflammatory cells in gingival tissue was higher in PD60 samples. The collagen content and the area occupied by birefringent collagen fibers were lower for PD60. Differential leukocyte counting showed that there was a significantly higher polymorphonuclear influx in group PD15, while PD60 showed a greater number of lymphocytes. PD60 showed higher RANTES, IP-10, MCP-1 gene transcripts, as well as a higher nitric oxide concentration. Clinical evaluation revealed that the PD60 group presented an increase in furcal area. In conclusion, in this animal model the increase of RANK/RANKL and HGF markers is related to a specific immune response, and probably contributed to the evolution of periodontal disease. Investigating the effect of these biomarkers can help in targeted therapy for bone resorption, since blocking these can inhibit bone loss.

Differing energy densities with laser 670nm InGaP controls inflammation and collagen reorganization in burns.

PURPOSE: This study compared different energy densities of laser on second degrees burns in rats aiming to determine the most effective dosimetry in stimulation of the healing process. METHODS: Burns were induced in the dorsal skin of 54 animals divided into three groups (n: 18): 1-without treatment; 2-irradiated lesions by the Indium Gallium Phosphide (InGaP) 670nm (4.93J/cm2) laser; 3-irradiated lesions by the InGaP-670nm (9.86J/cm2) laser. Samples were collected on the 2, 10 and 18 days after injury for structural, morphometry, biochemical analysis and Western blotting. RESULTS: The energy densities examined were effective in significantly increasing the total number of fibroblasts and blood vessels and reduce the number of inflammatory cells particularly in irradiated lesions with 9.86J/cm2. This same energy density significantly increased the amount of GAGs (Glycosaminoglycans), decreased the TGF-ß1 (Transforming Growth Factor ß1) and increased the VEGF (Vascular and Endothelial Growth Factor) during the experimental period. This energy density also significantly increased the Collagen type I and decreased Collagen type III and the active isoform of metalloproteinase 9 (MMP-9). CONCLUSIONS: The energy density of 9.86J/cm2 was more effective in promoting cellular responses related to neoangiogenesis, decreasing inflammation and collagen fibers reorganization.

Effects of microcurrent therapy on excisional elastic cartilage defects in young rats.

The effects of microcurrent application on the elastic cartilage defects in the outer ear of young animals were analyzed. Sixty male Wistar rats were divided into a control (CG) and a treated group (TG). An excisional lesion was created in the right outer ear of each animal. Daily treatment was started after 24h and consisted of the application of a low-intensity (20µA) continuous electrical current to the site of injury for 5min. The animals were euthanized after 7, 14 and 28 days of injury and the samples were submitted to analyses. In CG, areas of newly formed cartilage and intense basophilia were seen at 28 days, while in TG the same observations were made already at 14 days. The percentage of birefringent collagen fibers was higher in CG at 28 days. The number of connective tissue cells and granulocytes was significantly higher in TG. Ultrastructural analysis revealed the presence of chondrocytes in TG at 14 days, while these cells were observed in CG only at 28 days. Cuprolinic blue staining and the amount of glycosaminoglycans were significantly higher in TG at 14 days and 28 days. The amount of hydroxyproline was significantly higher in TG at all time points studied. The active isoform of MMP-2 was higher activity in TG at 14 days. Immunoblotting for type II collagen and decorin was positive in both groups and at all time points. The treatment stimulated the proliferation and differentiation of connective tissue cells, the deposition of glycosaminoglycans and collagen, and the structural reorganization of these elements during elastic cartilage repair.

Anti-hemorrhagic effect of hydro-alcoholic extract of the leaves of Mikania glomerata in lesions induced by Bothrops jararaca venom in rats.

PURPOSE: To investigate the effects of hydroalcoholic leaf extract of Mikania glomerata Spreng (Asteraceae) on the activity of Bothrops jararaca snake venom in Wistar rats. METHODS: Fifty four rats Wistar were divided into six groups of nine animals in each: control treated with saline; control treated with B. jararaca venom; control treated with M. glomerata extract; B. jararaca venom incubated with M. glomerata extract at proportions of 1:1, 1:2, and 1:4. RESULTS: Histopathological and morphometric analysis showed that intradermal administration of snake venom incubated with the hydroalcoholic extract at proportions of 1:1, 1:2 and 1:4 promoted a significant reduction in the number of inflammatory cells and a marked decrease in edema after the third hour. There was also a significant reduction in the intensity of the hemorrhagic halo in animals receiving the snake venom incubated with the extract, with the observation of a progressive and parallel inhibition with increasing proportion of M. glomerata. CONCLUSION: The Mikania glomerata hydroalcoholic extract exerted effective anti-inflammatory and antihemorrhagic activity against the effects induced by Bothrops jararaca snake venom.

Application of Jatropha curcas L. seed oil (Euphorbiaceae) and microcurrent on the healing of experimental wounds in Wistar rats.

PURPOSE: To investigate the effects of Jatropha curcas L. seed oil and microcurrent stimulation on the healing of wounds experimentally induced in Wistar rats. METHODS: Forty-eight animals were divided into four groups: (A) control; (B) treated with microcurrent (10 µA/2 min); (C) treated with J. curcas seed oil, and (D) treated with J. curcas seed oil plus microcurrent. Tissues samples were obtained two, six, ten and 14 days after injury and submitted to structural and morphometric analyses. RESULTS: The animals of groups A and C showed similar responses in terms of repair area, total number of cells, number of newly formed blood vessels, epithelial thickness, and percentage of area occupied by mature collagen fibers. Significant differences in all parameters analyzed were observed between animals of groups B and D and the control 10 and 14 days after experimentally induced injury. The morphometric data confirmed the structural findings CONCLUSIONS: The application of J. curcas seed oil alone was not effective on experimental wound healing when compared to control, but microcurrent application alone or combined with the oil exerted significant differences in the parameters studied. These findings suggest that the positive results were due to microcurrent stimulation.

Evolution of Periodontal Disease: Immune Response and RANK/RANKL/OPG System

Abstract The aim of this study was to evaluate markers of bone loss and immune response present in evolution of periodontal disease. One hundred and two Wistar rats were divided into three animals groups: PD0, without ligation and PD15 days and PD60 days, submitted to ligation placement with a sterile 3-0 silk cord in the cervical region of the upper first molar on both sides. Samples were obtained from the gingival tissue for histomorphometric analysis, immunohistochemical analysis of RANK, RANKL, OPG, characterization of the inflammatory infiltrate, quantification of nitric oxide, MCP-1, RANTES, IP10 chemokines, and expression of the TGF-b1, VEG, and bFGF. The number of inflammatory cells in gingival tissue was higher in PD60 samples. The collagen content and the area occupied by birefringent collagen fibers were lower for PD60. Differential leukocyte counting showed that there was a significantly higher polymorphonuclear influx in group PD15, while PD60 showed a greater number of lymphocytes. PD60 showed higher RANTES, IP-10, MCP-1 gene transcripts, as well as a higher nitric oxide concentration. Clinical evaluation revealed that the PD60 group presented an increase in furcal area. In conclusion, in this animal model the increase of RANK/RANKL and HGF markers is related to a specific immune response, and probably contributed to the evolution of periodontal disease. Investigating the effect of these biomarkers can help in targeted therapy for bone resorption, since blocking these can inhibit bone loss.

Resumo Este estudo avaliou marcadores de perda óssea e da resposta imune presentes na evolução da doença periodontal. Cento e dois ratos Wistar foram divididos em três grupos de animais: PD0, sem ligadura e PD15 dias e PD60 dias, submetidos a colocação de ligadura com um fio de seda estéril 3-0 na região cervical do primeiro molar superior em ambos os lados. Foram obtidas amostras de tecido gengival para análise histomorfométrica, análises imunohistoquímicas de RANK, RANKL, OPG, caracterização do infiltrado inflamatório, quantificação de óxido nítrico, expressão de quimiocinas MCP-1, RANTES, IP10 e do TGF-b1, VEGF e bFGF . O número de células inflamatórias no tecido gengival foi maior nas amostras PD60. O teor de colágeno na área ocupada pelas fibras de colágeno birrefringentes foram menores para PD60. A contagem diferencial de leucócitos mostrou que houve um influxo polimorfonuclear significativamente maior no grupo PD15, enquanto que PD60 mostrou número maior de linfócitos. PD60 apresentou transcritos de genes RANTES, IP-10, MCP-1 mais elevados, bem como uma maior concentração de óxido nítrico. A avaliação clínica revelou que o grupo PD60 apresentou aumento da área óssea exposta na região da furca. Em conclusão, neste modelo animal o aumento dos marcadores RANK/RANKL e HGF está relacionado a uma resposta imunológica específica e provavelmente contribuiu para a evolução da doença periodontal. Investigar o efeito destes biomarcadores pode ajudar na terapia dirigida para a reabsorção óssea, uma vez que bloquear estes pode inibir a perda óssea.

Effects of the application of Aloe vera (L.) and microcurrent on the healing of wounds surgically induced in Wistar rats.

PURPOSE: To investigate the effects of topical application of an Aloe vera gel combined or not with microcurrent application on the healing of skin wounds surgically induced in Wistar rats. METHODS: The animals were randomly divided into the following groups: control group, animals topically treated with Aloe vera, animals treated with a microcurrent, and animals receiving topical application of Aloe vera combined with microcurrent application. RESULTS: The results indicated differences in wound healing between the various treatments when compared to the control group. Tissue hyperplasia was lower in the control group compared to the other treated groups. Accelerated wound healing was observed in the group treated with Aloe vera compared to control. Animals submitted to microcurrent application only and the group treated with microcurrent plus Aloe vera presented an earlier onset of the proliferative phase compared to the control group and animals treated with Aloe vera gel alone. Morphometric data confirmed the structural findings. CONCLUSION: Simultaneous application of Aloe vera gel and microcurrent is an excellent choice for the treatment of open wounds thus indicating a synergistic action of these two applications.

Anti-hemorrhagic effect of hydro-alcoholic extract of the leaves of Mikania glomerata in lesions induced by Bothrops jararaca venom in rats

To investigate the effects of hydroalcoholic leaf extract of Mikania glomerata Spreng (Asteraceae) on the activity of Bothrops jararaca snake venom in Wistar rats. METHODS: Fifty four rats Wistar were divided into six groups of nine animals in each: control treated with saline; control treated with B. jararaca venom; control treated with M. glomerata extract; B. jararaca venom incubated with M. glomerata extract at proportions of 1:1, 1:2, and 1:4. RESULTS: Histopathological and morphometric analysis showed that intradermal administration of snake venom incubated with the hydroalcoholic extract at proportions of 1:1, 1:2 and 1:4 promoted a significant reduction in the number of inflammatory cells and a marked decrease in edema after the third hour. There was also a significant reduction in the intensity of the hemorrhagic halo in animals receiving the snake venom incubated with the extract, with the observation of a progressive and parallel inhibition with increasing proportion of M. glomerata. CONCLUSION: The Mikania glomerata hydroalcoholic extract exerted effective anti-inflammatory and antihemorrhagic activity against the effects induced by Bothrops jararaca snake venom.