Lycium barbarum polysaccharide fraction associated with photobiomodulation protects from epithelium thickness and collagen fragmentation in a model of cutaneous photodamage.

Ultraviolet radiation (UVR) is the major etiologic agent of cutaneous photoaging, and different strategies are used to prevent and treat this condition. The polysaccharide fraction (LBPF) isolated from Lycium Barbarum fruits (goji berry) contains several active ingredients with antioxidant, immune system modulation, and antitumor effects. In addition, the photobiomodulation (PBM) is widely applied in photoaging treatment. This study investigated the effects of LBPF and PBM against the UVR-induced photodamage in the skin of hairless mice. The mice were photoaged for 6 weeks in a chronic and cumulative exposure regimen using a 300-W incandescent lamp that simulates the UVR effects. From the third to the sixth week of photoaging induction, the animals received topical applications of LBPF and PBM, singly or combined, in different orders (first LBPF and then PBM and inversely), three times per week after each session of photoaging. After completion of experiments, the dorsal region skin was collected for the analysis of thickness, collagen content, and metalloproteinases (MMP) levels. A photoprotective potential against the increase of the epithelium thickness and the fragmentation of the collagen fibers was achieved in the skin of mice treated with LBPF or PBM singly, as well as their combination. All treatments maintained the skin collagen composition, except when PBM was applied after the LBPF. However, no treatment protected against the UVR-induced MMP increase. Taken together, we have shown that the LBPF and PBM promote a photoprotective effect in hairless mice skin against epidermal thickening and low collagen density. Both strategies, singly and combined, can be used to reduce the UVR-induced cutaneous photoaging.

Polysaccharide-rich hydrogel formulation combined with photobiomodulation repairs UV-induced photodamage in mice skin.

Prolonged skin exposure to ultraviolet radiation (UVR) induces premature aging in both the epidermis and the dermis. Chronic exposure to UVR induces the activation of mitogen-activated protein kinase (MAPK) signaling pathway, activating c-Jun, c-Fos expression, and transcription factor of AP-1 activating protein. AP-1 activation results in the positive induction of matrix metalloproteinase (MMP) synthesis, which degrade skin collagen fibers. Polysaccharides from the fruit of Lycium barbarum (LBP fraction) have a range of activities and have been demonstrate to repair the photodamage. In different approaches, laser application aims to recover the aged skin without destroying the epidermis, promoting a modulation, called photobiomodulation (PBM), which leads to protein synthesis and cell proliferation, favoring tissue repair. Here we developed a topical hydrogel formulation from a polysaccharide-rich fraction of Lycium barbarum fruits (LBP). This formulation was associated with PBM (red laser) to evaluate whether the isolated and combined treatments would reduce the UVR-mediated photodamage in mice skin. Hairless mice were photoaged for 6 weeks and then treated singly or in combination with LBP and PBM. Histological, immunohistochemistry, and immunofluorescence analyses were used to investigate the levels of c-Fos, c-Jun, MMP-1, -2, and -9, collagen I, III, and FGF2. The combined regimen inhibited UVR-induced skin thickening, decreased the expression of c-Fos and c-Jun, as well as MMP-1, -2, and -9 and concomitantly increased the levels of collagen I, III, and FGF2. The PBM in combination with LBP treatment is a promising strategy for the repair of photodamaged skin, presenting potential clinical application in skin rejuvenation.

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.

Activity of Porophyllum ruderale leaf extract and 670-nm InGaP laser during burns repair in rats.

BACKGROUND: In this study, we investigated the effects of an extract of the leaves of Porophyllum ruderale and laser irradiation on the healing of burns. METHODS: Seventy-two rats were divided in four groups: untreated controls, treated with laser irradiation, treated with P. ruderale and treated with both P. ruderale and laser irradiation. Burns were produced with a metal plate on the backs of the animals. Wound samples were collected for structural and morphometric analyses and to quantify the expression of TGF-ß1 and VEGF. RESULTS: Laser irradiation increased the number of fibroblasts, collagen fibers and newly formed vessels and decreased the number of granulocytes at the site of the wounds. Densitometric analysis revealed a significant increase in the expression of TGFß-1 in the wounds treated with laser irradiation and with the P. ruderale extract at the beginning of the healing process and a decreased during the experimental period. The expression of VEGF was highlighted in the lesions irradiated with laser alone. CONCLUSION: Inspite of not showing a beneficial effect on the laser combination with the P. ruderale extract, when the laser was used separately, a positive effects to enhance the healing of second-degree burns was promoted. P. ruderale was effective in decreasing the granulocytes during the repair process indicating a possible anti-inflammatory action of this extract of native flora, widely used in folk medicine, but little studied experimentally.

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.

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.

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.

Evaluation of the effects of electrical stimulation on cartilage repair in adult male rats.

This study describes the organization of mature hyaline xiphoid cartilage during repair in animals submitted to electrical current stimulation. Twenty male Wistar rats, 90 days old, were divided into a control group (CG) and a treated group (TG). A cylindrical full-thickness cartilage defects were created with a 3-mm punch in anesthetized animals. After 24h, TG received daily applications of a continuous electrical current (1Hz/20µA) for 5min. The animals were sacrificed after 7, 21 and 35 days for structural analysis. In CG, the repair tissue presented fibrous characteristics, with fibroblastic cells being infiltrated and permeated by blood vessels. Basophilic foci of cartilage tissue were observed on day 35. In TG, the repair tissue also presented fibrous characteristics, but a larger number of thick collagen fibers were seen on day 21. A large number of cartilaginous nests were observed on day 35. Cell numbers were significantly higher in TG. Calcification points were detected in TG on day 35. There was no difference in elastic fibers between groups. Ultrastructural analysis revealed the presence of chondrocyte-like cells in CG at all time points, but only on days 21 and 35 in TG. The amount of cuprolinic blue-stained proteoglycans was higher in TG on day 35. Microcurrent stimulation accelerates the repair process in non-articular hyaline cartilage.

Effects of microcurrent stimulation on hyaline cartilage repair in immature male rats (Rattus norvegicus).

BACKGROUND: In this study, we investigate the effects of microcurrent stimulation on the repair process of xiphoid cartilage in 45-days-old rats. METHODS: Twenty male rats were divided into a control group and a treated group. A 3-mm defect was then created with a punch in anesthetized animals. In the treated group, animals were submitted to daily applications of a biphasic square pulse microgalvanic continuous electrical current during 5 min. In each application, it was used a frequency of 0.3 Hz and intensity of 20 µA. The animals were sacrificed at 7, 21 and 35 days after injury for structural analysis. RESULTS: Basophilia increased gradually in control animals during the experimental period. In treated animals, newly formed cartilage was observed on days 21 and 35. No statistically significant differences in birefringent collagen fibers were seen between groups at any of the time points. Treated animals presented a statistically larger number of chondroblasts. Calcification points were observed in treated animals on day 35. Ultrastructural analysis revealed differences in cell and matrix characteristics between the two groups. Chondrocyte-like cells were seen in control animals only after 35 days, whereas they were present in treated animals as early as by day 21. The number of cuprolinic blue-stained proteoglycans was statistically higher in treated animals on days 21 and 35. CONCLUSION: We conclude that microcurrent stimulation accelerates the cartilage repair in non-articular site from prepuberal animals.

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.

Application of Jatropha curcas L. seed oil (Euphorbiaceae) and microcurrent on the healing of experimental wounds in Wistar rats / Aplicação do óleo das sementes de Jatropha curcas L. (Euphorbiaceae) e microcorrente no reparo de lesões experimentais em ratos Wistar

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.

OBJETIVO: Investigar os efeitos do óleo das sementes de Jatropha curcas L.e microcorrente em lesões experimentais em de ratos Wistar. MÉTODOS: Quarenta e oito animais foram divididos em quatro grupos: (A) controle, (B) tratado com aplicação de microcorrente, (C) tratado com óleo de sementes de J. curcas e (D) tratado com de óleo de sementes de J. curcas associado à microcorrente. Amostras de tecido foram obtidas no 2º, 6º, 10º e 14º dia após a lesão e submetidas às análises estrutural e morfométrica. RESULTADOS: Os animais dos grupos A e C apresentaram respostas semelhantes quanto a seus efeitos sobre as medidas da área de reparo, número total de células e de vasos sanguíneos neoformados, espessura do epitélio e porcentagem da área ocupada por fibras colágenas maduras. Os grupos de animais B e D apresentaram resultados diferenciados e significativos em todos os parâmetros analisados nos dez e 14 dias após a lesão experimental. Os dados morfométricos confirmaram os achados estruturais. CONCLUSÕES: A aplicação do óleo das sementes de J. curcas não promoveu respostas significativas no reparo das lesões experimentais quando comparadas ao controle, mas a microcorrente aplicada isolada ou combinada a este óleo apresentou diferenças significativos nos parâmetros estudados Este fato sugere que os resultados positivos se devem provavelmente a ação da aplicação da microcorrente.