Evaluation of platelet-rich plasma gel as an angiogenesis-inducing agent in canine advancement skin flaps / Avaliação do gel de plasma rico em plaquetas como agente indutor da angiogênese em flapes cutâneos de avanço em cães

This work aimed to evaluate the effect of platelet-rich plasma (PRP) on advancement skin flaps in dogs regarding improvement of vascularization, with focus on increasing its viable area, since there are reports that it is a potential angiogenesis stimulator. The experimental group was composed of eight adult bitches, in which two advancement skin flaps were made in the ventral abdominal region. No product was applied in the control flap (CF), while PRP was used in the contralateral flap, called treated flap (TF). The areas were clinically evaluated every two days until the 7th postoperative day regarding skin color and presence of necrosis. At 10 days, both flaps were removed and submitted to histological examination and blood vessel morphometry. The vessels counted in each group were statistically analyzed by the F-test at 1% probability. Results showed no significant difference in macroscopic changes in the wound, or CF and TF vascularization, thus suggesting that PRP gel did not improve advancement skin flap angiogenesis in bitches under the experimental conditions in which this research was developed.(AU)

Objetivou-se com o presente artigo avaliar a ação angiogênica do gel de plasma rico em plaquetas (PRP) em flapes cutâneos de avanço em animais da espécie canina, visando aumentar a viabilidade da pele após o procedimento, uma vez que existem relatos de que o produto é um potente estimulador da angiogênese. O grupo experimental foi composto por oito cadelas adultas, onde foram confeccionados dois flapes de avanço (de padrão subdérmico) na região abdominal ventral. Em um dos flapes, considerado controle (FC) não foi aplicado nenhum produto, enquanto que no flape contralateral, denominado tratado (FT), foi usado o gel de PRP. As áreas foram macroscopicamente avaliadas a cada dois dias até o 7º dia de pós-operatório em relação à coloração da pele e presença de área de necrose, e com 10 dias ambos os flapes foram coletados por biópsia e submetidos ao exame histológico e morfometria dos vasos sanguíneos. Os vasos contados em cada grupo foram estatisticamente analisados pelo teste de F ao nível de 1% de probabilidades. Os resultados demonstraram que não houve diferença significativa nas alterações macroscópicas das feridas e na morfometria vascular dos FC e FT, sugerindo dessa maneira que dentro das condições experimentais nas quais a pesquisa foi executada, que o gel de PRP não incrementou a angiogênese de flapes de avanço em cadelas.(AU)

Pterigión recidivante y sus alternativas terapéuticas / Recurrent pterygium and its therapeutic alternatives

RESUMEN El pterigión se trata de un crecimiento fibrovascular de morfología triangular que se extiende desde la conjuntiva hacia la córnea. Está clasificado dentro de las degeneraciones no involutivas o tumoraciones epiteliales benignas corneales y se presenta en el 10,2 por ciento de la población. El tratamiento quirúrgico es el más indicado, y tiene una tasa de recidiva independientemente de la técnica quirúrgica utilizada del 10,7 por ciento; esta tasa de recidiva se evidencia por la neovascularización y el tejido cicatrizal antes de los dos meses después de la cirugía. Lo anterior ha incentivado la investigación de nuevos tratamientos que disminuyan esta complicación, por lo que el objetivo de esta revisión bibliográfica fue la búsqueda de alternativas terapéuticas para el pterigión recidivante. Se realizó una búsqueda automatizada sobre el tema, utilizando la plataforma Infomed, cuya información fue resumida para la elaboración del informe final. Concluimos que existen diferentes tratamientos adyuvantes para disminuir la tasa de recurrencia, y es necesario realizar estudios donde se determine el tiempo y la frecuencia en la aplicación de estos para obtener resultados más efectivos en su uso(AU)

ABSTRACT Pterygium is a fibrovascular growth of triangular shape that extends from the conjunctiva to the cornea. It has been classified as a noninvolutionary degeneration or benign corneal epithelial tumor which affects 10.2 percent of the population. The treatment most commonly indicated is surgery, which has a recurrence rate of 10.7 percent, irrespective of the surgical technique used. Recurrence takes the form of neovascularization and scar tissue within two months after surgery. This has fostered research into new treatments to reduce this complication. The objective of the present bibliographic review was precisely to search for therapeutic alternatives for recurrent pterygium. An automated search was conducted about the topic on the platform Infomed. Data were summarized to write the final report. We concluded that there are several adjuvant treatments to reduce recurrence, and it is necessary to carry out studies determining the time and frequency of their application to obtain more effective results(AU)

Schinus terebinthifolius Raddi ( Aroeira) leaves oil attenuates inflammatory responses in cutaneous wound healing in mice

Abstract Purpose: To investigated the inflammatory, angiogenic and fibrogenic activities of the Schinus terebinthifolius Raddi leaves oil (STRO) on wound healing. Methods: The excisional wound healing model was used to evaluate the effects of STRO. The mice were divided into two groups: Control, subjected to vehicle solution (ointment lanolin/vaseline base), or STRO- treated group, administered topically once a day for 3, 7 and 14 days post-excision. We evaluated the macroscopic wound closure rate; the inflammation was evaluated by leukocytes accumulation and cytokine levels in the wounds. The accumulation of neutrophil and macrophages in the wounds were determined by assaying myeloperoxidase and N-acetyl-β-D-glucosaminidase activities. The levels of TNF-α, CXCL-1 and CCL-2 in wound were evaluated by ELISA assay. Angiogenesis and collagen fibers deposition were evaluated histologically. Results: We observed that macroscopic wound closure rate was improved in wounds from STRO-group than Control-group. The wounds treated with STRO promoted a reduction in leucocyte accumulation and in pro-inflammatory cytokine. Moreover, STRO treatment increased significantly the number of blood vessels and collagen fibers deposition, as compared to control group. Conclusion: Topical application of STRO display anti-inflammatory and angiogenic effects, as well as improvement in collagen replacement, suggesting a putative use of this herb for the development of phytomedicines to treat inflammatory diseases, including wound healing.

Low intensity ultrasound therapy induces angiogenesis and persistent inflammation in the chronic phase of the healing process of third degree burn wounds experimentally induced in diabetic and non-diabetic rats

ABSTRACT PURPOSE: To evaluate the effects of low intensity ultrasound on the healing process of third degree burn wounds in experimentally induced diabetic Wistar rats. METHODS: One hundred rats were divided into: control group; non-diabetic treated group; diabetic control group; diabetic treated group. The therapy was performed with a 3MHz ultrasound application, pulsed emission at 100Hz frequency, modulated at 20% with a dosage of 0.5W/cm2 during three minutes throughout 30 days. The surgical debridement of the wound was performed once at day 2. The wounds were morphometrically, macroscopically and microscopically evaluated at 3, 7, 14, 21 and 30 days. RESULTS: The wound contraction and collagen quantification were higher in all treated groups. Macroscopically, necrosis was higher in the diabetic control group. Granulation tissue was higher in treated groups during the proliferative and remodeling phase. Microscopically, there were greater mononuclear inflammatory infiltration, angiogenesis and fibroblast quantification in treated groups during the proliferative and remodeling phases. CONCLUSIONS: therapeutic ultrasound is beneficial in the inflammatory and proliferative phases of the healing process because it controlled the necrotic tissue, increased the granulation tissue and wound contraction. However in the remodeling phase it is not beneficial because of the continued angiogenesis and a mononuclear inflammatory infiltration.

Experimental study on the effects of bismuth subgallate on the inflammatory process and angiogenesis of the oral mucosa / Estudo experimental sobre os efeitos do subgalato de bismuto no processo inflamatório e de angiogênese em mucosa bucal

ABSTRACT INTRODUCTION: Bismuth subgallate is a salt derived from heavy metal. The aim of this study was to evaluate the effect of this salt on some phases of healing. OBJECTIVES: To assess the effect of subgallate on mucosa and to evaluate the association between the use of bismuth subgallate and neogenesis of vessels in oral mucosal wounds. METHODS: This was a prospective and experimental study. This study used sixty rats, which were divided into control and experimental groups. The animals were submitted to a surgical procedure, which caused oral mucosal injury. A saline solution was applied on the wound of the control group, and in the experimental group, a solution of bismuth subgallate was administrated. RESULTS: The experimental group showed greater inflammatory reaction with increasing monomorphic proliferation. There was increased vessel proliferation in the control group. CONCLUSION: Bismuth subgallate had a negative influence on the healing process, delaying the rate of new vessel formation and optimal wound healing.

RESUMO INTRODUÇÃO: O subgalato de bismuto é um sal derivado de metal pesado. A ideia desta pesquisa é avaliar sua interferência em alguma das fases da cicatrização. OBJETIVO: Delinear a ação do subgalato em mucosas. Avaliar a relação entre a utilização do subgalato de bismuto e a neoformação de vasos nas feridas em mucosa oral, para evidenciar o possível benefício resultante do seu uso. MÉTODO: Estudo experimental, prospectivo. Utilizou-se sessenta ratos, que foram divididos igualmente em grupo controle e experimento. Foram submetidos a um procedimento cirúrgico onde foi feito uma lesão na mucosa oral dos animais, após, uma solução de soro fisiológico foi aplicada sobre a lesão do grupo controle e sobre a ferida do grupo experimento foi aplicada uma solução de subgalato de bismuto. RESULTADOS: o grupo experimento apresentou maior reação inflamatória com crescente proliferação monomórfica. Vasos: houve maior proliferação no grupo controle. CONCLUSÕES: concluiu-se que o subgalato de bismuto teve uma ação negativa no processo de cicatrização, atrasando a velocidade de formação dos neovasos e a cicatrização ideal da ferida operatória.

Utilização de plasma rico em plaquetas para estimulação da angiogênese em flape de padrão axial toracodorsal em coelhos (Oryctolagus cuniculus) / Use of platelet rich plasma to stimulate angiogenesis in the thoracodorsal axial pattern flap in rabbits (Oryctolagus cuniculus)

Feridas de grandes extensões, com perda da viabilidade tecidual e retardo na cicatrização por segunda intenção são casos que se faz necessário o emprego de técnicas cirúrgicas reconstrutivas. O plasma rico em plaquetas (PRP) é um produto com maior concentração plaquetária, adjuvante no processo cicatricial de cirurgias reconstrutivas, auxiliando nos processos de hemostasia e estimulação da angiogênese. Dessa forma, delineou-se um estudo a fim de avaliar a eficácia do uso do gel produzido a partir do plasma rico em plaquetas (PRP) em flapes de avanço de padrão axial toracodorsal em coelhos, para avaliar a possibilidade de favorecer a integração do retalho no leito receptor. Utilizaram-se 30 coelhos da raça Nova Zelândia branco, separados em dois grupos de 15 animais, compreendendo os grupos plasma rico em plaquetas (GPRP), na qual empregou-se o gel antes da síntese da ferida cirúrgica, e controle (GC), na qual utilizou-se apenas solução fisiológica. Para obtenção do PRP, coletou-se sangue dos animais, e determinou-se a contagem plaquetária antes da preparação do gel. No início e término do experimento os animais foram pesados para posterior análise de ganho peso médio. Após o procedimento cirúrgico iniciou-se as avaliações macroscópicas no 3º, 7º e 14º dia, e avaliou-se presença ou ausência de exsudato, integridade da pele, edema, rubor e necrose. Após esta etapa, coletou-se o material da ferida cirúrgica para confecção das lâminas histológicas e posterior avaliação microscópica. Avaliou-se a proliferação vascular, presença de células mononucleares e polimorfonucleares, proliferação fibroblástica, colagenização, reepitelização e hemorragia. Os dados obtidos foram submetidos à análise estatística (Teste t Student, t emparalhado, e Kruskall Walis, sendo p<0,05)[...]

Wounds of large tracts with loss of tissue viability and delayed healing by secondary intention are cases where it is necessary the use of reconstructive surgical techniques. The platelet rich plasma (PRP) is a product with a higher platelet concentration, adjuvant in the healing process of reconstructive surgeries, assisting in the processes of hemostasis and stimulation of angiogenesis. Thus, a study was devised to assess the efficacy of the gel produced from the platelet rich plasma (PRP) in the forward axial flaps thoracodorsal pattern in rabbits to evaluate the ability to promote the integration of the flap the recipient bed. We used 30 white rabbits New Zealand, separated into two groups of 15 animals, comprising the platelet rich plasma groups (GPRP), in which we used the gel before the synthesis of the surgical wound , and control (CG), in which only saline was used. To obtain the PRP, the blood was collected from the animals, was determined and the platelet count before preparation of the gel. At the beginning and end of the experiment the animals were weighed for analysis of average weight gain. After surgery began macroscopic in the 3rd, 7th and 14th day reviews, and we assessed the presence or absence of exudate, skin integrity, edema, redness and necrosis. After this step, yielded the material for the manufacture of surgical wound, followed by microscopic histological slides. We evaluated vascular proliferation, presence of polymorphonuclear and mononuclear cells, fibroblast proliferation, collagen deposition, reepithelialization and hemorrhage. The data were subjected to statistical analysis (Student t test, t paired, and Kruskal Wallis test, and p<0.05)[...]

Effects of aroeira (Schinus terebinthifoliu Raddi) oil on cutaneous wound healing in rats

PURPOSE: To evaluate the effects of aroeira (Schinus terebinthifolius) ointment on skin wound healing in rats. METHODS: Adult male rats (n=20) were divided into four groups of five animals each, as follows: G4, G7, G14 and G21, which corresponds to 4th, 7th, 14th and 21th days postoperatively. Each animal were made two incisions on the skin, including the subcutaneous tissue, in the right and left sides of thoracic region, separated by a distance of two inches. The right lesion was treated with base ointment (vaseline, lanolin); the left one was treated with base ointment containing 5% of aroeira oil. At the end of each experimental period the lesions were evaluated for the contraction degree. Then held the collection of fragments that were fixed in 10% formalin and processed for paraffin embedding. In the histological sections (5μm) was evaluated the morphology and quantified the collagen and blood vessels. The data obtained were submitted to ANOVA test complemented by Tukey-Kramer test (p<0.05). RESULTS: The contraction of the lesions was higher in wounds treated with aroeira oil than in controls at 7th and 14th days (p<0.01), whereas in the 21st day all lesions were already completely healed. The morphology showed granulation tissue more developed, with fibroblasts more bulky and collagen fibers more arranged in the experimental group at 4th, 7th and 14th days. The morphometry showed a significant increase in the quantification of collagen fibers in the experimental group at 7th and 14th days (p<0.05). CONCLUSION: The aroeira oil accelerates the healing process of wounds as a macroscopic, morphological and morphometrical analysis.

Effects of angico extract (Anadenanthera colubrina var. cebil) in cutaneous wound healing in rats / Efeitos do extrato de angico (Anadenanthera colubrina var. cebil) em feridas cutâneas de ratos

PURPOSE: To study the effects of the angico extract (Anadenanthera colubrina var. cebil) on the healing of rat skin. METHODS: Twenty adult rats were divided into four groups of five animals each, the G4, G7, G14 and G21, which corresponds to the respective postoperative days. Each group received two incisions on skin and subcutaneous tissue in the right and left antimere of the thoracic region, separated by a distance of 2 cm. The right lesion was treated daily with saline and the left with the angico alcoholic extract (5%). At the end of each experimental period, animals were euthanized and fragments of the wound area, together with the edges were removed, fixed in 10% formaldehyde solution and processed for paraffin embedding. In the histological sections with 5 µm of thickness, were carried out immunohistochemical methods for detection of blood vessels (VEGF) and stained with hematoxylin and eosin for morphological analysis. Statistical analysis was done by ANOVA and Tukey test (p<0.05). RESULTS: Morphological analysis showed larger fibroblasts and a higher concentration of collagen fibers in days 7 and 14 in wounds treated with the angico extract. Morphometric analysis demonstrated a significant increase in the number of blood vessels in both the seventh and 14th days (p<0.01) in wounds treated with the angico extract. CONCLUSION: The angico alcoholic extract (Anadenanthera colubrina var. cebil) induces the acceleration of wound healing in skin wounds of rats.

OBJETIVO: Avaliar os efeitos do extrato de angico (Anadenanthera colubrina var. cebil) na cicatrização em pele de ratos. MÉTODOS: Ratos machos adultos (n=20) foram distribuídos em quatro grupos de cinco animais cada, a saber: G4, G7, G14 e G21, o que corresponde a quatro, sete, 14 e 21 dias de pós-operatório. Cada grupo recebeu duas incisões na pele compreendendo o tecido subcutâneo, nos antímeros direito e esquerdo da região torácica, separadas por uma distância de dois cm. A lesão esquerda com extrato alcoólico de angico (5%), iniciando-se logo após a cirurgia por 21 dias consecutivos. Ao final de cada período (4, 7, 14 e 21 de pós-operatório) experimental foram coletados fragmentos da área da ferida, fixada em formol a 10% e processadas para inclusão em parafina. Nos cortes histológicos com 5 µm de espessura, foram realizados métodos imunoistoquímicos para detecção dos vasos sanguíneos (VEGF) e coloração pela hematoxilina para análise morfológica. Os dados obtidos foram submetidos à análise estatística ANOVA complementada pelo teste de Tukey-Kramer (p<0,05). RESULTADOS: A análise morfológica mostrou fibroblastos mais volumosos e alta concentração de fibras colágenas no 7º e 14º dias nas feridas tratadas com extrato de angico. A análise morfométrica demonstrou aumento significativo no número de vasos sanguíneos no sétimo e 14º dias (p<0,01) de pós-operatório em feridas tratadas com extrato de angico. CONCLUSÃO: O extrato hidroalcoólico a 5% da casca e entrecasca do angico (Anadenanthera colubrina var. cebil) acelera a neoangiogênese em feridas cutâneas de ratos.

Fibroblastos geneticamente modificados para estimular angiogênese e vasculogênese em miocárdio isquêmico / Transplantation of genetically modified cardiac fibroblasts to induce angiogenesis and vasculogenesis in ischemic myocardium

Este trabalho avaliou o efeito de fibroblastos cardíacos (FC) modificados geneticamente para produzir VEGF (vascular endothelial growth factor) e/ou em conjunto com IGF-1(insulin -like growth factor) associados a um biopolímero de fibrina na indução de angiogênese, vasculogênese e melhora de função em miocárdio isquêmico. Em experimentos preliminares demonstramos que 106 FC modificados pelo AdRSVLacZ expressam o transgene na parede livre do ventrículo esquerdo de ratos Lewis por até 45 dias. Primeiro, o tratamento dos diversos grupos foi feito e 7 dias após, os animais foram submetidos à isquemia por 45 minutos seguida de reperfusão. 21 dias depois, a proteína humana VEGF e a densidade capilar apresentaram aumento no grupo VEGF (proteína humana VEGF: 1209,6±11,4 vs. Veículo 123,1±5,2; Célula 104,2±7,4 e Null 73,2±2,4 células positivas/campo, p< 0,01 e densidade capilar: 543,8 ± 52,1 vs. 349,2 ± 0,9, 288± 19,0 e 245 ± 2,6 capilares/mm2, p< 0,01). A imunofluorescência dupla-marcação para detecção de células endoteliais e células musculares lisas apresentou aumento no grupo VEGF sugerindo formação de vasos estruturados (45±3 vs. 10±2, 8±1 e 16±3, p<0,001) e a área de infarto foi reduzida no VEGF vs. VEÍCULO (3,0 ± 1,3% vs. 8,0 ± 0,8%, p< 0,05. Para testar o efeito terapêutico desta intervenção, um segundo estudo foi realizado com os grupos: VEÍCULO= controle, POLÍMERO = biopolímero de fibrina, CÉLULA, NULL, IGF-1, VEGF e IGF-1+VEGF. Os tratamentos foram realizados 24 horas após os animais terem sido submetidos à isquemia por ligadura permanente. Um mês depois, as proteínas humanas VEGF e IGF 1 apresentaram aumento significativo nos grupos VEGF, IGF-1 e IGF-1+VEGF, com *p=0,0001. Da mesma maneira, somente os grupos que receberam VEGF isoladamente ou associados a IGF-1 tiveram aumento do número de capilares e da densidade vascular e redução da porcentagem de colágeno...

The effect of modified cardiac fibroblasts (CF) expressing VEGF (vascular endothelial growth factor) and/or IGF-1 (insulin-like growth factor) associated to fibrin biopolymer to induce angiogenesis, vasculogenesis and improve cardiac function in ischemic cardiac tissue was tested. The direct injection of 106 CF genetically modified to express the reporter gene LACZ (AdRSVLACZ) indicated transgene expression up to 45 days. First, all groups were treated and 7 days later the animals were submitted to a 45 min cardiac ischemic injury. Twenty one days later VEGF protein and capillary density increased only in groups that received VEGF the groups: (VEGF protein: 1209.6±11.4 vs. VEHICLE: 123.1±5.2, Cell: 104.2±7.4 and Null: 73.2±2.4 positive cells/field, p< 0.01 and capillary: 543.8 ± 52.1 vs. 349.2 ± 0.9, 288± 19.0 and 245 ± 2.6 capillaries/mm2, p< 0.01). Merged image of immunoassaying for endothelial and smooth muscle cells specific markers, were significantly greater in VEGF group suggesting maturation of newly formed vessels (45±3 vs. 10±2, 8±1 and 16±3, p<0.001) and myocardial scar area was reduced in VEGF vs. VEHICLE (3.0 ± 1.3% vs. 8.0 ± 0.8%, p< 0.05). To test the therapeutic efficacy of this treatment, a second study was performed with groups: VEHICLE= control, POLYMER= fibrin biopolymer, CELL, NULL, IGF-1, VEGF and IGF-1+VEGF. Treatments were performed 24 hs following ligation of the descending coronary artery. After 4 weeks, VEGF and IGF-1 protein increased in IGF-1, VEGF and IGF-1+VEGF groups, p<0.0001. We observed only in VEGF groups an increase in capillary number and vascular density and reduction in myocardial collagen area (35,12 ± 7,05 vs. 31,28 ± 5,03 vs. 30,07 ± 6,21 vs. 25,89 ± 2,92 vs. 15,43 ± 2,02* vs. 16,07 ± 1,83%*, *p<0,05, to groups VEHICLE, POLYMER, CELL, NULL, IGF-1, VEGF, IGF-1+VEGF, respectively). The morphological and functional basal cardiac indices remained unchanged in all groups...

Expresión y efecto del gen de VEGF165 humano en músculo esquelético isquémico de conejos. ¿Necesidad de administración repetitiva para el tratamiento de la enfermedad vascular periféica? / Expression and effects after VEGF165 gene transfer: is it necessary the reiterative administration for the treatment of peripheral vascular disease?

Peripheral vascular disease is a disorder fifficult to treat. Due to the lack of an effective pharmacological therapy, the ischemia of an effective pharmacological therapy, the ischemia of the lower limbs pursues a relentless course, ending many times in the amputation...The authors present in this study a 6 years experience with this management, and recently the effects of an experimental model of peripheral vascular disease in rabbits is evaluated. The results obtained in this study are important because they could attribute to he trnasitority of the genetic expression the contradictory results founded in clinical assays of peripheral vascular disease