Preclinical study of experimental burns treated with photobiomodulation and Human Amniotic Membrane, both isolated and associated / Estudo pré-clínico de queimaduras experimentais tratadas com fotobiomodulação e membrana amniótica humana, isoladas e associadas / Estudio preclínico de quemaduras experimentales tratadas con fotobiomodulación y membrana amniótica humana, solas y combinadas

Abstract Objective: to evaluate the effect of photobiomodulation with low-level 660 nm laser alone or associated with Human Amniotic Membrane in the repair of partial-thickness burns in rats. Method: an experimental study conducted with 48 male Wistar rats, randomized into four groups: Control, Human Amniotic Membrane, Low-Level Laser Therapy, and Low-Level Laser Therapy associated with Human Amniotic Membrane. The histopathological characteristics of the skin samples were analyzed 7 and 14 days after the burn. The data obtained were submitted to the Kolmogorov-Smirnov and Mann-Whitney tests. Results: the histological analysis of the burn injuries showed a decrease in inflammation (p<0.0001) and an increase in proliferation of fibroblasts (p<0.0001) mainly at 7 days in all treatments related to the control group. At 14 days, the greater effectiveness in accelerating the healing process was significant (p<0.0001) in the Low-Level Laser Therapy group associated with the Human Amniotic Membrane. Conclusion: the association of photobiomodulation therapies with the Human Amniotic Membrane allowed verifying a reduction in the healing process time of the experimental lesions, stimulating its proposal as a treatment protocol in partial-thickness burns.

Resumo Objetivo: avaliar o efeito da fotobiomodulação com laser de baixa intensidade 660 nm isoladamente ou associada à membrana amniótica humana no reparo de queimaduras de espessura parcial em ratos. Método: estudo experimental com 48 ratos Wistar machos, randomizados em quatro grupos: Controle, Membrana Amniótica Humana, Terapia a Laser de Baixa Intensidade e Terapia a Laser de Baixa Intensidade associado à Membrana Amniótica Humana. As características histopatológicas das amostras de pele foram analisadas aos 7 e 14 dias após a queimadura. Os dados obtidos foram submetidos aos testes de Kolmogorov-Smirnov e Mann Whitney. Resultados: a análise histológica das lesões por queimadura mostrou a diminuição da inflamação (p <0,0001) e aumento da proliferação de fibroblastos (p <0,0001), principalmente nos 7 dias em todos os tratamentos relacionados ao grupo controle. Aos 14 dias, a maior efetividade na aceleração do processo cicatricial foi significativa (p<0,0001) no grupo Terapia a Laser de Baixa Intensidade associado à Membrana Amniótica Humana. Conclusão: a associação das terapias de fotobiomodulação à membrana amniótica humana permitiu comprovar redução no tempo do processo cicatricial das lesões experimentais, estimulando sua proposição como protocolo de tratamento em queimaduras de espessura parcial.

Resumen Objetivo: evaluar el efecto de la fotobiomodulación con láser de baja intensidad 660 nm de sola o combinada con la membrana amniótica humana en la reparación de quemaduras de espesor parcial en ratas. Método: estudio experimental con 48 ratas Wistar macho, aleatorizadas en cuatro grupos: Control, Membrana Amniótica Humana, Terapia con Láser de Baja Intensidad y Terapia con Láser de Baja Intensidad combinada con la Membrana Amniótica Humana. Las características histopatológicas de las muestras de piel fueron analizadas a los 7 y 14 días después de la quemadura. Los datos obtenidos fueron sometidos a las pruebas de Kolmogorov-Smirnov y Mann-Whitney. Resultados: el análisis histológico de las lesiones por quemadura mostró una disminución de la inflamación (p <0,0001) y un aumento de la proliferación de fibroblastos (p <0,0001) principalmente a los 7 días en todos los tratamientos en comparación con el grupo control; a los 14 días, en el grupo de Terapia con Láser de Baja Intensidad combinada con la Membrana Amniótica Humana la mayor efectividad en la aceleración del proceso de cicatrización fue significativa (p<0,0001). Conclusión: la asociación de terapias de fotobiomodulación con la membrana amniótica humana permitió comprobar que hubo una reducción en el tiempo del proceso de cicatrización de lesiones experimentales, lo cual favorece que se proponga como protocolo de tratamiento en quemaduras de espesor parcial.

Effects of the application of the amniotic membrane in the healing process of skin wounds in rats

Abstract Purpose: To evaluate the efficacy of the application of the human amniotic membrane (HAM) on the inflammatory process, fibroblast proliferation, formation of collagenand reduction of skin wound areas in rats. Methods: Thirty six rats were submitted to a surgical injury induction and divided into two groups (n = 18): group C (control) and T (treated with the HAM). The macroscopic evolution in the wound area and the histological characteristics of the skin samples were evaluated. Results: The regression of the wound area was greater in group T. The histological analysis revealed a significant reduction (p < 0.05) in the inflammatory infiltrate in group T at all experimental periods compared with that in the control group. Furthermore, the group T presented a significant increase in the proliferation of fibroblasts at 14 and 21 days compared with group C (p < 0.05). Regarding the deposition of mature collagen fibers, there was an increase in the replacement of type III collagen by type I collagen in group T (p < 0.05). Conclusion: Treatment with the HAM reduced the healing time as well as the inflammatory responses, increased the proliferation of fibroblasts, and induced a higher concentration of mature collagen fibers.

The low-level laser on acute myositis in rats

ABSTRACT PURPOSE: To analyze the effects of the low-level laser therapy in the acute myositis induced in rats. METHODS: Twelve rats were subjected to bilateral ovariectomy for inducing osteoporosis. After surgery, they were divided into two groups: Ovariectomy-control group (G1, n=6), receiving 0.5 ml distilled water by gavage for 30 days, and Ovariectomy plus mastruz group (G2, n=6), receiving 0.5 ml of the hydroalcoholic extract of mastruz at 10% concentration (50mg) daily, for the same period. Then, the blood of the animals was collected for further biochemical analysis (liver function) and tibia and liver were removed for histological and histomorphometric analyses. RESULTS: In the MT group there was a statistic significant decrease in the number of inflammatory cells, related to the MI group (p<0.05), increase in the fibroblastic proliferation, when compared to groups C and MI related to MT group (p<0.01) and statistic significant edema regression (p=0.0400) in the MT group CONCLUSION: The low-level laser therapy was efficient in the reduction of the inflammatory process, increase of the fibroblastic proliferation and the reduction of the edema.

Effects of the Low-Level Laser Therapy (LLLT) in the process of healing diabetic foot ulcers

ABSTRACT PURPOSE: To evaluate the effects of the low-level laser therapy applying Laser on the tissue repair in ulcer carriers due to diabetes. METHODS: Sixteen type II diabetic patients, ulcer carriers in the lower limbs, participated in the research from which eight were in the control group and eight were submitted to the low-level laser therapy with a pulsed wave form, visible ray, wave length of 632.8 nm, 30 mW peak power, (Laser - HTM). The application time was of 80 (4J/cm2) seconds. The application was punctual without contact (approximately 1mm of distance), the pen being held in a perpendicular position related to the wound, in equidistant points. There were 12 appointments, of which three were done weekly in alternated days. Photograph records and an application of the brief inventory of pain were done before and after 30 days of follow-up. RESULTS: There was a significant decrease in the size of the wound when compared to the control group (p<0.05). The pain was also reported as having an intense improvement in the treated group. CONCLUSION: The low-level laser treatment seems to be an efficient method, viable, painless and of low costs concerning the tissue repair ulcers in a diabetic foot.

Biochemical imaging of normal, adenoma, and colorectal adenocarcinoma tissues by Fourier transform infrared spectroscopy (FTIR) and morphological correlation by histopathological analysis: preliminary results

Introduction The colorectal cancer is a major health problem worldwide. Histology is considered the gold standard for differential diagnosis. However, it depends on the observer's experience, which can lead to discrepancies and poor results. Spectroscopic imaging by Fourier transform infrared (FTIR) is a technique that may be able to improve the diagnosis, because it is based on biochemical differences of the structural constituents of tissue. Therefore, the main goal of this study was to explore the use of FTIR imaging technique in normal colon tissue, colorectal adenoma, and adenocarcinoma in order to correlate their morphological structures with their biochemical imaging. Methods Samples were collected from normal (n = 4), adenoma (n = 4), and adenocarcinoma human colorectal tissue (n = 4) from patients undergoing colonoscopy or surgical resection of colon lesions. The samples were sectioned with a cryostat in sequential sections; the first slice was placed on CaF2 slide and the second slice was placed on glass slide for histological analysis (HE staining). The cluster analyses were performed by the software Cytospec (1.4.02)®. Results In normal samples, biochemical analysis classified six different structures, namely the lamina propria of mucous glands (epithelial cells and goblet cells), central lumen of the gland, mucin, and conjunctive tissue. In samples with adenoma and adenocarcinoma, altered regions could also be identified with high sensitivity and specificity. Conclusion The results of this study demonstrate the potential and viability of using infrared spectroscopy to identify and classify colorectal tissues.

Biomodulation effects of LED and therapeutic ultrasound combined with semipermeable dressing in the repair process of cutaneous lesions in rats

PURPOSE: To compare the biomodulatory effects of LED and ultrasound combined with semipermeable dressing in the repair of cutaneous lesions. METHODS: Eighty-four Wistar rats were submitted to surgical injury (2.5 cm) and divided into four groups (n=21): Group I (control), Group II (LED therapy, LED), Group III (LED therapy + dressing, LED+D) and Group IV (ultrasound + dressing, US+D). At seven, 14 and 21 days, the animals were euthanized, and the specimens of interest removed for histological analysis. RESULTS: Histological and histomorphometric analysis revealed a greater percent wound regression in animals receiving the dressing (group III: 55.97; group IV: 53.06), as well as a greater reduction in the inflammatory infiltrate (group III: 29.14; group IV: 31.71) since day 7. A later effect, with progression of the tissue repair process only after 14 and 21 days, was observed in the LED group intense fibroblast proliferation and greater collagen fiber production and organization were seen in the LED+D and US+D groups compared to the other groups. CONCLUSION: LED combined with a dressing was more effective at accelerating in the repair of cutaneous lesions. .

FT-Raman spectroscopic study of skin wound healing in diabetic rats treated with Cenostigma macrophyllum Tul

INTRODUCTION: Patients with diabetes mellitus exhibit a delay in the lesion repair process. The active components of Cenostigma macrophyllum may represent a viable alternative to facilitate the recovery of these lesions. The aim of this study was to evaluate the effects of emulsion oil-water Cenostigma macrophyllum in the repair process of lesions in rats with induced diabetes. METHODS: 63 male rats (Wistar, 200-250 g body weight, 30-40 days old) were distributed into the following groups: control (C), diabetic (D) and diabetic treated with Cenostigma macrophyllum (P), subdivided based on the experimental times, days 7, 14 and 28, with 21 animals per main group. Diabetes mellitus (DM) was induced by administration of streptozotocin (50 mg/kg via penile vein and 12-h fasting) and confirmed at day 21 (glycemic index > 240 mg/dL). In the animals of group P, 0.5 ml of the oil-water emulsion obtained from the plant seed was used. The samples were removed and hemisectioned, and one portion was used for the quantitative histological analysis of collagen using Masson's trichrome staining, while another portion was analyzed by FT-Raman spectroscopy. RESULTS: A higher percentage area of the volume of collagen fibers was observed for the experimental time Day 14 in group P compared with group D (p < 0.001). Regarding the ratio of areas of the amides I (1700‑1600 cm-1) and III (1245-1345 cm-1), the groups D and P show the opposite behavior. CONCLUSION: Cenostigma macrophyllum accelerated the repair process in skin of diabetic ratsfor14 days.

Cenostigma macrophyllum Tul. on the healing of skin wounds in rats with Diabetes mellitus

PURPOSE: To investigate the effect of Cenostigma macrophyllum Tul. in the tissue repair process of skin lesions in rats with induced Diabetes mellitus. METHODS: Sixty-three male Wistar rats were distributed into three groups: control (C), diabetic (D), and diabetic treated with an oil-in-water emulsion of the plant (DPL) subdivided according to time of observation (seven, 14, and 28 days). Diabetes was induced by administration of by injection of streptozotocin (50 mg/kg, after a 12-h fast) into the penile vein and confirmed by glucose levels exceeding 240 mg/dL. Held surgical lesion (2.5 cm in diameter) on the back of the animals. The DPL received topical application of group of the oil-in-water emulsion plant (0.5ml). To evaluate the levels of nitric oxide, was collected 5ml of blood from the abdominal aorta, after his euthanasia. The samples of interest were sent for routine histological processing. RESULTS:A reduction in the percentage of inflammatory cells and increased numbers of fibroblasts in the group DPL, seven days, compared to the other groups. At 14 days, the DPL group also showed a higher concentration of nitric oxide (p<0.01) than in groups C and D. CONCLUSION: The oil-in-water emulsion of C. macrophyllum Tul accelerated wound healing in diabetic rats.

Clinical and evolutionary characteristics of four patients with pulmonary histoplasmosis reported in the Paraíba Paulista Valley region / Características clínicas evolutivas de quatro pacientes com histoplasmose pulmonar encontrado na região do Vale do Paraíba

The type of pulmonary histoplasmosis presents limited lesions to the lungs, with symptoms that are clinically and radiological similar to chronic pulmonary tuberculosis. This paper describes the clinical features of four cases of pulmonary histoplasmosis. Aspects of diagnostic and clinical, epidemiological, laboratorial and imaging exams are discussed, in addition to the clinical status of the individuals five years after disease onset. The treatment of choice was oral medication, following which all the patients improved. It is important to understand the clinical status and the difficulties concerning the differential diagnosis of histoplasmosis, to assist the proper indication of cases, thus reducing potential confusion with other diseases.

A histoplasmose do tipo pulmonar apresenta lesões limitadas aos pulmões, cujos sintomas são clínica e radiologicamente similares à tuberculose pulmonar crônica. Esse trabalho descreve as características clínicas de quatro casos de histoplasmose. Os aspectos do diagnósticos clínicos, epidemiológicos, laboratoriais e exames de imagem são relacionados, além da situação clínica dos indivíduos, 5 após o início da doença. O tratamento de escolha foi atravésde medicação oral, aonde todos os pacientes evoluíram bem. É importante conhecer o quadro clínico e da dificuldade em diagnosticar a histoplasmose para ajudar a conduzir as boas indicações dos casos, reduzindo assim o malentendido com outras entidades patológicas.

Evaluation of sodium alendronate activity on bone repair under hormones absence

Objetivou-se avaliar a ação do alendronato sódico na reparação óssea de ratas ovariectomizadas. Foram utilizados 36 ratos fêmeas adultas ovariectomizadas sendo formados quatro grupos, com nove ratas em cada grupo, onde foi realizada uma lesão na tíbia. O grupo controle não recebeu medicamento, enquanto os outros três foram tratados com doses de 0,25 mg/kg, 0,5mg/kg e 0,75 mg/kg na água de beber, diariamente. Foram sacrificadas três ratas de cada grupo nos períodos de três, sete e 14 dias. A análise histológica das tíbias desses animais revelou uma deposição óssea mais marcante nos grupos tratados com as doses maiores de alendronato. O grupo de 14 dias com dose de 0,5 mg/kg, apresentou a deposição de trabéculas mais maduras e organizadas, sugerindo um processo de remodelação já bem desenvolvido, mostrando assim ser a ação do alendronato sódico dose-dependente.

Estudo do efeito do plumbum metallicum 30ch na reparação óssea em mandíbula de ratos sem barreira mecânica / Study of of plumbum metallicum 30ch effect in bone repair of rat mandibles without

Objetivos: O objetivo deste trabalho foi avaliar os efeitos da administração sistêmica do chumbo, preparadode acordo com a farmacotécnica homeopática Plumbum metallicum 30CH, na reparação óssea. Material emétodos: foram utilizados trinta ratos machos com três meses de idade, nos quais foram confeccionadosdefeitos cirúrgicos trans-ósseos com broca de aço nº 06 na região do ângulo da mandíbula do lado direito, semutilização de barreira mecânica. Cada animal do grupo experimental recebeu três gotas diárias de Plumbummetallicum 30CH acrescidas à mamadeira de água. Os animais do grupo controle receberam apenas água. Trêsanimais foram sacrificados nos seguintes períodos experimentais: um, sete, 14, 21 e 28 dias após a cirurgia.Após o sacrifício, as mandíbulas foram removidas, descalcificadas e submetidas ao preparo histológico derotina. Foram submetidas a cortes semi-seriados e coradas pelos métodos de hematoxilina-eosina e tricômicode Masson para análise histológica em microscópio de luz. Resultados: os resultados obtidos neste trabalhomostraram que o medicamento foi eficiente na estimulação da reparação óssea. A análise histológica qualitativamostrou evidência de efeito favorável do Plumbum mettalicum 30 CH. Apesar da interferência dotecido muscular, os animais mostraram, já a partir do período de 14 dias, neoformação óssea em praticamentetoda a extensão do defeito. Conclusão: a utilização do Plumbum metallicum 30CH favoreceu a reparação dedefeitos ósseos em mandíbula de ratos machos, sem a utilização de barreira mecânica.