A biocomplex to repair experimental critical size defects associated with photobiomodulation therapy

The association of scaffolds to repair extensive bone defects can contribute to their evolution and morphophysiological recomposition. The incorporation of particulate biomaterials into three-dimensional fibrin bioproducts together with photobiomodulation therapy (PBM) has potential and can improve regenerative medicine procedures. The objective of this experiment was to evaluate the effects of PBM therapy on critical size defects filled with xenogenic bone substitute associated with fibrin biopolymer. Methods: A critical defect of 8 mm was performed in 36 Wistar male adult rats that were divided into four groups. Groups BC and BC-PBM were defined as controls with defects filled by a clot (without or with PBM, respectively) and groups XS and XS-PBM that comprised those filled with biocomplex Bio-OssTM in association with fibrin biopolymer. PBM was applied immediately after the surgery and three times a week every other day, with the parameters: wavelength of 830 nm, energy density 6.2 J/cm2, output power 30 mW, beam area of 0.116 cm2, irradiance 0.258,62 W/cm2, energy/point 0.72 J, total energy 2.88 J. Fourteen and 42 days after the surgery, animals were euthanatized and subjected to microtomography, qualitative and quantitative histological analysis. Results: The BC-PBM and XS-PBM groups had a similar evolution in the tissue repair process, with a higher density of the volume of new formed bone in relation to the groups without PBM (p = 0.04086; p = 0.07093, respectively). Intense vascular proliferation and bone deposition around the biomaterial particles were observed in the animals of the groups in which biocomplex was applied (XS and XS-PBM). Conclusion: PBM therapy allowed an improvement in the formation of new bone, with a more organized deposition of collagen fibers in the defect area. Biocomplex favored the insertion and permanence of the particulate material in bone defects, creating a favorable microenvironment for accelerate repair process.(AU)

A biocomplex to repair experimental critical size defects associated with photobiomodulation therapy

Abstract Background: The association of scaffolds to repair extensive bone defects can contribute to their evolution and morphophysiological recomposition. The incorporation of particulate biomaterials into three-dimensional fibrin bioproducts together with photobiomodulation therapy (PBM) has potential and can improve regenerative medicine procedures. The objective of this experiment was to evaluate the effects of PBM therapy on critical size defects filled with xenogenic bone substitute associated with fibrin biopolymer. Methods: A critical defect of 8 mm was performed in 36 Wistar male adult rats that were divided into four groups. Groups BC and BC-PBM were defined as controls with defects filled by a clot (without or with PBM, respectively) and groups XS and XS-PBM that comprised those filled with biocomplex Bio-OssTM in association with fibrin biopolymer. PBM was applied immediately after the surgery and three times a week every other day, with the parameters: wavelength of 830 nm, energy density 6.2 J/cm2, output power 30 mW, beam area of 0.116 cm2, irradiance 0.258,62 W/cm2, energy/point 0.72 J, total energy 2.88 J. Fourteen and 42 days after the surgery, animals were euthanatized and subjected to microtomography, qualitative and quantitative histological analysis. Results: The BC-PBM and XS-PBM groups had a similar evolution in the tissue repair process, with a higher density of the volume of new formed bone in relation to the groups without PBM (p = 0.04086; p = 0.07093, respectively). Intense vascular proliferation and bone deposition around the biomaterial particles were observed in the animals of the groups in which biocomplex was applied (XS and XS-PBM). Conclusion: PBM therapy allowed an improvement in the formation of new bone, with a more organized deposition of collagen fibers in the defect area. Biocomplex favored the insertion and permanence of the particulate material in bone defects, creating a favorable microenvironment for accelerate repair process.

Laser photobiomodulation and antimicrobial photodynamic therapy in the treatment of ovine otitis

Background: Otitis is a severe inflammation of the skin of the auditory canal which can impact animals of all ages. Insheep, this disease can occur in isolated cases or in the entire flock. Laser photobiomodulation can be used in combination with medication or as single therapy and the effects are analgesia, modulation of the inflammatory process, edemareduction, tissue restoration and stimulation of local microcirculation. Antimicrobial photodynamic therapy stands out asa promising alternative to antimicrobial drugs for the treatment of localized infections. This study aimed to report the useof laser phototherapies on a sheep with bacterial otitis.Case: A case of bacterial otitis in a 4-year-old sheep, Dorper, was treated at the Veterinary Hospital of the State Universityof Santa Cruz. In the anamnesis, the owner reported that the patient presented crusts in the auricular pavilions, and hadbeen medicated with oxytetracycline 20%, administered intramuscularly, for two days and the crusts cleaned with iodine10%, but the animal did not present clinical improvement. On physical examination, the animal presented constant headswaying, edema of the ears, otalgia, enlarged parotid lymph nodes and ear wounds. No ear discharge was observed, andthe initial diagnosis was aural hematoma. The treatment prescribed was laser photobiomodulation for three consecutivedays. The lesions were irradiated with a diode laser with a power of 0.1 W, irradiance of 3.5 W/cm2, continuous emission,spot area of 0.028 cm2, wavelength of 808 nm, energy of 4J/point, and fluency/point of 142.8J/cm2, with seven pointson the external surface and four points on the internal surface of both ears. Two points were also targeted on the parotidlymph nodes with infrared laser (λ = 808 nm), with energy of 2J/point. On the fourth day of hospitalization, there was areduction in lesions, decreased edema and absence of pain and on the sixth day...

Laser photobiomodulation and antimicrobial photodynamic therapy in the treatment of ovine otitis

Background: Otitis is a severe inflammation of the skin of the auditory canal which can impact animals of all ages. Insheep, this disease can occur in isolated cases or in the entire flock. Laser photobiomodulation can be used in combination with medication or as single therapy and the effects are analgesia, modulation of the inflammatory process, edemareduction, tissue restoration and stimulation of local microcirculation. Antimicrobial photodynamic therapy stands out asa promising alternative to antimicrobial drugs for the treatment of localized infections. This study aimed to report the useof laser phototherapies on a sheep with bacterial otitis.Case: A case of bacterial otitis in a 4-year-old sheep, Dorper, was treated at the Veterinary Hospital of the State Universityof Santa Cruz. In the anamnesis, the owner reported that the patient presented crusts in the auricular pavilions, and hadbeen medicated with oxytetracycline 20%, administered intramuscularly, for two days and the crusts cleaned with iodine10%, but the animal did not present clinical improvement. On physical examination, the animal presented constant headswaying, edema of the ears, otalgia, enlarged parotid lymph nodes and ear wounds. No ear discharge was observed, andthe initial diagnosis was aural hematoma. The treatment prescribed was laser photobiomodulation for three consecutivedays. The lesions were irradiated with a diode laser with a power of 0.1 W, irradiance of 3.5 W/cm2, continuous emission,spot area of 0.028 cm2, wavelength of 808 nm, energy of 4J/point, and fluency/point of 142.8J/cm2, with seven pointson the external surface and four points on the internal surface of both ears. Two points were also targeted on the parotidlymph nodes with infrared laser (λ = 808 nm), with energy of 2J/point. On the fourth day of hospitalization, there was areduction in lesions, decreased edema and absence of pain and on the sixth day...(AU)

A utilização do laser de baixa intensidade na cicatrização de ferida em cão / The use of low intensity laser in wound healing in dog

Background: Low-intensity laser is effective in cellular metabolism, analgesia, and tissue repair. The bioelectric, bioenergetic, and biochemical effects of laser therapy stimulate local circulation, collagen formation, and epithelization. The objective of this study was to report the use of gallium-aluminum arsenide (GaAlAs) laser (830 nm) in healing two woundscaused by necrosis in a dog. Case: An 8-year-old bitch, a rescued victim of animal abuse, had tibiotarsal dislocation in the left posterior limb. Orthopedic surgery was performed with the placement of an external fixation device with six pins in the distal third of the left posterior limb. A radiographic examination performed 42 days after the surgery revealed the loss of the tibiotarsal ratio, decreased radiopacity of the carpal bones, edema, inflammation, muscle atrophy, and the rotation of the anatomical axis of the metatarsals and phalanges. It was then decided to perform a second orthopedic surgery, in which a bone graft and arthrodesis with an 8-screw titanium plate were performed. Four of the plate screws were placed proximally to the tibia and four distally to the tarsus in addition to a screw and a pin transfixed through the distal metaphyseal region of the tibia and calcaneus. Traction was felt during suturing in the dermis and epidermis of the limb, suggesting the possibility of dehiscence of the stitches. After 3 h of surgery, the limb was cold and edematous. After 5 days, the tissue of the medial region of the tarsus developed necrosis, exposing a wound 6 cm long, 1.5 cm wide, and 2 mm deep (wound 1). Another necrotic lesion was observed in the dorsal region of the tarsos - 6.5 cm long, 2 cm wide, 3 mm deep, and exposing 5 cm of the titanium plate (wound 2). Therapy with GaAlAs laser was then performed over the entire length of the wounds. In each therapeutic session, the laser was first used in punctual mode at 10 J/cm², 830 nm, 20 s at each point of the length of the lesion and then in scan mode at 10 J/cm², 830 nm, 1000 Hz, and 40 s continuously. Laser therapy sessions were conducted at 2-4 day intervals, with a 12-day interval between the eighth and ninth sessions. During the entire treatment, cleansing and debridement of the wounds were performed every 48 h with saline and chlorhexidine digluconate, using a compression bandage, as described earlier. Wound 1 healed completely after two laser therapy sessions. In wound 2, tissue repair stagnated after the ninth and last laser session, leaving 4 cm of the titanium plate still exposed. The limb could not support the body weight of the patient, and radiographic examination revealed that the anatomical axis of the metatarsals and the phalanges was rotated and bone conformation was poor. Given the anatomical conditions associated with the patient's clinical picture, it was decided to amputate the limb between the femur and tibia. Discussion: Therapeutic lasers act on mitochondrial respiration, increasing respiratory metabolism and stimulating DNA synthesis and tissue proliferation. In the present case, there was a decrease in inflammatory cells, edema, and the size of the wounds. The pathological conditions of the affected site, influence of trauma, and degree of tissue damage affected the results of the laser therapy. Despite the size difference between wounds 1 and 2, the exposed titanium plate was a determining factor for the partial healing of wound 2.

Epitelização de enxertos cutâneos em feridas recentes de coelhos tratados com membrana amniótica canina e/ou laserterapia / Skin graft epithelialization in rabbit fresh wounds treated with amniotic membrane and/or laser therapy

A enxertia cutânea é uma técnica cirúrgica simples e bastante útil para o reparo de feridas, principalmente aquelas onde existe dificuldade da aplicação do fechamento primário ou de outras técnicas reconstrutivas. Entretanto, para a sobrevivência do enxerto, é necessário que o leito da ferida esteja saudável e com presença de tecido de granulação exuberante. O objetivo do presente trabalho foi avaliar a aplicação da membrana amniótica e da laserterapia como potenciais estimulantes da cicatrização em enxertos aplicados em feridas sem tecido de granulação. Foram utilizados 42 coelhos, divididos em quatro grupos: grupo controle (GC), grupo membrana (GM), grupo laser (GL) e grupo membrana e laser (GML), submetidos a avaliações macro e microscópicas. Na avaliação macroscópica, foi possível notar que os pacientes dos grupos nos quais a membrana amniótica foi utilizada (GM e GML) apresentaram evidências associadas à acentuada reação inflamatória, à falha de integração do enxerto e à consequente necrose dele. Já os pacientes do GL apresentaram melhor aspecto do enxerto no último dia de avaliação. Na análise microscópica, observou-se intensa integração do enxerto à derme, reepitelização acentuada e escassas células inflamatórias no local do enxerto no GL. O contrário foi observado nos pacientes do GM e GML, nos quais aparentemente houve rejeição da membrana. A formação de colágeno não se correlacionou com outros fatores, como inflamação e necrose, em nenhum dos grupos de tratamento. Dessa forma, é possível afirmar que a laserterapia mostrou ser efetiva, contribuindo para o processo cicatricial e a integração do enxerto. Já a membrana amniótica canina não deve ser utilizada para esse fim, pois provoca intensa reação inflamatória, além de impedir a nutrição do enxerto.(AU)

Skin grafting is a simple surgical technique and useful to repair wounds, especially those where there is a difficulty to apply primary closure skin or other reconstructive techniques. However, for graft survival a healthy wound bed and the presence of an exuberant granulation tissue are necessary. This study aimed to evaluate the application of amniotic membrane and laser therapy as potential healing stimulants in grafts applied in wounds without granulation tissue. For this, we used 42 rabbits divided into four treatment groups, control group (CG), membrane group (MG), laser group (LG) and membrane and laser group (MLG), submitted to macroscopic and microscopic evaluation. The macroscopic examination showed that the patients of the groups where the amniotic membrane was used in order to stimulate re-epithelialization (MG and MLG) presented evidences associated with severe inflammatory reaction, graft integration failure and consequent necrosis. LG patients apparently had the best graph aspect in the last valuation date. Microscopic examination showed intense integration of the graft to the dermis, high re-epithelialization level, and scarce inflammatory cells in the graft site of LG patients. The opposite was observed in patients in the MG and MLG groups, where a rejection of the membrane was observed. Finally, collagen formation was not correlated with other factors such as inflammation and necrosis in any of the treatment groups. We can conclude that laser therapy was effective, contributing to the healing process and integration of the graft. Thus, canine amniotic membrane should not be used for this purpose because it causes intense inflammatory reaction besides avoiding graft nutrition.(AU)

Epitelização de enxertos cutâneos em feridas recentes de coelhos tratados com membrana amniótica canina e/ou laserterapia / Skin graft epithelialization in rabbit fresh wounds treated with amniotic membrane and/or laser therapy

A enxertia cutânea é uma técnica cirúrgica simples e bastante útil para o reparo de feridas, principalmente aquelas onde existe dificuldade da aplicação do fechamento primário ou de outras técnicas reconstrutivas. Entretanto, para a sobrevivência do enxerto, é necessário que o leito da ferida esteja saudável e com presença de tecido de granulação exuberante. O objetivo do presente trabalho foi avaliar a aplicação da membrana amniótica e da laserterapia como potenciais estimulantes da cicatrização em enxertos aplicados em feridas sem tecido de granulação. Foram utilizados 42 coelhos, divididos em quatro grupos: grupo controle (GC), grupo membrana (GM), grupo laser (GL) e grupo membrana e laser (GML), submetidos a avaliações macro e microscópicas. Na avaliação macroscópica, foi possível notar que os pacientes dos grupos nos quais a membrana amniótica foi utilizada (GM e GML) apresentaram evidências associadas à acentuada reação inflamatória, à falha de integração do enxerto e à consequente necrose dele. Já os pacientes do GL apresentaram melhor aspecto do enxerto no último dia de avaliação. Na análise microscópica, observou-se intensa integração do enxerto à derme, reepitelização acentuada e escassas células inflamatórias no local do enxerto no GL. O contrário foi observado nos pacientes do GM e GML, nos quais aparentemente houve rejeição da membrana. A formação de colágeno não se correlacionou com outros fatores, como inflamação e necrose, em nenhum dos grupos de tratamento. Dessa forma, é possível afirmar que a laserterapia mostrou ser efetiva, contribuindo para o processo cicatricial e a integração do enxerto. Já a membrana amniótica canina não deve ser utilizada para esse fim, pois provoca intensa reação inflamatória, além de impedir a nutrição do enxerto.(AU)

Skin grafting is a simple surgical technique and useful to repair wounds, especially those where there is a difficulty to apply primary closure skin or other reconstructive techniques. However, for graft survival a healthy wound bed and the presence of an exuberant granulation tissue are necessary. This study aimed to evaluate the application of amniotic membrane and laser therapy as potential healing stimulants in grafts applied in wounds without granulation tissue. For this, we used 42 rabbits divided into four treatment groups, control group (CG), membrane group (MG), laser group (LG) and membrane and laser group (MLG), submitted to macroscopic and microscopic evaluation. The macroscopic examination showed that the patients of the groups where the amniotic membrane was used in order to stimulate re-epithelialization (MG and MLG) presented evidences associated with severe inflammatory reaction, graft integration failure and consequent necrosis. LG patients apparently had the best graph aspect in the last valuation date. Microscopic examination showed intense integration of the graft to the dermis, high re-epithelialization level, and scarce inflammatory cells in the graft site of LG patients. The opposite was observed in patients in the MG and MLG groups, where a rejection of the membrane was observed. Finally, collagen formation was not correlated with other factors such as inflammation and necrosis in any of the treatment groups. We can conclude that laser therapy was effective, contributing to the healing process and integration of the graft. Thus, canine amniotic membrane should not be used for this purpose because it causes intense inflammatory reaction besides avoiding graft nutrition.(AU)

Wound-Healing potential of Sebastiania hispida (Mart.) Pax (Euphorbiaceae) ointment compared to low power laser in rats

Abstract Impaired wound healing represents a serious complication in some pathologies and the use of plant extracts has proved to improve tissue repair. The objectives of this study were to evaluate the healing potential of the ointment of Sebastiana hispida compared with Aluminum-Gallium Indium-Phosphide Laser (InGaAlP) in surgically induced wounds in rats and to perform the phytochemical analysis. The phytochemical analysis was performed in the classic way and also by HPLC. A controlled study was developed with 80 rats (200-250 g) in which a linear excision was performed in the dorsal region after shaving, measuring 2 cm × 1 cm (epidermis and dermis) exposing the muscle fascia. The rats were randomly divided into four groups of twenty animals each. The experimental groups (n = 5) were G1 (Saline); G2 (crude methanol plant extract 2% + Carbopol Gel 98%); G3 (crude methanol plant extract 2% + lanolin/vaseline) and G4 (laser). The incision healing processes were monitored during 3, 7, 14 and until 21 days after excision. The histologic parameters evaluated were Collagen fiber types, microscopic examination and neovascularization. There was a significant increase in the deposition of collagen fibers, as evidenced by a better organized epithelial tissue, keratinized and showing greater proliferation of new blood vessels in the inflammatory phase in the group treated with both the extract and laser. The results were correlated to the phenolic derivatives found after qualitative and quantitative analysis. These compounds were considered responsible for the healing process. The topical treatment with S. hispida leaves, in the two different formulations, was more effective than the application of the laser (Ingan ALP) 660 nm in the model used.

Resumo A cicatrização deficiente representa uma complicação grave em algumas patologias e o uso de extratos de plantas tem demonstrado melhora no processo de reparação tecidual. O objetivo deste estudo foi avaliar o potencial cicatrizante da pomada de Sebastiania hispida comparado ao Laser Indio Phosphide-Gallium-Aluminum (InGaAlP) em feridas cutâneas induzidas cirurgicamente em ratos e realizar analise fitoquímica. A análise fitoquímica foi via clássica e por CLAE. O experimento foi desenvolvido com 80 ratos (200-250 g), divididos aleatoriamente em quatro grupos (n = 5): G1 (salina); G2 (extrato bruto metanólico 2% + Carbopol Gel 98%); G3 (extrato bruto metanólico 2% + lanolina/vaselina) e G4 (laser). Após a tricotomia da região dorsal realizou uma excisão linear, medindo 2 cm × 1 cm (epiderme e derme), expondo a fáscia muscular. Em sequência os tratamentos e o monitoramento do processo de cicatrização das feridas cirúrgicas ocorreu nos tempos de: 3, 7, 14 e até 21 dias após a excisão. Os parâmetros histológicos avaliados foram: tipos de fibras colágenas, avaliação morfológica e neovascularização. Houve uma deposição significativa de fibras colágenas, evidenciado por um tecido epitelial mais bem organizado, queratinizado e mostrando uma maior proliferação de novos vasos sanguíneos na fase inflamatória do grupo tratado com o extrato e o laser. A eficiência do processo de cicatrização pode estar relacionada com a presença de compostos fenólicos e derivados detectados na análise qualitativa e quantitativa. A utilização do tratamento tópico com as duas formulações diferentes de S. hispida foram mais eficazes do que a aplicação do Laser (InGaAlP) no modelo utilizado.

Laserterapia Para Tratamento De Tendinopatia Induzida Em Ratos Wistar: Estudo Histomorfométrico / Laser Therapy For Treatment Of Tendinopathy Induced In Wistar Rats: An Histomorfometric Study / Terapia Con Láser Para Tratamiento De Tendinopatia Inducida En Ratas Wistar: Estudio Histomorfométrico

O objetivo desse estudo foi avaliar os efeitos da laserterapia na lesão do tendão calcanear comum de ratos Wistar. Foram utilizados 12 ratos machos adultos, distribuídos aleatoriamente em dois grupos (L=laser e C=controle). Todos foram submetidos à tendinopatia unilateral (escolha aleatória) mediante compressão transversal do tendão (10 segundos) com pinça Halstead mosquito, assim como escarificações (com bisturi). Após 24 horas da indução da lesão os animais do grupo L receberam laser (904 nm/3 J/cm²/9s) por 20 dias. Os do grupo C foram manipulados como se fossem receber a radiação. Após 3, 7, 14 e 21 dias da realização da lesão, três ratos de cada grupo foram submetidos à eutanásia, e os tendões obtidos para análise histomorfométrica. As amostras foram processadas como de rotina e os fragmentos corados com hematoxilina-eosina, tricrômico de Masson e Picrosirius Red. Os dados foram analisados por ANOVA, a 5% de probabilidade e análise de regressão. Não houve diferença entre grupos nem entre tempos para as características hemorragia, angiogênese, espessamento do epitendão. Independente do tratamento ocorreu diminuição (p=0,0129) da formação de aderência fibrinosa (do 3o ao 21o dias). Por outro lado, a avaliação morfométrica revelou maiores (p=0,0120) quantidades de fibroblastos no grupo que recebeu laserterapia, não havendo efeito de tempo. Avaliação semiquantitativa, revelou maiores (p=0,0000) quantidades de fibroblastos no grupo tratado, porém nessa análise, a quantidade dessas células aumentou com o tempo (p=0,0001) em ambos os grupos. Diferentemente, ANOVA revelou redução do infiltrado inflamatório do 3o ao 21o dia em ambos os grupos (histologia: p=0,0003; morfometria: p=0,0000).(AU)

The aim of this study was to evaluate the effects of laser therapy in tendinopathy of Achilles tendon of Wistar rats. Thus, 12 adult male rats which were divided into two groups (L=laser and C=control) were used. All animals were submitted to unilateral tendinopathy (random selection) by transverse compression of the tendon (10 seconds) with Halsted forceps, as well as 10 scarifications (using a scalpel). After 24 hours of induction of lesion all animals of Group L received laser (904 nm/3 J/cm²/ 9 s) for 20 days, while Group C were handled as if they would receive radiation. After 3, 7, 14 and 21 days lesion induction, three rats from each group were euthanized, and the tendons obtained for histomorphometric analysis. The samples were processed as routine and the sections stained with hematoxylin-eosin, Masson's trichrome and Picrosirius Red. Data were analyzed by ANOVA, at 5% probability and regression analysis. No differences between groups neither between time for hemorrhage characteristics, angiogenesis, thickening of epitendon. Independently of the treatment there was a decrease (p=0.0129) of fibrinous adhesion (3rd to 21st day). On the other hand, morphometric analysis revealed higher (p=0.0120) amounts of fibroblasts cells in the group receiving laser therapy, with no effect of time. Semiquantitative assessment also showed higher (p=0.0000) amounts of fibroblasts cells in the treated group, but in this analysis, the number of fibroblasts increased with time (p=0.0001) in both group. In contrast, ANOVA revealed a reduction of the inflammatory cells from 3rd to 21st day in both groups (histology: p=0.0003; morphometry: p=0.0000).(AU)

La finalidad de ese estudio fue evaluar los efectos del láser en la lesión del tendón calcáneo común de ratas Wistar. Fueron utilizadas 12 ratas machos adultos, distribuidos al azar en dos grupos (L=láser y C=control). Todos fueron sometidos a tendinopatía unilateral (selección aleatoria) por la compresión transversal del tendón (10 segundos) con una pinza Halstead-Mosquito, y escarificación (con bisturí). Después de 24 horas de la inducción de la lesión los animales del grupo L recibieron láser (904 nm/3J/ cm²/9s) durante 20 días. Los tendones del grupo C fueran manejados como si recibera la radiación. Después de 3, 7, 14 y 21 días de realización de la lesión, tres ratas de cada grupo fueron sacrificadas, y los tendones obtenidos para análisis histomorfométrico. Las muestras fueron procesadas como de rutina y los fragmentos se tiñeron con hematoxilina-eosina, tricrómico de Masson y Picrosirius Red. Los datos fueron analizados por ANOVA, con una probabilidad de 5%, además de análisis de regresión. No hubo diferencias entre los grupos o tiempos para las variables hemorragia, angiogénesis, engrosamiento del epitendón. Independientemente del tratamiento se redujo (p=0,0129) la formación de adherencia fibrinosa (del 3o al 21o días). Por otra parte, la evaluación morfométrica mostró cantidades más altas (p=0,0120) de los fibroblastos en el grupo que recibió el tratamiento con láser, sin efecto del tiempo. Evaluación semicuantitativa reveló mayor (p=0,0000) cantidad de fibroblastos en el grupo tratado, pero en esa análisis la cantidad de esas células aumentó con el tiempo (p=0,0001) en ambos grupos. En contraste, ANOVA reveló reducción del infiltrado inflamatorio entre el 3o y el 21o días, en ambos grupos (histología: p=0,0003; morfometría: p=0,0000). Aunque no hubo diferencias entre grupos en la cantidad de fibras de colágeno (I y III), la morfometría reveló que las ratas del grupo L presentaram mayores cantidades (p=0,0096) de fibras de colágeno.(AU)

Laserterapia Para Tratamento De Tendinopatia Induzida Em Ratos Wistar: Estudo Histomorfométrico / Laser Therapy For Treatment Of Tendinopathy Induced In Wistar Rats: An Histomorfometric Study / Terapia Con Láser Para Tratamiento De Tendinopatia Inducida En Ratas Wistar: Estudio Histomorfométrico

O objetivo desse estudo foi avaliar os efeitos da laserterapia na lesão do tendão calcanear comum de ratos Wistar. Foram utilizados 12 ratos machos adultos, distribuídos aleatoriamente em dois grupos (L=laser e C=controle). Todos foram submetidos à tendinopatia unilateral (escolha aleatória) mediante compressão transversal do tendão (10 segundos) com pinça Halstead mosquito, assim como escarificações (com bisturi). Após 24 horas da indução da lesão os animais do grupo L receberam laser (904 nm/3 J/cm²/9s) por 20 dias. Os do grupo C foram manipulados como se fossem receber a radiação. Após 3, 7, 14 e 21 dias da realização da lesão, três ratos de cada grupo foram submetidos à eutanásia, e os tendões obtidos para análise histomorfométrica. As amostras foram processadas como de rotina e os fragmentos corados com hematoxilina-eosina, tricrômico de Masson e Picrosirius Red. Os dados foram analisados por ANOVA, a 5% de probabilidade e análise de regressão. Não houve diferença entre grupos nem entre tempos para as características hemorragia, angiogênese, espessamento do epitendão. Independente do tratamento ocorreu diminuição (p=0,0129) da formação de aderência fibrinosa (do 3o ao 21o dias). Por outro lado, a avaliação morfométrica revelou maiores (p=0,0120) quantidades de fibroblastos no grupo que recebeu laserterapia, não havendo efeito de tempo. Avaliação semiquantitativa, revelou maiores (p=0,0000) quantidades de fibroblastos no grupo tratado, porém nessa análise, a quantidade dessas células aumentou com o tempo (p=0,0001) em ambos os grupos. Diferentemente, ANOVA revelou redução do infiltrado inflamatório do 3o ao 21o dia em ambos os grupos (histologia: p=0,0003; morfometria: p=0,0000).

The aim of this study was to evaluate the effects of laser therapy in tendinopathy of Achilles tendon of Wistar rats. Thus, 12 adult male rats which were divided into two groups (L=laser and C=control) were used. All animals were submitted to unilateral tendinopathy (random selection) by transverse compression of the tendon (10 seconds) with Halsted forceps, as well as 10 scarifications (using a scalpel). After 24 hours of induction of lesion all animals of Group L received laser (904 nm/3 J/cm²/ 9 s) for 20 days, while Group C were handled as if they would receive radiation. After 3, 7, 14 and 21 days lesion induction, three rats from each group were euthanized, and the tendons obtained for histomorphometric analysis. The samples were processed as routine and the sections stained with hematoxylin-eosin, Masson's trichrome and Picrosirius Red. Data were analyzed by ANOVA, at 5% probability and regression analysis. No differences between groups neither between time for hemorrhage characteristics, angiogenesis, thickening of epitendon. Independently of the treatment there was a decrease (p=0.0129) of fibrinous adhesion (3rd to 21st day). On the other hand, morphometric analysis revealed higher (p=0.0120) amounts of fibroblasts cells in the group receiving laser therapy, with no effect of time. Semiquantitative assessment also showed higher (p=0.0000) amounts of fibroblasts cells in the treated group, but in this analysis, the number of fibroblasts increased with time (p=0.0001) in both group. In contrast, ANOVA revealed a reduction of the inflammatory cells from 3rd to 21st day in both groups (histology: p=0.0003; morphometry: p=0.0000).

La finalidad de ese estudio fue evaluar los efectos del láser en la lesión del tendón calcáneo común de ratas Wistar. Fueron utilizadas 12 ratas machos adultos, distribuidos al azar en dos grupos (L=láser y C=control). Todos fueron sometidos a tendinopatía unilateral (selección aleatoria) por la compresión transversal del tendón (10 segundos) con una pinza Halstead-Mosquito, y escarificación (con bisturí). Después de 24 horas de la inducción de la lesión los animales del grupo L recibieron láser (904 nm/3J/ cm²/9s) durante 20 días. Los tendones del grupo C fueran manejados como si recibera la radiación. Después de 3, 7, 14 y 21 días de realización de la lesión, tres ratas de cada grupo fueron sacrificadas, y los tendones obtenidos para análisis histomorfométrico. Las muestras fueron procesadas como de rutina y los fragmentos se tiñeron con hematoxilina-eosina, tricrómico de Masson y Picrosirius Red. Los datos fueron analizados por ANOVA, con una probabilidad de 5%, además de análisis de regresión. No hubo diferencias entre los grupos o tiempos para las variables hemorragia, angiogénesis, engrosamiento del epitendón. Independientemente del tratamiento se redujo (p=0,0129) la formación de adherencia fibrinosa (del 3o al 21o días). Por otra parte, la evaluación morfométrica mostró cantidades más altas (p=0,0120) de los fibroblastos en el grupo que recibió el tratamiento con láser, sin efecto del tiempo. Evaluación semicuantitativa reveló mayor (p=0,0000) cantidad de fibroblastos en el grupo tratado, pero en esa análisis la cantidad de esas células aumentó con el tiempo (p=0,0001) en ambos grupos. En contraste, ANOVA reveló reducción del infiltrado inflamatorio entre el 3o y el 21o días, en ambos grupos (histología: p=0,0003; morfometría: p=0,0000). Aunque no hubo diferencias entre grupos en la cantidad de fibras de colágeno (I y III), la morfometría reveló que las ratas del grupo L presentaram mayores cantidades (p=0,0096) de fibras de colágeno.

Development of an ex vivo model of endovenous laser ablation of the great saphenous vein in a pilot study

PURPOSE:To develop an ex vivo model for the analysis of macroscopic, histological and immunohistochemical changes after experimental endovenous laser ablation (EVLA) of the great saphenous vein (GSV).METHODS:We describe a model produced with glass tubes and introducer sheaths to mimic the physiological conditions of EVLA procedures, such as tumescence and blood flow. A pilot study was conducted to evaluate an ex vivo procedure of EVLA of an incompetent GSV segment using a 1470-nm radial fiber diode laser (7 W power) and an automatic pull-back device. The vein segment was analyzed macroscopically and by hematoxylin & eosin staining, elastic fiber histochemistry, Gomori's trichrome staining, and alpha-smooth muscle actin immunohistochemistry.RESULTS:No perforations were observed macroscopically. No muscle cell adhesion was observed in the central part of the ablated vein, showing tissue disruption. There was low labeling for elastic fibers, disruption of muscle fibers, and a reduced expression of the specific marker for this cell type.CONCLUSION:This ex vivo endovenous laser ablation model is a low cost alternative to in vivo experiments, providing standardized experimental conditions.(AU)

Lower energy and pulse stacking. A safer alternative for skin tightening using fractional CO2 laser

PURPOSE: To evaluate the effect of different energies and stacking in skin shrinkage.METHODS: Three decreasing settings of a fractional CO2 laser were applied to the abdomen of Twenty five Wistar rats divided into three groups. Group I (n=5) was histologically evaluated for microthermal zones dimensions. Groups II and III (n=10 each) were macroscopic evaluated with freeware ImageJ for area contraction immediately and after 30 and 60 days.RESULTS: No statistical significance was found within microthermal zone histological dimensions (Group I) in all settings studied. (Ablation depth: 76.90 to 97.18µm; Coagulation depth: 186.01 to 219.84 µm). In Group II, macroscopic evaluation showed that all settings cause significant immediate skin contraction. The highest setting cause significant more intense tightening effect initially, contracting skin area from 258.65 to 179.09 mm2. The same pattern was observed in Group III. At 30 and 60 days, the lowest setting significantly sustained contraction.CONCLUSION: Lower fractional CO2 laser energies associated to pulse stacking could cause consistent and long lasting tissue contraction in rats.(AU)

LASERTERAPIA DE BAIXA INTENSIDADE NA AMPLIAÇÃO DAS DIMENSÕES DE RETALHOS SUBDÉRMICOS EM RATOS WISTAR

Atualmente houve aumento significativo de pacientes oncológicos atendidos na rotina Veterinária. A principal forma de controle de diversos neoplasmas é com a ressecção cirúrgica, e devido à excisão de grandes neoplasmas, é comum a confecção de extensos defeitos, aumentando a indicação de cirurgias reconstrutivas. Novas técnicas para síntese dos defeitos criados passaram a serem utilizadas na Medicina Veterinária, como o uso de retalhos ou enxertos. Na confecção de retalhos subdérmicos, uma das limitações é a proporção entre comprimento e largura do retalho, pois o comprimento não deve passar a proporção de 2,5:1, pois com o aumento desta proporção, eleva o risco de complicações no pós operatório. Sendo assim, novas maneiras para aperfeiçoar esta modalidade e melhorar a reparação tecidual no pós-operatório vem sendo pesquisadas, como por exemplo, a Laserterapia de Baixa Intensidade (LBI). Este trabalho teve como objetivo avaliar a eficácia da LBI em duas dosagens diferentes, afim de aumentar a proporção na extensão da confecção de retalhos subdérmicos, que atualmente é limitada à 2,5:1 (comprimento x largura). Foram utilizados 24 ratos (Rattus norvergicus albinus wistar), sendo cada um submetido à confecção de retalho cutâneo em seu dorso, distribuídos de forma aleatória em três grupos (Controle sem a terapia da LBI, grupo na dose de 4J/cm² e grupo na dose de 6J/cm²). Os dois grupos que foram irradiados pela terapia à laser sobre o retalho nos dias D0 (dia do procedimento cirúrgico), D2, D4 e D6. A avaliação macroscópica foi realizada aos dias três e sete não mostrou alteração de acordo com o tratamento estabelecido. A avaliação microscópica com a análise histopatológica não apontou diferença estatística entre os grupos. Conclui-se que a exposição de retalhos cutâneos à dose de energia 4J/cm² e 6J/cm² a cada 48 horas não interfere perceptivelmente sobre a cicatrização e a inflamação local, não contribuindo para o aumento da proporção do retalho.

Currently, there has been a significant increase in cancer patients treated in the veterinary routine. The main form of control of several neoplasms is with surgical resection, and due to the excision of large neoplasms, it is common to make extensive defects, increasing the indication for reconstructive surgery. New techniques for synthesizing the created defects started to be used in Veterinary Medicine, such as the use of flaps or grafts. In the manufacture of subdermal flaps, one of the limitations is the proportion between length and width of the flap, as the length should not exceed the proportion of 2.5:1, as the increase in this proportion increases the risk of complications in the postoperative period. Thus, new ways to improve this modality and improve tissue repair in the postoperative period have been researched, such as Low-level Laser Therapy (LLL). This study aimed to evaluate the effectiveness of LLLT at two different dosages, in order to increase the proportion in the extension of subdermal flaps, which is currently limited to 2.5:1 (length x width). Twenty-four rats (Rattus norvergicus albinus wistar) were used, each one submitted to a skin flap on its back, randomly distributed into three groups (Control - without LLL therapy, 4J/cm² dose group and no group dose of 6J/cm²). The two groups that were irradiated by laser therapy over the flap on days D0 (day of the surgical procedure), D2, D4 and D6. The macroscopic evaluation was carried out on days three and seven, showing no change in accordance with the established treatment. Microscopic evaluation with histopathological analysis showed no statistical difference between groups. It is concluded that the exposure of skin flaps to the dose of energy 4J/cm² and 6J/cm² every 48 hours does not perceptibly interfere with healing and local inflammation, not contributing to the increase in the proportion of the flap.

Use of low-power laser to assist the healing of traumatic wounds in rats

PURPOSE: To investigate the morphological aspects of the healing of traumatic wounds in rats using low-power laser. METHODS: Twenty four non isogenic, young adult male Wistar rats (Rattus norvegicus) weighing between 200 and 300g was used. The animals were randomly distributed into two groups: Control (GC) and Laser (GL), with 12 animals each. After shaving, anesthesia was performed in the dorsal region and then a surgical procedure using a scapel was carried out to make the traumatic wound. GL received five sessions of laser therapy in consecutive days using the following laser parameters: wavelength 660 nm, power 100 mW, dose 10 J/cm 2 . The wounds were evaluated through measurement of the area and depth of the wound (MW) and histological analysis (HA). RESULTS: When comparing the GC with the GL in MW there was a difference in area (p<0.001) and depth (p=0.003) measurement of the wounds in GL. The laser group presented more epithelization than GC (p=0.03). The other histological parameters were similar. CONCLUSION: The healing of wounds in rats was improved with the use of the laser.(AU)

Low-level laser therapy modulates musculoskeletal loss in a skin burn model in rats

PURPOSE: To investigate the effectiveness of low-level laser therapy (LLLT) on gastrocnemius muscle morphology and Myod imunoexpression in a model of dorsal burn in rats. METHODS: Sixteen male Wistar rats were distributed into two groups: control group (CG): rats submitted to scald burn injury without treatment and laser treated group (LG): rats submitted to scald burn injury and treated with laser therapy. Fourteen days post-surgery, gastrocnemius muscle was evaluated being the specimens stained with HE and morphometric data was evaluated. MyoD expression was assessed by immunohistochemistry. RESULTS: The results showed that laser treated animals presented more organized tissue morphology compared to the non-treated animals, with a higher number of nucleus in the fibers. Also, the cross sectional area of the fibers and the MyoD immunoexpression in the laser treated groups was higher. CONCLUSION: Low-level laser therapy had positive effects on gastrocnemius muscle, improving tissue muscle morphology, increasing cross sectional area and MyoD immunoexpression.(AU)

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

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.(AU)

The low-level laser on acute myositis in rats

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.(AU)

Analysis of the healing process of the wounds occurring in rats using lasertherapy in association with hydrocolloid

To evaluate wound healing in rats by using low-level laser therapy (LLLT) associated with hydrocolloid occlusive dressing . METHODS: Forty male, adult, Wistar rats were used, distributed into four groups: LG (received 2 J/cm² of laser therapy); HG (treated with hydrocolloid); LHG (treated with 2 J/cm² of laser therapy and hydrocolloid); and the CG (treated with 1 mL of 0.9% saline). The wound was evaluated at pre-determined periods 3rd and 7th days, considering the macroscopic and histological parameters (inflammatory cells, capillary neoformation, fibroblasts, collagen formation and reepithelialization). RESULTS: The LG group at seven days showed increased collagen formation, the LHG group at 3 days showed mild collagen formation. The HG group and the CG at 7 days showed complete reepithelialization. CONCLUSION: Low-level laser therapy as well as the hydrocolloid dressing have favored the wound-healing process in rats.(AU)

The influence of low-level laser irradiation on spinal cord injuries following ischemia- reperfusion in rats

To investigate if low level laser therapy (LLLT) can decrease spinal cord injuries after temporary induced spinal cord ischemia-reperfusion in rats because of its anti-inflammatory effects. Forty eight rats were randomized into two study groups of 24 rats each. In group I, ischemic-reperfusion (I-R) injury was induced without any treatment. Group II, was irradiated four times about 20 minutes for the following three days. The lesion site directly was irradiated transcutaneously to the spinal direction with 810 nm diode laser with output power of 150 mW. Functional recovery, immunohistochemical and histopathological changes were assessed. The average functional recovery scores of group II were significantly higher than that the score of group I (2.86 ± 0.68, vs 1.38 ± 0.09; p<0.05). Histopathologic evaluations in group II were showed a mild changes in compare with group I, that suggested this group survived from I-R consequences. Moreover, as seen from TUNEL results, LLLT also protected neurons from I-R-induced apoptosis in rats. Low level laser therapy was be able to minimize the damage to the rat spinal cord of reperfusion-induced injury.(AU)

Improvement of bone repair in diabetic rats subjected to 780 nm low-level laser therapy

To investigate the effect of low-level laser therapy on bone healing in diabetic rats. METHODS: Bone cavities (19 mm diameter) were performed in the femur of 72 alloxan-induced diabetic rats, which were assigned into four groups: CTR (non-diabetic control), DBT (diabetic) CTRL (non-diabetic irradiated) and DBTL (diabetic irradiated). Low-level laser therapy was performed every 48h for seven days. Animals were euthanized at seven, 18 and 30 days. Alkaline phosphatase serum levels and bone repair were analyzed. RESULTS: Low-level laser therapy significantly increased alkaline phosphatase in at seven and 18 days (p 0.001), and improved bone healing at seven (p 0.01), 18 (p 0.05) and 30 (p 0.01) in diabetic animals. In addition, bone healing in irradiated diabetic group was statistically similar to control group at 30 days (p>0.05). CONCLUSION: Low-level laser therapy increased the serum levels of alkaline phosphatase and improved bone healing in alloxan-induced diabetic rats.(AU)