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.

Blue and red light photoemitters as approach to inhibit Staphylococcus aureus and Pseudomonas aeruginosa growth / Fotoemissores de luz azul e vermelha como abordagem para inibir o crescimento de Staphylococcus aureus e Pseudomonas aeruginosa

The ability of pathogenic bacteria acquire resistance to the existing antibiotics has long been considered a dangerous health risk threat. Currently, the use of visible light has been considered a new approach to treat bacterial infections as an alternative to antibiotics. Herein, we investigated the antimicrobial effect of two range of visible light, blue and red, on Staphylococcus aureus and Pseudomonas aeruginosa, two pathogenic bacterial commonly found in healthcare settings-acquired infections and responsible for high rate of morbidity and mortality. Bacterial cultures were exposed to blue or red light (470 nm and 660 nm) provided by light-emitting diodes - LED. The fluencies and irradiance used for blue and red light were 284.90 J/cm², 13.19 mW/cm² and 603.44 J/cm², 27.93 mW/cm² respectively. Different experimental approaches were used to determine the optimal conditions of light application. Only exposure to blue light for 6 hours was able to inhibit about 75% in vitro growth of both bacterial species after 24 hours. The surviving exposed bacteria formed colonies significantly smaller than controls, however, these bacteria were able to resume growth after 48 hours. Blue light was able to inhibit bacterial growth upon inoculation in both saline solution and BHI culture medium. We can conclude that blue light, but not red light, is capable of temporarily retarding the growth of gram negative and gram positive bacteria.

A capacidade das bactérias patogênicas adquirirem resistência aos antibióticos existentes há muito tempo é considerada uma ameaça perigosa à saúde. Atualmente, o uso da luz visível tem sido considerado uma nova abordagem no tratamento de infecções bacterianas como alternativa aos antibióticos. Neste trabalho, investigamos o efeito antimicrobiano de duas faixas de luz visível, azul e vermelha, em Staphylococcus aureus e Pseudomonas aeruginosa, duas bactérias patogênicas comumente encontradas em infecções adquiridas em instituições de saúde e responsáveis por alta taxa de morbimortalidade. As culturas bacterianas foram expostas à luz azul ou vermelha(470 nm e 660 nm) fornecida por diodos emissores de luz - LED. As fluências e irradiâncias utilizadas para luz azule vermelha foram 284,90 J/cm², 13,19 mW/cm² e 603,44 J/cm², 27,93 mW/cm², respectivamente. Várias abordagens experimentais foram utilizadas para determinar as condições ótimas de aplicação da luz. Apenas a exposição à luz azul por 6 horas foi capaz de inibir cerca de 75% o crescimento in vitro de ambas as espécies bacterianas após24 horas. As bactérias expostas sobreviventes formaram colônias com um tamanho significativamente menor do que os controles, contudo, essas bactérias conseguiram retomar o crescimento normal após 48 horas. A luz azul foi capaz de inibir o crescimento das bactérias após sua inoculação em solução salina ou no meio de cultura rico em nutrientes BHI. Podemos concluir que a luz azul mas não a luz vermelha é capaz de retardar temporariamente o crescimento de bactérias Gram-negativas e Gram-positivas.

Blue and red light photoemitters as approach to inhibit Staphylococcus aureus and Pseudomonas aeruginosa growth / Fotoemissores de luz azul e vermelha como abordagem para inibir o crescimento de Staphylococcus aureus e Pseudomonas aeruginosa

The ability of pathogenic bacteria acquire resistance to the existing antibiotics has long been considered a dangerous health risk threat. Currently, the use of visible light has been considered a new approach to treat bacterial infections as an alternative to antibiotics. Herein, we investigated the antimicrobial effect of two range of visible light, blue and red, on Staphylococcus aureus and Pseudomonas aeruginosa, two pathogenic bacterial commonly found in healthcare settings-acquired infections and responsible for high rate of morbidity and mortality. Bacterial cultures were exposed to blue or red light (470 nm and 660 nm) provided by light-emitting diodes - LED. The fluencies and irradiance used for blue and red light were 284.90 J/cm², 13.19 mW/cm² and 603.44 J/cm², 27.93 mW/cm² respectively. Different experimental approaches were used to determine the optimal conditions of light application. Only exposure to blue light for 6 hours was able to inhibit about 75% in vitro growth of both bacterial species after 24 hours. The surviving exposed bacteria formed colonies significantly smaller than controls, however, these bacteria were able to resume growth after 48 hours. Blue light was able to inhibit bacterial growth upon inoculation in both saline solution and BHI culture medium. We can conclude that blue light, but not red light, is capable of temporarily retarding the growth of gram negative and gram positive bacteria.(AU)

A capacidade das bactérias patogênicas adquirirem resistência aos antibióticos existentes há muito tempo é considerada uma ameaça perigosa à saúde. Atualmente, o uso da luz visível tem sido considerado uma nova abordagem no tratamento de infecções bacterianas como alternativa aos antibióticos. Neste trabalho, investigamos o efeito antimicrobiano de duas faixas de luz visível, azul e vermelha, em Staphylococcus aureus e Pseudomonas aeruginosa, duas bactérias patogênicas comumente encontradas em infecções adquiridas em instituições de saúde e responsáveis por alta taxa de morbimortalidade. As culturas bacterianas foram expostas à luz azul ou vermelha(470 nm e 660 nm) fornecida por diodos emissores de luz - LED. As fluências e irradiâncias utilizadas para luz azule vermelha foram 284,90 J/cm², 13,19 mW/cm² e 603,44 J/cm², 27,93 mW/cm², respectivamente. Várias abordagens experimentais foram utilizadas para determinar as condições ótimas de aplicação da luz. Apenas a exposição à luz azul por 6 horas foi capaz de inibir cerca de 75% o crescimento in vitro de ambas as espécies bacterianas após24 horas. As bactérias expostas sobreviventes formaram colônias com um tamanho significativamente menor do que os controles, contudo, essas bactérias conseguiram retomar o crescimento normal após 48 horas. A luz azul foi capaz de inibir o crescimento das bactérias após sua inoculação em solução salina ou no meio de cultura rico em nutrientes BHI. Podemos concluir que a luz azul mas não a luz vermelha é capaz de retardar temporariamente o crescimento de bactérias Gram-negativas e Gram-positivas.(AU)

Effect of S53P4 bioactive glass and low-level laser therapy on calvarial bone repair in rats submitted to zoledronic acid therapy

ABSTRACT Purpose To evaluate the influence of bioactive glass and photobiomodulation therapy (PBMT) in calvarial bone repair process in rats submitted to zoledronic acid therapy. Methods Twenty-four rats were selected and treated with the dose of 0.035 mg/kg of zoledronic acid every two weeks, totalizing eight weeks, to induce osteonecrosis. After the drug therapy, surgical procedure was performed to create 5-mm diameter parietal bone defects in the calvarial region. The rats were then randomly assigned to groups according to the following treatments: AZC: control group, treated with blood clot; AZBIO: bone defect filled with bioactive glass; AZL: treated with blood clot and submitted to PBMT; and AZBIOL: treated with bioactive glass S53P4 and submitted to PBMT. Tissue samples were collected and submitted to histomorphometric analysis after 14 and 28 days. Results At 14 days, bone neoformation in the AZBIO (52.15 ± 9.77) and AZBIOL (49.77 ± 13.58) groups presented higher values (p ≤ 0.001) compared to the AZC (23.35 ± 10.15) and AZL groups (23.32 ± 8.75). At 28 days, AZBIO (80.24 ± 5.41)still presented significant higher bone recovery values when compared to AZC (59.59 ± 16.92)and AZL (45.25 ± 5.41) groups (p = 0.048). In the 28-day period, the AZBIOL group didn't show statistically significant difference with the other groups (71.79 ± 29.38). Conclusions The bioactive glass is an effective protocol to stimulate bone neoformation in critical defects surgically created in rats with drug induced osteonecrosis, in the studied periods of 14 and 28 days.

Effect of low-level laser therapy on the membrane induced by the Masquelet technique at an orthotopic site in rabbits

ABSTRACT Purpose: To evaluate the low-level laser therapy (LLLT) on the membrane induced by the Masquelet technique in rabbits. Methods: Twelve Norfolk rabbits at approximately 3 months of age were used. A 1-cm segmental defect was induced in both radii, which were filled with polymethylmethacrylate cylinder. LLLT was used postoperatively in the bone defect of one of the forelimbs every 48 hours for 15 days. Six rabbits were euthanatized on third and sixth postoperative weeks. Results: In both forelimbs, radiographs showed new bone growth from radius cut ends on the third postoperative week and more advanced stage on the sixth postoperative week. Ultrasound showed induced membrane one week after the surgery. Histologically, there were no significant differences in the semi-quantitative score of inflammation intensity, total number of blood vessels, bone metaplasia, and collagen. The average thicknesses were 2,050.17 and 1,451.96 μm for control membranes and 2,724.26 and 2,081.03 μm for irradiated membranes, respectively, on third and sixth postoperative weeks. Vascular endothelial growth factor A (VEGF-A) and platelet derived growth factor (PDGF) expression were present in the induced membranes of control and irradiated forelimbs, but there was no significant difference. Conclusions: Based on assessment methods, it was not possible to demonstrate the effect of LLLT on the induced membrane.

Antimicrobial photodinamic therapy combined with laser photobiomodulation in the treatment of skin wounds in a dog (Canis lupus familiaris)

Background: Laser photobiomodulation has been used in the treatment of various injuries and diseases. This promotes modulation of the inflammatory process, edema reduction and devitalized tissue regeneration. The advantages of Antimicrobial photodynamic therapy are its easy application and the absence of side effects. Other advantages are the cost of the therapy, minimal damage to animal tissue, the broad spectrum of action, and efficiency against strains resistant to antimicrobials. The aim of this study was to report the clinical and their resolution in a female dog with a traumatic, infected wound treated with laser phototherapy as an alternative therapy method. Case: A 3-year-old bitch Border Collie, weighing 18 kg, from the municipality of Ilhéus, Bahia, Brazil, waspresented for examination with a history of traumatic laceration of the left thoracic member. On the anamnesis, it was reported that the patient presented with laceration of the left thoracic member. The wound was cleaned and an antibiotic [30 mg/kg of 12/12 h] and anti-inflammatory [0.1 mg/kg every 24 h were prescribed, both for 5 days]. Twenty-four h after the surgical procedure, there was dehiscence of the sutures, with daily topical therapy based on zinc oxide for secondary intention healing. Upon physical examination, the wound was found to be contaminated with swollen and erythematous edges, an ulcerated area with devitalized tissue, serous exudate, and 8.8 cm2 of wounded area. Given the macroscopic characteristics of the lesion, phototherapy was associated with conventional therapy until complete healing of the wound, with three weekly applications at 48 h intervals. Initially, the wound was cleaned with saline solution at 0.9% and a single treatment with aPDT was scheduled due to the high degree of contamination. The dosimetry parameters of irradiation were calculated according to the wounded area with a diode laser of 0.1W of power, continuous emission, spot area of 0.028 cm2, and energy of 9 J per application point. A gauze imbibed with 1 mL of methylene blue aqueous solution (300 µM), which was the photosensitizer was applied to the lesion, with a pre-irradiation time of 5 min, after which it was irradiated with red laser (RL) (λ = 660nm) for 90 s per point, using the sweeping technique. The edge of the lesion was irradiated with infrared laser (IRL) (λ = 808 nm), total energy of 5 J, using the technique of specific points and 1 J of energy/point. After aPDT, low-intensity laser therapy (LILT) sessions were set up with RL and IRL, with energies of 0.5 J/point and 1 J/point, respectively. The wound was cleaned daily, protected with a bandage, and clinically evaluated until complete regression. Discussion: In the present case, methylene blue was used as a photosensitizer, but it is noteworthy that, apparently, each microorganism responds differently to photosensitizers. Thus, the therapy becomes specific for each application, for example: the type of photosensitizer, its concentration, pre-irradiation time, type of light used in photosensitization, wavelength, energy, power, mode delivery of light. Thus, for the best result, the specific protocol in each application should be used Low-intensity laser therapy is an easily executed technique with effective results. The use of PDT associated with photobiomodulation therapy enabled rapid healing of the cutaneous wound, in addition to an improvement in clinical signs and pain caused by the lesion. The technique proved to be an efficient alternative in the treatment of wounds, whether used in isolation or associated with conventional therapy.

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.

Staphylococcus aureus growth delay after exposure to low fluencies of blue light (470 nm) / Atraso no crescimento de Staphylococcus aureus após exposição à baixas fluências de luz azul (470 nm)

Antibiotic resistance is one of the greatest challenges to treat bacterial infections worldwide, leading to increase in medical expenses, prolonged hospital stay and increased mortality. The use of blue light has been suggested as an innovative alternative to overcome this problem. In this study we analyzed the antibacterial effect of blue light using low emission parameters on Staphylococcus aureus cultures. In vitro bacterial cultures were used in two experimental approaches. The first approach included single or fractionated blue light application provided by LED emitters (470 nm), with the following fluencies: 16.29, 27.16 and 54.32 J/cm2. For the second approach a power LED (470 nm) was used to deliver 54.32 J/cm2 fractionated in 3 applications. Our results demonstrated that bacterial cultures exposed to fractionated blue light radiation exhibited significantly smaller sizes colonies than the control group after 24 h incubation, however the affected bacteria were able to adapt and continue to proliferate after prolonged incubation time. We could conclude that the hypothetical clinical use of low fluencies of blue light as an antibacterial treatment is risky, since its action is not definitive and proves to be ineffective at least for the strain used in this study.

A resistência a antibióticos é um dos maiores desafios para o tratamento de infecções bacterianas em todo o mundo, levando ao aumento de despesas médicas, prolongamento da internação hospitalar e aumento da mortalidade. O uso da luz azul tem sido sugerido como uma alternativa inovadora para superar esse problema. Neste estudo, analisamos o efeito antibacteriano da luz azul usando parâmetros de baixa emissão em culturas de Staphylococcus aureus. Culturas bacterianas foram usadas em duas abordagens experimentais in vitro. A primeira abordagem incluiu o uso da aplicação única ou fracionada de luz azul fornecida por emissores de LED (470 nm), com as seguintes fluências: 16,29, 27,16 e 54,32 J/cm2. Para a segunda abordagem, um LED de potência (470 nm) foi usado para fornecer 54,32 J/cm2 fracionado em 3 aplicações. Nossos resultados demonstraram que as culturas bacterianas expostas à radiação de luz azul fracionada exibiram colônias de tamanhos significativamente menores do que o grupo controle após 24 h de incubação, no entanto, as bactérias afetadas foram capazes de se adaptar e continuar a proliferar após um tempo prolongado de incubação. Podemos concluir que o uso clínico hipotético de baixas fluências de luz azul como tratamento antibacteriano é arriscado, pois sua ação não é definitiva e mostra-se ineficaz, pelo menos para a cepa utilizada neste estudo.

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)

Copaiba oil-resin (Copaifera langsdorfii Desf: Caesalpiniaceae) associated with laser therapy for skin wound treatment in Wistar rats / Óleo resina de copaíba (Copaifera langsdorfii Desf: Caesalpiniaceae) associada a laserterapia para o tratamento de feridas cutâneas em ratos Wistar

Alternative protocols for the treatment of skin lesions have been developed with the use of techniques such as photobiomodulation and phytotherapy, aiming to optimize this process. To evaluate the effectiveness of copaiba (Copaiferalangsdorffii) oil-resin and low-level laser therapy for treating cutaneous wounds, 15 Wistar rats (Rattusnorvergicus) were used, in whom five 8-mm lesions were produced. The following protocols were applied: negative control group (T1); positive control group (T2); laser therapy with AsGa (904 nm), continuous, focal mode for 10 s, dosage of 4 J/cm² (T3); copaiba oil-resin (T4); and association group (copaiba and low-level laser) (T5). The efficacy of each technique was evaluated based on macroscopic aspects of the lesion, wound healing rate, and histopathological analysis (inflammatory infiltrate and collagen expression). The Kruskal-Wallis test was used for statistical analyses (P> 0.05). Copaiba treatment showed an advantage in type III collagen expression, whereas laser therapy demonstrated an enhanced capacity for tissue regeneration. The significant advantage obtained from the association treatment is the improvement of the macroscopic aspect of the wound, with a reduction in crust formation.

Protocolos alternativos para o tratamento de feridas cutâneas têm sido desenvolvidos com a utilização de técnicas como a fotobiomodulação e fitoterapia, objetivando acelerar esse processo. Com o objetivo de avaliar a efetividade do óleo--resina de copaíba (Copaifera langsdorffii) e do laser terapêutico no tratamento de feridas cutâneas foram selecionados 15 ratos Wistar (Rattus norvergicus), nos quais foram produzidas cinco lesões cutâneas de 8mm. Os tratamentos empregados foram: controle negativo (T1); controle positivo (T2); laserterapia com AsGa (9,04nm) no modo contínuo, de maneira focal, com duração de 10 segundos e dosimetria de 4 J/cm² (T3); óleo resina de copaiba (T4) e a associação dos tratamentos (copaiba e laserterapia) (T5). A eficácia de cada técnica foi avaliada com base nos aspectos macroscópicos das lesões, taxa de cicatriza-ção e análise histopatológica (infiltrado inflamatório e expressão do colágeno). Os dados obtidos foram analisados pelo teste de Kruskal-Wallis (P>0,05). O tratamento com a copaíba mostrou vantagem na expressão do colágeno tipo III, enquanto a laserterapia demonstrou aumento da capacidade de regeneração tecidual. A vantagem significativa obtida pela associação dos tratamentos foi a melhora do aspecto macroscópico da ferida, com redução na formação de crostas.

Copaiba oil-resin (Copaifera langsdorfii Desf: Caesalpiniaceae) associated with laser therapy for skin wound treatment in Wistar rats / Óleo resina de copaíba (Copaifera langsdorfii Desf: Caesalpiniaceae) associada a laserterapia para o tratamento de feridas cutâneas em ratos Wistar

Alternative protocols for the treatment of skin lesions have been developed with the use of techniques such as photobiomodulation and phytotherapy, aiming to optimize this process. To evaluate the effectiveness of copaiba (Copaiferalangsdorffii) oil-resin and low-level laser therapy for treating cutaneous wounds, 15 Wistar rats (Rattusnorvergicus) were used, in whom five 8-mm lesions were produced. The following protocols were applied: negative control group (T1); positive control group (T2); laser therapy with AsGa (904 nm), continuous, focal mode for 10 s, dosage of 4 J/cm² (T3); copaiba oil-resin (T4); and association group (copaiba and low-level laser) (T5). The efficacy of each technique was evaluated based on macroscopic aspects of the lesion, wound healing rate, and histopathological analysis (inflammatory infiltrate and collagen expression). The Kruskal-Wallis test was used for statistical analyses (P> 0.05). Copaiba treatment showed an advantage in type III collagen expression, whereas laser therapy demonstrated an enhanced capacity for tissue regeneration. The significant advantage obtained from the association treatment is the improvement of the macroscopic aspect of the wound, with a reduction in crust formation.(AU)

Protocolos alternativos para o tratamento de feridas cutâneas têm sido desenvolvidos com a utilização de técnicas como a fotobiomodulação e fitoterapia, objetivando acelerar esse processo. Com o objetivo de avaliar a efetividade do óleo--resina de copaíba (Copaifera langsdorffii) e do laser terapêutico no tratamento de feridas cutâneas foram selecionados 15 ratos Wistar (Rattus norvergicus), nos quais foram produzidas cinco lesões cutâneas de 8mm. Os tratamentos empregados foram: controle negativo (T1); controle positivo (T2); laserterapia com AsGa (9,04nm) no modo contínuo, de maneira focal, com duração de 10 segundos e dosimetria de 4 J/cm² (T3); óleo resina de copaiba (T4) e a associação dos tratamentos (copaiba e laserterapia) (T5). A eficácia de cada técnica foi avaliada com base nos aspectos macroscópicos das lesões, taxa de cicatriza-ção e análise histopatológica (infiltrado inflamatório e expressão do colágeno). Os dados obtidos foram analisados pelo teste de Kruskal-Wallis (P>0,05). O tratamento com a copaíba mostrou vantagem na expressão do colágeno tipo III, enquanto a laserterapia demonstrou aumento da capacidade de regeneração tecidual. A vantagem significativa obtida pela associação dos tratamentos foi a melhora do aspecto macroscópico da ferida, com redução na formação de crostas.(AU)

Healing of skin wounds in rats: influence of low-level laser therapy, alone or combined with natural products

Background: A great variety of natural products have been evaluated for the skin wound healing due to anti-inflammatory,antioxidant and antibacterial activities, and procollagen synthesis, of which may be mentioned the honey and propolis. Inturn, low-level laser therapy (LLLT) is considered an important tool in the treatment of skin wounds, because of ability toraise the ATP production, to stimulate the microcirculation, and formation of new blood vessels. Therefore, this study aimedto assess the influence of LLLT, alone or combined with natural products, in the healing of excisional skin wounds in rats.Materials, Methods & Results: One hundred twenty-six male Wistar rats, aged 14 weeks, were randomly divided intoseven groups (n = 18 per group) according to wound treatments: G1 (negative control): 0.9% saline solution; G2 (positive control): allantoin + zinc oxide ointment; G3: LLLT; G4: mixture of honey + 5% propolis hydroalcoholic solution;G5: LLLT + mixture of honey + hydroalcoholic solution of 5% propolis; G6: LLLT + honey in natura; G7: LLLT + 5%propolis hydroalcoholic solution. Six rats from each group were euthanized on the 7th, 14th, and 21th postoperative days.Macroscopic and histological evaluations of wound healing were performed. A decrease in wound area occurred in allgroups, but in general G4 had the higher reduction and G1 and G3 had the lower reductions. These findings correspondedto the percentage of wound contraction, since the largest contraction was G4 and the lowest contractions were G1 andG3. Histological analysis showed no significant difference among groups...

Healing of skin wounds in rats: influence of low-level laser therapy, alone or combined with natural products

Background: A great variety of natural products have been evaluated for the skin wound healing due to anti-inflammatory,antioxidant and antibacterial activities, and procollagen synthesis, of which may be mentioned the honey and propolis. Inturn, low-level laser therapy (LLLT) is considered an important tool in the treatment of skin wounds, because of ability toraise the ATP production, to stimulate the microcirculation, and formation of new blood vessels. Therefore, this study aimedto assess the influence of LLLT, alone or combined with natural products, in the healing of excisional skin wounds in rats.Materials, Methods & Results: One hundred twenty-six male Wistar rats, aged 14 weeks, were randomly divided intoseven groups (n = 18 per group) according to wound treatments: G1 (negative control): 0.9% saline solution; G2 (positive control): allantoin + zinc oxide ointment; G3: LLLT; G4: mixture of honey + 5% propolis hydroalcoholic solution;G5: LLLT + mixture of honey + hydroalcoholic solution of 5% propolis; G6: LLLT + honey in natura; G7: LLLT + 5%propolis hydroalcoholic solution. Six rats from each group were euthanized on the 7th, 14th, and 21th postoperative days.Macroscopic and histological evaluations of wound healing were performed. A decrease in wound area occurred in allgroups, but in general G4 had the higher reduction and G1 and G3 had the lower reductions. These findings correspondedto the percentage of wound contraction, since the largest contraction was G4 and the lowest contractions were G1 andG3. Histological analysis showed no significant difference among groups...(AU)

Use of phytotherapics, low power laser and ozone for biting wound in dog

Background: Skin wounds are among the most common cases in small animals, highlighting the importance of alternativetherapies as an aid to the healing process. These include ozone, which has a bactericidal and immunomodulatory action; thelow power laser that determines photodynamic changes triggering metabolism elevation; and the use of medicinal plants,in which several herbal medicines has been mentioned in research relating their benefits to wound healing in humans andanimals. Thus, it aims to report the case of a dog submitted to these treatments due to chronic wound due to bite.Case: A 6-year-old canine, female, mixed-breed, 8 kg, with a perforating wound in the left anterior limb in the scapularand humeral region, caused by trauma from the bite of another dog, previously treated, without clinical improvement,has been attended presenting apathy, increased body temperature (40.1°C), difficulty in walking and inability to supportthe injured limb, which was swollen, hyperemic, with elevated temperature, absence of hair, elevated pain sensitivity andpresence of 2 circular lesions, approximately 2 cm in diameter, caused by trauma, and others of size between 0.5 and 1 cmin diameter, which appeared during the course of the disease, characterized as fistulas, through which there was drainageof characteristic fluid exudative and purulent discharge. It was collected blood sample for blood count and purulent material through sterile swab for culture and antibiogram. Systemic treatment with ceftiofur, dipyrone, meloxican and topicalcleaning of the wounds with hydrogen peroxide and physiological solution; dimethyl sulfoxide ointment, and hot compresses was chosen. After seven days, the animal showed significant general clinical...

Use of phytotherapics, low power laser and ozone for biting wound in dog

Background: Skin wounds are among the most common cases in small animals, highlighting the importance of alternativetherapies as an aid to the healing process. These include ozone, which has a bactericidal and immunomodulatory action; thelow power laser that determines photodynamic changes triggering metabolism elevation; and the use of medicinal plants,in which several herbal medicines has been mentioned in research relating their benefits to wound healing in humans andanimals. Thus, it aims to report the case of a dog submitted to these treatments due to chronic wound due to bite.Case: A 6-year-old canine, female, mixed-breed, 8 kg, with a perforating wound in the left anterior limb in the scapularand humeral region, caused by trauma from the bite of another dog, previously treated, without clinical improvement,has been attended presenting apathy, increased body temperature (40.1°C), difficulty in walking and inability to supportthe injured limb, which was swollen, hyperemic, with elevated temperature, absence of hair, elevated pain sensitivity andpresence of 2 circular lesions, approximately 2 cm in diameter, caused by trauma, and others of size between 0.5 and 1 cmin diameter, which appeared during the course of the disease, characterized as fistulas, through which there was drainageof characteristic fluid exudative and purulent discharge. It was collected blood sample for blood count and purulent material through sterile swab for culture and antibiogram. Systemic treatment with ceftiofur, dipyrone, meloxican and topicalcleaning of the wounds with hydrogen peroxide and physiological solution; dimethyl sulfoxide ointment, and hot compresses was chosen. After seven days, the animal showed significant general clinical...(AU)

Effect of low intensity photobiomodulation associated with norbixin-based poly (hydroxybutyrate) membrane on post-tenotomy tendon repair: in vivo study

Purpose: To evaluate the in vivo response of photobiomodulation therapy associated with norbixin-based poly(hydroxybutyrate) membrane (PHB) in tenotomized calcaneal tendon. Methods: Thirty rats were randomly allocated to six groups (n=5 each): LED groups (L1, L2 and L3) and membrane + LED groups (ML1, ML2 and ML3). The right calcaneal tendons of all animals were sectioned transversely and were irradiated with LED daily, one hour after surgery every 24 hours, until the day of euthanasia. At the end of the experiments the tendons were removed for histological analysis. Results: The histological analysis showed a significant reduction in inflammatory cells in the ML1, ML2 and ML3 groups (p=0.0056, p=0.0018 and p 0.0001, respectively) compared to those in the LED group. There was greater proliferation of fibroblasts in the ML1 (p 0.0001) and L3 (p 0.0001) groups. A higher concentration of type I collagen was also observed in the ML1 group (p=0.0043) replacing type III collagen. Conclusion: Photobiomodulation in association with norbixin-based PHB membrane led to control of the inflammatory process. However, it did not favor fibroblast proliferation and did not optimize type I collagen formation in the expected stage of the repair process.(AU)

Effect of low power laser in biomodulation of cultured osteoblastic cells of Wistar rats

Purpose:To analyze aspects of the biomodulating effect of light in biological tissues, bone cells from surgical explants of the femur of rats were irradiated with low intensity laser.Methods:Bone cells were cultured and irradiated with LASER light (GaAlAs). Growth, cell viability, mineralized matrix formation, total protein dosage, immunostimulatory properties, cytochemical analysis, gene expression of bone proteins were examined using live cell imaging and cell counting by colorimetric assay. The gene expression of: alkaline phosphatase (ALP), type 1 collagen, osteocalcin and osteopontin through the real-time polymerase chain reaction.Results:At 8 days, the viability of the irradiated culture was 82.3% and 72.4% in non-irradiated cells. At 18 days, the cellular viability (with laser) was 77.42% and 47.62% without laser. At 8 days, the total protein concentration was 21.622 mg / mol in the irradiated group and 16, 604 mg / mol in the non-irradiated group and at 18 days the concentration was 37.25 mg / mol in the irradiated group and 24, 95 mg / mol in the non-irradiated group.Conclusion:The laser interfered in the histochemical reaction, cell viability, matrix mineralization, and maintained the cellular expression of proteins.(AU)

Influence of low-level laser irradiation on osteocalcin protein and gene expression in bone tissue1

Purpose: To evaluate osteocalcin gene and protein expression in vitro and in an in vivo model of ostectomy. Methods: Twenty Wistar rats were assigned into two groups A (n=10, laser) and B (n=10, control). Ostectomy was performed in the femur diaphysis; the twenty fragments removed, composed in vitro groups named as in vivo (A and B) and cultivated in CO2 atmosphere for thirteen days. Low-level laser irradiation was performed in groups A (in vivo and in vitro) by an GaAlAs device (λ=808 nm, dose of 2J/cm2, power of 200mW, power density of 0.2W/cm2, total energy of 1.25J, spot diameter of 0.02mm) for 5 seconds, at one point, daily. It was performed immunocytochemistry assays in vivo and in vitro groups. In vitro groups were also submitted to RNA extraction, cDNA synthesis and gene expression by quantitative PCR. Statistical analysis was realized with p<0.05. Results: Immunocytochemistry scores showed no significant differences between control and laser groups either in vivo and in vitro. Gene expression also showed no statistical differences. Conclusion: Low-level laser irradiation did not alter osteocalcin protein and gene expression in vivo and in vitro in the studied period but it may have been expressed them in an earlier period.(AU)