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)

Meningioma intracraniano em cão - tratamento com radioterapia / Intracranial meningioma in a dog - treatment with radiotherapy

Background: The most common primary brain neoplasm is meningioma. Dolichocephalic breeds are predisposed and there is no sexual predilection. Clinical signs depend on the location and size of the tumor and have a progressive course. Primary treatement includes surgery, radiotherapy or both. This study aimed to describe the treatment of a dog with suspected intracranial meningioma with definitive radiotherapy, which resulted in significant clinical improvement and prolonged survival. Case: A 9-year-old Shetland Shepherd bitch was diagnosed with a head tilt to the left side that progressed over a few weeks. She previously received corticosteroid therapy, which resulted a clinical improvement that worsened after treatment was discontinued. Computed tomography revealed an extra-axial brain mass in the caudal fossa, lateralized to the right, welldelimited, and measuring approximately 1.5 × 1.4 × 1.7 cm, suggestive of intracranial meningioma. The patient was treated with radiotherapy using Cobalt-60 equipment, with 18 daily fractions of 2.5 Gy at a total dose of 45 Gy using parallel and opposite technique fields. A new tomography performed 30 days after treatment showed a remission of 85% of the initial brain mass measuring approximately 0.9 × 1.0 × 0.5 cm, as well as complete resolution of the clinical signs initially presented. After 14 months, the patient presented with signs of lethargy and ataxia and was medicated with hydroxyurea at a dose of 50 mg/kg 3 times a week and corticosteroid therapy. However, the patient's neurological condition deteriorated, and she was subjected to reirradiation using the same protocol used previously, which resulted in clinical improvement and a 54% reduction in tumor volume on magnetic resonance. As a late side effect, only permanent alopecia in the irradiated region was observed. The patient died of disease 330 days after the second course of radiotherapy, with a total survival time of 1087 days. Discussion: Meningiomas are extra-axial neoplasms of the central nervous system that grow inside the dura mater. The literature shows that meningiomas are more common in dolichocephalic races with a mean age of 9 years, which supports our findings. Meningiomas most commonly affect the cortical thalamus and cerebellopontine region in dogs, which are normally associated with vestibular symptoms, as seen in this case. Diencephalic damage can result in vestibular signals since the thalamus functions as a relay station for vestibular afferent stimuli that are relayed to the cerebral cortex. In addition to the vestibular syndrome, common clinical signs associated with meningiomas in dogs include seizures, behavioral changes, and walking in circles, which are frequently misinterpreted due to tumor-induced side effects, such as cerebral edema, obstructive hydrocephalus, and cerebral hernia. Advanced imaging techniques should be used to diagnose intracranial neoplasms. In this case, computed tomography was critical for diagnosis and treatment planning. Meningioma treatment may comprise palliative measures, surgery, and radiotherapy. Radiotherapy as a single treatment can improve the quality of life with a decrease in clinical signs and a median survival time of approximately 250-536 days, as reported in the literature. Hydroxyurea can be a therapeutic option in inoperable cases and for patients with clinical limitations to undergo successive anesthesia during radiotherapy. Its most serious side effect is progressive myelosuppression. It can cause temporary partial tumor remission and improvement in clinical signs. As previously stated, radiotherapy can be an effective primary treatment option for treating intracranial meningiomas in dogs, with significant improvement in neurological clinical signs and mild side effects.

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.

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.

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.

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.

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)

Perfil hematológico e bioquímico de roedores de laboratório alimentados com rações contendo diferentes fontes lipídicas e métodos de esterilização / Hematological and biochemical profile of laboratory rodents feeded with rations containing different lipid sources and sterilization methods

Objetivou-se com esse estudo avaliar os efeitos de diferentes métodos de esterilização e fontes de óleo (salmão e linhaça) em formulação de rações sobre os parâmetros bioquímicos e hematológicos de ratos Wistar e camundongos NIH em diferentes idades. Foram avaliados 336 animais, sendo 168 de cada biomodelo. Houve efeito significativo das fontes de óleo sobre os parâmetros bioquímicos plasmáticos em camundongos NIH e ratos Wistar (P 0,05). Conclui-se que o óleo de salmão e linhaça podem ser utilizados nas rações para camundongos NIH e ratos Wistar, independente do método de esterilização adotado.

The objective of this study was to evaluate the effects of different methods of sterilization and oil sources (salmon and flaxseed) in the formulation of diets on the biochemical and hematological parameters of Wistar rats and NIH mice at different ages. 336 animals were evaluated, 168 from each biomodel. There was a significant effect of oil sources on plasma biochemical parameters in NIH mice and Wistar rats (P 0.05). It was concluded that salmon and linseed oil can be used in diets for NIH mice and Wistar rats, regardless of the sterilization method adopted.

Perfil hematológico e bioquímico de roedores de laboratório alimentados com rações contendo diferentes fontes lipídicas e métodos de esterilização / Hematological and biochemical profile of laboratory rodents feeded with rations containing different lipid sources and sterilization methods

Objetivou-se com esse estudo avaliar os efeitos de diferentes métodos de esterilização e fontes de óleo (salmão e linhaça) em formulação de rações sobre os parâmetros bioquímicos e hematológicos de ratos Wistar e camundongos NIH em diferentes idades. Foram avaliados 336 animais, sendo 168 de cada biomodelo. Houve efeito significativo das fontes de óleo sobre os parâmetros bioquímicos plasmáticos em camundongos NIH e ratos Wistar (P 0,05). Conclui-se que o óleo de salmão e linhaça podem ser utilizados nas rações para camundongos NIH e ratos Wistar, independente do método de esterilização adotado.(AU)

The objective of this study was to evaluate the effects of different methods of sterilization and oil sources (salmon and flaxseed) in the formulation of diets on the biochemical and hematological parameters of Wistar rats and NIH mice at different ages. 336 animals were evaluated, 168 from each biomodel. There was a significant effect of oil sources on plasma biochemical parameters in NIH mice and Wistar rats (P < 0.05). There were no significant effects of the sterilization methods on any evaluated parameter (P > 0.05). It was concluded that salmon and linseed oil can be used in diets for NIH mice and Wistar rats, regardless of the sterilization method adopted.(AU)

Types of sterilization in feed containing different lipidic sources for golden hamster (Mesocricetus auratus) / Tipos de esterilização em rações com diferentes fontes lipídicas para hamster golden (Mesocricetus auratus)

The Golden hamster has been gaining significance as a new experimental biomodel, finding use as a reliable diagnostic tool in biomedical research and for zoonosis. Authentic data in terms of digestibility, interactions among raw materials and essential nutrients, besides the influence exerted by various sterilization processes on animal behavior remain unclear. We aimed to assess the influence of sterilization, via autoclaving and irradiation, of pellet feeds prepared using salmon or linseed oil on the digestibility and plasma biochemical parameters in Golden hamsters. Randomized evaluations were conducted on 36 adult male Golden hamsters (Mesocricetus auratus), distributed in six treatments and six replications, namely: common salmon oil; radiated salmon; autoclaved salmon; common linseed oil; radiated linseed and autoclaved linseed. A remarkable effect of the sterilization was evident on the digestibility and protein solubility of the feed, which was lower for autoclaved diets. There was also a significant effect on blood parameters. Animals fed diets containing linseed oil showed lower blood glucose compared to the others. Thus, the inference reached was that while salmon and linseed oil can be used in laboratory hamster feeds, autoclaving disturbs the nutritional quality of the rations.

O hamster Golden é um importante biomodelo utilizado na pesquisa biomédica e como meio de diagnóstico para zoonoses. Informações de qualidade, digestibilidade, interações das matérias primas e exigências nutricionais, assim como os efeitos decorrentes dos diferentes processos de esterilização sobre o desempenho animal ainda são desconhecidos. Objetivou-se com esse estudo avaliar os efeitos da esterilização por autoclavagem e irradiação de rações peletizadas, formuladas com óleo de salmão ou linhaça, sobre a digestibilidade e parâmetros bioquímicos plasmáticos de hamsters Golden. Foram avaliados 36 hamsters Golden (Mesocricetus auratus), machos adultos distribuídos em delineamento experimental inteiramente casualizado com seis tratamentos e seis repetições, a saber: óleo de salmão comum; salmão irradiada; salmão autoclavada; óleo de linhaça comum; linhaça irradiada e linhaça autoclavada. Houve efeito significativo da esterilização sobre a digestibilidade e solubilidade proteica da ração, que foi pior para as rações autoclavadas. Também houve efeito significativo sobre os parâmetros sanguíneos. Animais alimentados com rações contendo óleo de linhaça apresentaram glicemia mais baixa em comparação aos demais. Conclui-se que o óleo de salmão e linhaça pode ser utilizado em rações para hamsters de laboratório, porém a autoclavação interfere na qualidade nutricional das dietas.

Types of sterilization in feed containing different lipidic sources for golden hamster (Mesocricetus auratus) / Tipos de esterilização em rações com diferentes fontes lipídicas para hamster golden (Mesocricetus auratus)

The Golden hamster has been gaining significance as a new experimental biomodel, finding use as a reliable diagnostic tool in biomedical research and for zoonosis. Authentic data in terms of digestibility, interactions among raw materials and essential nutrients, besides the influence exerted by various sterilization processes on animal behavior remain unclear. We aimed to assess the influence of sterilization, via autoclaving and irradiation, of pellet feeds prepared using salmon or linseed oil on the digestibility and plasma biochemical parameters in Golden hamsters. Randomized evaluations were conducted on 36 adult male Golden hamsters (Mesocricetus auratus), distributed in six treatments and six replications, namely: common salmon oil; radiated salmon; autoclaved salmon; common linseed oil; radiated linseed and autoclaved linseed. A remarkable effect of the sterilization was evident on the digestibility and protein solubility of the feed, which was lower for autoclaved diets. There was also a significant effect on blood parameters. Animals fed diets containing linseed oil showed lower blood glucose compared to the others. Thus, the inference reached was that while salmon and linseed oil can be used in laboratory hamster feeds, autoclaving disturbs the nutritional quality of the rations.(AU)

O hamster Golden é um importante biomodelo utilizado na pesquisa biomédica e como meio de diagnóstico para zoonoses. Informações de qualidade, digestibilidade, interações das matérias primas e exigências nutricionais, assim como os efeitos decorrentes dos diferentes processos de esterilização sobre o desempenho animal ainda são desconhecidos. Objetivou-se com esse estudo avaliar os efeitos da esterilização por autoclavagem e irradiação de rações peletizadas, formuladas com óleo de salmão ou linhaça, sobre a digestibilidade e parâmetros bioquímicos plasmáticos de hamsters Golden. Foram avaliados 36 hamsters Golden (Mesocricetus auratus), machos adultos distribuídos em delineamento experimental inteiramente casualizado com seis tratamentos e seis repetições, a saber: óleo de salmão comum; salmão irradiada; salmão autoclavada; óleo de linhaça comum; linhaça irradiada e linhaça autoclavada. Houve efeito significativo da esterilização sobre a digestibilidade e solubilidade proteica da ração, que foi pior para as rações autoclavadas. Também houve efeito significativo sobre os parâmetros sanguíneos. Animais alimentados com rações contendo óleo de linhaça apresentaram glicemia mais baixa em comparação aos demais. Conclui-se que o óleo de salmão e linhaça pode ser utilizado em rações para hamsters de laboratório, porém a autoclavação interfere na qualidade nutricional das dietas.(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)

Detection of differentially methylated regions of irradiated fig tree selections

Fig tree (Ficus carica L.) breeding programs using conventional methods, such as directed crosses, to obtain new cultivars, are unworkable in many countries, including Brazil. Consequently, genetic breeding through mutagenesis has emerged as an important line of research that can improve this crop, and be a significant source of information about this species and assist in the implementation of propagation projects and appropriate management. The aim of this study was to verify the existence of epigenetic variability attributable to DNA methylation in irradiated fig selections when compared both to each other and to the main commercial cultivar, Roxo-de-Valinhos, which had previously used methylation-sensitive amplified polymorphism (MSAP) and DNA sequencing to detect the position of polymorphic regions, analyzable by bioinformatic tools. The sequencing of DNA, isolated from the differentially methylated sites, makes it possible to observe different patterns of methylation by sequencing the treated DNA with sodium bisulfite in the coding regions of regulatory genes active in the development, and fruit ripening stages. Furthermore, they have been found in the mitochondrial DNA of treatments which regulate the supply of energy in Adenosine triphosphate (ATP) form in plants. Closely related to their development, they justify the different phenotypes found in both fruit and plant growth that have suffered stress due to exposure to gamma radiation. Thus, future studies on gene expression in treatments have emerged as an extremely important strategy for understanding these complex regulatory systems, which may lead to the identification of genes of agricultural interest for the fig tree crop, and allow for manipulation and subsequent propagation of improved crops for commercial purposes.

Detection of differentially methylated regions of irradiated fig tree selections

Fig tree (Ficus carica L.) breeding programs using conventional methods, such as directed crosses, to obtain new cultivars, are unworkable in many countries, including Brazil. Consequently, genetic breeding through mutagenesis has emerged as an important line of research that can improve this crop, and be a significant source of information about this species and assist in the implementation of propagation projects and appropriate management. The aim of this study was to verify the existence of epigenetic variability attributable to DNA methylation in irradiated fig selections when compared both to each other and to the main commercial cultivar, Roxo-de-Valinhos, which had previously used methylation-sensitive amplified polymorphism (MSAP) and DNA sequencing to detect the position of polymorphic regions, analyzable by bioinformatic tools. The sequencing of DNA, isolated from the differentially methylated sites, makes it possible to observe different patterns of methylation by sequencing the treated DNA with sodium bisulfite in the coding regions of regulatory genes active in the development, and fruit ripening stages. Furthermore, they have been found in the mitochondrial DNA of treatments which regulate the supply of energy in Adenosine triphosphate (ATP) form in plants. Closely related to their development, they justify the different phenotypes found in both fruit and plant growth that have suffered stress due to exposure to gamma radiation. Thus, future studies on gene expression in treatments have emerged as an extremely important strategy for understanding these complex regulatory systems, which may lead to the identification of genes of agricultural interest for the fig tree crop, and allow for manipulation and subsequent propagation of improved crops for commercial purposes.(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)