Patricia Brassolatti

Comparative effects of two different doses of low‐level laser therapy on wound healing third‐degree burns in rats

Burns are injuries caused by direct or indirect contact to chemical, physical, or biological agents. Low‐level laser therapy (LLLT) is a promising treatment since it is low‐cost, non‐invasive, and induces cell proliferation. This study aimed to investigate the effects of LLLT (660 nm) at two different fluences (12.5 J/cm2 and 25 J/cm2) per point of application on third‐degree burns in rats. Thirty rats (Wistar) divided into GC, GL12.5, and GL25 were used in the study, and submitted to burn injury through a soldering iron at 150°C, pressed on their back for 10 s. LLLT was applied immediately, and 2, 4, 6, and 8 days after wound induction. Histological analysis revealed a decreased inflammatory infiltrate in the group treated with 25 J/cm2, and intense inflammatory infiltrate in the control group and in the group treated with 12.5 J/cm2. The immunostaining of COX‐2 was more intense in the control groups and in the group treated with 12.5 J/cm2 than in the group treated with 25 J/cm2. Conversely, VEGF immunomarking was more expressive in the group treated with 25 J/cm2 than it was in the other two groups. Therefore, our findings suggest that the use of 25 J/cm2 and 1 J of energy was more effective in stimulating the cellular processes involved in tissue repair on third‐degree burns in rats by reducing the inflammatory phase, and stimulating angiogenesis, thus restoring the local microcirculation which is essential for cell migration. Microsc. Res. Tech. 79:313–320, 2016.

Photobiomodulation mechanisms in the kinetics of the wound healing process in rats

OBJECTIVES The healing process of cutaneous lesions is considered a complex event divided into distinct and overlapping phases, which responds satisfactorily to photobiomodulation (PBM). PBM is indicated as a therapeutic resource capable of assisting tissue repair. The present study aimed to analyze the kinetics of cutaneous wounds healing process after application of the GaAlAs laser for treating wounds on the dorsum of rats. MATERIALS AND METHODS This study was approved by the Animal Ethics Committee of UFSCar. The animals were divided into 2 groups (n = 10); control group (CG) used 0.9% saline solution and the laser group (LG) used GaAlAs, 670 nm continuous pulse, 30 mW power, 14.28 J/cm2 energy density, irradiating 1 point per wound for 30s, totaling 15 consecutive days of treatment. Samples were collected on the 4th, 11th and 16th days for histological analysis of HE, Picrosirius-Red, immunohistochemistry (Collagen1, TNF-α, VEGF). Statistical analyzes used the one-way ANOVA test for intra and inter group evaluations, and the Tukey post-test. Level of significance was set at p < 0.05. RESULTS The histopathological analysis (HE) showed a statistically significant difference for lower values of inflammatory infiltrate in LG versus CG on the 16th day; and for the increase of collagen in the 11th and 16th days of treatment. There was a statistically significant difference in the increase of VEGF on the 11th day for LG; decrease of TNF-α on the 4th and 11th day for LG, and increase of collagen type 1 on the 4th and 16th days for LG. The birefringence analysis of the percentage of collagen fibers presented on the 11th day of treatment revealed a greater quantity and significant statistical difference. Collagen fibers showed improved organization and arrangement on the 11th day for LG. CONCLUSION Our results show that PBM is effective in helping the kinetics of the cutaneous wound healing process in rats and promotes the necessary stimuli for the satisfactory evolution of healing process, ultimately leading to structurally desirable tissue.

Evaluation of the low-level laser therapy application parameters for skin burn treatment in experimental model: a systematic review

Burn is defined as a traumatic injury of thermal origin, which affects the organic tissue. Low-level laser therapy (LLLT) has gained great prominence as a treatment in this type of injury; however, the application parameters are still controversial in the literature. The aims of this study were to review the literature studies that use LLLT as a treatment in burns conducted in an experimental model, discuss the main parameters used, and highlight the benefits found in order to choose an appropriate therapeutic window to be applied in this type of injury. The selection of the studies related to the theme was carried out in the main databases (PubMed, Cochrane Library, LILACS, Web of Science, and Scopus in the period from 2001 to 2017). Subsequently, the articles were then chosen that fell within the inclusion criteria previously established. In the end, 22 were evaluated, and the main parameters were presented. The analyzed studies presented both LLLT use in continuous and pulsed mode. Differences between the parameters used (power, fluence, and total energy) were observed. In addition, the protocols are distinct as to the type of injury and the number of treatment sessions. Among the results obtained by the authors are the improvements in the local microcirculation and cellular proliferation; however, a study reported no effects with LLLT as a treatment. LLLT is effective in accelerating the healing process. However, there is immense difficulty in establishing the most adequate protocol, due to the great discrepancy found in the applied dosimetry values.

Photobiomodulation on critical bone defects of rat calvaria: a systematic review

Bone defects following trauma represent a high impact on the quality of life of millions of people around the world. The aim of this study was to review photobiomodulation (PBM) action in the treatment of bone critical defects in rat calvaria, related to evaluation of the current protocols applied. One hundred and forty-seven articles related to the subject were found by searching the main databases (Pubmed, Lilacs, Web of Science, and Scopus) considering the period of publication until the year 2017, and only 14 corresponded the inclusion criteria established for this systematic review. The main parameters of the PBM were expressed in Table 1. In addition, it was possible to observe the use of two different wavelengths (red and infrared), which are considered therapeutic. Most of the evaluated articles presented positive results that describe a greater amount of neoformed bone, an increase in collagen synthesis, and a contribution to microvascular reestablishment. However, two studies report no effect on the repair process when the PBM was used. In addition, we observed considerable variations between the values of power, fluence, and total energy, which make it difficult to compare the results presented between the selected studies. It was possible to conclude that the infrared laser was more effective in positively stimulating the bone repair process of critical defects. Furthermore, a discrepancy was found in the parameter values used, which made it difficult to choose the best protocol for the treatment of this type of lesion.

Mitochondrial dynamics (fission and fusion) and collagen production in a rat model of diabetic wound healing treated by photobiomodulation: comparison of 904 nm laser and 850 nm light-emitting diode (LED)

OBJECTIVE Mitochondrial dysfunction has been associated with the development of diabetes mellitus which is characterized by disorders of collagen production and impaired wound healing. This study analyzed the effects of photobiomodulation (PBM) mediated by laser and light-emitting diode (LED) on the production and organization of collagen fibers in an excisional wound in an animal model of diabetes, and the correlation with inflammation and mitochondrial dynamics. METHODS Twenty Wistar rats were randomized into 4 groups of 5 animals. Groups: (SHAM) a control non-diabetic wounded group with no treatment; (DC) a diabetic wounded group with no treatment; (DLASER) a diabetic wounded group irradiated by 904 nm pulsed laser (40 mW, 9500 Hz, 1 min, 2.4 J); (DLED) a diabetic wounded group irradiated by continuous wave LED 850 nm (48 mW, 22 s, 1.0 J). Diabetes was induced by injection with streptozotocin (70 mg/kg). PBM was carried out daily for 5 days followed by sacrifice and tissue removal. RESULTS Collagen fibers in diabetic wounded skin were increased by DLASER but not by DLED. Both groups showed increased blood vessels by atomic force microscopy. Vascular endothelial growth factor (VEGF) was higher and cyclooxygenase (COX2) was lower in the DLED group. Mitochondrial fusion was higher and mitochondrial fusion was lower in DLED compared to DLASER. CONCLUSION Differences observed between DLASER and DLED may be due to the pulsed laser and CW LED, and to the higher dose of laser. Regulation of mitochondrial homeostasis may be an important mechanism for PBM effects in diabetes.

Alternative animal model for studies of total skin thickness burns

PURPOSE To present an alternative experimental model of third degree burn of easy reproducibility. METHODS Eighteen male Wister rats were randomly divided into three groups, 6 of which were allocated to each group. A soldering iron coupled to an aluminum plate was used to produce burn, at a temperature of 150ºC, with different exposure times per group. Group 5 (G5) animals were burned at 150°C with exposure time of 5 seconds; Group 10 (G10) the animals were burned at 150°C with exposure time of 10 seconds and group 15 (G15) the animals were burned at 150°C with exposure time of 15 seconds. RESULTS Histopathological analyzes showed that all three groups had similar morphological characteristics, with total thickness involvement. CONCLUSION The technique is effective to reproduce a third degree burn and suggests the temperature of 150ºC with 5 seconds of exposure in order to minimize the risks to the animals.