Extensive burn injury causes bone collagen network alteration and growth delay related to RANK-L immunoexpression change.

PURPOSE: Extensive burn injury mainly affects children, and hypermetabolic state can lead to growth delay. This study aimed to investigate bone histopathological and morphometric aspects, collagen fibers network and the immunoexpression of biological markers related to bone development in a young experimental model for extensive burn. MATERIALS AND METHODS: A total of 28 male Wistar rats were distributed into Control (C) and subjected to scald burn injury (SBI) groups. Sham or injured animals were euthanized 4 or 14 days post-lesion and proximal epiphyses of the femur were submitted to histological, morphometric (thickness epiphyseal plate), and RUNX-2 and receptor activator of nuclear factor kappa- ß ligand (RANK-L) immunoexpression methods. RESULTS: Histopathological femoral findings showed delayed appearance of the secondary ossification center in SBI, 14 days post-injury. Collagen fibers 4 days after injury were observed in articular cartilage as a pantographic network with a transversally oriented lozenge-shaped mesh, but this network was thinner in SBI. Fourteen days after the injury, the pantographic network of collagen presented square-shaped mesh in C, but this aspect was changed to a wider mesh in SBI. Morphometric analysis of epiphyseal plate revealed that the SBI group had less thickness than the respective controls (p<0.05). RUNX-2 showed no difference between groups, but RANK-L score was higher in all SBI groups. CONCLUSIONS: Extensive burn injury causes delayed bone growth and morphological changes. Alterations in collagen network and enhancement in immunoreactivity of RANK-L result in increased osteoclastogenesis.

Temporal study following burn injury in young rats is associated with skeletal muscle atrophy, inflammation and altered myogenic regulatory factors.

INTRODUCTION: Burn injury (BI) greater than 40% has been associated with protein catabolism and it is characterized by a hypermetabolic response followed for muscle loss. OBJECTIVE: The purpose of this study was to investigate the temporal effects of extensive experimental BI in the skeletal muscle distant from lesion, through morphological analysis, expression of genes related to muscle atrophy, inflammation and the myogenic regulatory factors. MATERIALS AND METHODS: A total of 60 young male wistar rats were distributed into two groups, control (C) and subjected to scald burn injury (SBI). The animals were euthanized 1, 4 and 14 days post-sham or 45% of the total body surface BI. The medial head of gastrocnemii muscles were submitted to histopathological, morphometric (muscle fibers area and density), MyoD and myogenin immunoexpression, and gene expression for TNF-α, iNOS and E3 ubiquitin ligases (MuRF1 and MAFbx). RESULTS: Histopathological findings were consistent with increased amount of connective tissue and inflammatory process. Muscle fiber area of SBI groups was smaller than C and no differences were found in fiber muscle density. TNF-α was higher in SBI groups, one and 14 days post-injury; iNOS expression was higher on the first and fourth day post-injury. MuRF-1 was higher on the day four and MAFbx on the day 14. CONCLUSION: In conclusion, BI causes inflammation, atrophy and myogenesis stimulation in muscle as a result of systemic host response.

Cyclooxygenase-2 expression in skeletal muscle of knockout mice suffering Duchenne muscular dystrophy.

The purpose of the present study was to investigate the role of cyclooxygenase-2 (COX-2) expression in fibrotic lesion in mdx mice. A total of six male C57BL/10 mice and six C57BL/10-DMD/mdx were distributed into two groups: control and animals with Duchenne muscular dystrophy (DMD). The medial part of gastrocnemius muscle was evaluated being the specimens stained with hematoxylin and eosin (H&E) and Sirius Red under normal and polarized light to differentiate type I (red and yellow) and III (green) collagen. COX-2 expression was assessed by immunohistochemistry. The results revealed histopathological changes in C57BL/10-DMD/mdx as depicted by regenerating fibers. Sirius Red stain showed a substantial increase in the amount of type I collagen of mdx mice. DMD induced a strong COX-2 immunoexpression in intercellular space. Taken together, our results are consistent with the notion that necrotic and fibrotic lesions are able to increase COX-2 expression in DMD.

Protein Malnutrition Pre- and Postnatal and Nutritional Rehabilitation Modulates the Morphology of Muscle Fibers in Wistar Rats.

This study investigated the effects of pre- and postnatal conditions of protein deficiency followed to nutritional rehabilitation in the morphology of skeletal muscle. Twelve Wistar male rats were distributed in two groups: nourished (N), with normal protein diet and undernourished (U), with low protein diet. The respective diet was maintained until animals completed 21 days of life. After that, part of group U (n = 4) received normal protein diet, forming a third group, renourished group (R). Forty-two-day-old animals were euthanized and we performed histopathological and morphometric analysis of the soleus muscle. Analysis stained in hematoxylin and eosin (H&E) of the group N revealed polygonal and equidistant muscle fibers, with normal distribution in muscle fascicles. However, D group had rounded and disorganized fibers with different distances between them in muscle fascicles. R group presented muscle fibers with several formats, polygonal and rounded, and some muscle fascicles starting the reorganization process. In N group, analysis of the connective tissue showed predominance of type I collagen and a lower amount collagen type III, both well organized. Whereas U group had a predominance of disorganized type III collagen, in R group, there was return of type I collagen, but partially organized. Muscle fiber area of U (163.18 ± 52.55 µm2) and R (381.79 ± 26.62 µm2) groups was smaller than N (1229.2 µm2 ± 61.12 µm2). Muscle fibers density of groups U (3369 ± 1226 fibers/mm2) and R (1979 ± 28 fibers/mm2) was larger than N (830 ± 113 fibers/mm2). The nutritional rehabilitation in the present study showed an attempt of reorganization of the muscle tissue.

Burn injury induces histopathological changes and cell proliferation in liver of rats.

AIM: To investigate effects of severe burn injury (BI) in rat liver through the histopathological and inflammatory markers analysis. METHODS: Forty-two male Wistar rats were distributed into two groups, control (C) and subjected to scald BI (SBI). The animals were euthanized one, four and 14 d post sham or 45% of the total body surface BI. Liver fragments were submitted to histopathological, morphoquantitative (hepatocyte area and cell density), ciclooxigenase-2 (COX-2) immunoexpression, and gene expression [real-time polymerase chain reaction for tumor necrosis factor (TNF)-α, inducible nitric oxide synthase (iNOS) and caspase-3] methods. RESULTS: Histopathological findings showed inflammatory process in all periods investigated and hepatocyte degeneration added to increased amount of connective tissue 14 d post injury. Hepatocyte area, the density of binucleated hepatocytes and density of sinusoidal cells of SBI groups were increased when compared with control. COX-2 immunoexpression was stronger in SBI groups. No differences were found in TNF-α, iNOS and caspase-3 gene expression. CONCLUSION: BI induces histopathological changes, upregulation of COX-2 immunoexpression, and cell proliferation in liver of rats.

Burn injury induces skeletal muscle degeneration, inflammatory host response, and oxidative stress in wistar rats.

Burn injuries (BIs) result in both local and systemic responses distant from the site of thermal injury, such as skeletal muscle. The purpose of this study was to investigate the expression of cyclooxygenase-2 (COX-2) and hydroxy-2'-deoxyguanosine (8-OHdG) as a result of inflammation and reactive oxygen species production, respectively. A total of 16 male rats were distributed into two groups: control (C) and submitted to BI. The medial part of gastrocnemius muscle formed the specimens, which were stained with hematoxylin and eosin and were evaluated. COX-2 and 8-OHdG expressions were assessed by immunohistochemistry, and cell profile area and density of muscle fibers (number of fibers per square millimeter) were evaluated by morphometric methods. The results revealed inflammatory infiltrate associated with COX-2 immunoexpression in BI-gastrocnemius muscle. Furthermore, a substantial decrease in the muscle cell profile area of BI group was noticed when compared with the control group, whereas the density of muscle fibers was higher in the BI group. 8-OHdG expression in numerous skeletal muscle nuclei was detected in the BI group. In conclusion, the BI group is able to induce skeletal muscle degeneration as a result of systemic host response closely related to reactive oxygen species production and inflammatory process.

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

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

Systemic host response following skin burn injury in rats: cytotoxicity and genotoxicity evaluation.

AIM: The aim of the present study was to investigate whether skin burn injury (BI) can induce cellular changes in skeletal muscle, liver, kidney and blood by means of DNA damage and cytotoxicity. MATERIALS AND METHODS: Thirty male Wistar rats were distributed into two groups: control (C) and submitted to scald burn (SB), subdivided into three subgroups: 1, 4 or 14 days post-injury. The gastrocnemius muscle and liver were dissected for histopathological evaluation and the single-cell gel (comet) assay was used to investigate damage in skeletal muscle, liver, kidney and blood cells. RESULTS: Histopathological analysis of the muscle in the SB group revealed congested vessels containing inflammatory cells for all periods evaluated post-injury. In liver, the one day post-injury SB group showed sinusoidal congestion, while that of 14 days post-injury exhibited an increased number of Kupffer cells. CONCLUSION: Despite the histophatological evidence, none of the groups showed any signs of genotoxicity in these target tissues.

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

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