Laser-photobiomodulation on experimental cancer pain model in Walker Tumor-256.

CONTEXT: Cancer Pain is considered a common and significant clinical problem in malignant neoplasms, comprising 20% to 50% of all patients with tumor progression. Laser photobiomodulation (L-PBM) has been used in a multitude of pain events, ranging from acute trauma to chronic articular. However, L-PBM has never been tested in cancer pain. OBJECTIVES: Evaluate hyperalgesia, edema, COX-1, COX-2, IL-10, and Bdkrb1 mRNA in low-level laser irradiated Walker-256 tumor-bearing rats. METHODS: Rat hind paw injected with Walker Tumor-256 (W-256) and divided into six groups of 6 rats: G1 (control) - W-256 injected, G2- W-256 + Nimesulide, G3- W-256 + 1 J, G4- W-256 + 3 Jand G5- W256 + 6 J. Laser parameters: λ = 660 nm, 3.57 W/cm2, Ø = 0.028 cm2. Mechanical hyperalgesia was evaluated by Randall-Selitto test. Plethysmography measured edema; mRNA levels of COX-1, COX-2, IL-10, and Bdkrb1were analyzed. RESULTS: It was found that the W-256 + 1 J group showed a decrease in paw edema, a significant reduction in pain threshold. Higher levels of IL-10 and lower levels of COX-2 and Bdkrb1 were observed. CONCLUSION: Results suggest that 1 J L-PBM reduced the expression of COX-2 and Bdkrb1 and increasing IL-10 gene expression, promoting analgesia to close levels to nimesulide.

Antitumor effect of cell therapy with mesenchymal stem cells on murine melanoma B16-F10.

In this study, the antitumor and immunomodulatory effects of mesenchymal stem cells (MSC) obtained from bone marrow in the treatment of dorsal melanoma B16-F10. The MSC cells were obtained from the bone marrow of isogenic C57BL/6J mice, characterized and inoculated by two routes, intratumor (it) and intravenous (iv). The hematological profile, expression markers and receptors, phases of the cell cycle and mitochondrial electrical potential were evaluated by flow cytometry. The dorsal tumor mass showed a significant reduction after treatment by the two routes of administration with a significant effect by the intravenous route. MSC showed immunomodulatory potential and did not induce an increase in the markers involved in tumor control and progression. The number of cells in the sub-G1 phase increased significantly after treatments compared to the control group. The percentage of cells in phases G0/G1, S and G2/M decreased, with only the group (it) showing a significant reduction. The intratumor group showed a significant decrease in the G2/M phase. Treatment with MSC provided a significant decrease in the percentage of metabolically active tumor cells, demonstrating its intrinsic effect in the control of cell proliferation. Regarding the mechanism of cell death, MSCs modulated the expression of proteins involved in the regulation of the cell cycle, angiogenesis receptors and pro-apoptotic proteins by intrinsic and extrinsic routes. Therefore, the use of undifferentiated MSC, administered intratumor and intravenous is possibly a promising treatment for melanoma.

Laser-photobiomodulation on experimental cancer pain model in walker tumor-256

Context Cancer Pain is considered a common and significant clinical problem in malignant neoplasms, comprising 20% to 50% of all patients with tumor progression. Laser photobiomodulation (L-PBM) has been used in a multitude of pain events, ranging from acute trauma to chronic articular. However, L-PBM has never been tested in cancer pain. Objectives Evaluate hyperalgesia, edema, COX-1, COX-2, IL-10, and Bdkrb1 mRNA in low-level laser irradiated Walker-256 tumor-bearing rats. Methods Rat hind paw injected with Walker Tumor-256 (W-256) and divided into six groups of 6 rats: G1 (control) - W-256 injected, G2- W-256 + Nimesulide, G3- W-256 + 1 J, G4- W-256 + 3 Jand G5- W256 + 6 J. Laser parameters: λ = 660 nm, 3.57 W/cm2, Ø = 0.028 cm2. Mechanical hyperalgesia was evaluated by Randall–Selitto test. Plethysmography measured edema; mRNA levels of COX-1, COX-2, IL-10, and Bdkrb1were analyzed. Results It was found that the W-256 + 1 J group showed a decrease in paw edema, a significant reduction in pain threshold. Higher levels of IL-10 and lower levels of COX-2 and Bdkrb1 were observed. Conclusion Results suggest that 1 J L-PBM reduced the expression of COX-2 and Bdkrb1 and increasing IL-10 gene expression, promoting analgesia to close levels to nimesulide.

Antitumor effect of cell therapy with mesenchymal stem cells on murine melanoma B16-F10

In this study, the antitumor and immunomodulatory effects of mesenchymal stem cells (MSC) obtained from bone marrow in the treatment of dorsal melanoma B16-F10. The MSC cells were obtained from the bone marrow of isogenic C57BL/6J mice, characterized and inoculated by two routes, intratumor (it) and intravenous (iv). The hematological profile, expression markers and receptors, phases of the cell cycle and mitochondrial electrical potential were evaluated by flow cytometry. The dorsal tumor mass showed a significant reduction after treatment by the two routes of administration with a significant effect by the intravenous route. MSC showed immunomodulatory potential and did not induce an increase in the markers involved in tumor control and progression. The number of cells in the sub-G1 phase increased significantly after treatments compared to the control group. The percentage of cells in phases G0/G1, S and G2/M decreased, with only the group (it) showing a significant reduction. The intratumor group showed a significant decrease in the G2/M phase. Treatment with MSC provided a significant decrease in the percentage of metabolically active tumor cells, demonstrating its intrinsic effect in the control of cell proliferation. Regarding the mechanism of cell death, MSCs modulated the expression of proteins involved in the regulation of the cell cycle, angiogenesis receptors and pro-apoptotic proteins by intrinsic and extrinsic routes. Therefore, the use of undifferentiated MSC, administered intratumor and intravenous is possibly a promising treatment for melanoma.

Antitumor effect of cell therapy with mesenchymal stem cells on murine melanoma B16-F10

In this study, the antitumor and immunomodulatory effects of mesenchymal stem cells (MSC) obtained from bone marrow in the treatment of dorsal melanoma B16-F10. The MSC cells were obtained from the bone marrow of isogenic C57BL/6J mice, characterized and inoculated by two routes, intratumor (it) and intravenous (iv). The hematological profile, expression markers and receptors, phases of the cell cycle and mitochondrial electrical potential were evaluated by flow cytometry. The dorsal tumor mass showed a significant reduction after treatment by the two routes of administration with a significant effect by the intravenous route. MSC showed immunomodulatory potential and did not induce an increase in the markers involved in tumor control and progression. The number of cells in the sub-G1 phase increased significantly after treatments compared to the control group. The percentage of cells in phases G0/G1, S and G2/M decreased, with only the group (it) showing a significant reduction. The intratumor group showed a significant decrease in the G2/M phase. Treatment with MSC provided a significant decrease in the percentage of metabolically active tumor cells, demonstrating its intrinsic effect in the control of cell proliferation. Regarding the mechanism of cell death, MSCs modulated the expression of proteins involved in the regulation of the cell cycle, angiogenesis receptors and pro-apoptotic proteins by intrinsic and extrinsic routes. Therefore, the use of undifferentiated MSC, administered intratumor and intravenous is possibly a promising treatment for melanoma.

Proinflammatory effects of photoactivated methylene blue on rat model of Walker 256 carcinosarcoma

Photodynamic therapy (PDT) is an anticancer therapy that associates the photosensitizer (PS), oxygen and light to destroy cancer cells. Methylene blue (MB) is considered a second generation phenothiazine dye with excellent photochemical properties. Aim: To evaluate whether MB-mediated PDT can induce oxidative stress and inflammation, therefore, interfering tumor growth. Materials and Methods: The study was conducted on Wistar rats transplanted with Walker 256 carcinosarcoma (W256). The proinflammatory interleukins levels (IL-1ß, IL-6, IL-10, TNF-a) were determined by ELISA, mRNA expression of COX-1, COX-2, iNOS and eNOS by RT-PCR, lipid peroxidation was measured by the TBARS method. Moreover, myeloperoxidase (MPO) activity in neutrophils was determined by MPO activity assay. All indices mentioned above were determined in tumor tissue. Kaplan — Meier and Gehan — Breslow — Wilcoxon tests were used for survival analysis. Results: We found that the treatment of W256 with 0.1% MB + 1 J/cm2 provoked a significant increase in the interleukins levels (IL-1ß, IL-6, IL-10, TNF-a), prostaglandin E2, the mRNA expression of COX-2, iNOS, lipid peroxidation and MPO activity in tumor tissue, which were statistically different (p < 0.05) compared to other experimental and control groups. The results of the estimation of survival curves show a greater probability of survival in 0.1% MB + 1 J/cm2 (total energy dose =142.8 J/cm2 ) treated group. Conclusion: Our results suggest that treatment of W256 with 0.1% MB + 1 J/cm2 was able to promote cytotoxic effects in tumor tissue by the generation of reactive oxygen species causing inflammation and thus interfering in the tumor growth

Laser photobiomodulation of pro-inflammatory mediators on Walker Tumor 256 induced rats.

Laser photobiomodulation or low-level laser therapy (LLLT) is recognized worldwide for its expansive use in medicine. LLLT has been reported to increase enzymatic activity, increasing the mitochondrial transmembrane potential, leading to an increased energy availability and signal transduction. Nevertheless, an inhibitory effect is also observed by the production of excessive ROS which can result the shutdown of mitochondrial energy production, and finally to apoptosis. However, the mechanism of apoptosis induced by LLLT is still not well understood. The main objective of the present study was to investigate the hypothesis that LLLT induces oxidative stress and stimulates the generation of pro-inflammatory markers interfering in tumor progression. METHODS: Seventy-two female Walker Tumor induced Wistar rats (eight weeks of age, 200g body weight) were used for this study. TW-256 cells were suspended in phosphate buffered saline and then subcutaneously inoculated at 1×107viabletumorcells/ml per rat into the right flank (tumor-bearing rats). After a period of 14days in order to assess the development of the solid tumor mass, the animals were randomized and distributed in four groups (n=8 animals/group): (1) Control or irradiated by LLLT (2) Laser 1J - 35,7J/cm2, (3) Laser 3J - 107,14J/cm2 and (4) Laser 6J - 214,28J/cm2; (Thera Laser - 660nm, 100mW DMC®, São Carlos, Brazil) at four equidistant points according to their respective treatment groups, conducted three times on alternate days. The regulation and expression of inflammatory mediators IL-1ß, IL-6, IL-10, TNF-α was assessed by ELISA and gene expression of COX-1, COX-2, iNOS, eNOS was analyzed by RT-PCR. RESULTS: We found that the 1Joule (J) treated group promoted a significant increase in the levels of different inflammatory markers IL-1ß, the gene expression of COX-2, iNOS, which was statistically different (p<0.05) when compared among different treatment and control groups. With Respect IL-6, IL-10, TNF-α levels statistically significant reduce was observed in 1Joule treated group when comparing to different energies groups and control group. CONCLUSION: Our results suggest the evidence 1J-35,7J/cm2 treatment was able to produce cytotoxic effects by generation of ROS causing acute inflammation and thus may be employed as the best energy dose associated with Photodynamic Therapy.

Laser photobiomodulation of pro-inflammatory mediators on walker tumor 256 induced rats

Laser photobiomodulation or low-level laser therapy (LLLT) is recognized worldwide for its expansive use in medicine. LLLT has been reported to increase enzymatic activity, increasing the mitochondrial transmembrane potential, leading to an increased energy availability and signal transduction. Nevertheless, an inhibitory effect is also observed by the production of excessive ROS which can result the shutdown of mitochondrial energy production, and finally to apoptosis. However, the mechanism of apoptosis induced by LLLT is still not well understood. The main objective of the present study was to investigate the hypothesis that LLLT induces oxidative stress and stimulates the generation of pro-inflammatory markers interfering in tumor progression. Methods Seventy-two female Walker Tumor induced Wistar rats (eight weeks of age, 200 g body weight) were used for this study. TW-256 cells were suspended in phosphate buffered saline and then subcutaneously inoculated at 1 × 107 viable tumor cells/ml per rat into the right flank (tumor-bearing rats). After a period of 14 days in order to assess the development of the solid tumor mass, the animals were randomized and distributed in four groups (n = 8 animals/group): (1) Control or irradiated by LLLT (2) Laser 1J – 35,7 J/cm2, (3) Laser 3 J – 107,14 J/cm2 and (4) Laser 6 J – 214,28 J/cm2; (Thera Laser - 660 nm, 100 mW DMC®, São Carlos, Brazil) at four equidistant points according to their respective treatment groups, conducted three times on alternate days. The regulation and expression of inflammatory mediators IL-1ß, IL-6, IL-10, TNF-a was assessed by ELISA and gene expression of COX-1, COX-2, iNOS, eNOS was analyzed by RT-PCR. Results We found that the 1 Joule (J) treated group promoted a significant increase in the levels of different inflammatory markers IL-1ß, the gene expression of COX-2, iNOS, which was statistically different (p < 0.05) when compared among different treatment and control groups. With Respect IL-6, IL-10, TNF-a levels statistically significant reduce was observed in 1 Joule treated group when comparing to different energies groups and control group. Conclusion Our results suggest the evidence 1 J–35,7 J/cm2 treatment was able to produce cytotoxic effects by generation of ROS causing acute inflammation and thus may be employed as the best energy dose associated with Photodynamic Therapy.

Effects of low-level laser therapy (LLLT) and diclofenac (topical and intramuscular) as single and combined therapy in experimental model of controlled muscle strain in rats.

Muscle injuries represent ca 30% of sports injuries and excessive stretching of muscle causes more than 90% of injuries. Currently the most used treatments are nonsteroidal anti-inflammatory drugs (NSAIDs), however, in last years, low-level laser therapy (LLLT) is becoming an interesting therapeutic modality. The aim of this study was to evaluate the effect of single and combined therapies (LLLT, topical application of diclofenac and intramuscular diclofenac) on functional and biochemical aspects in an experimental model of controlled muscle strain in rats. Muscle strain was induced by overloading tibialis anterior muscle of rats. Injured groups received either no treatment, or a single treatment with topical or intramuscular diclofenac (TD and ID), or LLLT (3 J, 810 nm, 100 mW) 1 h after injury. Walking track analysis was the functional outcome and biochemical analyses included mRNA expression of COX-1 and COX-2 and blood levels of prostaglandin E2 (PGE2 ). All treatments significantly decreased COX-1 and COX-2 gene expression compared with injury group (P < 0.05). However, LLLT showed better effects than TD and ID regarding PGE2 levels and walking track analysis (P < 0.05). We can conclude that LLLT has more efficacy than topical and intramuscular diclofenac in treatment of muscle strain injury in acute stage.