CAROLINA GOUVEA DE SOUZA CONTATORI

CAROLINA GOUVEA DE SOUZA CONTATORI

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Responses of melanoma cells to photobiomodulation depend on cell pigmentation and light parameters
2022 - CONTATORI, CAROLINA G. de S.; SILVA, CAMILA R.; PEREIRA, SAULO de T.; RODRIGUES, MARIA F.S.D.; LUNA, ARTHUR C. de L.; MARQUES, MARCIA M.; RIBEIRO, MARTHA S.
Melanoma is a highly aggressive skin cancer that requires new approaches for its management. Low-level laser therapy, currently named photobiomodulation therapy (PBM), has been used to improve different conditions but its effects and safe use on melanoma remain unexplored. Herein, we investigated the PBM impact on melanoma cells differing by pigmentation using near-infrared (NIR) and red lasers in vitro. In vivo, we evaluated the effects of the red laser on melanoma-bearing mice. Amelanotic (SK-MEL-37) and melanotic (B16F10) cells were exposed in vitro to a NIR (780 nm, 40 mW) or a red laser (660 nm, 40 mW) in 3 different light doses: 30, 90, and 150 J/cm2 and responses were assessed regarding mitochondrial activity, invasiveness, migration, and VEGF production. In vivo, melanoma-bearing mice received the red laser delivering 150 J/cm2 directly to the tumor on 3 consecutive days. Mice were monitored for 15 days regarding tumor progression and mouse survival. We noticed that amelanotic cells were unresponsive to NIR light. In contrast, NIR irradiation at 30 J/cm2 promoted an increase in the invasiveness of pigmented cells, even though all light doses have inhibited cell migration. Regarding the red laser on pigmented cells, the highest light dose (150 J/cm2) decreased the VEGF production and migration. In vivo, melanoma-bearing mice treated with red laser showed smaller tumor volume and longer survival than controls. We conclude that PBM appears to be safe for amelanotic non-pigmented melanoma but triggers different responses in melanotic pigmented cells depending on light parameters. Additionally, a high dose of red laser impairs the invasive behavior of melanoma cells, probably due to the decrease in VEGF synthesis, which may have contributed to tumor arrest and increased mouse survival. These findings suggest that red laser therapy could be a new ally in the supportive care of melanoma patients.
Efeitos da fotobiomodulação pré-exposição à radiação ionizante em células de câncer de mama
2021 - SILVA, CAMILA R.; PEREIRA, SAULO T.; CONTATORI, CAROLINA G.S.; PINTO, MAYARA S.; SALVEGO, CAMILA A.; RIBEIRO, MARTHA S.
Melanoma cell migration in response to red and near-infrared low-level light
2021 - CONTATORI, CAROLINA G. de S.; PINTO, MAYARA S.; RIBEIRO, MARTHA S.
INTRODUCTION Cell migration plays an important role in tissue formation and cancer progression. In vitro scratch assay has been used for many years to study cell migration to mimic the migration of in vivo cells, and, thus, to evaluate cancer growth. Low-level red and near-infrared light (LLL) can increase normal cell migration. However, the impact of LLL on tumor cells remains unclear. OBJECTIVES In this work, we aimed to evaluate the effects of a single LLL dose on melanoma cell migration. MATERIALS AND METHODS B16F10 (murine melanoma) cells were cultivated in RPMI medium with 10% of fetal bovine serum until they reached 80% confluency. The cell line was seeded in a 6-well plate at a density of 2x10 5 cells/well in triplicate at two different moments. A wound scratch was performed to disrupt the confluent cellmonolayerwith a 10 μL pipette tip. Immediately after the injury, the cells were submitted to the LLL at two distinct wavelengths (660 and 780 nm) provided by a LED and a laser, respectively, delivering 3 different energies (1.3, 3.6, and 6 J) at an irradiance of 4.2 mW/cm 2 . The control group was not irradiated. Cells were photographed immediately and at 3, 12, 24, and 36 h after the scratch. The wound closure was measured using ImageJ software. To evaluate the overall migration, we calculated the areas under the curve for each group. DISCUSSION AND RESULTS Cells exposed to the red laser at 6 J migrated slower than control. In contrast, LLL at 780 nm promoted faster cell migration when irradiated with 3.6 J. CONCLUSION These results suggest that low-level LEDs at 660 nm could prevent melanoma progression in higher energies. However, 780 nm should be avoided at middle energies.
Melanoma cell migration in response to red and near-infrared low-level light
2021 - CONTATORI, CAROLINA G. de S.; PINTO, MAYARA S.; RIBEIRO, MARTHA S.
Cell migration plays an important role in tissue formation and cancer progression. In vitro scratch assay has been used for many years to study cell migration to mimic the migration of in vivo cells, and, thus, to evaluate cancer growth. Low-level red and near-infrared light (LLL) can increase normal cell migration. However, the impact of LLL on tumor cells remains unclear. In this work, we aimed to evaluate the effects of a single LLL dose on melanoma cell migration. B16F10 (murine melanoma) cells were cultivated in RPMI medium with 10% of fetal bovine serum until they reached 80% confluency. The cell line was seeded in a 6-well plate at a density of 2x10 5 cells/well in triplicate at two different moments. A wound scratch was performed to disrupt the confluent cell monolayer with a 10 μL pipette tip. Immediately after the injury, the cells were submitted to the LLL at two distinct wavelengths (660 and 780 nm) provided by a LED and a laser, respectively, delivering 3 different energies (1.3, 3.6, and 6 J) at an irradiance of 4.2 mW/cm2. The control group was not irradiated. Cells were photographed immediately and at 3, 12, 24, and 36 h after the scratch. The wound closure was measured using ImageJ software. To evaluate the overall migration, we calculated the areas under the curve for each group. Cells exposed to the red laser at 6 J migrated slower than control. In contrast, LLL at 780 nm promoted faster cell migration when irradiated with 3.6 J. These results suggest that low-level LEDs at 660 nm could prevent melanoma progression in higher energies. However, 780 nm should be avoided at middle energies.
Effects of low-level laser irradiation on VEGF expression of melanoma cell lines
2020 - CONTATORI, C.G. de S.; SILVA, C.R.; YOSHIMURA, T.M.; RIBEIRO, M.S.
Impact of low-level laser irradiation on tumor cell lines remains controversial. Vascular endothelial growth factor (VEGF) is a key molecule to form new blood vessels, which contribute for cancer development and growth. The aim of this study was to evaluate the effects of different light fluences on human melanoma SKMEL 37 cells and murine melanoma B16F10 cells using a near infrared laser (λ= 780 nm) with output power of 40 mW delivering energies of 1.2, 3.6 and 6 J (fluences of 30, 90 and 150 J/cm2, respectively). The cell lines were irradiated 24 h after they were seeded in a 96-well plate at a density of 5x103 cells per well, in triplicate at three different days. Following irradiation, both cell line supernatants were stored in Eppendorf tubes at - 20°C until VEGF-A expression measurement. Specific ELISA kits were used according to cell line (murine or human). Samples and standard solutions were added in a 96-well plate antibody-coated and incuba ted over night at 4°C. Reagent dilution and set time followed fabricant instructions. The stop solution was added and the absorbance was read in a microplate reader at 450 nm. Results showed a non-statistically significant difference among treated and control groups for both cell lines. These findings indicate that irradiation with near infrared laser does not influence VEGF expression on melanoma cell lines regardless the fluence used and should be tested to prevent cancer growth in preclinical assays.
Photobiomodulation can delay tumor progression in breast cancer bearing-mice
2018 - SILVA, CAMILA R.; CONTATORI, CAROLINA G. de S.; PEREIRA, SAULO de T.; RIBEIRO, MARTHA S.
Cancer is a worldwide health problem and new therapeutic strategies are necessary. Photobiomodulation is a noninvasive and cost-effective therapy, but its use in cancer cells is still controversial. In this study, we explore the effects of PBM on breast tumor bearing-mice.
Efeitos do laser de baixa potência de emissão infravermelha (λ=780nm) em células de melanoma murino e humano
2019 - CONTATORI, CAROLINA G. de S.
O câncer de pele pode ser do tipo melanoma ou não melanoma, sendo comum em pessoas acima de 40 anos, de pele clara ou com doenças cutâneas prévias. A incidência do melanoma é baixa, porém, é considerado o mais agressivo e mortal devido ao seu alto poder metastático. A cirurgia ainda é a forma de tratamento mais empregada para a doença, sendo muito invasiva e, portanto, terapias coadjuvantes estão sendo empregadas a fim de melhorar a qualidade de vida dos pacientes, como o laser de baixa potência (LBP). Sabe-se que o LBP pode desencadear efeito bioestimulatório em culturas celulares crescidas sob déficit nutricional, porém em linhagens tumorais sua ação é controversa. Dessa forma, o objetivo desse estudo consiste em investigar os efeitos inibitórios do LBP no comportamento de células de melanoma murino B16F10 e humano SKMEL 37 utilizando um laser de emissão infravermelha (λ = 780 nm) com diferentes densidades de energia. Foram adotados 4 grupos experimentais: G0 (grupo controle), G30 (30 J.cm-2), G90 (90 J.cm-2) e G150 (150 J.cm-2) a fim de verificar a viabilidade celular, através do ensaio de MTT e vermelho neutro; o comportamento de invasão celular, obtido através do ensaio de invasão transwell; e o papel do LBP na expressão do fator de crescimento endotelial vascular (VEGF), verificado através do ensaio imunoenzimático ELISA. Os resultados mostraram que a densidade de energia de 30 J.cm-2 estimulou o comportamento de invasão da linhagem celular B16F10. Por outro lado, o LBP não exerceu influência na expressão do fator de crescimento endotelial vascular, na viabilidade celular, e na atividade mitocondrial de ambas as linhagens celulares, em nenhuma das densidades de energia utilizadas, em comparação ao controle.
Effects of near-infrared low level laser irradiation on melanoma cells
2019 - CONTATORI, CAROLINA G. de S.; SILVA, CAMILA R.; RIBEIRO, MARTHA S.
Low-level laser (LLL) therapy promotes biostimulating effects in cell cultures growing in nutritional deficit. However, the effects of LLLs on tumor cell lines remain controversial. Studies indicate stimulatory, inhibitory or even no influence in this type of cells. Therefore, the aim of this study was to evaluate the influence of LLL irradiation on the cell viability (with and without nutritional deficit) of human melanoma SKMEL 37 cells and murine melanoma B16F10 cells using an infrared laser (k = 780 nm) with different radiant exposures. The cell lines were subjected to the LLL 24 h after they were seeded in a 96-well plate at a density of 5 104 cells per well. The analysis of cell proliferation by mitochondrial activity occurred at intervals of 24 and 72 h after laser irradiation. At each time, culture medium was removed and 180 μL of PBS and 20 μL of MTT were added. The plates were incubated for 4 h and the absorbance was read in a microplate reader at 570 nm. Results showed a non-significant statistical difference among the groups for both cell lines regardless the nutritional medium. The metabolic pattern was similar among the groups. It is concluded that irradiation with 780 nm laser light at radiant exposures of 30, 90 and 150 J/cm2 and an output power of 40 mW does not promote cell proliferation on melanoma cell lines.
Low level light therapy on breast tumor. In vitro and in vivo studies
2018 - PEREIRA, SAULO; CONTATORI, CAROLINA; RIBEIRO, MARTHA S.
Low Level Light Therapy (LLLT) has been gaining prominence in health areas due to search for less invasive and cost-effective treatments. However, its use is not indicated to treat patients with cancer due to a few studies in vivo. The aim of this work was to evaluate the effects of LLLT on breast tumor 4T1 cells in vitro and in a murine model. For the in vitro assay, 4T1 cells were submitted to a single session of LLLT with red light (660 ± 20 nm) delivering energies of 1.2 J and 6 J. The cell viability was measured by MTT assay. For in vivo study, nine BALB/c female mice (6 weeks old) received 4T1 cells transfected with luciferase (4T1-Luc) into the mammary fat pad. After two weeks, the animals were divided into control (n=3) and test groups were submitted to LLLT with same parameters described above. The tumor progression was monitored by a caliper and bioluminescence. The anti-fatigue effect of LLLT was assessed by a forced swimming test. Our results showed that LLLT was not able to increase cell viability regardless the energy used. In vivo, the lower energy promoted an increase of the tumor volume compared to control group. On the other hand, the higher energy was able to arrest tumor progression as well as increase the swimming time. Taken together, our results suggest that LLLT triggers opposite effects on breast tumor depending on the type of assay.