TANIA MATEUS YOSHIMURA

TANIA MATEUS YOSHIMURA

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Red LED radiation as a potential radioprotector using a zebrafish embryo model
2024 - GUSMÃO, KARINE R. dos A.; RODRIGUES, MURILO A.V.; YOSHIMURA, TANIA M.; ROGERO, JOSE R.; RIBEIRO, MARTHA S.; FREITAS, ANDERSON Z. de
Radiation therapy (RT) is crucial for cancer treatment but often causes severe side effects that impact quality of life, emphasizing the need for better tissue radioprotection strategies. This study investigated the radioprotective potential of red LED irradiation using zebrafish embryos. After determining the median lethal dose of gamma radiation (LD50) to be 16.7 Gy, embryos were exposed to red LED before gamma radiation. While red LED alone did not affect survival rates, pre-treatment with red light increased survival to over 50% after LD50 gamma radiation. These results suggest that red LED could help reduce the acute effects of RT.
Combined effects of optical therapy and stem cells for tendon rehabilitation
2024 - SAUTER, ISMAEL P.; YOSHIMURA, TANIA M.; RAPOSO, BRUNO L.; RIBEIRO, MARTHA S.
Tendon injuries often fail to fully regenerate after surgery, leading to impaired function and possible graft rejection. Advances in regenerative medicine include tendon engineering with stem cells. Herein, we demonstrate that adipose-derived mesenchymal stem cells respond differently to culture media, and red LED enhances metabolism and collagen synthesis.
Photodynamic therapy mediated by a red LED and methylene blue inactivates resistant leishmania amazonensis
2023 - CABRAL, FERNANDA V.; YOSHIMURA, TANIA M.; SILVA, DANIELA de F.T. da; CORTEZ, MAURO; RIBEIRO, MARTHA S.
Cutaneous leishmaniasis is a neglected parasitic disease that leads to destructive lesions. The emergence of drug resistance has been a global concern over the past years. Photodynamic therapy (PDT) mediated by a red LED and methylene blue (MB) involves the overproduction of oxidative stress, which oxidizes several cellular biomolecules and prevents the selection of resistant strains. Herein, we investigated the potential of PDT mediated by MB against wild-type and miltefosine-resistant strains of Leishmania amazonensis. As a result, both strains were susceptible to PDT, thus encouraging us to seek the best conditions to overcome the drug resistance problem in cutaneous leishmaniasis.
Could light-based technologies improve stem cell therapy for skin wounds?
2023 - YOSHIMURA, TANIA M.; CABRAL, FERNANDA V.; SELLERA, FABIO P.; POZZO, LORENA; RIBEIRO, MARTHA S.
Several diseases or conditions cause dermatological disorders that hinder the process of skin repair. The search for novel technologies has inspired the combination of stem cell (SC) and light-based therapies to ameliorate skin wound repair. Herein, we systematically revised the impact of photobiomodulation therapy (PBM) combined with SCs in animal models of skin wounds and quantitatively evaluated this effect through a meta-analysis. For inclusion, SCs should be irradiated in vitro or in vivo, before or after being implanted in animals, respectively. The search resulted in nine eligible articles, which were assessed for risk of bias. For the meta-analysis, studies were included only when PBM was applied in vivo, five regarding wound closure, and three to wound strength. Overall, a positive influence of SC + PBM on wound closure (mean difference: 9.69; 95% CI: 5.78–13.61, P < 0.00001) and strength (standardized mean difference: 1.7, 95% CI: 0.68–2.72, P = 0.001) was detected, although studies have shown moderate to high heterogeneity and a lack of information regarding some bias domains. Altogether, PBM seems to be an enabling technology able to be applied postimplantation of SCs for cutaneous regeneration. Our findings may guide future laboratory and clinical studies in hopes of offering wound care patients a better quality of life.
Methylene blue-mediated antimicrobial photodynamic therapy on chicken semen
2023 - NOVAES, G.A.; BLANK, M.H.; YOSHIMURA, T.M.; RIBEIRO, M.S.; PEREIRA, R.J.G.
Background: Artificial insemination is widely employed in poultry, but high degrees of bacterial contamination are often observed in semen because of its passage through the cloaca. Consequently, most semen extenders for birds have antibiotics that could aggravate bacterial resistance. Methods: We evaluated the potential of antimicrobial photodynamic therapy (PDT) as an alternative to the use of antibiotics, and assessed whether changes in concentration and incubation time with methylene blue (MB), radiant exposure, and irradiance of light affect spermatozoa activity and bacteria in chicken semen. Results: Incubation with MB (< 25 µM) did not alter sperm motility, regardless of the pre-irradiation time (PIT, 1 or 5 min). Following 1 min of PIT with MB at 10 µM, samples were irradiated for 30, 60, 120, and 180 s at irradiances of 44, 29, and 17 mW/ cm² (660 nm LedBox). MB and light alone did not interfere with the analyzed parameters. However, when both factors were associated, increases in light dose led to greater reductions in sperm parameters, regardless of the irradiance used. Besides, PDT conditions that were less harmful to spermatozoa were not able to significantly reduce bacterial colonies in chicken semen. Conclusions: A failure in MB selectivity could explain unsuccessful bacterial reduction following PDT. Further research involving other photosensitizers or conjugating molecules to MB to target microbial cells is needed for PDT application in poultry breeders.
Plasma proteome responses in zebrafish following λ-carrageenan-Induced inflammation are mediated by PMN leukocytes and correlate highly with their human counterparts
2022 - CHARLIE-SILVA, IVES; FEITOSA, NATALIA M.; PONTES, LETICIA G.; FERNANDES, BIANCA H.; NOBREGA, RAFAEL H.; GOMES, JULIANA M.M.; PRATA, MARIANA N.L.; FERRARIS, FAUSTO K.; MELO, DANIELA C.; CONDE, GABRIEL; RODRIGUES, LETICIA F.; ARACATI, MAYUMI F.; CORREA-JUNIOR, JOSE D.; MANRIQUE, WILSON G.; SUPERIO, JOSHUA; GARCEZ, AGUINALDO S.; CONCEICAO, KATIA; YOSHIMURA, TANIA M.; NUNEZ, SILVIA C.; ETO, SILAS F.; FERNANDES, DAYANNE C.; FREITAS, ANDERSON Z.; RIBEIRO, MARTHA S.; NEDOLUZHKO, ARTEM; LOPES-FERREIRA, MONICA; BORRA, RICARDO C.; BARCELLOS, LEONARDO J.G.; PEREZ, ANDREA C.; MALAFAIA, GUILHEME; CUNHA, THIAGO M.; BELO, MARCO A.A.; GALINDO-VILLEGAS, JORGE
Regulation of inflammation is a critical process for maintaining physiological homeostasis. The λ-carrageenan (λ-CGN) is a mucopolysaccharide extracted from the cell wall of red algae (Chondrus crispus) capable of inducing acute intestinal inflammation, which is translated into the production of acute phase reactants secreted into the blood circulation. However, the associated mechanisms in vertebrates are not well understood. Here, we investigated the crucial factors behind the inflammatory milieu of λ-CGN-mediated inflammation administered at 0, 1.75, and 3.5% (v/w) by i.p. injection into the peritoneal cavity of adult zebrafish (ZF) (Danio rerio). We found that polymorphonuclear leukocytes (neutrophils) and lymphocytes infiltrating the ZF peritoneal cavity had short-term persistence. Nevertheless, they generate a strong pattern of inflammation that affects systemically and is enough to produce edema in the cavity. Consistent with these findings, cell infiltration, which causes notable tissue changes, resulted in the overexpression of several acute inflammatory markers at the protein level. Using reversed-phase high-performance liquid chromatography followed by a hybrid linear ion-trap mass spectrometry shotgun proteomic approach, we identified 2938 plasma proteins among the animals injected with PBS and 3.5% λ-CGN. First, the bioinformatic analysis revealed the composition of the plasma proteome. Interestingly, 72 commonly expressed proteins were recorded among the treated and control groups, but, surprisingly, 2830 novel proteins were differentially expressed exclusively in the λ-CGN-induced group. Furthermore, from the commonly expressed proteins, compared to the control group 62 proteins got a significant (p < 0.05) upregulation in the λ-CGN-treated group, while the remaining ten proteins were downregulated. Next, we obtained the major protein-protein interaction networks between hub protein clusters in the blood plasma of the λ-CGN induced group. Moreover, to understand the molecular underpinnings of these effects based on the unveiled protein sets, we performed a bioinformatic structural similarity analysis and generated overlapping 3D reconstructions between ZF and humans during acute inflammation. Biological pathway analysis pointed to the activation and abundance of diverse classical immune and acute phase reactants, several catalytic enzymes, and varied proteins supporting the immune response. Together, this information can be used for testing and finding novel pharmacological targets to treat human intestinal inflammatory diseases.
Dosimetria e modo de aplicação dos lasers de baixa potência
2021 - RIBEIRO, MARTHA S.; SILVA, CAMILA R.; YOSHIMURA, TANIA M.
Low power light triggers opposite effects on stem cells
2021 - YOSHIMURA, TANIA M.; SAUTER, ISMAEL P.; RIBEIRO, MARTHA S.
INTRODUCTION Photobiomodulation (PBM) has been gaining importance in a wide range of medical fields in the past few years, particularly in stem cell-based regenerative medicine. Improving in vitro cell proliferation, differentiation and viability are ways where PBM could play a pivotal role optimizing biotechnological and bioengineering applications. OBJECTIVES Here we investigated whether different wavelengths (blue, green and red) would promote distinct outcomes in human adipose-derived stem cells (hADSCs) cultured in regular and supplemented media for tenocyte differentiation. MATERIALS AND METHODS Freshly isolated hADSCs were cultured in a specific stem cell medium (MSCGM, Lonza), DMEM or a tenogenic medium (TEN-M: DMEM supplemented with growth factors and ascorbic acid). Cells were irradiated every 48 h (23.28 mW/cm 2 , 17 min 10 s delivering 24 J/cm 2 per session) using a LED irradiator (LEDbox, BioLambda). MTT and crystal violet assays were used to evaluate cell metabolic activity and proliferation. DISCUSSION AND RESULTS Red wavelength (660 nm) significantly increased metabolic activity after five irradiations, but only for cells cultured in TEN-M. Oppositely, blue (450 nm) and green (520 nm) light decreased both cell proliferation and metabolic rate, with more pronounced effects for blue light in TEN-M. Considering these findings, we examined whether irradiating only the media would generate toxic compounds that could impair cell viability. We therefore assessed reactive oxygen species (ROS) production by p-nitrosodimethylaniline/histidine assay while irradiating the three different media under the same conditions as mentioned above. Immediately after blue and green light exposure, an increment in ROS production was observed for DMEM and TEN-M, that continuously increased until reaching between 4.5 and 7.1 μM one-hour after irradiation – with higher values for TEN-M exposed to blue light. CONCLUSION Since no significant ROS formation was observed following red light exposure, we concluded that medium composition was responsible for the different effects on metabolic activity and proliferation observed after irradiation with different wavelengths.
Low power light triggers opposite effects on stem cells
2021 - YOSHIMURA, TANIA M.; SAUTER, ISMAEL P.; RIBEIRO, MARTHA S.
Photobiomodulation (PBM) has been gaining importance in a wide range of medical fields in the past few years, particularly in stem cell-based regenerative medicine. Improving in vitro cell proliferation, differentiation and viability are ways where PBM could play a pivotal role optimizing biotechnological and bioengineering applications. Here we investigated whether different wavelengths (blue, green and red) would promote distinct outcomes in human adipose-derived stem cells (hADSCs) cultured in regular and supplemented media for tenocyte differentiation. MATERIALS AND METHODS. Freshly isolated hADSCs were cultured in a specific stem cell medium (MSCGM, Lonza), DMEM or a tenogenic medium (TEN-M: DMEM supplemented with growth factors and ascorbic acid). Cells were irradiated every 48 h (23.28 mW/cm 2, 17 min 10 s delivering 24 J/cm 2 per session) using a LED irradiator (LEDbox, BioLambda). MTT and crystal violet assays were used to evaluate cell metabolic activity and proliferation. Red wavelength (660 nm) significantly increased metabolic activity after five irradiations, but only for cells cultured in TEN-M. Oppositely, blue (450 nm) and green (520 nm) light decreased both cell proliferation and metabolic rate, with more pronounced effects for blue light in TEN-M. Considering these findings, we examined whether irradiating only the media would generate toxic compounds that could impair cell viability. We therefore assessed reactive oxygen species (ROS) production by p-nitrosodimethylaniline/ histidine assay while irradiating the three different media under the same conditions as mentioned above. Immediately after blue and green light exposure, an increment in ROS production was observed for DMEM and TEN-M, that continuously increased until reaching between 4.5 and 7.1 μM one-hour after irradiation – with higher values for TEN-M exposed to blue light. Since no significant ROS formation was observed following red light exposure, we concluded that medium composition was responsible for the different effects on metabolic activity and proliferation observed after irradiation with different wavelengths.
A novel superparamagnetic nanoplatform assisted by light against nonlocal bacterial infections
2020 - RIBEIRO, M.S.; YOSHIMURA, T.M.; TOLEDO, V.H.; HADDAD, P.S.
The increasing resistance to antimicrobials worldwide requires new strategies to fight infectious diseases. Antimicrobial photodynamic therapy (APDT) emerges in this scenario as a promising approach. APDT is a light-based therapy combined to a photosensitizer drug and oxygen that kills microorganisms via oxidative stress. Due to its mechanism of action, i.e., generalized oxidation of all cell structures, bacterial resistance to APDT is very improbable. Currently, however, APDT is limited to local treatments. Thus, a platform to deliver the photosensitizer drug in internal organs is urgently necessary. Superparamagnetic iron oxide nanoparticles (SPIONs) can be easily directed to target sites using an external magnetic field, which made them excellent drug delivery systems. In addition, their large surface to volume ratio allows the functionalization of therapeutic molecules on their surface. In this work, we synthesized and characterized SPIONs covered by methylene blue (MB)-entrapped silica (SPIONs-silica-MB) and investigated their potential combined to APDT to kill Escherichia coli, the most common bacteria found in urinary tract infections. We synthesized SPIONs by the co-precipitation of iron (II) and (III) chlorides in the presence of a weak base and covered them with a double layer of silica leading to the hybrid material magnetite-silica-MB. SPIONS were characterized by Fourier transform infrared spectroscopy, powder X-ray diffraction, and magnetic measurements. Kinetics of MB release and production of singlet oxygen from SPION-silica-MB were also obtained. The average crystallite size of magnetite was found to be 14 nm. Infrared spectra showed characteristic bands of iron-oxygen and others associated with silicate groups. At room temperature, the nanoplatform presented magnetic behavior due to the magnetite core. MB release increased exponentially until 18 h, when it reached a plateau. Singlet oxygen was spontaneously released from SPIONs-silica-MB. To verify the photodynamic activity of SPIONs-silica-MB on bacterial cells, 1x10^7 colony forming units of E. coli were suspended in SPIONs-silica-MB PBS solution (50 microM of MB) and were irradiated using a red LED (625 ± 20 nm) of 235 mW over an orbital shaker to prevent precipitation during 5, 10, 15 and 20 min at three different MB release times: 6, 12 and 22 h. Proper controls were established and showed no killing. In contrast, SPIONs-silica-MB-mediated APDT promoted bacteria inactivation depending on both the time of MB release and irradiation time. Our results show that bacteria are completely eradicated following 22 h of MB release and 20 min of irradiation. These findings motivate the use of SPIONs-silica-MB to mediate APDT against nonlocal infectious diseases.