Publications by authors named "Natalia Mayumi Inada"

36 Publications

Total mouth photodynamic therapy mediated by red LED and porphyrin in individuals with AIDS.

Lasers Med Sci 2021 Aug 13. Epub 2021 Aug 13.

São Carlos Institute of Physics, University of São Paulo, São Carlos, Brazil.

Due to the immune changes resulting from HIV/AIDS infection, systemic and local infections throughout the body are common. The use of high activity antiretroviral therapy has been widely used during treatment, which, added to the use of antibiotics, antifungals, and the patients' own immunocompromised state, cause important changes in the oral microbiota. The emergence of pathological microorganisms and with high resistance to drug therapies are frequent and cause serious damage to the oral health of these patients. In this sense, antimicrobial photodynamic therapy (aPDT) appears as a promising alternative in the control of these oral infections. The aim of the study was to test the effectiveness of a therapeutic protocol for total oral aPDT mediated by a 660-nm red LED (light-emitting diode) associated with porphyrin in individuals with AIDS. Patients were selected by exclusion criteria and randomly distributed into groups to test the effectiveness of antimicrobial aPDT with 50 µg/ml porphyrin associated with the red LED. Before and after the treatments, saliva samples were collected and processed in duplicate in selective culture media. Colonies were counted and the results obtained in Log10 CFU/ml and tested statistically. It was concluded that aPDT was effective in reducing oral enterobacteria, in addition to reducing Streptococcus spp. and general count of microorganisms, when considering the numbers of TCD4 and TCD8 lymphocytes.
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http://dx.doi.org/10.1007/s10103-021-03377-zDOI Listing
August 2021

HPV-induced condylomata acuminata treated by Photodynamic Therapy in comparison with trichloroacetic acid: A randomized clinical trial.

Photodiagnosis Photodyn Ther 2021 Sep 29;35:102465. Epub 2021 Jul 29.

Sao Carlos Institute of Physics, University of Sao Paulo, Sao Carlos, SP, Brazil.

Objectives: This is a randomized controlled clinical trial comparing Photodynamic Therapy (PDT) and the application of trichloracetic acid (TAA) in the treatment of HPV condyloma in the perianal and vulva regions. Design, Randomised controlled, open label, trial. They were allocated to each treatment following randomization by a computer program.

Setting: Women Health Ambulatory in São Carlos city, São Paulo State in the Brazil.

Participants: 36 patients evaluated. 31 patients fulfilled the study requirements.

Intervention: Photodynamic Therapy (PDT) versus trichloracetic acid (TAA). The PDT protocol used the prodrug methyl aminolevulinate incubated for 3 hours and irradiation at 630 nm (100 J/cm²). In the treatment using TAA, warts received a small amount of acid using a cotton swab. Both treatments were repeated weekly until the lesions disappeared completely or until 10 sessions were completed.

Main Outcome Measure: Clinical analysis. Follow-up between 12 and 30 months after the complete treatment.

Results: A total of 16 patients were treated with PDT and 15 patients with TAA. A complete response rate of 60% for TAA and 63% for PDT, with a recurrence rate of 33% for TAA and 0% for PDT.

Conclusion: PDT appears not only to treat lesions due to physical destruction of condyloma and subclinical lesions, but also to modulate the immune system and/or also to decrease the local viral load, suggesting a lower recurrence compared to the TAA group.
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http://dx.doi.org/10.1016/j.pdpdt.2021.102465DOI Listing
September 2021

One-Pot Microwave-Assisted Synthesis of Carbon Dots and and Antimicrobial Photodynamic Applications.

Front Microbiol 2021 21;12:662149. Epub 2021 Jun 21.

São Carlos Institute of Physics, University of São Paulo, São Carlos, Brazil.

Carbon-based photosensitizers are more attractive than the other ones based on their low cost, high stability, broadband of light absorption, tunable emission spectra, high quantum yield, water solubility, high resistance to metabolic degradation, and selective delivery. These properties allow multiple applications in the field of biology and medicine. The present study evaluated and the antimicrobial photodynamic effect of a one-pot microwave produced C-DOTS based on citric acid. The assays assessed the effectiveness of illuminated C-DOTS (C-DOTS + light) against suspension and biofilm. The concentrations of 6.9 and 13.8 mg/mL of C-DOTS and light doses of 20 and 40 J/cm were able to reduce significantly the microorganisms. Based on these parameters and results, the experiments were conducted in mice, evaluating this treatment on wounds contaminated with The viability test showed that C-DOTS-mediated photodynamic inactivation reduced 10 log of the bacteria present on the skin lesions. These results, altogether, showed that antibacterial photodynamic therapy using C-DOTS is a promising and viable treatment for Gram-positive bacteria-infected wounds.
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http://dx.doi.org/10.3389/fmicb.2021.662149DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8255795PMC
June 2021

Follow-up of pressure ulcer treatment with photodynamic therapy, low level laser therapy and cellulose membrane.

J Wound Care 2021 Apr;30(4):304-310

Institute of Physics, USP, São Carlos, SP, Brazil.

Objective: A pressure ulcer (PU) is an area of tissue trauma caused by continuous and prolonged pressure, often associated with hospitalised patients immobilised due to neurological problems, negatively affecting their quality of life, and burdening the public budget. The aim of this study was to report the follow-up, for 45 weeks, of three patients with neurological lesions due to trauma who subsequently developed PUs, and who were treated with a combination of photodynamic therapy (PDT), low level laser therapy (LLLT) and cellulose membrane (CM).

Method: PDT was mediated by the photosensitiser curcumin on a 1.5% emulsion base. Blue LED light at 450 nm was delivered continuously for 12 minutes at an irradiance of 30mW/cm and total energy delivered to the tissue was 22J/cm. LLLT was performed with 660 nm laser, punctuated and continuous, twice a week with parameters: spot size 0.04cm, power of 40mW, 10 seconds per point, fluence of 10J/cm and irradiance of 1000mW/cm.

Results: All PUs had a significant reduction (range: 95.2-100%) of their area after 45 weeks of follow-up and two PUs had complete healing at 20 weeks and 30 weeks. All of the PUs showed a reduction in contamination with the PDT treatments in different proportions.

Conclusion: From the results obtained, we conclude that the combination of PDT, LLLT and CM is a promising treatment for PU healing.
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http://dx.doi.org/10.12968/jowc.2021.30.4.304DOI Listing
April 2021

Strategies to Improve the Antimicrobial Efficacy of Photodynamic, Sonodynamic, and Sonophotodynamic Therapies.

Lasers Surg Med 2021 Oct 28;53(8):1113-1121. Epub 2021 Jan 28.

Optics Group from São Carlos Institute of Physics (IFSC), University of São Paulo (USP), Av. Trabalhador São-carlense, 400-Centro, CEP 13560-970, São Carlos, SP, Brazil.

Background And Objectives: This work evaluated antimicrobial photodynamic therapy (PDT), sonodynamic therapy (SDT), and the association of both therapies (sonophotodynamic therapy [SPDT]), mediated by curcumin (Cur) against Staphylococcus aureus biofilm. Next, additional strategies for these treatments were assessed.

Materials And Methods: S. aureus biofilms received PDT, SDT, and SPDT, mediated by Cur (80 µM), LED light (450 nm), and 1 MHz ultrasound. The same treatments were also performed adding a strategy: Cur with sodium dodecyl sulfate (SDS), Cur with potassium iodide (KI) or a pre-treatment with ultrasound. Cell viability was determined and biofilm architecture was evaluated under confocal microscopy.

Results: SPDT was more effective to inactivate the bacteria than PDT and SDT. SDS achieved the greatest viability reductions, followed by KI and ultrasound pre-treatment. Confocal images revealed biofilm disruption and a reduced number of cells in all treatments. However, SPDT exhibited a pronounced effect and it was greater using SDS.

Conclusion: SPDT was more effective and additional strategies potentiated its effectiveness. Lasers Surg. Med. © 2021 Wiley Periodicals LLC.
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http://dx.doi.org/10.1002/lsm.23383DOI Listing
October 2021

Curcumin/d-mannitol as photolarvicide: induced delay in larval development time, changes in sex ratio and reduced longevity of Aedes aegypti.

Pest Manag Sci 2021 May 21;77(5):2530-2538. Epub 2021 Feb 21.

São Carlos Institute of Physics, University of São Paulo, São Carlos, Brazil.

Background: Resistant populations of Ae. aegypti have been a major problem in arboviruses epidemic areas, generating a strong demand for novel methods of vector control. In this regard, our group has demonstrated the use of curcumin as an efficient photoactive larvicide to eliminate Ae. aegypti larvae. This work was aimed to evaluate the Ae. aegypti (Rockefeller) development under sublethal conditions, using a curcumin/d-mannitol (DMC) formulation. The photolarvicidal efficacy under semi-field and field conditions (wild populations) was also analyzed, as well as the photobleaching and residual activity of DMC.

Results: A delay in development time when larvae were exposed to sublethal concentrations of DMC was observed, followed by significant changes in sex ratio and reduction in longevity. DMC also presented a low residual activity when compared to usual larvicides, and had a substantial photolarvicidal activity against wild populations in field trials, achieving 71.3% mortality after 48 h.

Conclusions: Overall, these findings are of great biological importance for the process of enabling the implementation of DMC as a new product in the control of Ae. aegypti larvae, and contributes to the improvement of new plant-based larvicides. © 2021 Society of Chemical Industry.
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http://dx.doi.org/10.1002/ps.6286DOI Listing
May 2021

MAL-associated methyl nicotinate for topical PDT improvement.

J Photochem Photobiol B 2020 Dec 2;213:112071. Epub 2020 Nov 2.

Sao Carlos Institute of Physics, University of Sao Paulo, Brazil; Hagler Fellow, Texas A&M University, College Station, Texas, USA.

Photosensitization of all tissue in sufficient quantity to generate damage is one of the limiting factors for Photodynamic Therapy (PDT) efficiency. Methyl nicotinate (MN) is a thermogenic and vasodilating substance that facilitates the topical tissue penetration of some compounds. The topical MAL (methyl aminolevulinate) PDT is commonly used as a precursor of protoporphyrin IX (PpIX). This study investigates the safety of topical use in NM, as well as its ability to improve the efficiency of topical PDT. For this, we investigate the cytotoxicity of MN, as well as its actions in increasing cellular metabolism and vasodilation. Besides, its ability to optimize the formation of PpIX in the tissue when associated with MAL cream was investigated, besides assessing the severity of necrosis obtained by treatments. The cytotoxicity of MN was tested for concentrations of 0, 0.1, 0.25, 0.5, 0.75 and 1% in cell culture. For the concentration of 0.5%, the cellular metabolism was evaluated using confocal microscopy to calculate the redox rate. In the Chorioallantoic Membrane Model, vasodilation was evaluated for concentrations of 0.5 and 1% MN during 1 h of incubation. In the animal model, the healthy skin of Wistar rat was used to evaluate the production of PpIX in the tissue and the degree of necrosis obtained by Photodynamic therapy when using NM associated with methyl aminolevulinate. It was observed the non-cytotoxicity in vitro of MN in the concentration used (0.5%) and its ability to increase cellular metabolism. In a chorioallantoic model, the MN vasodilation power was demonstrated for different caliber of vessels. In vivo studies are showing that the incorporation of MN in the MAL cream increases the amount of PpIX produced in the tissue causing a higher effect on the epidermis after PDT. This improvement of the protocol may make the procedure more effective both in the destruction of tumor tissue and in the treatment of deeper cells decreasing possible recurrence, in addition to allowing improvements in the protocol, such as reducing the cream's incubation time.
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http://dx.doi.org/10.1016/j.jphotobiol.2020.112071DOI Listing
December 2020

Tuning the properties of carboxymethylchitosan-based porous membranes for potential application as wound dressing.

Int J Biol Macromol 2021 Jan 28;166:459-470. Epub 2020 Oct 28.

Sao Carlos Institute of Chemistry/University of Sao Paulo, Av. Trabalhador sao-carlense, 400, 13566-590 Sao Carlos, São Paulo, Brazil. Electronic address:

Wound repair is a complex process that calls for strategies to allow a rapid and effective regeneration of injured skin, which has stimulated the research of advanced wound dressings. Herein, highly porous membranes of N,O-carboxymethylchitosan (CMCh), and poly (vinyl alcohol) (PVA) were successfully prepared via a green and facile freeze-drying method of blend solutions containing CMCh/PVA at weight ratio 25/75. Membranes composed only by CMCh were also prepared and genipin was used for crosslinking. Different contents of TiO nanoparticles were incorporated to both type of membranes, which were characterized in terms of morphology, porosity (Φ), swelling capacity (S.C.), mechanical properties, susceptibility to lysozyme degradation and in vitro cytotoxicity toward human fibroblast (HDFn) and keratinocytes (HaCaT) cells. Larger apparent pores were observed in the surface of the genipin-crosslinked CMCh membrane, which resulted in higher porosity (Φ ≈ 76%) and swelling capacity (S.C. ≈ 1720%) as compared to CMCh/PVA membrane (Φ ≈ 68%; S.C. ≈ 1660%). The porosity of both types of membranes decreased upon the addition of TiO nanoparticles while swelling capacity increased. Due to their high porosity and swelling capacity, adequate mechanical properties, controlled degradability, and cytocompatibility, such carboxymethylchitosan-based membranes are potentially useful as wound dressings.
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http://dx.doi.org/10.1016/j.ijbiomac.2020.10.204DOI Listing
January 2021

Evolution of surviving Streptoccocus pyogenes from pharyngotonsillitis patients submit to multiple cycles of antimicrobial photodynamic therapy.

J Photochem Photobiol B 2020 Sep 1;210:111985. Epub 2020 Aug 1.

São Carlos Institute of Physics, University of São Paulo, Box 369, 13566-590, São Carlos, SP, Brazil.

It is estimated over 600 million pharyngotonsillitis (PT) cases worldwide per year and 30% of this total are caused by Streptococcus pyogenes with standard antibiotic treatment. Antimicrobial Photodynamic Therapy (aPDT) has been studied for the clinical research in infectious diseases. The study aim was to analyze the evolution of aPDT on inactivation of clinical strains of multiple cycles. S. pyogenes and clinical strains isolated from patients with PT were incubated with curcumin in formulation (2.25 mg/ml) and irradiated at 450 nm in Light fluence rates. A mortality was a measure of the counting colony forming units per milliliter (CFU/ml) surviving. Parameters of bacterial biofilm formation, uptake of photosensitizer (PS) and efficacy of antibiotics on survival of bacteria of each cycle were tested. The bacteria profile remains unchanged between 10 aPDT cycles was observed. The bacterial colony survival presented a reduction in capacity to form biofilm due adhesion of strains and PS uptake rate. The antibiotic remained efficient after aPDT cycles. Our in vitro results suggested a low-level of development of PDT resistance, however a decrease of photosensitizer uptake was observed. Furthermore, there is no cross effect on aPDT cycles and the first application of antibiotics.
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http://dx.doi.org/10.1016/j.jphotobiol.2020.111985DOI Listing
September 2020

High-risk HPV clearance and CIN 3 treated with MAL-PDT: A case report.

Photodiagnosis Photodyn Ther 2020 Sep 30;31:101937. Epub 2020 Jul 30.

Sao Carlos Institute of Physics, University of Sao Paulo, Brazil. Electronic address:

High-grade cervical intraepithelial neoplasia (CIN) is the precursor to cervical cancer. HPV (human papillomavirus) infection is strongly related with this disease. The CIN treatment is generally excision of the transformation zone (ETZ). Photodynamic therapy (PDT) has also shown to be a promising treatment. We are reporting a case of a 33-years-old patient with high-grade CIN 3 treated with topical MAL (methyl aminolevulinate) PDT. Was applied 2.5 g of 20 % (w/w) MAL cream overnight and the cervix was illuminated twice, with three weeks apart, using a probe with LEDs simultaneously with a cylindrical laser fiber emitting both at 630 nm, with a fluency of 150 J/cm. CIN 3 and the presence of high-risk HPV virus was eliminated 120 days after the second procedure. There was no recurrence at 6 months follow-up. This case report using MAL-PDT and a different light arrangement with LEDs and laser fiber associated both cured the patient with CIN 3 and eliminated low and high-risk HPV in just two PDT sessions.
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http://dx.doi.org/10.1016/j.pdpdt.2020.101937DOI Listing
September 2020

Curcumin in formulations against Aedes aegypti: Mode of action, photolarvicidal and ovicidal activity.

Photodiagnosis Photodyn Ther 2020 Sep 30;31:101840. Epub 2020 May 30.

São Carlos Institute of Physics, University of São Paulo, São Carlos 13560-970, São Paulo, Brazil.

Combating the Aedes aegypti vector is still the key to control the transmission of many arboviruses, such as Dengue, Zika, and Chikungunya. As few products are efficient for Aedes aegypti control, the search for new strategies have become pivotal., t Substances with photodynamic activity, such as curcumin and their formulations, are strongly encouraged, due to their multi-target mechanism of action. In this study, we evaluated the photolarvicidal and ovicidal activity of curcumin in the presence of sucrose (named SC) and d-mannitol (named DMC). To support the understanding of the larvicidal action of these formulations, Raman micro-spectroscopy was employed. We also studied the morphological changes in Danio rerio (Zebrafish) gills, a non-target organism, and demonstrate that this is an environmentally friendly approach. Both SC and DMC presented a high photo-larvicidal potential. DMC showed the highest larval mortality, with LC values between 0.01 and 0.02 mg.L. DMC also significantly decreased egg hatchability, reaching a hatching rate of 10 % at 100 mg.L. The analysis of molecular mechanisms via Raman micro-spectroscopy showed that DMC is highly permeable to the peritrophic membrane of the larva, causing irreversible damage to the simple columnar epithelium of the digestive tube. Histological changes found in the D. rerio gills were of minimal or moderate pathological importance, indicating an adaptive trait rather than detrimental characteristics. These findings indicate that curcumin in sugar formulations is highly efficient, especially DMC, proving it to be a promising and safe alternative to control Aedes mosquitoes. Moreover, Raman micro-spectroscopy demonstrated high potential as an analytical technique to understand the mechanism of action of larvicides.
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http://dx.doi.org/10.1016/j.pdpdt.2020.101840DOI Listing
September 2020

Environmental safety and mode of action of a novel curcumin-based photolarvicide.

Environ Sci Pollut Res Int 2020 Aug 20;27(23):29204-29217. Epub 2020 May 20.

Optical Group, São Carlos Institute of Physics, University of São Paulo, Sao Carlos, São Paulo, 13560-970, Brazil.

Aedes aegypti is the vector of important diseases like dengue, zika, chikungunya, and yellow fever. Vector control is pivotal in combating the spread of these mosquito-borne illnesses. Photoactivable larvicide curcumin obtained from Curcuma longa Linnaeus has shown high potential for Ae. aegypti larvae control. However, the toxicity of this photosensitizer (PS) might jeopardize non-target aquatic organisms. The aim of this study was to evaluate the toxicity of this PS to Daphnia magna and Danio rerio, besides assessing its mode of action through larvae biochemical and histological studies. Three PS formulations were tested: PS in ethanol+DMSO, PS in sucrose, and PS in D-mannitol. The LC of PS in ethanol+DMSO to D. rerio was 5.9 mg L, while in D. magna the solvents were extremely toxic, and LC was not estimated. The PS formulations in sugars were not toxic to neither of the organisms. Reactive oxygen species (ROS) were generated in D. magna exposed to 50 mg L of PS in D-mannitol, and D. rerio did not elicit this kind of response. D. magna feeding rates were not affected by the PS in D-mannitol. Concerning Ae. aegypti larvae, there were changes in reduced glutathione and protein levels, while catalase activity remained stable after exposure to PS in D-mannitol and sunlight. Histological changes were observed in larvae exposed to PS in sucrose and D-mannitol, most of them irreversible and deleterious. Our results show the feasibility of this photolarvicide use in Ae. aegypti larvae control and its safety to non-target organisms. These data are crucial to this original vector control approach implementation in public health policies.
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http://dx.doi.org/10.1007/s11356-020-09210-4DOI Listing
August 2020

HPV condylomatosis region treated with multiple sessions of MAL-PDT: A case report.

Photodiagnosis Photodyn Ther 2020 Sep 7;31:101812. Epub 2020 May 7.

Sao Carlos Institute of Physics, University of Sao Paulo, Brazil. Electronic address:

Condyloma or genital warts affect the tissues of the genital area due to infections induced by Human papillomavirus. There are some topical therapies indicated for condyloma lesions treatment but all of them present a high rate of recurrence. Photodynamic Therapy is proving to be an interesting strategy for treating these lesions, capable also to treat virus-infected cells on subclinical lesions. A patient with a significative condylomatosis region was treated with six MAL-PDT sessions. Due to its large thickness, other topical procedures probably would have low effectiveness. After the treatment, the tissue presented clinical absence of lesion and no apparent scar, with good aesthetic result and preserving the sensitivity of the region and a follow up of eleven months without recurrence.
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http://dx.doi.org/10.1016/j.pdpdt.2020.101812DOI Listing
September 2020

Total mouth photodynamic therapy mediated by blue led and curcumin in individuals with AIDS.

Expert Rev Anti Infect Ther 2020 07 26;18(7):689-696. Epub 2020 Apr 26.

São Carlos Institute of Physics, University of São Paulo , São Carlos, Brazil.

Objectives: To test the effectiveness of an efficient therapeutic protocol for the total mouth antimicrobial photodynamic therapy (aPDT) mediated by 450 nm blue LED associated with curcumin in individuals with AIDS.

Methods: Patients were selected by exclusion criteria and randomly distributed in groups to test the effectiveness of antimicrobial aPDT with curcumin 0.75 mg/mL associated with the blue LED (67 mW/cm, 20.1 J/cm). Before and after the treatments, samples were collected from the saliva being processed in duplicate in selective culture media. The colonies were counted and the results obtained in log10 CFU/mL were statistically tested (T-paired statistical test, 5%).

Results: The log10 CFU/mL of spp., spp., and total count of microorganisms showed statistically significant (p = 0.023; p = 0.001 and p = 0.017, respectively) reduction after treatment in patients with aPDT.

Conclusion: aPDT was effective in reducing spp. in addition to reducing spp., enterobacteria and the total count of microorganisms when considering the numbers of TCD4 and TCD8 lymphocytes. The aPDT in the studied protocol was able to control clinically important intraoral microorganisms for AIDS patients, both those with TCD4 lymphocytes above or below 25% of normal and those with TCD8 lymphocytes above 25% of normal.
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http://dx.doi.org/10.1080/14787210.2020.1756774DOI Listing
July 2020

Graphene Oxide Mediated Broad-Spectrum Antibacterial Based on Bimodal Action of Photodynamic and Photothermal Effects.

Front Microbiol 2019 15;10:2995. Epub 2020 Jan 15.

São Carlos Institute of Physics, University of São Paulo, São Carlos, Brazil.

Graphene oxide (GO) with their interesting properties including thermal and electrical conductivity and antibacterial characteristics have many promising applications in medicine. The prevalence of resistant bacteria is considered a public health problem worldwide, herein, GO has been used as a broad spectrum selective antibacterial agent based on the photothermal therapy (PTT)/photodynamic therapy (PDT) effect. The preparation, characterization, determination of photophysical properties of two different sizes of GO is described. light dose and concentration-dependent studies were performed using Gram-negative and Gram-positive bacteria based on the PTT/PDT effect used ultra-low doses (65 mW cm) of 630 nm light, to achieve efficient bacterial decontamination. The results show that GO and nanographene oxide (nGO) can sensitize the formation of O and allow a temperature rise of 55°C to 60°C together nGO and GO to exert combined PTT/PDT effect in the disinfection of gram-positive and gram-negative bacteria. A complete elimination of and bacteria based on GO and nGO is obtained by using a dose of 43-47 J cm for high concentration used in this study, and a dose of around 70 J cm for low dose of GO and nGO. The presence of high concentrations of GO allows the bacterial population of and to be more sensitive to the use of PDT/PTT and the efficiency of and bacteria disinfection in the presence of GO is similar to that of nGO. In human neonatal dermal fibroblast, HDFs, no significant alteration to cell viability was promoted by GO, but in nGO is observed a mild damage in the HDFs cells independent of nGO concentration and light exposure. The unique properties of GO and nGO may be useful for the clinical treatment of disinfection of broad-spectrum antimicrobials. The antibacterial results of PTT and PDT using GO in gram-positive and gram-negative bacteria, using low dose light, allow us to conclude that GO and nGO can be used in dermatologic infections, since the effect on human dermal fibroblasts of this treatment is low compared to the antibacterial effect.
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http://dx.doi.org/10.3389/fmicb.2019.02995DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6974586PMC
January 2020

Optimization for microbial incorporation and efficiency of photodynamic therapy using variation on curcumin formulation.

Photodiagnosis Photodyn Ther 2020 Mar 7;29:101652. Epub 2020 Jan 7.

São Carlos Institute of Physics, University of São Paulo, Box 369, 13566-970, São Carlos, SP, Brazil.

Introduction: A mixture of curcuminoids: curcumin, desmethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC) are named natural curcumin. It is a lipophilic photosensitizer (PS) highly soluble in an organic solvent such as dimethyl sulfoxide (DMSO). Curcumin is a PS used for microbial inactivation using photodynamic action. However, this solvent has high cytotoxicity and is unavailable in formulations for clinical use. This study aimed to investigate the interactions of curcuminoids syrup with Streptococcus sp., a gram-positive coccus and one of the major pharyngeal pathogens, responsible for diseases such as pharyngitis.

Methods: Bacteria were incubated with curcuminoids (natural curcumin, synthetic, DMC, BDMC) at 37 °C in formulations: 1) syrup (water + sucrose) 2) solution alcohol + DMSO. Was centrifuged, and the supernatant collected for absorbance analysis. The results obtained correlating the absorbance with the supernatant to the absorbance of the default concentration. A study of microbial metabolism by growth curve was carried out to justify the result.

Results: The incorporation of curcumin in syrup is superior to alcohol/DMSO solution by microorganisms. Curcumin incorporation by S. mutans, S. pyogenes, isolated bacteria was 24, 26, 27 % in syrup and 10, 13, 5 % in alcohol/DMSO, respectively. Also, the presence of carbohydrate in a solution can activate the bacterial metabolism, getting better uptake results and photodynamic inactivation to natural curcumin and DMC. Such finds care optimizes the use of curcumin without complications generated by the solvent.
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http://dx.doi.org/10.1016/j.pdpdt.2020.101652DOI Listing
March 2020

Luminescent nanohybrids based on silica and silylated Ru(II)-Yb(III) heterobinuclear complex: new tools for biological media analysis.

Nanotechnology 2019 Nov 8;31(8):085709. Epub 2019 Nov 8.

Institute of Chemistry, São Paulo State University, UNESP, CP355-Araraquara-SP, Brazil. CIRIMAT Université de Toulouse, CNRS, INPT, UPS, Toulouse, France. 118 route de Narbonne, F-31062, Toulouse Cedex 9, France.

Lanthanide (Ln) complexes emitting in the near-infrared (NIR) region have fostered great interest as upcoming optical tags owing to their high spatial and temporal resolution emission as well deeper light penetration in biological tissues for non-invasive monitoring. For use in live-cell imaging, lanthanide complexes with long-wavelength absorption and good brightness are especially critical. Light-harvesting ligands of Ln complexes are typically excited in the ultraviolet region, which in turn trigger simultaneously autofluorescence and long-exposition damage of living systems. The association of d-metalloligands rather than organic chromophores enables the excitation of NIR-emitting Ln complex occurs in the visible region. Taking advantage of the long-lived excited states and intense absorption band in the ultraviolet (UV) to NIR region of Ru(II), we successfully design a dual-emitting (in the visible and NIR region) d-f heterobinuclear complex based on Ru(II) metalloligand and Yb(III) complex. In addition, we developed luminescent nanohybrids by grafting of Ru(II)-Yb(III) heterobinuclear complexes containing silylated ligands on the surface of mesoporous and dense silica matrix. The nanomarkers were successfully applied for imaging of murine melanoma B16-F10 and neonatal human dermal fibroblast HDFn cell cultures by one-photon or two-photon absorption using laser scanning confocal microscopy. Great cellular uptake, low cytotoxicity and the possibility to achieve visible and NIR emission via two-photons excitation show that the nanohybrids are remarkable markers for in vitro and a potential tool for in vivo applications.
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http://dx.doi.org/10.1088/1361-6528/ab55c3DOI Listing
November 2019

Photodynamic inactivation for in vitro decontamination of Staphylococcus aureus in whole blood.

Photodiagnosis Photodyn Ther 2019 Dec 11;28:58-64. Epub 2019 Aug 11.

São Carlos Institute of Physics, University of São Paulo, PO Box 369, 13560-970, São Carlos, São Paulo, Brazil.

Background: Blood can be the target of microbial cells in the human body. Erythrocytes, platelets, and plasma concentrates in blood bags used in hemotherapy for blood transfusion are contamination targets, which can trigger serious diseases in blood. These infections can cause septicemia that can lead to death if not recognized rapidly and treated adequately. The aim of this study was to evaluate the photodynamic inactivation in the in vitro decontamination of Staphylococcus aureus in whole blood, erythrocytes and platelet-rich plasma.

Methods: Photodynamic inactivation using light doses of 10, 15 and 30 J/cm at 630 nm and an hematoporphyrin-derivative photosensitizer (Photogem®) solutions at 25 and 50 μg/mL were evaluated. Toxicity of treatment was determined by hemolysis and cell viability assays.

Results: The S. aureus reduction in phosphate buffered saline (PBS), whole blood, erythrocytes and platelet-rich plasma at 15 J/cm and 50 μg/mL were 7.2, 1.0, 1.3 and 0.4 log CFU/mL, respectively. Quantitative and qualitative analyses were performed in whole blood samples, and Photogem® showed a low risk of hemolysis (10.7%) in whole blood. However, 100% of erythrocytes suffered hemolysis in the absence of plasma. The cell viability assay showed 13.9% of apoptosis in erythrocytes, but normal platelet viability.

Conclusion: S. aureus inactivation of whole blood samples using 50 μg/mL Photogem® and 15 J/cm resulted in better outcomes, providing promising indications for treatment of bacterial contamination of blood, and in this work, alternative possibilities to apply the technique for blood decontamination are discussed.
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http://dx.doi.org/10.1016/j.pdpdt.2019.08.013DOI Listing
December 2019

Long Term Effectiveness of Photodynamic Therapy for CIN Treatment.

Pharmaceuticals (Basel) 2019 Jul 12;12(3). Epub 2019 Jul 12.

São Carlos Institute of Physics, University of Sao Paulo, São Carlos 13566-590, Brazil.

(1) Background: Cervical cancer is the third most commonly diagnosed cancer and the fourth leading cause of cancer death in women worldwide. The highest incidence rates are in Africa, followed by South-Central Asia and South America. According to the Brazilian National Institute of Cancer (INCA), 16,370 new cases of cervical cancer were estimated for each year of the biennium of 2018-2019. About 90% of cervical cancers originate from the malignant progression of cervical intraepithelial neoplasia (CIN) which is classified based on cytohistological characteristics (low- and high-grade lesions). The present study reports the long-term effectiveness of topical photodynamic therapy (PDT) for CIN grades 1 and 2/3 with up to two years of follow up. (2) Methods: A total of 56 patients with CIN 1, ten with CIN 2, and 14 patients for the placebo group were enrolled in this study. (3) Results: 75% ( = 42) of CIN 1 patients presented a complete response to PDT and only 23.2% ( = 13) of recurrence, progression, and/or lesions remaining two years after PDT. For CIN 2/3 patients, 90% were observed to be cured after one and two years of follow up. (4) Conclusions: PDT presented best results two years after a non-invasive, fast, and low-cost procedure and in comparison with the placebo group, preventing the progression of cervical intraepithelial neoplasia and preserving the cervix.
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http://dx.doi.org/10.3390/ph12030107DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6789515PMC
July 2019

Overall Results for a National Program of Photodynamic Therapy for Basal Cell Carcinoma: A Multicenter Clinical Study to Bring New Techniques to Social Health Care.

Cancer Control 2019 Jan-Dec;26(1):1073274819856885

1 São Carlos Institute of Physics, University of São Paulo (USP), São Carlos, São Paulo, Brazil.

Along the past years, a national program to implement photodynamic therapy (PDT) for nonmelanoma skin cancer (NMSC) was performed over the Brazilian territory. Using a strategy involving companies, national bank, and medical partners, equipment, medication, and protocols were tested in a multicenter study. With results collected over 6 years, we could reach a great deal of advances concerning the use of PDT for skin cancer. We present the overall reached results of the program and discuss several aspects about it, including public politics of treatment. A discussion about advantages of this technique within conditions of health care is placed, comparing PDT with surgery, including an analysis about the implementation of PDT in countries in development as Brazil, considering not only technical but social aspects, as the distribution of medical doctor in the Brazilian territory. The program resulted in a huge dissemination of PDT in Brazil and many countries in Latin America, in a partnership among public politics, universities, companies, and hospitals and clinics and in the insertion of national technologies as option to treat NMSC. Consequence of the program is mainly the continuation of the use of PDT in Brazil and many countries in Latin America.
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http://dx.doi.org/10.1177/1073274819856885DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6598334PMC
December 2019

Single visit PDT for basal cell carcinoma - A new therapeutic protocol.

Photodiagnosis Photodyn Ther 2019 Jun 16;26:375-382. Epub 2019 Apr 16.

Skin Department, Foundation Hospital Amaral Carvalho, Jau, São Paulo, Brazil. Electronic address:

Non-melanoma skin cancer is the most prevalent type of cancer in Brazil and worldwide. Topical Photodynamic Therapy is a technique that offers advantages as: excellent aesthetic result, possibility of application for outpatients in ambulatory setting, and presenting a minimum functional impact of the treated anatomic site. Fractionated Photodynamic Therapy is a modification of the usual technique in which the full dose of light is delivered in steps separated by a periods of time ("dark intervals"). In Brazil, no studies using this technique for treatment of BCC have been published. Thus, we proposed to evaluate the complete and partial response to the four different protocols of fractional Photodynamic Therapy, when evaluated 30 days after treatment. The study showed a complete response of 65.8%, 67.6%, 72.7% and 95.4% in the groups 1, 2, 3 and 4, respectively. We observed that the dark interval and the irradiated light dose are parameters of great importance for the final response to the treatment. Our results suggest that Fractionated Photodynamic Therapy is a technique with excellent aesthetic result and complete response when evaluated 30 days after treatment. However, a longer follow-up will be necessary for better understanding of the behavior of the lesions treated.
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http://dx.doi.org/10.1016/j.pdpdt.2019.04.016DOI Listing
June 2019

Raman Microspectroscopy as a Tool to Elucidate the Efficacy of Topical Formulations Containing Curcumin.

Pharmaceuticals (Basel) 2019 Mar 23;12(1). Epub 2019 Mar 23.

São Carlos Institute of Physics, University of São Paulo, Trabalhador Sao-carlense Av., 400, São Carlos, SP 13566-590, Brazil.

The success of the onychomycosis treatment is directly associated with factors such as the choice of the medication, the administration route, and the pharmaceutical formulation. Photodynamic therapy (PDT) is an emerging and promising technique indicated for onychomycosis treatment. For this application, the main challenge is the efficient delivery of the photosensitizer (PS). Curcumin is widely used as a PS, however it is an unstable molecule and it is a challenge to develop a formulation with good penetration into the nail plate, maintaining the stability of curcumin. In this study, the molecular mechanisms underlying the efficacy of two topical formulations containing curcumin used in a clinical trial for onychomycosis treatment were analyzed by Raman microspectroscopy. It is shown that curcumin is present in both formulations in aggregated and non-aggregated states, and in aggregates it is present in different conformations, depending on the interaction with the solvent. This proves to be critical for efficient and uniform PS delivery to the nail and its complete use during the treatment. These analyses are showing how promising Raman microspectroscopy is in understanding the molecular mechanisms of the efficiency of photosensitizers and are helping to improve the development of pharmaceutical formulations.
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http://dx.doi.org/10.3390/ph12010044DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6469165PMC
March 2019

The use of light-emitting diode imaging as exclusion criterion for melanoma diagnosis.

J Am Acad Dermatol 2019 03 3;80(3):e49-e50. Epub 2018 Aug 3.

São Carlos Institute of Physics, University of São Paulo, São Carlos, Brazil.

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http://dx.doi.org/10.1016/j.jaad.2018.07.031DOI Listing
March 2019

Optical techniques for the diagnosis and treatment of lesions induced by the human papillomavirus - A resource letter.

Photodiagnosis Photodyn Ther 2018 Sep 11;23:106-110. Epub 2018 Apr 11.

Federal Medical Center, Butner North Carolina, 27509, USA.

Human papillomaviruses (HPV) are the most common sexually-transmitted virus, and carcinogenic HPV strains are reported to be responsible for virtually all cases of cervical cancer and its precursor, the cervical intraepithelial neoplasia (CIN). About 30% of the sexually active population are considered to be affected by HPV. Around 600 million people are estimated to be infected worldwide. Diseases related to HPV cause significant impact from both the personal welfare point of view and public healthcare perspective. This resource letter collects relevant information regarding HPV-induced lesions and discusses both diagnosis and treatment, with particular attention to optical techniques and the challenges involved to the implementation of those approaches.
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http://dx.doi.org/10.1016/j.pdpdt.2018.04.004DOI Listing
September 2018

A threshold dose distribution approach for the study of PDT resistance development: A threshold distribution approach for the study of PDT resistance.

J Photochem Photobiol B 2018 May 30;182:85-91. Epub 2018 Mar 30.

São Carlos Institute of Physics, University of Sao Paulo, Av. Trabalhador São-carlense, 400, São Carlos, SP 13566-590, Brazil.

Photodynamic therapy (PDT) is a technique with well-established principles that often demands repeated applications for sequential elimination of tumor cells. An important question concerns the way surviving cells from a treatment behave in the subsequent one. Threshold dose is a core concept in PDT dosimetry, as the minimum amount of energy to be delivered for cell destruction via PDT. Concepts of threshold distribution have shown to be an important tool for PDT results analysis in vitro. In this study, we used some of these concepts for demonstrating subsequent treatments with partial elimination of cells modify the distribution, which represents an increased resistance of the cells to the photodynamic action. HepG2 and HepaRG were used as models of tumor and normal liver cells and a protocol to induce resistance, consisted of repeated PDT sessions using Photogem® as a photosensitizer, was applied to the tumor ones. The response of these cells to PDT was assessed using a standard viability assay and the dose response curves were used for deriving the threshold distributions. The changes in the distribution revealed that the resistance protocol effectively eliminated the most sensitive cells. Nevertheless, HepaRG cell line was the most resistant one among the cells analyzed, which indicates a specificity in clinical applications that enables the use of high doses and drug concentrations with minimal damage to the surrounding normal tissue.
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http://dx.doi.org/10.1016/j.jphotobiol.2018.03.022DOI Listing
May 2018

Nanostructured electrospun nonwovens of poly(ε-caprolactone)/quaternized chitosan for potential biomedical applications.

Carbohydr Polym 2018 Apr 31;186:110-121. Epub 2018 Jan 31.

Sao Carlos Institute of Chemistry/University of Sao Paulo, Av. Trabalhador sao-carlense, 400-13566-590, Sao Carlos, São Paulo, Brazil. Electronic address:

Blend solutions of poly(ε-caprolactone) (PCL) and N-(2-hydroxy)-propyl-3-trimethylammonium chitosan chloride (QCh) were successfully electrospun. The weight ratio PCL/QCh ranged in the interval 95/5-70/30 while two QCh samples were used, namely QCh1 (DQ¯ = 47.3%; DPv¯ = 2218) and QCh2 (DQ¯ = 71.1%; DPv¯ = 1427). According to the characteristics of QCh derivative and to the QCh content on the resulting PCL/QCh nonwoven, the nanofibers displayed different average diameter (175 nm-415 nm), and the nonwovens exhibited variable porosity (57.0%-81.6%), swelling capacity (175%-425%) and water vapor transmission rate (1600 g m 24 h-2500 g m 24 h). The surface hydrophilicity of nonwovens increases with increasing QCh content, favoring fibroblast (HDFn) adhesion and spreading. Tensile tests revealed that the nonwovens present a good balance between elasticity and strength under both dry and hydrated state. Results indicate that the PCL/QCh electrospun nonwovens are new nanofibers-based biomaterials potentially useful as wound dressings.
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http://dx.doi.org/10.1016/j.carbpol.2018.01.045DOI Listing
April 2018

Manual Operated Ultraviolet Surface Decontamination for Healthcare Environments.

Photomed Laser Surg 2017 Dec 12;35(12):666-671. Epub 2017 Oct 12.

2 Department of Physics and Material Science, São Carlos Institute of Physics, University of São Paulo , São Carlos, Brazil .

Objective: The aim of this study was to evaluate the effectiveness of a new handheld equipment based on a mercury low-pressure vapor lamp. The Surface UV device was tested in Staphylococcus aureus, Streptococcus mutans, Streptococcus pneumoniae, two strains of Escherichia coli, Pseudomonas aeruginosa, Candida albicans, and other clinical microorganisms isolated from different surfaces of a public health hospital.

Background Data: The incidence of hospital infections has increased in recent years. Despite the variety of available chemicals to reduce the microorganisms, the search for antimicrobial agents and the characterization of novel targets are a continued need. Also, the minimization of chemical procedures is a constant need, and the use of ultraviolet (UV) light as a germicidal device for microorganisms' inactivation has been an alternative and one possible approach for the reduction of contamination.

Materials And Methods: The in vitro decontamination was performed by application of Surface UV in different species of microorganisms (study 1). The surface decontamination was carried out by application of Surface UV on each surface of hospital environment (study 2). The device presents ultraviolet C (UV-C) light at 254 nm and produces an irradiance of 13 mW/cm at a distance of 1 cm of the surfaces. The light dose was 0.78 J/cm for 60 sec of application in both studies.

Results: The results for in vitro decontamination indicated a log reduction factor of 6.5 for S. aureus, 6.7 for S. mutans, 6.2 for S. pneumoniae, 5.4 for E. coli, 5.2 for E. coli (ATCC 8739), 5.4 for P. aeruginosa, and 6.7 for C. albicans. The hospital level of microorganisms decreases more by 75% after the procedure.

Conclusions: The study highlights the development and successful application of a new portable device that can reduce the risk of contamination in health settings. Our results suggest that Surface UV is efficient and may be an alternative decontamination method.
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http://dx.doi.org/10.1089/pho.2017.4298DOI Listing
December 2017

Determination of the threshold dose distribution in photodynamic action from in vitro experiments.

J Photochem Photobiol B 2016 Sep 23;162:168-175. Epub 2016 Jun 23.

São Carlos Institute of Physics, University of São Paulo, PO Box 369, 13560-970, São Carlos, SP, Brazil.

The concept of threshold in photodynamic action on cells or microorganisms is well observed in experiments but not fully explored on in vitro experiments. The intercomparison between light and used photosensitizer among many experiments is also poorly evaluated. In this report, we present an analytical model that allows extracting from the survival rate experiments the data of the threshold dose distribution, ie, the distribution of energies and photosensitizer concentration necessary to produce death of cells. Then, we use this model to investigate photodynamic therapy (PDT) data previously published in literature. The concept of threshold dose distribution instead of "single value of threshold" is a rich concept for the comparison of photodynamic action in different situations, allowing analyses of its efficiency as well as determination of optimized conditions for PDT. We observed that, in general, as it becomes more difficult to kill a population, the distribution tends to broaden, which means it presents a large spectrum of threshold values within the same cell type population. From the distribution parameters (center peak and full width), we also observed a clear distinction among cell types regarding their response to PDT that can be quantified. Comparing data obtained from the same cell line and used photosensitizer (PS), where the only distinct condition was the light source's wavelength, we found that the differences on the distribution parameters were comparable to the differences on the PS absorption. At last, we observed evidence that the threshold dose distribution matches the curve of apoptotic activity for some PSs.
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http://dx.doi.org/10.1016/j.jphotobiol.2016.06.041DOI Listing
September 2016

Sclerodermiform BCC treated with multiple PDT sessions.

Photodiagnosis Photodyn Ther 2016 Jun 4;14:91-2. Epub 2016 Feb 4.

Biophotonics Laboratory, Institute of Physics of São Carlos, University of São Paulo (USP), São Carlos, São Paulo 13566-590, Brazil.

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http://dx.doi.org/10.1016/j.pdpdt.2016.01.010DOI Listing
June 2016

Development and comparison of two devices for treatment of onychomycosis by photodynamic therapy.

J Biomed Opt 2015 Jun;20(6):061109

Onychomycosis is the most common nail disorder. The treatment for this type of infection is one of the main difficult ones in clinical practice, due to the fact that the nails are nonvascularized structures, which compromise the penetration of drugs delivered systemically and favor slow nail growth. We present two devices based on light-emitting diode arrays as light sources for the treatment of onychomycosis by photodynamic therapy (PDT). PDT is an emerging technique that uses a photosensitizer (PS) activated by light in the presence of oxygen. The PS absorbs energy from light and transfers it to oxygen, producing reactive oxygen species such as hydroxyl radicals, superoxide, and singlet oxygen which inactivate fungi and bacteria. Our proposal is the use of a portable and secure light source device in patients with onychomycosis. Additional advantages are the low cost involved, the possibility of topical treatment rather than systemic and the simplicity of operation. These advantages are important to ensure the implementation of this technology for the treatment of an impacting health problem.
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http://dx.doi.org/10.1117/1.JBO.20.6.061109DOI Listing
June 2015
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