Marcos Pinotti

In vitro photodynamic inactivation of Candida spp. growth and adhesion to buccal epithelial cells.

In this study, photodynamic inactivation (PDI) was used to inhibit in vitro growth and adhesion of different Candida isolates to buccal epithelial cells (BEC). Experimental conditions were optimized and 25muM toluidine blue O (TBO) and 15min of irradiation time by light emitting diode (LED) (energy density of 180J/cm(2)) were selected due to higher reductions in cellular viability obtained after treatment. Reduction media of Log(10) 3.41 in viable cellular growth and media of 55% in the inhibition of adhesion to buccal epithelial cells were obtained. Two fluconazole resistant isolates were susceptible to PDI (Log(10) 3.54 in IB05 and Log(10) 1.95 in CG09) and a second session of this treatment for CG09 isolate inhibited cellular viability in 100%, without producing heat. The results permit to conclude that photodynamic inactivation under these experimental conditions would be a possible alternative approach to inhibit Candida spp. cellular growth and adhesion to buccal epithelial cells.

Light Transmission through a Translucent Fiber Post

This study is a quantitative assessment of the luminous energy transmitted through different translucent fiber posts. After embedding the posts in black resin, the blocks were submitted to sequential cuts in a precision machine, and depths of 16 mm, 12 mm, 8 mm, and 4 mm were assessed for light transmission with a digital photometer. The quantitative analysis showed significant differences between different posts and depths. Furthermore, the values obtained revealed that the quantity of luminous energy transmitted depends on the type of post and that for all of them there was a significant reduction of the quantity of light transmitted as the depth increased. Even without the post, the luminous intensity inside the canal seems to decrease to levels that are insufficient for polymerization, especially in the apical third.

Effects of low-power light therapy on wound healing: LASER x LED

Several studies demonstrate the benefits of low-power light therapy on wound healing. However, the use of LED as a therapeutic resource remains controversial. There are questions regarding the equality or not of biological effects promoted by LED and LASER. One objective of this review was to determine the biological effects that support the use of LED on wound healing. Another objective was to identify LED´s parameters for the treatment of wounds. The biological effects and parameters of LED will be compared to those of LASER. Literature was obtained from online databases such as Medline, PubMed, Science Direct and Scielo. The search was restricted to studies published in English and Portuguese from 1992 to 2012. Sixty-eight studies in vitro and in animals were analyzed. LED and LASER promote similar biological effects, such as decrease of inflammatory cells, increased fibroblast proliferation, stimulation of angiogenesis, granulation tissue formation and increased synthesis of collagen. The irradiation parameters are also similar between LED and LASER. The biological effects are dependent on irradiation parameters, mainly wavelength and dose. This review elucidates the importance of defining parameters for the use of light devices.

Photodynamic inhibition of Trichophyton rubrum: in vitro activity and the role of oxidative and nitrosative bursts in fungal death

OBJECTIVES Antimicrobial photodynamic inhibition (aPI) is based on the use of a light source and a photosensitizer to kill pathogens. Little is known about aPI of dermatophytic fungi and its mechanism of action. We aimed to evaluate aPI of Trichophyton rubrum. METHODS We performed tests using toluidine blue (TBO) as a photosensitizer and a 630 nm light-emitting diode (LED) as a source of light to target 12 T. rubrum isolates. Susceptibility testing with cyclopiroxolamine, time-kill curves and quantification of reactive oxygen species (ROS), peroxynitrite (ONOO·) and nitric oxide (NO·) were performed. RESULTS The optimal conditions for in vitro aPI were 10 mg/L for TBO and 48 J/cm(2) for LED; these conditions were fungicidal or inhibited >98% of fungal growth depending on the strain tested. LED or TBO treatment alone did not inhibit growth. The MICs of cyclopiroxolamine were 2.0 mg/L for 90% of the strains. Analysis of time-kill curves revealed that pathogen death occurred 24 h post-treatment. Quantification of ROS, ONOO· and NO· revealed improvement after aPI. CONCLUSIONS Photodynamic inhibition was more efficient in promoting cell death than the antifungal cyclopiroxolamine against T. rubrum. ROS, ONOO· and NO· were important in the fungicidal activity of aPI. A suggested mechanism for this activity is that TBO is excited by LED light (630 nm), reacts with biomolecules and increases the availability of transition electrons and substrates for nitric oxide synthase, thereby increasing the oxidative and nitrosative bursts in the fungal cell.

Hip orthosis powered by pneumatic artificial muscle: voluntary activation in absence of myoelectrical signal.

Powered orthosis is a special class of gait assist device that employs a mechanical or electromechanical actuator to enhance movement of hip, knee, or ankle articulations. Pneumatic artificial muscle (PAM) has been suggested as a pneumatic actuator because its performance is similar to biological muscle. The electromyography (EMG) signal interpretation is the most popular and simplest method to establish the patient voluntary control of the orthosis. However, this technique is not suitable for patients presenting neurological lesions causing absence or very low quality of EMG signal. For those cases, an alternative control strategy should be provided. The aim of the present study is to develop a gait assistance orthosis for lower limb powered by PAMs controlled by a voluntary activation method based on the angular behavior of hip joint. In the present study, an orthosis that has been molded in a patient was employed and, by taking her anthropometric parameters and movement constraints, the adaptation of the existing orthosis to the powered orthosis was planned. A control system was devised allowing voluntary control of a powered orthosis suitable for patients presenting neurological lesions causing absence or very low quality of EMG signal. A pilot clinical study was reported where a patient, victim of poliovirus, successfully tested a hip orthosis especially modified for the gait test evaluation in the parallel bar system. The hip orthosis design and the control circuitry parameters were able to be set to provide satisfactory and comfortable use of the orthosis during the gait cycle.

Investigation of Mast Cells in Human Gingiva Following Low-Intensity Laser Irradiation

OBJECTIVE The aims of the present study were to investigate the effect of low-intensity laser irradiation on the total number of mast cells as well as the percentage of degranulation in human gingiva. Blood vessel dilation was also evaluated. BACKGROUND DATA It has been proposed that low-intensity laser irradiation can ameliorate pain, swelling, and inflammation. In periodontal tissue, mast cells may influence either the destructive events or the defense mechanism against periodontal disease via secretion of cytokines and through cellular migration to improve the healing process. Mast cells play an important role in the inflammatory process. METHODS Twenty patients with gingival enlargement indicated for gingivectomy were selected. Gingival fragments were obtained from each patient and divided into three different groups before surgery. One fragment was removed without any irradiation. The two others were submitted to punctual irradiation with an energy density of 8 J/cm(2) at an output power of 50 mW at 36 Hz for 36 sec before gingivectomy. Nondegranulated and degranulated mast cells were counted in five areas of the gingival fragment connective tissue. Major and minor diameters of the blood vessels were also measured. RESULTS Both red and infrared radiation promoted a significant increase in mast cell degranulation compared to controls; however, no statistically significant differences (p > 0.05) were observed between the irradiated groups. No significant differences among the groups were observed regarding blood vessel size. CONCLUSION The results suggests that red and infrared wavelengths promote mast cell degranulation in human gingival tissue, although no dilation of blood vessels was observed. The effects of premature degranulation of mast cells in human tissue and the laser radiation protocol applied in this study encourage further investigations to extend these results into clinical practice.

Cryptococcus gattii: In Vitro Susceptibility to Photodynamic Inactivation

Cryptococus gattii is an emergent primary human pathogen that causes meningismus, papilledema, high intracranial pressure and focal involvement of the central nervous system in immunocompetent hosts. Prolonged antifungal therapy is the conventional treatment, but it is highly toxic, selects for resistant strains, contributes to therapy failure and has a poor prognosis. Photodynamic inactivation (PDI) offers a promising possibility for the alternative treatment of cryptococcosis. The aim of this study was to test the effectiveness of toluidine blue O (TBO) and light‐emitting diode (LED) against C. gattii strains with distinct susceptibility profile to antifungal drugs (amphotericin B: 0.015–1.0 μg mL−1; itraconazole: 0.015–2 μg mL−1; fluconazole: 4–64 μg mL−1). Using 25 μm (6.76 μg mL−1) TBO and LED energy density of 54 J cm−2 these fungal isolates presented variable susceptibility to PDI. The production of reactive oxygen species (ROS)/peroxynitrite was determined, and the catalase and peroxidase activities were measured. After PDI, high amounts of ROS/peroxynitrite are produced and higher catalase and peroxidase activities could be correlated with a lower susceptibility of C. gattii isolates to PDI. These results indicate that PDI could be an alternative to C. gattii growth inhibition, even of isolates less susceptible to classical antifungal drugs, also pointing to mechanisms related to their variable susceptibility behavior.

An approach on determining the displacements of the pelvic floor during voluntary contraction using numerical simulation and MRI.

The present study was conducted in order to establish a methodology based on the finite element method to simulate the contraction of the pelvic floor (PF) muscles. In the generated finite element model, a downward pressure of 90 cm H2O was applied, while actively contracting the PF muscles with different degrees of muscular activation (10, 50 and 100%). The finite element methodology of the active contraction behaviour proposed in this study is adequate to simulate PF muscle contraction with different degrees of muscular activation. In this case, in particular, for an activation of 100%, the numerical model was able to displace the pubovisceral muscle in a range of values very similar to the displacement found in the magnetic resonance imaging data. In the analysed case study, it would be possible to conclude that an intensity contraction of 50% would be necessary to produce enough stiffness to avoid possible urine loss.

Melanin Protects Paracoccidioides brasiliensis from the Effects of Antimicrobial Photodynamic Inhibition and Antifungal Drugs

ABSTRACT Paracoccidioidomycosis (PCM) is a public health concern in Latin America and South America that when not correctly treated can lead to patient death. In this study, the influence of melanin produced by Paracoccidioides spp. on the effects of treatment with antimicrobial photodynamic inhibition (aPI) and antifungal drugs was evaluated. aPI was performed using toluidine blue (TBO) as a photosensitizer and a 630-nm light-emitting diode (LED) light. The antifungals tested were itraconazole and amphotericin B. We evaluated the effects of each approach, aPI or antifungals, against nonmelanized and melanized yeast cells by performing susceptibility tests and by quantifying oxidative and nitrosative bursts during the experiments. aPI reduced nonmelanized cells by 3.0 log units and melanized cells by 1.3 log units. The results showed that melanization protects the fungal cell, probably by acting as a scavenger of nitric oxide and reactive oxygen species, but not of peroxynitrite. Melanin also increased the MICs of itraconazole and amphotericin B, and the drugs were fungicidal for nonmelanized and fungistatic for melanized yeast cells. Our study shows that melanin production by Paracoccidioides yeast cells serves a protective function during aPI and treatment with itraconazole and amphotericin B. The results suggest that melanin binds to the drugs, changing their antifungal activities, and also acts as a scavenger of reactive oxygen species and nitric oxide, but not of peroxynitrite, indicating that peroxynitrite is the main radical that is responsible for fungal death after aPI.

LED Phototherapy Improves Healing of Nipple Trauma: A Pilot Study

OBJECTIVE The purpose of this study was to evaluate the clinical effectiveness of a LED phototherapy prototype apparatus in the healing of nipple trauma in breastfeeding women. BACKGROUND DATA There is no scientific evidence of an effective treatment for nipple trauma. METHODS The experimental group was treated with orientation on nipple care and adequate breastfeeding techniques in addition to active LED phototherapy. The control group was treated with orientation on nipple care and adequate breastfeeding techniques in addition to placebo LED phototherapy. Participants were treated twice a week, for a total of eight sessions. Healing of the nipple lesions was measured by a reduction in their area, and decrease in pain intensity was measured in accordance with an 11-point Pain Intensity Numerical Rating Scale and a standard 7-point patient global impression of change. RESULTS Statistically significant reductions in measured nipple lesion area (p<0.001) were observed for both the experimental and control groups with an increase in the number of treatment sessions. A significant difference between the experimental and control groups was observed for the healing of nipple lesions (p<0.001). The pain intensity was significantly reduced only in the experimental group (p<0.001). CONCLUSIONS Preliminary results demonstrated the prototype apparatus for LED phototherapy to be an effective tool in accelerating the healing of nipple trauma.