Papers by Antonio Pinheiro

Photomedicine and laser surgery, 2014
Objective: The aim of this study was to assess, immunohistochemically, the impact of hypothyroidi... more Objective: The aim of this study was to assess, immunohistochemically, the impact of hypothyroidism and the effect of laser phototherapy on the expression of type I and III collagen during wound healing. Background data: Hypothyroidism has been associated with the disruption of the body's metabolism, including the healing process. Laser phototherapy has been shown to be effective in improving wound healing, but its usefulness on enhancing wound healing under hypothyroid condition remains unknown. Materials and methods: Using general anesthesia, a standard surgical wound (1 cm 2 ) was created on the dorsa of 48 Wistar rats divided into four groups of 12 animals each: control euthyroid (EC), euthyroid plus laser (EL), control hypothyroid (HC), and hypothyroid plus laser (HL). The irradiation with laser GaAlAs [k660 nm, 40 mW, 1 W/cm 2 , continuous wave (CW), ø = 0.04 cm 2 ] started immediately after surgery and was repeated every other day until end-point of study was reached, and animals were euthanized (i.e., 7 and 14 days). Laser light was applied on four different points (6 J, 150 sec and 150 J/cm 2 per point). Hypothyroidism was induced in rats with propylthiouracil (0.05 g/100 mL) administered orally for 4 weeks and maintained until the end of the experiment. Immunohistochemistry for collagen I and III was performed with EnVision TM in the specimens removed. Results: Seven days after the surgery EC, EL, and HL groups showed higher immunoexpression of collagen I and lower immunoexpression of collagen III in the newly formed tissue. There was increased immunoexpression of collagen I in EC when compared with HC ( p = 0.019). The immunoexpression of collagen III was significantly lower in EL than in EC ( p = 0.047) and HL ( p = 0.019). No significant difference was found in the experimental period of 14 days among the groups. Conclusions: Laser light therapy performed with the parameters of this investigation increased immunoexpression of collagen type I during tissue repair, and improved the quality of newly formed tissue in the presence of hypothyroidism.

Lasers in medical science, 2014
The treatment of bone loss is difficult. Many techniques are proposed to improve repair, includin... more The treatment of bone loss is difficult. Many techniques are proposed to improve repair, including biomaterials and, recently, phototherapies. This work studied bone mineralization by Raman spectroscopy assessing intensities of Raman peaks of both inorganic (∼960, ∼1,070 cm−1) and organic (∼1,454 cm−1) contents in animal model. Six groups were studied: clot, laser, light-emitting diode (LED), biomaterial (HA + β-tricalcium phosphate), laser + biomaterial, and LED + biomaterial. Defects at right tibia were performed with a drill. When indicated, defects were further irradiated at a 48-h interval during 2 weeks. At the 15th and 30th days, the tibias were withdrawn and analyzed. The ∼960-cm−1 peak was significantly affected by phototherapy on both clot- and biomaterial-filled defects. The ∼1,070-cm−1 peak was affected by both time and the use of the LED light on clot-filled defects. On biomaterial-filled defects, only the use of the laser light significantly influenced the outcome. No significant influence of either the time or the use of the light was detected on clot-filled defects as regards the ∼1,454-cm−1 peak. Raman intensities of both mineral and matrix components indicated that the use of laser and LED phototherapies improved the repair of bone defects grafted or not with biphasic synthetic microgranular HA + β-tricalcium phosphate.
British Journal of Oral & Maxillofacial Surgery, 1993
UK SUMMARY.

Parasitic diseases represent a major public health problems in Latin America, in particular, Chag... more Parasitic diseases represent a major public health problems in Latin America, in particular, Chagas disease or American trypanosomiasis, caused by the protozoan parasite Trypanosoma cruzi, infects more than 18 million people in all countries of Latin America. Visible light induces a photochemical reaction, that induces the activation of enzymes used mainly in the respiratory chain, and that light has the primary targets lysosomes and mitochondria of cells, increasing, the mitochondrial ATP production. The purpose of this study was to assess the morpho-structural generated in the epimastigote form of Trypanosoma cruzi, after irradiation with a semiconductor laser InGaAlP, at a wavelength (λ) equal to 660 nm+/-10 nm, 40 mW optical Power, emitting red light in the visible spectrum, with a dose of 6 J/cm2 in continuous mode. Then the parasites that have undergone irradiation were analyzed by optical microscopy and compared to untreated. It found the increase in size of the kinetoplast (structure with high concentration of extracellular DNA-kDNA, whose main function is to encode the respiratory chain enzymes such as ATPase and citocromoxidase), the cell nucleus and the cell volume of the parasite, leaving the more rounded.

Policies to combat Chagas disease presents a considerable degree of negligence and is classified ... more Policies to combat Chagas disease presents a considerable degree of negligence and is classified at level III by TDR, where the focus of research is based on the improvement and wider dissemination of existing tools and strategies for combating them. The PACT is based on topical or systemic administration of a nontoxic dye sensitive to light, followed by low dose irradiation with visible light of wavelength appropriate. In the presence of oxygen found in the cells, the photosensitizer (FS) enabled may react with molecules in its vicinity by electron transfer or hydrogen, leading to production of free radicals (type I reaction) or by energy transfer to oxygen (type II reaction), leading to production of singlet oxygen. Both paths can lead to cell death and the destruction of diseased tissue. In this work, we verify the effectiveness of PACT associated with a semiconductor laser InGaAlP, a wavelength (λ) equal to 660nm ± 10nm, 30 mW optical power, emitting red light in the visible spectrum, with a dose of 4J/cm 2 in continuous mode, using methylene blue in five differents concentrations on the infective trypomastigote forms of Trypanosoma cruzi. To determine the viability of the parasites, one sample from each treatment group at each concentration was removed and analyzed in a hemocytometer, observing the decrease in the number of live parasites for the solution without treatment. The results demonstrated significant percentage of parasite lysis (up to 86% lethality), what can not be observed in the groups treated with laser or with the FS.

Policies to combat Chagas disease presents a considerable degree of negligence and is classified ... more Policies to combat Chagas disease presents a considerable degree of negligence and is classified at level III by TDR, where the focus of research is based on the improvement and wider dissemination of existing tools and strategies for combating them. The PACT is based on topical or systemic administration of a nontoxic dye sensitive to light, followed by low dose irradiation with visible light of wavelength appropriate. In the presence of oxygen found in the cells, the photosensitizer (FS) enabled may react with molecules in its vicinity by electron transfer or hydrogen, leading to production of free radicals (type I reaction) or by energy transfer to oxygen (type II reaction), leading to production of singlet oxygen. Both paths can lead to cell death and the destruction of diseased tissue. In this work, we verify the effectiveness of PACT associated with a semiconductor laser InGaAlP, a wavelength (λ) equal to 660 nm+/-10 nm, 30 mW optical power, emitting red light in the visible spectrum, with a dose of 4 J/cm2 in continuous mode, using methylene blue in five differents concentrations on the infective trypomastigote forms of Trypanosoma cruzi. To determine the viability of the parasites, one sample from each treatment group at each concentration was removed and analyzed in a hemocytometer, observing the decrease in the number of live parasites for the solution without treatment. The results demonstrated significant percentage of parasite lysis (up to 86% lethality), what can not be observed in the groups treated with laser or with the FS.

Photomedicine and Laser Surgery, 2010
The aim of this study was to analyze the effect of two phototherapies, laser and polarized light,... more The aim of this study was to analyze the effect of two phototherapies, laser and polarized light, on diode laser (970lambda nm) wounds. Lasers have been used in surgery, and some wavelengths may cause thermal damage to the tissue and affect healing. Several studies have shown that some wavelengths are effective in improving healing. Coherent and noncoherent light have been successfully used on the modulation of biological phenomena of several origins. Thirty-one Wistar rats were divided into 3 groups (GI to GIII). A 20-mm x 2-mm wound was created on the dorsum of each animal with a diode laser (Sirolaser, Sirona, Bensheim, Germany). Group GI acted as control. On GII, laser light (lambda655 nm, 30 mW, phi approximately 3 mm, 12 J/cm(2)) was used and on GIII illumination with polarized light (lambda400-2000 nm, 40 mW, phi approximately 5.5 cm, 12 J/cm(2)) was used, every other day (GII) or daily (GIII) for 7 days. The animals were killed at 0, 7, and 14 days after surgery. Specimens were taken, routinely processed, stained and imunnomarked [HE (hematoxylin-eosin), sirius red, alpha-smooth muscle actin (SMA)], and underwent histological analysis. GII showed better response at day 14 when re-epithelialization was in a more advanced stage. The number of myofibroblasts was significantly different over the healing time (7 to 14 days); this number was smaller than that observed on G1. On GIII at day 7, the number of myofibroblasts was significantly higher than for GII. At day 14, a more pronounced deposition of collagen matrix was also seen, and inflammation was discrete and more advanced for GIII. The results of the present study showed that the effect of the use of laser light was more evident at early stages of healing and that the use of polarized light improved the resolution of the inflammatory reaction, increased the deposition of collagen, increased the number of myofibroblasts, and quickened re-epithelialization during the experimental time.

Parasitic diseases represent a major public health problems in Latin America, in particular, Chag... more Parasitic diseases represent a major public health problems in Latin America, in particular, Chagas disease or American trypanosomiasis, caused by the protozoan parasite Trypanosoma cruzi, infects more than 18 million people in all countries of Latin America. Visible light induces a photochemical reaction, that induces the activation of enzymes used mainly in the respiratory chain, and that light has the primary targets lysosomes and mitochondria of cells, increasing, the mitochondrial ATP production. The purpose of this study was to assess the morpho-structural generated in the epimastigote form of Trypanosoma cruzi, after irradiation with a semiconductor laser InGaAlP, at a wavelength (λ) equal to 660nm ± 10nm, 40 mW optical Power, emitting red light in the visible spectrum, with a dose of 6J/cm 2 in continuous mode. Then the parasites that have undergone irradiation were analyzed by optical microscopy and compared to untreated. It found the increase in size of the kinetoplast (structure with high concentration of extracellular DNA -kDNA, whose main function is to encode the respiratory chain enzymes such as ATPase and citocromoxidase), the cell nucleus and the cell volume of the parasite, leaving the more rounded.
Photomedicine and Laser Surgery, 2009

Bell's palsy is defined as a peripheral facial nerve palsy, idiophatic, and sudden onset and... more Bell's palsy is defined as a peripheral facial nerve palsy, idiophatic, and sudden onset and is considered the most common cause of this pathology. It is caused by damage to cranial nerves VII, resulting in complete or partial paralysis of the facial mimic. May be associated with taste disturbances, salivation, tearing and hyperacusis. It is diagnosed after ruling out all possible etiologies, because its cause is not fully understood.Some researches shows that herpes virus may cause this type of palsy due to reactivation of the virus or by imunnomediated post-viral nerve demielinization. Physical therapy, corticosteroids and antiviral therapy have become the most widely accepted treatments for Bell's palsy. Therapy with low-level laser (LLLT) may induce the metabolism of injured nerve tissue for the production of proteins associated with its growth and to improve nerve regeneration. The success of the treatment of Bell's palsy by using laser phototherapy isolated or in association with other therapeutic approach has been reported on the literature. In most cases, the recovery occurs without uneventfully (complications), the acute illness is not associated with serious disorders. We will present a clinical approach for treating this condition.
International Congress Series, 2003

International Congress Series, 2003
The aim of this study was to assess histologically the effect of LLLT (λ 830 nm) on the repair of... more The aim of this study was to assess histologically the effect of LLLT (λ 830 nm) on the repair of standardized bone defects on the femur of Wistar albinus rats which were grafted with inorganic bovine bone Gen-ox ® . Three randomized groups were studied: group I (control, n=6); group II (Gen-ox ® , n=9) and group III (Gen-ox ® + LLLT, n=9). The animals were irradiated every 48 h during 15 days; the first irradiation was performed immediately after the procedure. The animals were irradiated transcutaneuosly at four points around the defect. At each point a dose of 4 J/cm 2 was given (Ø~0.6 mm, 40 mW) and the total dose per session was 16 J/cm 2 . The animals were killed by an overdose of general anesthetic 15, 21 and 30 days after surgery. The specimens were routinely processed by embedding in paraffin, serially cut and stained with H&E and Picrosirius and analyzed under light microscopy. The results showed evidence of a more advanced repair in the irradiated group when compared to the non-irradiated groups. The repair of the irradiated group was characterized by both increased bone formation and amount of collagen fibers around the graft within the cavity from the 15th day after surgery, also considering the osteoconductive capacity of the Gen-ox ® . We conclude that LLLT had a positive effect on the repair of bone defects implanted with inorganic bovine bone.
Quimica Nova, 2005
Eighteen circular blocks of resins cured either by a LED or a halogen lamp (20, 40 and 60 s), had... more Eighteen circular blocks of resins cured either by a LED or a halogen lamp (20, 40 and 60 s), had their top (T) and bottom (B) surfaces studied using a FT-Raman spectrometer. Systematic changes in the intensity of the methacrylate C=C stretching mode at 1638 cm-1 as a function of exposure duration were observed. The calculated degree of conversion (DC) ranged from 45.0% (B) to 52.0% (T) and from 49.0% (B) to 55.0% (T) for the LED and halogen lamp, respectively. LED and halogen light produced similar DC values with 40 and 60 s of irradiation.
International Congress Series, 2003
International Congress Series, 2003

Photomedicine and laser surgery, 2008
The aim of this investigation was to compare by light microscopy the effects of laser photobiomod... more The aim of this investigation was to compare by light microscopy the effects of laser photobiomodulation at wavelengths of 660 and 780 nm on third-degree burns in Wistar rats. Burns are severe injuries that result in the loss of fluid and destruction of tissue, infection, and shock that may result in death. Laser energy has been suggested as an effective method to improve wound healing. Fifty-five animals were used in this study. A third-degree burn measuring 1.5 x 1.5 cm was created on the dorsum of each animal. The animals were divided into three subgroups according the type of laser photobiomodulation they received (wavelength of 660 or 780 nm, 35 mW, theta = 2 mm, and 20 J/cm2). In the animals receiving treatment, it was begun immediately post-burn at four points around the burn (5 J/cm2) and repeated at 24-h intervals for 21 d. The animals were humanely killed after 3, 5, 7, 14, and 21 d by an intraperitoneal overdose of general anesthetic. The specimens were routinely cut and stained, and then were analyzed by light microscopy. The results showed more deposition of collagen fibers, larger amounts of granulation tissue, less edema, a more vigorous inflammatory reaction, and increased revascularization on all laser-treated animals. These features were more evident at early stages when the 660-nm laser was used, and were more evident throughout the experimental period for the animals receiving 780-nm laser therapy. We concluded that laser photobiomodulation using both wavelengths improved healing of third-degree burns on Wistar rats.

Lasers in medical science, 2012
We studied peaks of calcium hydroxyapatite (CHA) and protein and lipid CH groups in defects graft... more We studied peaks of calcium hydroxyapatite (CHA) and protein and lipid CH groups in defects grafted with mineral trioxide aggregate (MTA) treated or not with LED irradiation, bone morphogenetic proteins and guided bone regeneration. A total of 90 rats were divided into ten groups each of which was subdivided into three subgroups (evaluated at 15, 21 and 30 days after surgery). Defects were irradiated with LED light (wavelength 850 ± 10 nm) at 48-h intervals for 15 days. Raman readings were taken at the surface of the defects. There were no statistically significant differences in the CHA peaks among the nonirradiated defects at any of the experimental time-points. On the other hand, there were significant differences between the defects filled with blood clot and the irradiated defects at all time-points (p < 0.001, p = 0.02, p < 0.001). There were significant differences between the mean peak CHA in nonirradiated defects at all the experimental time-points (p < 0.01). The mean peak of the defects filled with blood clot was significantly different from that of the defects filled with MTA (p < 0.001). There were significant differences between the defects filled with blood clot and the irradiated defects (p < 0.001). The results of this study using Raman spectral analysis indicate that infrared LED light irradiation improves the deposition of CHA in healing bone grafted or not with MTA.

Photomedicine and laser surgery, 2008
The aim of the present investigation was to histologically assess the effect of laser photobiomod... more The aim of the present investigation was to histologically assess the effect of laser photobiomodulation (LBPM) on the repair of autologous bone grafts in a rodent model. A major problem in modern dentistry is the recovery of bone defects caused by trauma, surgical procedures, or pathologies. Several types of biomaterials have been used to improve the repair of these defects. These materials are often associated with procedures of guided bone regeneration (GBR). Twenty four animals were divided into four groups: group I (control); group II (LPBM of the bone graft); group III (bone morphogenetic proteins [BMPs] + bone graft); and group IV (LPBM of the bed and the bone graft + BMPs). When appropriate the bed was filled with lyophilized bovine bone and BMPs used with or without GBR. The animals in the irradiated groups received 10 J/cm(2) per session divided over four points around the defect (4 J/cm(2)), with the first irradiation immediately after surgery, and then repeated seven times every other day. The animals were humanely killed after 40 d. The results showed that in all treatment groups, new bone formation was greater and qualitatively better than the untreated subjects. Control specimens showed a less advanced repair after 40 d, and this was characterized by the presence of medullary tissue, a small amount of bone trabeculi, and some cortical repair. We conclude that LPBM has a positive biomodulatory effect on the healing of bone defects, and that this effect was more evident when LPBM was performed on the surgical bed intraoperatively, prior to the placement of the autologous bone graft.

Lasers in medical science, 2014
This study aimed to assess bone repair in defects grafted or not with hydroxyapatite (HA) on heal... more This study aimed to assess bone repair in defects grafted or not with hydroxyapatite (HA) on healthy and iron-deficiency anemia (IDA) rats submitted or not to LED phototherapy (LED-PT) by Raman spectroscopy. The animals were divided in eight groups with five rats each: Clot; Clot + LED; IDA + Clot; IDA + LED; Graft; Graft + LED; IDA + Graft; and IDA + Graft + LED. When appropriated, irradiation with IR LED (λ850 ± 10 nm, 150 mW, CW, Φ = 0.5 cm2, 16 J/cm2, 15 days) was carried out. Raman shifts: ∼960 [symmetric PO4 stretching (phosphate apatite)], ∼1,070 [symmetric CO3 stretching (B-type carbonate apatite)], and ∼1,454 cm−1 [CH2/CH3 bending in organics (protein)] were analyzed. The mean peak values for ∼960, ∼1,070, and ∼1,454 cm−1 were nonsignificantly different on healthy or anemic rats. The group IDA + Graft + LED showed the lowest mean values for the peak ∼960 cm−1 when compared with the irradiated IDA group or not (p ≤ 0.001; p ≤ 0.001). The association of LED-PT and HA-graft showed lowest mean peak at ∼1,454 cm−1 for the IDA rats. The results of this study indicated higher HA peaks as well as a decrease in the level of organic components on healthy animals when graft and LED phototherapy are associated. In the other hand, IDA condition interfered in the graft incorporation to the bone as LED phototherapy only improved bone repair when graft was not used.

Lasers in medical science, 2013
Osteomyelitis is an acute or chronic inflammation in the marrow spaces in the superficial or cort... more Osteomyelitis is an acute or chronic inflammation in the marrow spaces in the superficial or cortical bone, and can be associated with bacterial or fungal infections. Chronic osteomyelitis represents a major health problem due to its difficult treatment and increased morbidity. Photodynamic antimicrobial therapy (PAmT) is a treatment based on a cytotoxic photochemical reaction in which a bright light produced by a laser system and an active photosensitizer absorbed by cells leads to a process of activation that induces a series of metabolic reactions that culminates a bacterial killing. The aim of the present randomized study was to evaluate, by in vitro and in vivo microbiological analysis, the effects of PAmT on tibial surgical bone defects in rats infected by Staphylococcus aureus using bacterial counts carried out immediately and after 30 days after treatment as outcome measure. In the preliminary in vitro study, a diode laser (λ660 nm; 40 mW; ϕ = 0.4 cm2; 5 or 10 J/cm2) and 5, 10, and 15 μg/mL toluidine blue were tested, and the best parameter was chosen for the in vivo study. The concentration of 5 μg/mL was selected to perform the decontamination of S. aureus-infected tibial bone defects in rats. The findings were subjected to statistical analysis. For all PAmTs groups, with the different concentrations, treatment showed significant reductions (p < 0.001) in the amount of bacteria. The in vivo study PAmT group presented a bacterial reduction of 97.4 % (p < 0.001). The PAmT using toluidine blue was effective in reducing the number of S. aureus in both in vitro and in vivo studies.
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Papers by Antonio Pinheiro