The use of photobiomodulation therapy or LED and mineral trioxide aggregate improves the repair of complete tibial fractures treated with wire osteosynthesis in rodents.

The repair of large bone defects is lengthy and complex. Both biomaterials and phototherapy have been used to improve bone repair. We aimed to describe histologically the repair of tibial fractures treated by wiring (W), irradiated or not, with laser (λ780 nm, 70 mW, CW, spot area of 0.5 cm2, 20.4 J/cm2 (4 × 5.1 J/cm2, Twin Flex Evolution®, MM Optics, Sao Carlos, SP, Brazil) per session, 300 s, 142.8 J/cm2 per treatment) or LED (λ850 ± 10 nm, 150 mW, spot area of 0.5 cm2, 20.4 J/cm2 per session, 64 s, 142.8 J/cm2 per treatment, Fisioled®, MM Optics, Sao Carlos, Sao Paulo, Brazil) and associated or not to the use of mineral trioxide aggregate (MTA, Angelus®, Londrina, PR, Brazil). Inflammation was discrete on groups W and W + LEDPT and absent on the others. Phototherapy protocols started immediately before suturing and repeated at every other day for 15 days. Collagen deposition intense on groups W + LEDPT, W + BIO-MTA + LaserPT and W + BIO-MTA + LEDPT and discrete or moderate on the other groups. Reabsorption was discrete on groups W and W + LEDPT and absent on the other groups. Neoformation varied greatly between groups. Most groups were partial and moderately filed with new-formed bone (W, W + LaserPT, W + LEDPT, W + BIO-MTA + LEDPT). On groups W + BIO-MTA and W + BIO-MTA + LaserPT bone, neoformation was intense and complete. Our results are indicative that the association of MTA and PBMT (λ = 780 nm) improves the repair of complete tibial fracture treated with wire osteosynthesis in a rodent model more efficiently than LED (λ = 850 ± 10 nm).

Raman spectroscopic study of the effect of the use of laser/LED phototherapy on the repair of complete tibial fracture treated with internal rigid fixation.

This study aimed to assess the repair of complete surgical tibial fractures fixed with internal rigid fixation (IRF) associated or not to the use of mineral trioxide aggregate (MTA) cement and treated or not with laser (λ = 780 nm, infrared) or LED (λ = 850 ±â€¯10 nm, infrared) lights, 142.8 J/cm2 per treatment, by means of Raman spectroscopy. Open surgical tibial fractures were created on 18 rabbits (6 groups of 3 animals per group, ∼8 months old) and fractures were fixed with IRF. Three groups were grafted with MTA. The groups of IRF and IRF + MTA that received laser or LED were irradiated every other day during 15 days. Animals were sacrificed after 30 days, being the tibia surgically removed. Raman spectra were collected via the probe at the defect site in five points, resulting in 15 spectra per group (90 spectra in the dataset). Spectra were collected at the same day to avoid changes in laser power and experimental setup. The ANOVA general linear model showed that the laser irradiation of tibial bone fractures fixed with IRF and grafted with MTA had significant influence in the content of phosphate (peak ∼960 cm-1) and carbonated (peak ∼1,070 cm-1) hydroxyapatites as well as collagen (peak 1,452 cm-1). Also, peaks of calcium carbonate (1,088 cm-1) were found in the groups grafted with MTA. Based on the Raman spectroscopic data collected in this study, MTA has been shown to improve the repair of complete tibial fractures treated with IRF, with an evident increase of collagen matrix synthesis, and development of a scaffold of hydroxyapatite-like calcium carbonate with subsequent deposition of phosphate hydroxyapatite.

Freeze-dried versus fresh platelet-rich plasma in acute wound healing of an animal model.

Aim: To compare freeze-dried and fresh platelet-rich plasma (PRP) preparations, in a pre-clinical study. Materials & methods: 30 Wistar male rats were used to compare and characterize human PRP which was applied at the perilesional area in an acute wound model, evaluated by macroscopical and histological analysis. Results: Despite the increased growth factor concentration after the freeze-drying process, no change in the healing kinetics was observed in vivo. Nevertheless, a significant increased number of myofibroblasts was demonstrated in comparison with the fresh PRP group. We also demonstrated a significant increased percentage of blood vessels in comparison with controls in both the superficial and deep epidermis. Conclusion: These results encourage randomized clinical trials to evaluate the effectiveness of freeze-dried PRP for skin ulcer treatment.

Treatment of male pattern alopecia with platelet-rich plasma: A double-blind controlled study with analysis of platelet number and growth factor levels.

BACKGROUND: Promising results with platelet-rich plasma (PRP) in androgenetic alopecia that could be associated with platelet number and growth factor levels were described. OBJECTIVE: Analyze the platelet countand growth factor levels in PRP and their correlation with hair growth parameters evaluated by using the TrichoScan (Tricholog GmbH, Freiburg, Germany). METHODS: A total of 26 patients were randomized to receive 4 subcutaneous injections of PRP or saline. Hair growth, hair density, and percentage of anagen hairs were evaluated by using the TrichoScan method before injection, 15 days after the last injection, and again 3 months after the last injection. Growth factors (platelet-derived growth factor, epidermal growth factor, and vascular endothelial growth factor) were measured by the Luminex method (Millipore, Bedford, MA). RESULTS: We demonstrated a significant increase in hair count (P = .0016), hair density (P = .012) and percentage of anagen hairs (P = .007) in the PRP group versus in the control group, without correlation with platelet counts or quantification of the growth factors in PRP. LIMITATIONS: Other growth factors that could be related to response to PRP were not evaluated. CONCLUSION: Our data favor the use of PRP as a therapeutic alternative in the treatment of androgenetic alopecia. The lack of association between platelet count, platelet-derived growth factor, epidermal growth factor, and vascular endothelial growth factor levels and clinical improvement suggest that other mechanisms could be involved in this response.

Laser/LED phototherapy on the repair of tibial fracture treated with wire osteosynthesis evaluated by Raman spectroscopy.

The aim of the present study was to assess, by means of Raman spectroscopy, the repair of complete surgical tibial fractures fixed with wire osteosynthesis (WO) treated or not with infrared laser (λ780 nm) or infrared light emitting diode (LED) (λ850 ± 10 nm) lights, 142.8 J/cm2 per treatment, associated or not to the use of mineral trioxide aggregate (MTA) cement. Surgical tibial fractures were created on 18 rabbits, and all fractures were fixed with WO and some groups were grafted with MTA. Irradiated groups received lights at every other day during 15 days, and all animals were sacrificed after 30 days, being the tibia removed. The results showed that only irradiation with either laser or LED influenced the peaks of phosphate hydroxyapatite (~ 960 cm-1). Collagen (~ 1450 cm-1) and carbonated hydroxyapatite (~ 1070 cm-1) peaks were influenced by both the use of MTA and the irradiation with either laser or LED. It is concluded that the use of either laser or LED phototherapy associated to MTA cement was efficacious on improving the repair of complete tibial fractures treated with wire osteosynthesis by increasing the synthesis of collagen matrix and creating a scaffold of calcium carbonate (carbonated hydroxyapatite-like) and the subsequent deposition of phosphate hydroxyapatite.

Heme and iron induce protein aggregation.

Heme is an essential molecule expressed in many tissues where it plays key roles as the prosthetic group of several proteins involved in vital physiological and metabolic processes such as gas and electron transport. Structurally, heme is a tetrapyrrole ring containing an atom of iron (Fe) in its center. When released into the extracellular milieu, heme exerts several deleterious effects, which make it an important player in infectious and noninfectious hemolytic diseases where large amounts of free heme are observed such as malaria, dengue fever, ß-thalassemia, sickle cell disease and ischemia-reperfusion. Our recent work has uncovered an unappreciated cellular response triggered by heme or Fe, one of its degradation products, on macrophages, which is the formation of protein aggregates known as aggresome-like induced structres (ALIS). This response was shown to be fully dependent on ROS production and the activation of the transcription factor NFE2L2/NRF2. In addition, we have demonstrated that heme degradation by HMOX1/HO-1 (heme oxygenase 1) is required and that Fe is essential for the formation of ALIS, as heme analogs lacking the central atom of Fe are not able to induce these structures. ALIS formation is also observed in vivo, in a model of phenylhydrazine (PHZ)-induced hemolysis, indicating that it is an integral part of the host response to excessive free heme and that it may play a role in cellular homeostasis.

Protein aggregation as a cellular response to oxidative stress induced by heme and iron.

Hemolytic diseases include a variety of conditions with diverse etiologies in which red blood cells are destroyed and large amounts of hemeproteins are released. Heme has been described as a potent proinflammatory molecule that is able to induce multiple innate immune responses, such as those triggered by TLR4 and the NLRP3 inflammasome, as well as necroptosis in macrophages. The mechanisms by which eukaryotic cells respond to the toxic effects induced by heme to maintain homeostasis are not fully understood, however. Here we describe a previously uncharacterized cellular response induced by heme: the formation of p62/SQTM1 aggregates containing ubiquitinated proteins in structures known as aggresome-like induced structures (ALIS). This action is part of a response driven by the transcription factor NRF2 to the excessive generation of reactive oxygen species induced by heme that results in the expression of genes involved in antioxidant responses, including p62/SQTM1. Furthermore, we show that heme degradation by HO-1 is required for ALIS formation, and that the free iron released on heme degradation is necessary and sufficient to induce ALIS. Moreover, ferritin, a key protein in iron metabolism, prevents excessive ALIS formation. Finally, in vivo, hemolysis promotes an increase in ALIS formation in target tissues. Our data unravel a poorly understood aspect of the cellular responses induced by heme that can be explored to better understand the effects of free heme and free iron during hemolytic diseases such as sickle cell disease, dengue fever, malaria, and sepsis.

Correlation between Volume of Apical Periodontitis Determined by Cone-beam Computed Tomography Analysis and Endotoxin Levels Found in Primary Root Canal Infection.

INTRODUCTION: This clinical study was conducted to correlate the levels of endotoxins and bacterial counts found in primary endodontic infection with the volume of periapical bone destruction determined by cone-beam computed tomography (CBCT) analysis. Moreover, the levels of bacteria and endotoxins were correlated with the development of clinical features. METHODS: Twenty-four root canals with primary endodontic disease and apical periodontitis were selected. Clinical features such as pain on palpation, pain on percussion, and previous episode of pain were recorded. The volume (cubic millimeters) of periapical bone destruction was determined by CBCT analysis. Endotoxins and bacterial samplings were collected by using sterile/apyrogenic paper points. Endotoxins were quantified by using limulus amebocyte lysate assay (KQCL test), and bacterial count (colony-forming units [CFU]/mL) was determined by using anaerobic culture techniques. Data were analyzed by Pearson correlation and multiple logistic regression (P < .05). RESULTS: Endotoxins and bacteria were detected in 100% of the root canal samples (24 of 24), with median values of 10.92 endotoxin units (EU)/mL (1.75-128 EU/mL) and 7.5 × 10(5) CFU/mL (3.20 × 10(5)-8.16 × 10(6) CFU/mL), respectively. The median volume of bone destruction determined by CBCT analysis was 100 mm(3) (10-450 mm(3)). The multiple regression analysis revealed a positive correlation between higher levels of endotoxins present in root canal infection and larger volume of bone destruction (P < .05). Moreover, higher levels of endotoxins were also correlated with the presence of previous pain (P < .05). CONCLUSIONS: Our findings revealed that the levels of endotoxins found in root canal infection are related to the volume of periapical bone destruction determined by CBCT analysis. Moreover, the levels of endotoxin are related to the presence of previous pain.

Genistein prevents ultraviolet B radiation-induced nitrosative skin injury and promotes cell proliferation.

Nitric oxide (NO) levels increase considerably after 24h of exposure of skin to ultraviolet B (UVB) radiation, which leads to nitrosative skin injury. In addition, increased NO levels after exposure to UVB radiation are associated with inhibition of cell proliferation. Compared to the UV-control group, UV-genistein at 10 mg/kg (UV-GEN10) group showed tissue protection, decreased lipid peroxide and nitrotyrosine formation, and low CAT activity. Furthermore, NO levels and iNOS labeling remained high. In this group, the reduction in lipid peroxides and nitrotyrosine was accompanied by upregulation of cell proliferation factors (Ki67 and PCNA), which indicated that prevention of nitrosative skin injury promoted cell proliferation and DNA repair. Genistein also prevented nitrosative events, inhibited ONOO(-) formation, which leads to tissue protection and cell proliferation. The UV-GEN15 group did not result in a greater protective effect compared to that with UV-GEN10 group. In the UV-GEN15 group, histological examination of the epidermis showed morphological alterations without efficient protection against lipid peroxide formation, as well as inhibition of Ki67 and PCNA, and VEGF labeling, which suggested inhibition of cell proliferation. These results help to elucidate the mechanisms underlying the photoprotective effect of genistein and reveal the importance of UVB radiation-induced nitrosative damage.

The immune response of hemocytes of the insect Oncopeltus fasciatus against the flagellate Phytomonas serpens.

The genus Phytomonas includes parasites that are etiological agents of important plant diseases, especially in Central and South America. These parasites are transmitted to plants via the bite of an infected phytophagous hemipteran. Despite the economic impact of these parasites, many basic questions regarding the genus Phytomonas remain unanswered, such as the mechanism by which the parasites cope with the immune response of the insect vector. In this report, using a model of systemic infection, we describe the function of Oncopeltus fasciatus hemocytes in the immune response towards the tomato parasite Phytomonas serpens. Hemocytes respond to infection by trapping parasites in nodular structures and phagocytizing the parasites. In electron microscopy of hemocytes, parasites were located inside vacuoles, which appear fused with lysosomes. The parasites reached the O. fasciatus salivary glands at least six hours post-infection. After 72 hours post-infection, many parasites were attached to the salivary gland outer surface. Thus, the cellular responses did not kill all the parasites.

Time-dependent reactive species formation and oxidative stress damage in the skin after UVB irradiation.

This study provides evidence that skin oxidative stress injury caused by UVB irradiation is mediated predominantly by reactive oxygen species immediately after irradiation and by reactive nitrogen species at later time points. Animals were pre-treated with free radical scavengers (deferrioxamine, histidine), α-tocopherol, or inhibitors of nitric oxide synthase (NOS) (L-NAME or aminoguanidine) or left untreated and subjected to UVB irradiation. α-Tocopherol inhibited the increase in lipid peroxidation, as evaluated by chemiluminescence at 0 h and 24 h after UVB irradiation. Immediately after UVB irradiation, lipid peroxidation increased moderately and was abolished by free radical scavengers but not by NOS inhibitors. Likewise, the reduction of antioxidant capacity was not reversed by NOS inhibitors. Nitric oxide augmentation was not observed at this time point. Twenty-four hours after irradiation, increased lipid peroxidation levels and nitric oxide elevation were observed and were prevented by NOS inhibitors. Low concentrations of GSH and reduced catalase activity were also observed. Altogether, these data indicate that reactive oxygen species (singlet oxygen and hydroxyl radicals) are the principal mediators of immediate damage and that reactive nitrogen species (*NO and possibly ONOO(-)) seem to be involved later in skin oxidative injury induced by UVB radiation. The reduced catalase activity and low level of GSH suggest that *NO and H(2)O(2) may react to generate ONOO(-), a very strong lipid peroxidant species.

Mechanism of anticlastogenicity of Agaricus blazei Murill mushroom organic extracts in wild type CHO (K(1)) and repair deficient (xrs5) cells by chromosome aberration and sister chromatid exchange assays.

Agaricus blazei Murill is a medicinal mushroom native to Brazil. The present work assessed the clastogenic and anticlastogenic potential of organic extracts (ethanol and chloroform/methanol) from the lineage AB97/11 in chinese hamster CHO-K(1) (wild type) and CHO-xrs5 (repair deficient) cells using the chromosome aberration (CA) and sister chromatid exchange (SCE) assays. In these experimental conditions were observed: (a) anticlastogenic effect at concentrations of 0.06 and 0.09% of the EtOH extract and at the 0.03 and 0.06% concentrations of the C/MetOH extract in CHO-K(1); (b) absence of protector effect on CHO-xrs5 cells; and (c) absence of protector effect in the SCE assay. These results indicate that organic extracts of A. blazei lineage AB97/11 present bio-antimutagenic type protective activity.

Miocardite primaria aguda.Estudo a longo prazo de sete criancas. / Acute primary myocarditis. Long-term study of seven children

Os autores fizeram uma revisao da miocardite primaria na crianca e destacam que desde 1973 tiveram a oportunidade de observar 19 criancas portadoras desta patologia, sendo que somente sete foram incluidas nesta revisao, pois as demais tiveram o inicio da doenca, ou seja, sua fase aguda, em epoca anterior ao inicio deste trabalho. Comentam, tambem, os aspectos clinicos, radiologicos, eletrocardiograficos e terapeuticos e discutem as diferentes opinioes quanto ao uso, ou nao, da corticoterapia; dao enfase especial a evolucao e relatam que duas criancas evoluiram para a cura, quatro para uma forma cronica e uma para o obito. Todos os pacientes com evolucao cronica foram observados durante quatro anos e, progressivamente, no ECG, passaram a evidenciar sinais de acentuada hipertrofia ventricular esquerda com ondas Q importantes em V5 e V6. Frente a estes achados os autores comecaram os recentes trabalhos que indicam a possibilidade da fibroelastose endocardica ser determinada por uma previa miocardite a virus