Sperm selection by birefringence: a promising non-invasive tool to improve ICSI outcomes.

OBJECTIVE: Despite higher sperm DNA fragmentation may affect intracytoplasmic sperm injection (ICSI) outcomes, sperm selection protocols do not evaluate this parameter. Therefore, sperm's head birefringence has been suggested as an adjuvant of seminal processing to select viable sperm for couples with severe male factor. Considering men with normal seminal parameters may also curse with DNA fragmentation, the aim of this study was to evaluate the impact of sperm selection by birefringence on ICSI outcomes in couples with different infertility factors compared to those submitted to conventional sperm selection. METHODS: In this case-control study, medical records from 181 couples who underwent ICSI from January 2018 to August 2020 (107 from the Conventional and 74 from the Birefringence group) were included in the study. Clinical characteristics and ICSI outcomes were compared between the groups using Student's t test or Chi-square test (p<0.05) and a multivariate logistic regression model was applied regarding clinical pregnancy. RESULTS: Despite the Birefringence group showed higher female age (p=0.01), lower seminal sperm concentration (p<0.01) and higher sperm DNA fragmentation (p<0.01), those patients cursed with both higher cleavage rate (p=0.04), clinical pregnancy rate per transfer (p=0.03) and clinical pregnancy rate per initiated cycle (p=0.02). The logistic regression showed a positive group effect on clinical pregnancy. CONCLUSIONS: The findings suggest a positive clinical impact of this cheap and easily reproducible adjuvant technique on ICSI outcomes in couples with different infertility factors. If confirmed by further methodologically appropriate studies, the sperm's head birefringence could be considered to improve the reproductive chances of those patients.

Variability in the Epithelial Expression of Ki67 and Polarization Characteristics of Collagen in Odontogenic Cysts and Tumours

ABSTRACT Objective: To assess the proliferation of epithelium (using the Ki67 index) and the polarization pattern of collagen in selected odontogenic cysts and tumours. In addition, an exploratory analysis of the effect of inflammation on the proliferation rate was done. Material and Methods: Following immunohistochemical staining, the labelling/proliferation index of Ki67 was calculated. The thickness and corresponding polarization colour of 100 juxta-epithelial picrosirius red-stained collagen fibers were assessed using linear micrometry with an eyepiece reticule under × 1000 magnification. Inflammation was graded subjectively as mild, moderate, and severe. Results: Overall Ki-67 expression was higher in the radicular cyst, Odontogenic Keratocyst, Ameloblastoma, while suprabasal Ki-67 positivity was maximum in Odontogenic Keratocyst. The stromal collagen fibers in Ameloblastoma showed predominantly green birefringence, whereas Odontogenic Keratocyst had orange birefringence. There was no significant association of inflammation with Ki-67 expression or birefringence patterns. Conclusion: The highest Ki67 expression in the radicular cyst, followed by Odontogenic Keratocyst and Ameloblastoma. Differences in the collagen maturation pattern were noted innately in five lesions studied and were further influenced by inflammatory changes. Epithelial proliferation and concomitant expression of thickness and maturity of the stromal collagen are innate features of the lesion further influenced by inflammation in various odontogenic cysts and tumours and may, in turn, guide the clinical behavior.

Ultrasonic Assessment of the Influence of Cold Rolling and Recrystallization Annealing on the Elastic Constants in a TWIP Steel.

The evolution of the elastic constants, C33, C44 and C55, Poisson's ratio and acoustic birefringence of a Fe-0.5 wt% C-21.5 wt% Mn twinning-induced plasticity (TWIP) steel with reduction by cold rolling and recrystallization annealing was assessed from measurements of the times of flight of ultrasonic waves propagating along the thickness of the rolled plates. As the reduction increased, changes in the elastic constants resulted in a steadily increasing orthotropy, which was clearly shown by Poisson's ratio and acoustic birefringence. Although optical metallography and hardness measurements showed that partial or full recrystallization is attained after annealing at 600 °C and 700 °C, the ultrasonic measurements revealed that a high level of orthotropy remains.

Shear-Coated Linear Birefringent and Chiral Cellulose Nanocrystal Films Prepared from Non-Sonicated Suspensions with Different Storage Time.

Nanocelluloses are very attractive materials for creating structured films with unique optical properties using different preparation techniques. Evaporation-induced self-assembly of cellulose nanocrystals (CNC) aqueous suspensions produces iridescent films with selective circular Bragg reflection. Blade coating of sonicated CNC suspensions leads to birefringent CNC films. In this work, fabrication of both birefringent and chiral films from non-sonicated CNC suspensions using a shear-coating method is studied. Polarization optical microscopy and steady-state viscosity profiles show that non-sonicated CNC suspensions (concentration of 6.5 wt%) evolve with storage time from a gel-like shear-thinning fluid to a mixture of isotropic and chiral nematic liquid crystalline phases. Shear-coated films prepared from non-sonicated fresh CNC suspensions are birefringent, whereas films prepared from suspensions stored several weeks show reflection of left-handed polarized light. Quantification of linear and circular birefringence as well circular dichroism in the films is achieved by using a Mueller matrix formalism.

Sperm Selection for micro TESE-ICSI in Non-Obstructive Azoospermia, a Case Report.

TESE-ICSI (testicular sperm extraction associated with intracytoplasmic sperm injection) represents a technique to attain pregnancy in couples with non-obstructive azoospermia (NOA) and other unlikely situations. Because of the poor pregnancy outcomes obtained by this procedure, we need new sperm selection techniques to improve the livebirth rate of NOA patients. Here we describe a successful micro TESE-ICSI cycle performed with sperm selected through high magnification and polarized light microscopy in a couple with two previous ICSI failures.

Acmella oleracea extract increases collagen content and organization in partially transected tendons.

Acmella oleracea contains spilanthol as the main active compound, which possesses analgesic and anti-inflammatory effects that can favor tendon reorganization. To analyze the effect of A. oleracea on the content and organization of collagen in injured tendons, the calcaneal tendon of male Lewis rats was partially transected and treated at the site of injury with a topical application of 20% A. oleracea ointment (AO group) or with the ointment base without the plant extract (B group). The animals were euthanized 21 days after partial transection. Higher collagen concentration was observed in the AO group than in the B group, and morphological analysis using polarization microscopy showed higher birefringence in the AO group than in the B group, indicating higher collagen organization. No difference was observed in the number of fibroblasts, blood vessels, proteoglycan distribution, and maximum load between the B and AO groups. In conclusion, topical application of 20% A. oleracea ointment increased the molecular organization and content of collagen, thus indicating a potential application in tendon repair. Studies on the later phases of the tendon healing process are necessary to demonstrate the possible biomechanical changes after the application of A. oleracea ointment.

Transected Tendon Treated with a New Fibrin Sealant Alone or Associated with Adipose-Derived Stem Cells.

Tissue engineering and cell-based therapy combine techniques that create biocompatible materials for cell survival, which can improve tendon repair. This study seeks to use a new fibrin sealant (FS) derived from the venom of Crotalus durissus terrificus, a biodegradable three-dimensional scaffolding produced from animal components only, associated with adipose-derived stem cells (ASC) for application in tendons injuries, considered a common and serious orthopedic problem. Lewis rats had tendons distributed in five groups: normal (N), transected (T), transected and FS (FS) or ASC (ASC) or with FS and ASC (FS + ASC). The in vivo imaging showed higher quantification of transplanted PKH26-labeled ASC in tendons of FS + ASC compared to ASC on the 14th day after transection. A small number of Iba1 labeled macrophages carrying PKH26 signal, probably due to phagocytosis of dead ASC, were observed in tendons of transected groups. ASC up-regulated the Tenomodulin gene expression in the transection region when compared to N, T and FS groups and the expression of TIMP-2 and Scleraxis genes in relation to the N group. FS group presented a greater organization of collagen fibers, followed by FS + ASC and ASC in comparison to N. Tendons from ASC group presented higher hydroxyproline concentration in relation to N and the transected tendons of T, FS and FS + ASC had a higher amount of collagen I and tenomodulin in comparison to N group. Although no marked differences were observed in the other biomechanical parameters, T group had higher value of maximum load compared to the groups ASC and FS + ASC. In conclusion, the FS kept constant the number of transplanted ASC in the transected region until the 14th day after injury. Our data suggest this FS to be a good scaffold for treatment during tendon repair because it was the most effective one regarding tendon organization recovering, followed by the FS treatment associated with ASC and finally by the transplanted ASC on the 21st day. Further investigations in long-term time points of the tendon repair are needed to analyze if the higher tissue organization found with the FS scaffold will improve the biomechanics of the tendons.

Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency / Fenótipos clínicos e anisotropias ópticas da córnea em coelhos com deficiência de células tronco limbais

ABSTRACT Purposes: To investigate the intra-laboratory reproducibility of clinical features and to evaluate corneal optical anisotropies in a rabbit model of limbal stem cell deficiency. Methods: Limbal injury was induced in the right eye of 23 adult New Zealand White rabbits using a highly aggressive protocol that combined 360 degrees limbal peritomy, keratolimbectomy, alkaline chemical burn, and mechanical removal of the epithelium. Clinical evaluation of the injured eyes was performed for 28 days and included corneal impression cytology. Corneas with a severe clinical outcome set typical of limbal stem cell deficiency were then collected, subjected to a histopathological examination, and examined for optical anisotropies. Corneas from healthy rabbit eyes were used as controls. Differences in optical path due to stromal collagen birefringence, as well as linear dichroism related to the expression and spatial orientation of glycosaminoglycan chains from proteoglycans, were measured from cross-sections under a quantitative polarized light microscope. Results: One eye showed signs of hypopyon and was excluded. Signs of ocular inflammation were observed in all eyes studied (n=22). Corneal impression cytology did not detect goblet cells. Twelve of the 22 corneas presented a clinical outcome set typical of limbal stem cell deficiency, which is characterized by the presence of epithelial defects, inflammatory cells, moderate-to-severe opacity, and neovascularization. Microscopic studies under polarized light revealed that relative to controls, limbal stem cell deficiency caused a 24.4% increase in corneal optical path differences. Further, corneas with limbal stem cell deficiency were less dichroic than controls. Conclusions: These results suggest that rabbit models of limbal stem cell deficiency must be rigorously screened for use in preclinical studies to ensure experimental homogeneity because protocols used to create limbal stem cell deficiency could be not associated with good intra-laboratory reproducibility of clinical features. Limbal stem cell deficiency, as induced herein, altered the optical anisotropic properties of the corneal stroma. Such alterations are indicative of changes in collagen packing and the spatial orientation of glycosaminoglycan chains from proteoglycans. Knowledge of these changes is important to potentiate strategies aimed at restoring the morphofunctional integrity of the corneal stroma affected by limbal stem cell deficiency.

RESUMO Objetivos: Investigar a reprodutibilidade intra-la­boratorial dos fenótipos clínicos e avaliar anisotropias ópticas em córneas de coelhos com deficiência de células tronco limbais. Métodos: Lesões ao limbo foram feitas no olho direito de 23 coelhos adultos da Nova Zelândia Branco, usando um protocolo altamente agressivo, que envolveu peritomia limbal em 360 graus, ceratolimbectomia, cauterização por álcali, e remoção mecânica de epitélio remanescente. Os olhos foram clinicamente avaliados por 28 dias, inclusive por citologia de impressão corneal. As córneas que manifestaram um conjunto de alterações típicas de deficiência de células tronco limbais foram coletadas e submetidas à estudos em histopatologia e em anisotropias ópticas. Córneas saudáveis foram usadas como controles. Diferenças de caminho óptico de birrefringência relacionada à organização do colágeno estromal, e dicroísmo linear relacionado à expressão e à orientação das cadeias de glicosaminoglicanos dos proteoglicanos estromais, foram quantificados por microscopia de luz polarizada. Resultados: Um olho apresentou hipópio e foi excluído do estudo. Todos os olhos estudados (n=22) apresentaram sinais de inflamação ocular. A citologia de impressão não detectou células caliciformes na superfície corneal. Doze de 22 córneas manifestaram alterações clínicas típicas de deficiência de células tronco limbais, caracterizado por defeitos epiteliais, infiltrados inflamatórios, opacidade de moderada à severa, e neovascularização. Estudos por microscopia de luz polarizada mostraram que a deficiência de células tronco limbais aumentou a diferenças de caminho óptico corneal em 24,4% (versus controles). As cór­neas com deficiência de células tronco limbais foram menos dicroicas do que as córneas controle. Conclusões: Coelhos com deficiência de células tronco limbais, para aplicações em estudos pré-clínicos, devem ser rigorosamente selecionados para assegurar homogeneidade experimental, pois há evidências de que protocolos utilizados para indução de deficiência de células tronco limbais não estão associados com boa reprodutibilidade intra-laboratorial de fenótipos clínicos. A deficiência de células tronco limbais, como induzida aqui, alterou as propriedades ópticas anisotrópicas do estroma corneal. Tais alterações são indicativas de mudanças no empacotamento de colágeno e na orientação das cadeias de glicosaminoglicanos dos proteoglicanos. Conhecimentos nessas alterações são importantes para potencializar estratégias que visam a restabelecer a integridade morfofuncional do estromal corneal acometido pela deficiência de células tronco limbais.

Ciprofloxacin-lidocaine-based hydrogel: development, characterization, and in vivo evaluation in a second-degree burn model.

The purpose of this work was to develop an effective carbomer hydrogel to be used to treat second-degree burns that combined ciprofloxacin and lidocaine (CbCipLid hydrogel). Its antibiotic and anesthetic efficacy and the physical and chemical properties of the CbCipLid hydrogel (release rate and kinetics, rheology, appearance, and drug content) were evaluated both before and after a sterilization cycle and also after 6 months of storage. For the in vivo studies, second-degree burns were developed in a rat model. Animals were divided into three groups: CbCipLid hydrogel, silver sulfadiazine cream (reference), and carbomer hydrogel (as control). The treatments were applied daily for 21 days, and the healing was monitored by macroscopic observation and histologic evaluation. The anesthetic effect was evaluated through the corneal touch threshold in a rabbit eye model. The CbCipLid hydrogel obtained is transparent and allows the loading of ciprofloxacin above its solubility at a neutral pH, with a rheology which is convenient for topical administration. Its physical and chemical properties remained unchanged after sterilization and for at least six additional months. Both ciprofloxacin and lidocaine are reversibly released from the CbCipLid hydrogel with a kinetics fitting the Higuchi model. The presence of a biologic-like fluid increased the rate of drug delivery through an ionic exchange mechanism. Treatment with the CbCipLid hydrogel decreased the wound-healing period, compared with the reference, and was associated with a greater number of fibroblasts and a faster rate of epithelialization and dermis reconstruction. These differences were assigned to the moist environment provided by the hydrogel and also to the presence of a therapeutic concentration of ciprofloxacin. Moreover, CbCipLid hydrogel provides an immediate anesthetic effect, which is significantly more intense than that of the reference. Based on these results, it is believed that the CbCipLid hydrogel could be a potential candidate in the prophylaxis/treatment of second-degree burns.

Collagen Fibers in the Healing Process of Rat Achilles Tendon Rupture Using Different Times of Ultrasound Therapy.

Objective: The aim of this study was to evaluate the organization of collagen fibers in the healing process of rat Achilles tendon rupture using different times of ultrasound therapy (TUS). Approach: Forty Wistar rats were selected. Among these, 32 were submitted to total tenotomy of the calcaneous tendon and divided into 5 groups: control group (CG, n = 8), without tenotomy or any treatment; tenotomy group (n = 8), with tenotomy and without treatment; TUS groups-TUS3 (n = 8), TUS4 (n = 8), and TUS5 (n = 8)-submitted to tenotomy and treated with TUS for 3, 4, and 5 min per effective radiating area (ERA), respectively. The animals were sacrificed on the 12th postoperative day. The tendons were surgically removed for analysis of the collagen fiber organization using the birefringence technique (OR, optical retardation). Results: The collagen fibers exhibited better aggregation and organization in the UST3, TUS4, and TUS5 groups compared with CG (p < 0.05). The TUS5 group had better response rates in intergroup comparison. Innovation: The dose response of therapeutic TUS is influenced by many variables. The scientific evidence to support the dosimetry is insufficient. The application time is an important variable to be considered in TUS. In this study, the longer the application time, the better for organization and aggregation of collagen fibers in the rat tendon. Conclusion: TUS applied for 5 min per ERA presented higher dose response to the organization of collagen fibers in the healing process of rat Achilles tendon rupture.

Linear Birefringent Films of Cellulose Nanocrystals Produced by Dip-Coating.

Transparent films of cellulose nanocrystals (CNC) are prepared by dip-coating on glass substrates from aqueous suspensions of hydrolyzed filter paper. Dragging forces acting during films' deposition promote a preferential alignment of the rod-shaped CNC. Films that are 2.8 and 6.0 µm in thickness show retardance effects, as evidenced by placing them between a linearly polarized light source and a linear polarizer sheet in the extinction configuration. Transmission Mueller matrix spectroscopic ellipsometry measurements at normal incidence as a function of sample rotation were used to characterize polarization properties. A differential decomposition of the Mueller matrix reveals linear birefringence as the unique polarization parameter. These results show a promising way for obtaining CNC birefringent films by a simple and controllable method.

Supra-organization and optical anisotropies of the extracellular matrix in the amniotic membrane and limbal stroma before and after explant culture.

In this research we evaluated the supramolecular organizations and the optical anisotropical properties of the de-epithelialized human amniotic membrane and rabbit limbal stroma, before and after explant culture. Birefringence, monochromatic light spectral absorption and linear dichroism of the main extracellular matrix biopolymers, that is, the fibrillar collagens and proteoglycans, were investigated by polarized light microscopy combined with image analysis. Our results demonstrated that the culture procedure-induced stimuli altered the supra-organizational characteristics (in terms of collagens/proteoglycans spatial orientation and ordered-aggregational state) of the amniotic and limbal extracellular matrix, which led to changes in optical anisotropical properties.

Efeito da aplicação do ultrassom terapêutico durante 4 e 5 minutos por área do transdutor no processo de reparação de tendão de ratos / Effect of application times 3, 4 and 5 minutes ERA of therapeutic ultrasound in tendon injury of calcaneal rats

O objetivo deste estudo foi avaliar o efeito dos tempos de aplicação 3, 4 e 5 minutos por ERA do ultrassom terapêutico (UST) na organização das fibras de colágeno em lesão do tendão do calcâneo de ratos. Foram utilizados quarenta ratos machos Wistar, dos quais 32 sofreram tenotomia total do tendão do calcâneo e foram divididos em 5 grupos: GC, sem tenotomia e tratamento; GT, com tenotomia e sem tratamento; UST3, UST4 e UST5 submetidos à tenotomia e tratados com UST nos tempos de 3, 4 e 5 minutos por área de radiação efetiva respectivamente. Os animais foram submetidos à primeira aplicação do UST foi 24 horas após a cirurgia de tenotomia. A irradiação ultrassônica foi aplicada com os seguintes parâmetros: 1 MHz, modo pulsado com 20% do ciclo de trabalho (2 ms de emissão / 8 ms de intervalo), frequência de 100 Hz, 0,5 W / cm² de intensidade e ERA de 0,5 cm². A aplicação foi realizada 1x/dia. Os animais foram sacrificados após a 10ª sessão de tratamento, no 12º dia pós-operatório. Os tendões foram retirados cirurgicamente para análise da organização das fibras colágenas através do método de birrefringência (retardo óptico - OR). As fibras colágenas mostraram melhor agregação e organização no grupo UST3, UST4 e UST5 quando comparado ao GT (p<0.05) e o UST5 apresentou melhor resposta na comparação intergrupos. Conclui-se que o UST, aplicado no tempo de 5 minutos por área de radiação efetiva, apresentou a melhor dose-resposta quanto à organização das fibras colágenas no reparo tecidual de tendões de ratos...

The aim of this study was to evaluate the effect of application times 3, 4 and 5 minutes ERA of therapeutic ultrasound in the organization of the collagen fibers in rat calcaneal tendon injury. Forty male Wistar rats were used, of which 32 underwent complete tenotomy of the calcaneal tendon and were divided into 5 groups: GC without tenotomy and treatment; GT tenotomy with and without treatment; UST3, UST4 UST5 and submitted to tenotomy treated with therapeutic ultrasound at times 3, 4 and 5 minutes per effective radiating area respectively. The animals were submitted to the first application of therapy US tenotomy 24 hours after surgery. Ultrasonic irradiation was applied with the following parameters: 1 MHz, pulsed mode at 20% duty cycle (2ms transmission / 8 ms interval), frequency 100 Hz, 0.5 W / cm² intensity and ERA 0.5 cm². The application was performed 1x / day. The animals were sacrificed after the 10th treatment session, on the 12th postoperative day. The tendons were surgically removed for analysis of the organization of the collagen fibers through birefringence method (optical delay - OR). The collagen fibers showed better aggregation and organization in group UST3, UST4 and UST5 when compared to the GT (p <0.05) and UST5 showed better response in the intergroup comparison. We conclude that the UST, applied in time of 5 minutes for effective radiation area, presented the best dose-response as the organization of the collagen fibers in tissue repair of rat tendons...

Optical anisotropy reveals molecular order in a mouse enthesis.

Entheses are specialized biological structures that functionally anchor tendons to bones. The complexity, mechanical characteristics and properties of the entheses, particularly those related to exercise, mechanical load and pathologies, have been extensively analyzed; however, the macromolecular organization of the enthesis fibers, as assessed by polarization microscopy, has not yet been investigated. Morphological and optical anisotropy characteristics, such as birefringence, linear dichroism (LD) and differential interference contrast (DIC-PLM) properties, are thus analyzed in this study of a healthy adult mouse calcaneal tendon-bone enthesis. The molecular and supramolecular order of collagen and GAGs was determined for the collagen bundles of this enthesis. Based on a birefringence plot pattern as well as on metachromasy and linear dichroism after toluidine blue staining at pH 4.0, a similarity between the calcaneal tendon-bone enthesis and cartilage during ossification may be assumed. This similarity is assumed to favor the adequacy of this enthesis to support a compressive load. Considering that the collagen-proteoglycan complexes and the enthesis fibers themselves have a chiral nature, these structures could be acting via reciprocal signaling with the cellular environment of the enthesis.

Influencia de un enjuague a base de fluoruro y xilitol en la remineralización in vitro del esmalte en dientes temporales / Influence exerted by a xylitol and fluoride based mouthwash on the in vitro enamel remineralization of primary teeth

El propósito del estudio fue evaluar la eficacia de un enjuague bucal con fluoruro de sodio y xilitol, en la remineralización del esmalte de dientes temporales. Se utilizaron 40 dientes sin caries, aplicando ácido fosfórico al 35% durante 20 segundos, y fueron inmersos en el enjuague por 0, 15, 30, 45 y 60 días. Se observaron cortes longitudinales de 150-250 µm de cada muestra y se valoró la remineralización, de acuerdo a la birrefringencia observada después de aplicarles solución de Thoulet (1.47 IR) bajo luz polarizada en un fotomicroscopio. A los 15 días tuvo una media de 0.444 (± 0.527), a los 30 días de 0.778 (± 0.441), a los 45 de 1.444 (± 0.527) y a los 60 días de 1.47 (± 0.483). El análisis de varianza permitió establecer diferencias estadísticamente significativas entre los grupos (p < 0.001), y comparándolos entre sí (p < 0.05). Puede concluirse que el enjuague empleado tuvo un ligero efecto remineralizante en el esmalte.

The purpose of the present study was to assess the effectiveness on primary teeth of a fluoride and xylitol based mouthwash. 40 caries-free teeth were used. 35% phosphoric acid was applied during 20 seconds. Teeth were then immersed in the mouthwash for 0, 15, 30, 45 and 60 days. 150-250μm longitudinal slices were taken of each sample. Re-mineralization was assessed according to bi-refringence observed after applying Thoulet solution (1.47 IR). Assessment was conducted under polarized light in a photo-microscope. At 15 days, a mean of 0.444 (± 0.527) was observed. After 30 days the observed mean was 0.778 (± 0.441). At 45 days, observed mean was 1.444 (± 0.527), and at 60 days, observed mean was 1.47 (± 0.483). Variance analysis established statistically significant differences among groups (p < 0.001) as well as when comparisons among groups were established (p < 0.05). After conducting the aforementioned tests it could be concluded that the employed mouthwash exerted a slight re-mineralizing effect upon the enamel of treated teeth.

Identificación de colágeno I y III con picrosirius red/ polarización en el estroma de tumores salivales / Identification of type I and III collagen by picrosirius red/polarization of tumoral salivary stroma

El estroma juega un rol importante en los procesos tumorales de invasión y metástasis. Las fibras de colágeno tipo I son el principal componente estructural del estroma en distintos tumores. Sin embargo, hay muy pocos estudios en los tumores de glándulas salivales. Basándonos en estos antecedentes el objetivo de la presente comunicación fue estudiar las características del colágeno con Picrosirius red/polarización en tumores benignos y malignos de glándulas salivales para evaluar su posible rol en los mecanismos de progresión tumoral. Cortes histológicos de adenoma pleomórfico, carcinoma adenoide quístico y carcinoma epitelial mioepitelial se colorearon con H/E y Picrosirius red y se examinaron con microscopio de polarización. La birrefringencia del colágeno con Picrosirius/polarización resultó diferente en el estroma de los tumores malignos (carcinoma adenoide quístico y carcinoma epitelial mioepitelial), con predominio de colágeno I, en comparación con el tumor benigno (adenoma pleomórfico), con predominio de colágeno III. El diferente perfil de coloración en las fibras colágenas producidas en el estroma de los tumores analizados podría relacionarse con diferentes mecanismos de expansión tumoral, los que fueron poco estudiados en los tumores de glándulas salivales. Más estudios son necesarios para obtener resultados más concluyentes que contribuyan al diagnóstico, pronóstico y tratamiento.

The stroma plays an important rol in tumoral invasion and metastasis. Type I collagen is the main structural component of the stroma in several tumors. However, there are few studies on salivary gland tumors. Based on this background the objective of the present communication was to study collagen characteristics with picrosirius red/polarization on malignant and benign tumors of salivary glands to evaluate its posible rol in the tumoral progression mechanism. Histological sections of pleomorphic adenoma, adenoid cystic carcinoma and epithelial/myoepithelial carcinoma were stained with H/E and picrosirius red and were studied with polarization microscope. Collagen birefringence with Picrosirius/polarization was different in the malignant tumor stroma (adenoid cystic carcinoma and epithelialmyoepithelial carcinoma), with predominance of type I collagen, compared with a benign tumor (pleomorphic adenoma), with predominance of type III collagen. The different staining profile in collagen fibers produced in the benign and malignant stroma tumors analized could be related with different tumoral expansion mechanism, which were scarce studied on the salivary glands tumors. More studies are needed to obtain more conclusive results to contribute to diagnosis, prognosis and treatment.

Magnetic field measurements based on Terfenol coated photonic crystal fibers.

A magnetic field sensor based on the integration of a high birefringence photonic crystal fiber and a composite material made of Terfenol particles and an epoxy resin is proposed. An in-fiber modal interferometer is assembled by evenly exciting both eigenemodes of the HiBi fiber. Changes in the cavity length as well as the effective refractive index are induced by exposing the sensor head to magnetic fields. The magnetic field sensor has a sensitivity of 0.006 (nm/mT) over a range from 0 to 300 mT with a resolution about ±1 mT. A fiber Bragg grating magnetic field sensor is also fabricated and employed to characterize the response of Terfenol composite to the magnetic field.

Hydrostatic pressure sensing with high birefringence photonic crystal fibers.

The effect of hydrostatic pressure on the waveguiding properties of high birefringence photonic crystal fibers (HiBi PCF) is evaluated both numerically and experimentally. A fiber design presenting form birefringence induced by two enlarged holes in the innermost ring defining the fiber core is investigated. Numerical results show that modal sensitivity to the applied pressure depends on the diameters of the holes, and can be tailored by independently varying the sizes of the large or small holes. Numerical and experimental results are compared showing excellent agreement. A hydrostatic pressure sensor is proposed and demonstrated using an in-fiber modal interferometer where the two orthogonally polarized modes of a HiBi PCF generate fringes over the optical spectrum of a broad band source. From the analysis of experimental results, it is concluded that, in principle, an operating limit of 92 MPa in pressure could be achieved with 0.0003% of full scale resolution.

Influência de diferentes comprimentos de onda da laserterapia de baixa intensidade na regeneração tendínea do rato após tenotomia / Influence of different low-intensity laser therapy wavelengths for rat tendon regeneration following tenotomy

OBJETIVO: Comparar a influência da terapia laser baixa intensidade (TLBI) na organização das fibras de colágeno, na dose de 3J/cm² em dois comprimentos de onda (lambda) (670nm e 904nm) e na associação de ambos, durante o reparo do tendão calcâneo de ratos. MÉTODO Foram utilizadas 37 ratas da raça Wistar divididas em 5 grupos, submetidas a tenotomia total por meio de cisalhamento transversal do tendão na porção média entre a inserção calcânea e a junção miotendínea, com exceção do grupo GP (n= 6) que não houve lesão. O grupo GA (n= 8) foi submetido à irradiação laser lambda= 904nm, diodo de GaAs, emissão pulsada, duração do pulso de 180ns, freqüência de pulso de 2000Hz e potência de pico de 15W e dose de 3J/cm². O grupo GB (n= 8) foi submetido à irradiação laser lambda= 670nm, diodo de AlGaInP, emissão contínua, potência 30mW e dose de 3J/cm². O grupo GAB (n= 8) recebeu irradiação de ambos os lasers 670nm e 904nm associados. O grupo GCL (n= 7) foi submetido a um tratamento placebo, assim como um grupo padrão. Após um total de 12 sessões de laserterapia, os animais foram submetidos à eutanásia no 14º dia pós-operatório. Após processamento histológico padrão, foi realizada a quantificação do retardo óptico pela birrefringência. RESULTADOS: A análise estatística mostra que todos os grupos irradiados apresentaram valores de retardo óptico maiores quando comparados ao grupo placebo (p= 0,00001), o que indica que apresentaram melhor organização das fibras colágenas ao longo do eixo longitudinal do tendão, evidenciando melhor qualidade no reparo. O grupo GAB foi o que apresentou melhores resultados (p= 0,00001) quando comparado aos demais grupos, excetuando-se o GP, que é o referencial de retardo óptico para esta faixa etária.

OBJECTIVE: The aim of this study was to compare the effect of the low level laser therapy in the organization of the collagen fibers with 3 J/cm² of dose in two wavelengths (670nm and 904nm) and in the combination of both on the tissue repair of the calcaneus tendon. METHODS: We used 37 female Wistar rats divided in 5 groups which had been submitted to the total tenotomy. Group GA (n= 8) was submitted to the laser irradiation in lambda= 904nm, GaAS diode, pulsated emission, peak power 15W and dose of 3J/cm², group GB (n= 8) was submitted to laser irradiation lambda= 670nm AlGaInP diode, continuous emission, power 30 mW and dose 3J/cm², group GAB (n= 8) was submitted to association of both of lasers and group GCL (n= 7) which the calcaneum tendon was submitted to placebo treatment. The group GP (n= 6) was a standard group without lesion and treatment. The animals received 12 sessions of lasertherapy and submitted to euthanasia on 14th day after surgery. The tendons were submitted to histological procedure and were analyzed to evaluate the optic retardation (in nm) for the birefringence measurement. RESULTS: The statistic analysis showed that the irradiated groups have higher values of optic retardation statistically significant than placebo group (p= 0,00001). It indicates that all the treated groups show better organization of collagen fibers in the longitudinal axis of the tendon, evidencing better quality in the tissue repair. The group GAB was that shown best results (p= 0,00001) when compared to another groups.