Exercício resistido promove reparo no musculoesquelético em modelo experimental de artrite reumatoide / Resisted exercise promotes repair in musculoskeletal structure in experimental model of rheumatoid arthritis

Distúrbios decorrentes da artrite reumatoide (AR) resultam em incapacitação funcional, com reflexos econômicos e sociais aos sistemas de saúde. Uma alternativa terapêutica que tem sido proposta é a atividade física na modalidade resistida. O presente estudo analisou os efeitos do exercício resistido de subida em escada (ERSE) na histomorfologia do músculo extensor longo dos dedos (EDL) de ratos Wistar, em modelo experimental de AR. Ratos machos (n=20) foram distribuídos aleatoriamente em quatro grupos. A lesão da AR foi induzida por administração intra-articular de Adjuvante Completo de Freund (CFA). Os resultados revelaram que a AR alterou a histomorfometria das fibras do EDL e que o ERSE promoveu reparo muscular, sugerindo sua eficiência na restauração da funcionalidade muscular. Ainda, o ERSE pode ser uma opção de tratamento voltado à melhoria na qualidade de vida dos portadores de AR.

Disorders from rheumatoid arthritis (RA) result in functional disability, with economic and social impacts on health systems. A therapeutic alternative that has been proposed is physical activity in the resisted modality. Current study analyzed the effects of resistance exercise on stairs (ERSE) on the histomorphology of the extensor digitorum longus muscle (EDL) of Wistar rats in an experimental model of RA. Male rats (n=20) were randomly assigned to four groups. RA injury was induced by intra-articular administration of Freund's Complete Adjuvant (CFA). Results revealed that RA altered the histomorphometry of EDL fibers and that ERSE promoted muscle repair, suggesting its efficiency in restoring muscle functionality. Furthermore, ERSE may be a treatment option to improve the life quality of people with RA.

Split-Block Graft Versus Cortico-cancellous Block Graft for Horizontal Ridge Augmentation: Cone Beam Computed Tomography and Histomorphometric Study / Enxerto de bloco dividido versus enxerto de bloco córtico-esponjoso para aumento horizontal do rebordo: imagens de tomografia computadorizada de feixe cônico e estudo histomorfométrico

Objective: The aim of this study was to compare the horizontal dimensional changes of split-bone block and cortico-cancellous block graft in horizontal ridge augmentation using Cone Beam Computed Tomography (CBCT). The quality of the regenerated bone in both groups was compared histologically and histomorphometrically. Material and methods: Twenty patients were randomly divided in two equal groups (n= 10): Split-bone block group which was harvested from the external oblique ridge or cortico-cancellous block graft group which was harvested from the mandibular symphysis. Pre-augmentation crestal ridge width was measured using bone caliper. CBCT scans were taken on the 2nd week and 4th month post-operatively to measure crestal and total horizontal ridge dimensions. A biopsy was collected from the regenerated ridge immediately before implant insertion on the 4thmonth post-operatively. Results: Pre-augmentation crestal bone widths of both groups were comparable (P= 0.870). On the 2nd week and 4th month post-operatively, split-bone block showed a significantly wider crestal (P= 0.028 and P= 0.001 respectively) and total horizontal ridge dimension (P= 0.025 and P= 0.002 respectively), and on the 4th month post-operatively, it showed significantly lesser resorption at crest (P= 0.040) and in total horizontal ridge dimension (P= 0.017) than cortico-cancellous block. Histologically, the regenerated bone quality was similar in both groups. Histomorphometric analysis showed a non-significant difference in percentage of mature (P= 0.365) and immature collagen (P= 0.531) between both groups. Conclusion: Split-bone block maintained a significantly wider ridge and experienced less resorption after 4 months than the cortico-cancellous block graft, with no difference in regenerated bone quality between both groups. (AU)

Objetivo: O objetivo deste estudo foi comparar as mudanças dimensionais horizontais do enxerto de bloco ósseo dividido e do enxerto de bloco córtico-esponjoso no aumento de rebordo horizontal por meio da Tomografia computadorizada de feixe cônico (TCFC). A qualidade do osso regenerado em ambos os grupos foi comparada histológica e histomorfometricamente. Material e Métodos:Vinte pacientes foram divididos aleatoriamente em dois grupos iguais (n=10): Grupo de blocos de osso dividido retirado da crista oblíqua externa ou Grupo de enxerto em bloco córtico-esponjoso retirado da sínfise mandibular. A largura da crista pré-aumentada foi medida usando calibrador ósseo. Os exames de TCFC foram realizados na segunda semana e quarto mês de pós operatório para medir as dimensões da crista e dimensões horizontais totais do rebordo. Uma biopsia foi coletada da crista regenerada imediatamente antes da inserção do implante no quarto mês pós-operatório. Resultados: As larguras do osso crestal pré-aumentado de ambos os grupos eram compatíveis (p = 0,870). Na segunda semana e quarto mês pós-operatórios, o bloco ósseo dividido mostrou uma crista significativamente mais larga (p = 0,28 e p = 0,001 respectivamente) e a dimensão da crista horizontal total (p = 0,025 e p = 0,002 respectivamente), e no quarto mês pós-operatório, apresentou reabsorção significativamente menor na crista (p = 0,040) e na dimensão da crista horizontal total (p = 0,017) em comparação ao bloco córtico-esponjoso. Histologicamente, a qualidade do osso regenerado foi semelhante em ambos os grupos. A análise histomorfométrica mostrou uma diferença não significativa na porcentagem de colágeno maduro (p = 0.365) e colágeno imaturo (p = 0,531) entre ambos os grupos. Conclusão: O bloco ósseo dividido manteve uma crista significativamente mais larga e apresentou menos reabsorção após 4 meses, em comparação ao enxerto em bloco córtico-esponjoso, sem diferença na qualidade do osso regenerado entre ambos os grupos. (AU)

Estudio histomorfométrico de folículos tiroideos humanos con microscopía holográfica digital / Histomorphometric study of human thyroid follicles by digital holographic microscopy

Introducción: La microscopía holográfica digital ha permitido a la microscopía óptica hacer uso de herramientas numéricas y computacionales; y esto, a su vez, ha favorecido múltiples avances en el estudio de las células y los tejidos en diferentes campos de la medicina y otras ciencias afines. Objetivo: Describir las características histológicas y morfométricas de los folículos tiroideos humanos con la microscopía holográfica digital. Métodos: Se realizó, desde el punto de vista histomorfométrico, un estudio descriptivo y transversal de folículos tiroideos humanos utilizando una instalación de microscopía holográfica digital. Se empleó la técnica de inclusión en parafina y tinción de hematoxilina-eosina para el procesamiento de las muestras. Se realizaron de 10 a 12 capturas de hologramas por muestra y el método de doble propagación para la reconstrucción de los hologramas. Se calculó el área, el perímetro, el diámetro mayor y menor de los folículos y cavidades foliculares y se realizaron reconstrucciones de imágenes holográficas en tres dimensiones. Se determinó como medida de tendencia central la media aritmética y como medida de dispersión la desviación típica o estándar. Resultados: Parámetros foliculares: área (5140,31 ± 1126,71 µm2); perímetro (2961,54 ± 71,2 µm); diámetro mayor:(921,17 ± 24,34 µm); diámetro menor: (746,67 ± 18,08 µm); altura del epitelio (7,92 ± 0,96). Cavidades foliculares: área (3686,18 ±1023,52 µm2); diámetro mayor: (698,86 ± 19,55 µm) y diámetro menor: (581,15 ± 13,82 µm). Conclusiones: Existen parámetros foliculares, determinados mediante la microscopía holográfica digital, no reportados por la literatura consultada, que resultan de interés en el estudio histológico de los folículos tiroideos humanos(AU)

Introduction: Digital holographic microscopy has made it possible to incorporate the use of numerical and computer tools into optical microscopy. This in turn has led to great progress in the study of cells and tissues in several fields of medicine and related sciences. Objective: Describe the histological and morphometric characteristics of human thyroid follicles using digital holographic microscopy. Methods: A descriptive cross-sectional histomorphometric study was conducted of human thyroid follicles using a digital holographic microscopy facility. Sample processing was based on inclusion technique by paraffin and hematoxylin-eosin staining. Ten to twelve holographic captures were made per sample, and the double propagation method was used for holographic reconstruction. Estimation was carried out of the area, perimeter, and greatest and smallest diameter of follicles and follicular cavities, and tri-dimensional reconstructions were made of holographic images. Arithmetic mean was determined as the measure of central tendency, and typical or standard deviation as the measure of dispersion. Results: Follicular parameters: area (5 140.31 ± 1 126.71 µm2); perimeter (2 961.54 ± 71.2 µm); greatest diameter (921.17 ± 24.34 µm); smallest diameter (746.67 ± 18.08 µm); epithelial height (7.92 ± 0.96). Follicular cavities: area (3 686.18 ± 1 023.52 µm2); greatest diameter (698.86 ± 19.55 µm); smallest diameter (581.15 ± 13.82 µm). Conclusions: A number of follicular parameters determined by digital holographic microscopy have not been reported by the literature consulted, and they are of interest to the histological study of human thyroid follicles(AU)

Histomorphometric evaluation of the effects of mandibular advancement appliance and low level laser therapy (LLLT) with different doses on condylar cartilage and subchondral bone in rats / Evaluación histomorfométrica de los efectos del aparato de avance mandibular y la terapia con láser de bajo nivel (TLBN) con diferentes dosis en cartílago condilar y hueso subcondral en ratas

The aim of this study was to evaluate the effects of mandibular advancement appliance and low level laser therapy (LLLT) with different doses on cellular hypertrophic changes in the mandibular condyle of rats. Forty-eight 8-week-old male Wistar albino rats weighing between 260 and 280 g were randomly divided into four experimental and control groups. Group I was the control group; group II was the mandibular advancement appliance group; group III was the 8 J/cm2 (0.25 W, 20 s) laser irradiation with mandibular advancement appliance group; and group IV was the 10 J/cm2 (0.25 W, 25 s) laser irradiation with mandibular advancement appliance group. Mandibular condyle cartilage and subchondral bone changes with different LLLT dose and mandibular advancement appliance were evaluated by histomorphometrical analysis. Subchondral bone fraction results showed that there were no significant differences between groups (p<0.05). The statistically significant differences found between control group and experimental groups in anterior and posterior cartilage layers thickness (p<0.05) and (p<0.01). Posterior and anterior condylar cartilage layers of rats react differentially to LLLT and mandibular advancement application. Maximum changes in condylar cartilage layers were found in 8 J/cm2 laser irradiation with mandibular appliance group.

El objetivo de este estudio fue evaluar los efectos del aparato de avance mandibular y la terapia con láser de bajo nivel (TLBN) con diferentes dosis sobre los cambios hipertróficos celulares, en el cóndilo mandibular de ratas. Cuarenta y ocho ratas albinas macho Wistar de 8 semanas de edad con un peso de 260 y 280 g se dividieron aleatoriamente en cuatro grupos experimentales y control. El grupo I control; grupo II, dispositivos de avance mandibular; grupo III de irradiación con láser de 8 J / cm2 (0.25 W, 20 s) con el grupo dispositivos de avance mandibular; y grupo IV con irradiación láser de 10 J / cm2 (0,25 W, 25 s) con el grupo de dispositivos de avance mandibular. El cartílago del cóndilo mandibular y los cambios en el hueso subcondral con diferentes dosis de TLBN y dispositivo de avance mandibular, se evaluaron mediante análisis histomorfométrico. Los resultados de la fracción ósea subcondral indicaron que no hubo diferencias significativas entre los grupos (p <0,05). Las diferencias estadísticamente significativas encontradas entre el grupo control y los grupos experimentales, en el grosor del cartílago anterior y posterior (p<0,05) y (p<0,01). Las capas de cartílago condilar posterior y anterior de las ratas reaccionan de manera diferencial a la aplicación de TLBN y avance mandibular. Se encontraron cambios significativos en las capas de cartílago condilar con irradiación láser de 8 J /cm2 con el grupo de dispositivos mandibulares.

Histomorphometry of human vermiform appendix

Background: Acute Appendicitis is one of the most common acute abdominal conditions. Advances in modern radiographic imaging have improved diagnostic accuracy, however the diagnosis of appendicitis remains essentially an enigmatic challenege. Though much work has been done on morphometry but there is less work done on variation of histomorphometric features of appendix. Hence the study was carried out by keeping the following objectives in mind. Aim was to study the general histomorphology and measure the histomorphometric parameters of human vermiform appendix. And also, to study diagnostic significance of histomorphology and histomorphometric parameters in causing appendicitis.Methods: The study was conducted in the Department of Anatomy. Specimens of appendix were obtained from embalmed cadavers. Serial sections were taken at different levels and slides were prepared. The slides were then examined using Nikon Trinocular Research microscope under magnification power of 40x for various parameters, after staining with H&E.Results: The mean luminal diameter varied from 1.32±0.65mm at base to 1.22±0.72mm from base. Diffuse lymphatic tissue was seen in both mucosa and sub-mucosa.Conclusions: Definitely there is a relationship between lymphoid follicle diameter and mucosal-serosal thickness on one hand and that between the luminal diameter and lymphoid follicle diameter on the other hand. Since our study was restricted to geriatric age group so studies need to be done in different age groups to highlight any further relationship.

Histomorphometric reference data of transiliac bone biopsy in children from 8 to 17 years old / Datos histomorfométricos de referencia de la biopsia ósea en niños de 8 a 17 años

Abstract: Background: Histomorphometric analysis of bone samples is a key tool for studying bone metabolism; however, only a few pediatric reference data exist. The aim of the present study is to report more reference data and to investigate if histomorphometric differences exist between age and gender. Methods: We obtained 19 transiliac bone samples previously marked with tetracycline, from children between 8 and 17 years (13 were male), with normal blood test results and urine biochemical bone markers. We evaluated bone histomorphometric parameters using a digitalizing table with osteomeasure to obtain normative data of means and standard deviations, as well as median and range. Due to the small sample, a Monte Carlo simulation was applied. Structural, static, dynamic, and resorptic histomorphometric parameters were evaluated by age and gender following the American Society for Bone and Mineral Research recommendations. Results: Bone volume (in the older children) and mineral apposition rate (in the younger children), the eroded surface (in boys), and the new bone wall thickness (in girls) were significantly increased. On the trabecular area of mineralization front, the modeling and the remodeling bone formation were similar (16 and 18%). The rest of the histomorphometric bone parameters by age and gender showed no significant difference. Conclusion: In healthy children, these bone histomorphometric findings, with these techniques and for this ages could be used as reference values.

Resumen: Introducción: El análisis histomorfométrico del tejido óseo para el estudio de las enfermedades metabólicas óseas, cuando se correlacionan los hallazgos clínicos, sigue siendo la herramienta con mayor sensibilidad y especificidad para la mayoría de los diagnósticos. En los niños existen pocos reportes histomorfométricos del tejido óseo metabólico normal, por lo que nuestro propósito es reportar más datos de referencia e investigar si hay diferencias histomorfométricas entre edades y sexos. Métodos: Estudio realizado en 19 niños de 8 a 17 años (13 masculinos) sin anomalías clínicas ni bioquímicas evidentes. Se tomaron muestras de tejido óseo transilíaco marcadas con tetraciclina. Se obtuvieron medias, desviaciones y rangos histomorfométricos totales, y correlación por edad y sexo, siguiendo las recomendaciones para la histomorfometría de la American Society for Bone and Mineral Research. Se realizó una simulación Montecarlo. Resultados: El volumen óseo (en niños mayores), la velocidad de agregación del mineral (en niños menores), la erosión trabecular periférica (en niños) y el grosor de la pared ósea nueva (en niñas) exhibieron aumentos significativos. En el área trabecular del frente de mineralización, el modelado y el remodelado de la formación ósea fueron similares (16 y 18%). El resto de los parámetros histomorfométricos óseos no mostraron diferencias significativas. Conclusiones: Estos hallazgos histomorfométricos del tejido óseo de niños normales con estas técnicas y para estas edades pueden ser utilizados como valores de referencia.

Histomorphometric study of rabbit's maxillary sinus augmentation with various graft materials / 대한해부학회지

The purpose of this animal study is to evaluate, by histomorphometric analysis, bone regeneration in rabbit's maxillary sinuses with blood clots alone, Bio-Oss, β-tricalcium phosphate (β-TCP), and demineralized tooth dentin (DTD) grafting. Bilateral sinus augmentation procedures were performed in 18 adult male rabbits. Rectangular replaceable bony windows were made with a piezoelectric thin saw insert. In the group 1, blood clots were filled; group 2, anorganic bovine graft (Bio-Oss) was grafted; group 3, β-TCP was grafted; group 4, DTD was grafted, and covered by replaceable bony windows. Animals were sacrificed at 2, 4, and 8 weeks after surgical procedure. The augmented sinuses were evaluated by histomorphometric analysis using hematoxylin and eosin and Masson's trichrome stains. Histologically, new bone formation was revealed along the elevated sinus membrane and all graft materials. The new bone area of the group 2 was significantly greater than the group 1, and of the group 3 was significantly greater than the group 2, and of the group 4 was significantly greater than the group 3 at 8 weeks with P < 0.05. The bone marrow area of group 1 was significantly greater than other groups at 8 weeks. The DTD area was significantly lesser than Bio-Oss or β-TCP particles area at 8 weeks. This present study suggests that DTD can be effective graft materials for bone regeneration of the maxillary sinus augmentation.

Alveolar bone repair with strontium- containing nanostructured carbonated hydroxyapatite

ABSTRACT Objective: This study aimed to evaluate bone repair in rat dental sockets after implanting nanostructured carbonated hydroxyapatite/sodium alginate (CHA) and nanostructured carbonated hydroxyapatite/sodium alginate containing 5% strontium microspheres (SrCHA) as bone substitute materials. Methods: Twenty male Wistar rats were randomly divided into two experimental groups: CHA and SrCHA (n=5/period/group). After one and 6 weeks of extraction of the right maxillary central incisor and biomaterial implantation, 5 μm bone blocks were obtained for histomorphometric evaluation. The parameters evaluated were remaining biomaterial, loose connective tissue and newly formed bone in a standard area. Statistical analysis was performed by Mann-Withney and and Wilcoxon tests at 95% level of significance. Results: The histomorphometric results showed that the microspheres showed similar fragmentation and bio-absorbation (p>0.05). We observed the formation of new bones in both groups during the same experimental periods; however, the new bone formation differed significantly between the weeks 1 and 6 (p=0.0039) in both groups. Conclusion: The CHA and SrCHA biomaterials were biocompatible, osteoconductive and bioabsorbable, indicating their great potential for clinical use as bone substitutes.

Estudio histomorfométrico de la corteza cerebelosa con microscopia holográfica digital / Histomorphometric study of the cerebellar cortex with digital holographic microscopy

Se realizó un estudio descriptivo y transversal de la corteza cerebelosa humana, desde el punto de vista histomorfométrico, desde enero hasta septiembre del 2015, con el empleo de la microscopia holográfica digital instalada en el Departamento de Holografía Digital de la Universidad de Oriente en Santiago de Cuba, con vistas efectuar mediciones que permitieran establecer comparaciones con otros estudios. Los cálculos mostraron el grosor de las capas molecular y granulosa, el área, el perímetro, los diámetros mayores y menores del cuerpo y el núcleo de las células de Purkinje. Asimismo, se tomaron imágenes holográficas en tres dimensiones, que posibilitaron concluir la existencia de parámetros determinados mediante este procedimiento, los que no habían sido notificados y que resultan de interés en el estudio histológico de la corteza cerebelosa

A descriptive and cross-sectional study of the human cerebellar cortex, from the histomorphometric point of view, was carried out from January to September, 2015, using the digital holographic microscopy installed in the Digital Holography Department of Oriente University in Santiago de Cuba, aimed at making measurings that allowed to establish comparisons with other studies. The calculations showed the thickness of the molecular and granular layers, the area, perimeter, greatest and smallest diameters of the body and Purkinje cells nucleus. Also, holographic images in three dimensions were taken, that facilitated to conclude the existence of certain parameters by means of this procedure, those that had not been notified and are of interest in the histological study of the cerebellar cortex

Accelerated Healing Period with Hydrophilic Implant Placed in Sheep Tibia

Abstract The objective of this study was to evaluate the early osseointegration of two different implants surfaces, a sandblasted and acid-etched surface (TN) compared with same geometry and surface roughness modified to be hydrophilic/wettable by conditioning in an isotonic solution of 0.9% sodium-chloride (TA) through histological and histomorphometric analysis after sheep tibia implantation. Forty dental implants, divided in two groups (TN and TA) were placed in the left tibia of twenty healthy, skeletally mature Santa Ines sheep (n=5/experimental period). After 7, 14, 21 and 28 days post-implantation, the samples were removed and the sheep were kept alive. Analysis of resin sections (30 μm) allowed the quantification of bone area (BA) and bone-to-implant contact (BIC). TA group presented nearly 50% increase in BA at 14 days (p<0.001, ANOVA - Tukey's post test) compared with 7 days. The TA presented higher values than the TN for BA and BIC at 14, 21, and 28 days after placement, stabilizing bone healing. TA hydrophilic surface promoted early osseointegration at 14 and 21 days compared to TN, accelerating bone healing period post-implant placement in sheep tibia.

Resumo O objetivo deste estudo foi avaliar a osseointegração precoce de duas superficies diferentes de implantes, uma superfície tratada com jateamento e ataque ácido (TN) comparada com outra superfície com a mesma geometria e rugosidade e modificada com hidrofilicidade / molhabilidade através do condicionamento em solução isotônica de 0,9% de cloreto de sódio (TA) através de análises histológica e histomorfométrica após a implantação em tíbia de ovelhas. Quarenta implantes dentários, divididos em dois grupos (TN e TA), foram implantados na tíbia esquerda de vinte ovelhas Santa Inês (n=5/período experimental) saudáveis e esqueleticamente maduras. Após 7, 14, 21 e 28 dias pós-implantação, as amostras foram coletadas e as ovelhas foram mantidas vivas. A análise dos cortes em resina (30 μm) permitiu a quantificação da área de osso (BA) e do contato osso-implante (BIC). O grupo TA apresentou aumento de quase 50% na BA aos 14 dias (p <0,001, ANOVA - pós teste de Tukey) em comparação com 7 dias. O grupo TA apresentou valores maiores do que o TN para BA e BIC aos 14, 21 e 28 dias após a implantação, estabilizando a cicatrização óssea. A superfície hidrofílica TA promoveu uma osseointegração precoce aos 14 e 21 dias em relação à TN, acelerando o período de cicatrização óssea após a instalação dos implantes em tíbias de ovelha.

Physico-chemical and Histomorphometric Evaluation of Zinc-containing Hydroxyapatite in Rabbits Calvaria

Abstract The aim of this study was to characterize the physico-chemical properties and bone repair after implantation of zinc-containing nanostructured porous hydroxyapatite scaffold (nZnHA) in rabbits' calvaria. nZnHA powder containing 2% wt/wt zinc and stoichiometric nanostructured porous hydroxyapatite (nHA - control group) were shaped into disc (8 mm) and calcined at 550 °C. Two surgical defects were created in the calvaria of six rabbits (nZnHA and nHA). After 12 weeks, the animals were euthanized and the grafted area was removed, fixed in 10% formalin with 0.1 M phosphate buffered saline and embedded in paraffin (n=10) for histomorphometric evaluation. In addition, one sample from each group (n=2) was embedded in methylmethacrylate for the SEM and EDS analyses. The thermal treatment transformed the nZnHA disc into a biphasic implant composed of Zn-containing HA and Zn-containing β-tricalcium phosphate (ZnHA/βZnTCP). The XRD patterns for the nHA disc were highly crystalline compared to the ZnHA disc. Histological analysis revealed that both materials were biologically compatible and promoted osteoconduction. X-ray fluorescence and MEV-EDS of nZnHA confirmed zinc in the samples. Histomorphometric evaluation revealed the presence of new bone formation in both frameworks but without statistically significant differences (p>0.05), based on the Wilcoxon test. The current study confirmed that both biomaterials improve bone repair, are biocompatible and osteoconductive, and that zinc (2wt%) did not increase the bone repair. Additional in vivo studies are required to investigate the effect of doping hydroxyapatite with a higher Zn concentration.

Resumo O objetivo deste estudo foi caracterizar físico-químicamente e avaliar o reparo ósseo de discos de hidroxiapatita porosa contendo zinco após a implantação em calvária de coelhos (nZnHA). O pó de nZnHA e o pó hidroxiapatita porosa estequiométrica nanoestruturada (controle - nHA) foram confeccionados em discos (8 mm) e calcinados a 550˚C. Dois defeitos cirúrgicos foram criados na calvária de seis coelhos para a implantação dos discos. Após 12 semanas, os animais foram eutanasiados e as áreas enxertadas foram removidas, fixadas em formol a 10% e embebidas em parafina (n=10) para avaliação histomorfométrica. Além disso, uma amostra de cada grupo (n=2) foi embebida em metilmetacrilato para análise de MEV e EDS. O tratamento térmico dos discos de nZnHA transformou-os em implantes bifásicos compostos por HA contendo Zinco e β fosfato tricálcico com Zinco (ZnHA/βZnTCP). Os discos de nHA, apresentaram-se altamente cristalinos e com baixa solubilidade quando comparados aos discos de ZnHA. A análise histológica revelou que ambos os materiais foram biologicamente compatíveis e promoveram a osteocondução. As análises de FRX e MEV-EDS confirmaram a presença do zinco nas amostras de nZnHA. A avaliação histomorfométrica revelou a presença de neoformação óssea em ambos os grupos, porém sem diferenças estatísticas entre eles, com base no teste de Wilcoxon (p>0,05). O presente estudo confirmou que ambos os biomateriais otimizaram o reparo ósseo, foram biocompatíveis e osseocondutivos e a presença do zinco não favoreceu o reparo ósseo. Estudos adicionais in vivo devem ser conduzidos a fim de investigar o efeito de maiores concentrações de zinco.

Utilidad de la microscopia holográfica digital para el estudio histomorfométrico de eritrocitos humanos / Usefulness of the digital holographic microscopy for the histomorphometric study of human red blood cells

Se realizó una investigación descriptiva y transversal para determinar la utilidad de la microscopia holográfica digital en el estudio histomorfométrico de eritrocitos humanos, mediante el análisis comparativo con la microscopia óptica, en el Departamento de Holografía Digital de la Universidad de Oriente de Santiago de Cuba, de abril del 2014 a igual mes del 2015. A tal efecto se seleccionaron muestras de eritrocitos diluidos en las soluciones amortiguadoras Hepes y Hepa, se emplearon el programa Holodig® y el sistema Matlab® para las mediciones en la técnica holográfica, y en el procesamiento estadístico se aplicó la prueba de la t de Student para la comparación de medias entre ambas técnicas microscópicas. Se obtuvo que la microscopia holográfica digital permitiera observar los límites y contornos de los eritrocitos estudiados y las mínimas variaciones en su forma, además de realizar estudios morfométricos, cuyos resultados mostraron similitud a los obtenidos con la microscopia óptica, y de otros parámetros, como el volumen

A descriptive and cross-sectional investigation to determine the usefulness of the digital holographic microscopy in the histomorphometric study of human red blood cells, by means of the comparative analysis with the optic microscopy, was carried out in the Digital Holography Department of Oriente University in Santiago de Cuba, from April, 2014 to the same month in 2015. To such an effect, red blood cells samples diluted in the Hepes and Hepa shock-absorbing solutions were selected, Holodig program and Matlab® system were used for the measurement in the holographic technique, and in the statistical processing the Student t test was applied for the comparison of means between both microscopic techniques. It was concluded that the digital holographic microscopy allowed to observe the limits and contours of the studied red blood cells and the minimum variations in their form, besides carrying out morphometric studies which results showed similarity to those obtained with the optic microscopy, and of other parameters, as volume

Análise histomorfométrica dos cardiomiócitos e deposição de colágeno no músculo cardíaco de ratas ooforectomizadas / Histomorphometric analysis of cardiomyocytes and collagen deposition in the heart muscle of ovariectomized rats

Esse estudo objetivou avaliar o efeito da ooforectomia sob os parâmetros histomorfométricos dos cardiomiócitos e quantificação de colágeno em ratas Wistar. Vinte ratas foram utilizadas e separadas em dois grupos: GI - ooforectomizada e GII - sham-ooforectomizada. Após a recuperação anestésica os animais foram acomodados em gaiolas e separados nas condições de ciclo claro/escuro 12/12h por um período de 6 meses. Passado o período experimental, os animais foram anestesiados, o coração retirado e mergulhado em formaldeído a 10%. Os fragmentos dos ventrículos foram submetidos ao processamento histológico e corados com hematoxilina e eosina. As análises histomorfométricas (área celular e área e volume nuclear) foram realizadas pelo microscópio de luz e software ImageJ versão 1.44. Os dados foram submetidos à análise ANOVA e quando significantes, complementados pelo teste t de student (p<0,001). Observou-se uma diminuição significativa das áreas celular e nuclear dos cardiomiócitos do grupo GI quando comparado ao grupo GII, bem como a área ocupada, pelo colágeno, no músculo cardíaco foi maior em GI, quando comparado a GII. Diante disso, conclui-se que as alterações observadas nos cardiomiócitos do grupo GI, sugerem uma maior atividade apoptótica nesse grupo, devido à diminuição dos níveis séricos do estrógeno provocado pela ooforectomia e o aumento da área ocupada pelo colágeno, no grupo ooforectomizado, está associada ao efeito cardioprotetor do estrógeno.

This study aimed to evaluate the effect of oophorectomy on the morphometric variables of cardiomyocytes and quantification collagen in Wistar rats. Twenty rats were used and divided into two groups: GI - ovariectomized and GII - sham-ovariectomized. After recovery from anesthesia the animals were housed in separate cages and under the conditions of light / dark cycle 12/12h for a period of six months. After the experimental period, the animals were anesthetized, the heart removed and immersed in 10% formaldehyde. The fragments of the ventricles were subjected to histological analysis and stained with hematoxylin and eosin. Histomorphometric analysis (cell area and nuclear area and volume) were performed by light microscopy and ImageJ software version 1.44. The data were submitted to ANOVA and when significant, complemented by Student's t test (p<0.001). There was a significant decrease in cellular and nuclear areas of the GI cardiomyocytes compared to GII and the area occupied by collagen was greater in cardiac muscle GI, when compared to GII. Therefore, it is concluded that the changes observed in the GI group cardiomyocytes, suggest a higher apoptotic activity in this group, due to decreased serum estrogen levels caused by ovariectomy and increased area occupied by collagen in oophorectomized group is associated the cardioprotective effect of estrogen.

Demineralized dentin matrix combined with recombinant human bone morphogenetic protein-2 in rabbit calvarial defects

OBJECTIVES: The aim of this study was to compare the osteogenic effects of demineralized dentin matrix (DDM) combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) in rabbit calvarial defects with DDM and anorganic bovine bone (ABB) combined with rhBMP-2. MATERIALS AND METHODS: Four round defects with 8-mm diameters were created in each rabbit calvaria. Each defect was treated with one of the following: 1) DDM, 2) ABB/rhBMP-2, or 3) DDM/rhBMP-2. The rhBMP-2 was combined with DDM and ABB according to a stepwise dry and dip lyophilizing protocol. Histological and microcomputed tomography (µCT) analyses were performed to measure the amount of bone formation and bone volume after 2- and 8-week healing intervals. RESULTS: Upon histological observation at two weeks, the DDM and ABB/rhBMP-2 groups showed osteoconductive bone formation, while the DDM/rhBMP-2 group showed osteoconductive and osteoinductive bone formation. New bone formation was higher in DDM/rhBMP-2, DDM and ABB decreasing order. The amounts of bone formation were very similar at two weeks; however, at eight weeks, the DDM/rhBMP-2 group showed a two-fold greater amount of bone formation compared to the DDM and ABB/rhBMP-2 groups. The µCT analysis showed markedly increased bone volume in the DDM/rhBMP-2 group at eight weeks compared with that of the DDM group. Notably, there was a slight decrease in bone volume in the ABB/rhBMP-2 group at eight weeks. There were no significant differences among the DDM, ABB/rhBMP-2, and DDM/rhBMP-2 groups at two or eight weeks. CONCLUSION: Within the limitations of this study, DDM appears to be a suitable carrier for rhBMP-2 in orthotopic sites.

The effects of high-fat diet on implant osseointegration: an experimental study

OBJECTIVES: In this study, we investigated whether a high-fat diet (HFD) affected the bone implant connection (BIC) in peri-implant bone. MATERIALS AND METHODS: Four male rabbits were used in this study. Dental implant surgery was introduced into each tibia, and four implants were integrated into each animal. In both the normal diet (ND) group (n=2) and HFD group (n=2), 8 implants were integrated, for a total of 16 integrated implants. The animals continued with their respective diets for 12 weeks post-surgery. Afterward, the rabbits were sacrificed, and the BIC was assessed histomorphometrically. RESULTS: Histologic and histomorphometric analyses demonstrated that BIC was not impaired in the HFD group compared to the ND group. CONCLUSION: Within the limitations of this study, we found that HFD did not decrease the BIC in rabbit tibias.

Histomorphometric adaptation of yak (Bos grunniens) abomasum to the Qinghai-Tibetan plateau environment / Adaptación histomorfométrica del abomaso de yak (Yos grunniens) al medio ambiente de la meseta tibetana Qinghai

Six abomasums of yaks (Bosgrunniens) were studied with gross dissection and histological methods. It was found that the mucosa of the yak abomasum was covered with simple columnar epithelium. There were lots of spiral folds (10) in the fundic glandular area. The developed membrane of lamina propria was occupied by high density glands. According to the morphological characteristics of the glands, the abomasum was divided into the cardiac, gastric and pyloric glands. Cardiac glands were curved tubular glands with the intumescent bottom and small glandular cavity. Fundic glands were simple tubular glands or branched tubular glands, where the chief, parietal and mucous neck cells can be observed clearly. Pyloric glands were curled tubular glands, the closer to the deep of the lamina propria, the more obvious the glands curl. Staining of glycoconjugate revealed that the mucosal epithelium of the cardiac gastric and pyloric glands and gastric pits epithelium mainly secreted neutral glycoconjugate, but other portions of cardiac and gastric glands secreted mixed and acid glycoconjugate respectively. By Gordon-Sweet's reticular fiber staining, it was found that the mucous neck cells possessed the characteristic of argyrophilic phenomenon. There was a large number of argyrophilic granules in the supranuclear cytoplasm in contrast with the chief cells. Furthermore, there were isolated lymphoid nodules and diffuse lymphoid tissue in the abomasum glands, especially in corpus abomasi. Grimelius silver staining showed that the argyrophil cells were located in the glandular epithelium and lamina propria of glands, which can also be observed in connective tissue. These endocrine cells dispersed individually in epithelial cells, occasionally in 3­5 cell groups. Therefore, the yaks were grazed throughout the year on diverse natural grassland and had evolved morphological characteristics of the abomasum enabling them to consume a wide variety of plant species, thereby better adapting them to harsh plateau environment.

Seis abomaso yak (Bosgrunniens) fueron estudiados con disección y métodos histológicos. Se encontró que la mucosa del abomaso yak estaba cubierta de epitelio columnar simple. Se observaron pliegues en espiral (10) en la zona glandular fúndica. La membrana desarrollada de la lámina propia contenía glándulas de alta densidad. De acuerdo con las características morfológicas de las glándulas, el abomaso se dividió en las glándulas cardíacas, gástricas y pilórica. Las glándulas cardíacas se curvan en glándulas tubulares con la parte inferior intumescente y una pequeña cavidad glandular. Las glándulas fúndicas eran glándulas tubulares simples o glándulas tubulares ramificadas, donde se pueden observar con claridad las células principales, parietales y mucosas del cuello. Las glándulas pilóricas fueron glándulas tubulares curvadas, cuanto más cercanas a la lámina propia, más evidente fue su forma ondulada. La tinción glucoconjugada reveló que el epitelio de la mucosa de las glándulas gástricas cardiacas, pilóricas y el epitelio de las fosas gástricas secretaron principalmente un glucoconjugado neutro, pero otras porciones cárdicas y de las glándulas gástricas secretaron un glucoconjugado mixto y ácido, respectivamente. A la tinción de fibras reticulares, se encontró que las células mucosas del cuello poseían características argirófilas. Se observó un gran número de gránulos en el citoplasma supranuclear en contraste con las células principales. Además, no fueron aislados los nódulos linfoides y presentaba tejido linfoide difuso en las glándulas de abomaso, especialmente en el cuerpo del abomaso. La tinción Gordon Sweet indicó que las células argirofílicas se localizaron en el epitelio y lámina propia glandular, lo que también se observó en el tejido conectivo. Estas células endocrinas se dispersan individualmente en las células epiteliales, de vez en cuando en grupos celulares de 3-5. De esta forma, los yak pastorean durante todo el año, en diversos pastizales naturales, y han evolucionado sus características morfológicas que les permiten consumir una amplia variedad de especies de plantas, con lo que se adaptan mejor a las condiciones inhóspitas.

Histomorphometric characterization of forestomach of yak (Bos grunniens) in the Qinghai-Tibetan Plateau / Caracterización histomorfométrica del preestómago del yak (Bos grunniens) en la Meseta Qinghai-Tíbet

Six forestomachs of yaks (Bos grunniens) were studied with gross dissection and histological methods. It was found that the forestomach of yak consisted of the following three parts, rumen, reticulum and omasum, which were composed of the mucosa, submucosa, muscularis and serosa. In addition, the mucosal epithelium was covered with stratified squamous epithelium, with part of keratinized the shallow cells. Rumen, the mucosa of which formed ligulate papillae varying in size and shape, was no muscularis mucosa. Reticulum, consisted of a surface epithelium that invaginated to various extent into the lamina propria, formed various folds in shape, namely, grid-like small rooms. Furthermore, there are many secondary folds densely covered with keratinized papillae. The most striking feature of the omasum was to be formed the laminae omasi varying in length, with short and rough papillae distributing on both sides. Taken together, there was no glands within the mucosa and lamina propria of forestomach of yak, where diffuse lymphoid tissues can be observed clearly. It is, therefore, believed that the yak forestomach may have evolved those specific structural characteristics in response to the unique living environment and dietary habits impose on the Qinghai-Tibetan Plateau.

Seis preestómagos de yaks (Bos grunniens) fueron estudiados mediante disección macroscópica y métodos histológicos. Se encontró que el preestómago del yak constaba de tres partes: rumen, retículo y omaso, compuestas de mucosa, submucosa, muscular y serosa. Además, el epitelio de la mucosa se conformó con epitelio escamoso estratificado, con parte de células cornificadas superficiales. En el rumen, la mucosa formó papilas linguladas que variaron en tamaño y forma. El retículo, consistió en una superficie epitelial que se invaginó en distinta medida en la lámina propia, conformando varias formas de pliegues, es decir, cuadrículas como pequeños cubículos. Además, existían muchos pliegues secundarios densamente cubiertos con papilas cornificadas. La característica más llamativa del omaso, fue formar láminas que variaron en longitud, con papilas cortas y ásperas distribuidas en ambos lados. Tomados en conjunto, no hubo glándulas dentro de la mucosa y la lámina propia del preestómago del yak, donde los tejidos linfoides difusos se pueden observar claramente. Por lo tanto, creemos que esas características estructurales específicas del preestómago del yak pudieron haber evolucionado en respuesta a las condiciones de vida únicas y hábitos dietéticos que se presentan en la meseta de Qinghai-Tíbet.

NanoSynt: avaliação histológica e histomorfométrica de um novo substituto ósseo. O uso da nanotecnologia na conquista de um melhor padrão de osteocondução / NanoSynt: histological and histomorphometric analyses of a new bone substitute. The use of nanotechnology towards a better osteoconductive pattern

Objetivo: apresentar os resultados histológicos e histomorfométricos do uso de três substitutos ósseos disponíveis comercialmente, com relação à sua capacidade de regeneração óssea. Material e métodos: foram utilizados três biomateriais para preenchimento de defeitos críticos na calvária de coelhos, e mantidos por quatro e oito semanas in vivo. Resultados: do ponto de vista morfológico, um padrão de cicatrização semelhante foi observado para os três diferentes materiais, em que a formação do osso foi observada a partir das margens para o centro do defeito, nos tempos decorridos in vivo. De um ponto de vista quantitativo, o material NanoSynt apresentou maiores níveis médio de formação óssea, quantidade de biomaterial e de manutenção de espaço do defeito in vivo, quando comparado aos outros dois biomateriais. Conclusão: todos os substitutos ósseos utilizados apresentaram características de biocompatibilidade, não interferindo negativamente no processo de reparo. Entretanto, o biomaterial sintético NanoSynt apresentou maior nível de formação óssea e manutenção do espaço do defeito, quando comparado aos materiais Boneceramic e Bio-Oss durante os dois períodos in vivo do presente estudo (quatro e oito semanas).

Objective: to present the histological and histomorphometric results on the use of three commercially available bone substitutes with respect to its capacity of bone regeneration. Material and methods: three biomaterials were used to fill critical defects in the calvaria of rabbits kept 4 and 8 weeks in vivo. Results: from the morphological point of view, a similar pattern of healing was observed for three different materials, in which bone formation was observed from the margins to the center of the deffect, according to evaluated periods. From a quantitative point of view, the NanoSynt material exhibited higher mean levels of bone formation, amount of biomaterial, and space maintenance defect in vivo when compared to the other two biomaterials. Conclusion: all bone substitutes used showed biocompatibility and does not interfere negatively in the repair process. However, the synthetic biomaterial NanoSynt showed higher levels of bone formation and maintenance of the area of the defect when compared to materials BoneCeramic and Bio-Oss during the two periods in vivo in the present study (4 and 8 weeks).

The influence of systemically administered oxytocin on the implant-bone interface area: an experimental study in the rabbit

PURPOSE: The purpose of this study was to assess the effect of systemically administered oxytocin (OT) on the implant-bone interface by using histomorphometric analysis and the removal torque test. MATERIALS AND METHODS: A total of 10 adult, New Zealand white, female rabbits were used in this experiment. We placed 2 implants (CSM; CSM Implant, Daegu, South Korea) in each distal femoral metaphysis on both the right and left sides; the implants on both sides were placed 10 mm apart. In each rabbit, 1 implant was prepared for histomorphometric analysis and the other 3 were prepared for the removal torque test (RT). The animals received intramuscular injections of either saline (control group; 0.15 M NaCl) or OT (experimental group; 200 microg/rabbit). The injections were initiated on Day 3 following the implant surgery and were continued for 4 subsequent weeks; the injections were administered twice per day (at a 12-h interval), for 2 days per week. RESULTS: While no statistically significant difference was observed between the two groups (P=.787), the control group had stronger removal torque values. The serum OT concentration (ELISA value) was higher in the OT-treated group, although no statistically significant difference was found. Further, the histomorphometric parameter (bone-toimplant contact [BIC], inter-thread bone, and peri-implant bone) values were higher in the experimental group, but the differences were not significant. CONCLUSION: We postulate that OT supplementation via intramuscular injection weakly contributes to the bone response at the implant-bone interface in rabbits. Therefore, higher concentrations or more frequent administration of OT may be required for a greater bone response to the implant. Further studies analyzing these aspects are needed.

Uso do plasma rico em plaquetas no reparo de úlceras de córnea profundas induzidas em coelhos. Avaliação clínica e histomorfométrica / Use of platelet-rich plasma in the treatment of deep corneal ulcers induced in rabbits. Clinical and histomorphometric evaluation

Avaliou-se e comparou-se, clinicamente, por meio de histopatologia e morfometria, o processo de reparação corneana de úlceras experimentais induzidas em coelhos, com o uso de plasma rico em plaquetas (PRP) sob a forma de colírio ou tampão. Foram utilizados 60 coelhos, constituindo-se quatro grupos experimentais de 15 animais cada, designados grupo plaqueta (GP), grupo tampão (GT), grupo controle (GC) e grupo controle amniótica (GA). Os grupos experimentais foram subdivididos em três subgrupos (M4, M7, M30), de acordo com o período final de avaliação, aos quatro, sete e 30 dias, respectivamente. Não houve diferenças entre os tratamentos utilizados quanto aos sinais relacionados à sensibilidade ocular, quemose e secreção ocular. Os grupos tratados com PRP, na forma de tampão ou colírio, apresentaram menor opacidade corneana do que os animais tratados apenas com membrana amniótica no momento final de avaliação. Quanto à presença da úlcera corneana, os grupos tratados com PRP apresentaram menor ulceração corneana em relação aos demais grupos. No exame histomorfométrico, verificou-se maior epitelização corneana na fase inicial da lesão no tratamento à base de colírio de plasma rico em plaquetas. O uso da membrana amniótica promoveu espessamento do epitélio e estroma corneano, com sinergismo dela quando associada ao plasma rico em plaquetas.

The repair process induced corneal ulcer in rabbits using platelet-rich plasma in the form of eyedrop or clot was clinically evaluated and compared. Sixty rabbits were divided into four groups of 15 animals, denominated platelet group (PG), clot group (CLG), control group (CG), and amniotic control group (AG). Experimental groups were then subdivided into three groups (M4, M7, M30), corresponding to the end of the evaluation period. There were no differences between treatments regarding ocular sensitivity, chemosis and ocular secretion. The groups treated with PRP either as eyedrop or a clot showed less opacity than the animals treated only with amniotic membrane at the moment of the final evaluation. The presence of corneal ulcers in the groups treated with PRP showed lower intensity than the other groups. Histomorphometric examination showed that corneal epithelization in the initial phase of the lesion was greater when using PRP. The use of amniotic membrane promoted corneal epithelial and stromal thickness, as well as synergism when associated to PRP.