Supervivencia de células fibroblásticas humanas en ausencia de suplementación / Survival of Human Fibroblastic Cells in the Absence of Supplementation

Resumen Introducción. Los fibroblastos gingivales (FGs) son células del tejido conjuntivo gingival que han tomado en los últimos años una relevancia promisoria por su probable utilización en la terapia celular, dadas sus capacidades de multipotencialidad y de autorrenovación. Objetivo. Conocer y describir el impacto de la ausencia en la suplementación de Suero Fetal Bovino (SFB) en la supervivencia de fibroblastos gingivales en cultivos. Materiales y métodos. Fibroblastos gingivales fueron aislados de tejido gingival de pacientes sanos y cultivados en medios de cultivos DMEM (Dulbecco's Modified of Eagle Medium) en ausencia y suplementados con 0.2% de SFB a 37°C en una atmósfera húmeda con 5% de CO2. Se llevó a cabo una evaluación morfológica, de supervivencia y proliferación de los FGs, así como la identificación mediante la técnica de inmunofluorescencia de marcadores del citoesqueleto celular como la actina y mitocondrias. Resultados. Los FGs cultivados en ausencia y con suplementación de 0.2% de SFB evidenciaron una forma fusiforme, con núcleos ovalados y numerosas prolongaciones citoplasmáticas durante el tiempo de cultivo. Un leve aumento en la proliferación de FGs fue observado en aquellas células en contacto con el medio DMEM+0.2% de SFB comparadas con el medio donde estuvo ausente la suplementación. El inmunomarcaje de la actina y las mitocondrias dejó en evidencia que la ausencia y suplementación a 0.2% de SFB no afectó su localización en los FGs evaluados. Conclusión. Los fibroblastos gingivales sobreviven y proliferan en ausencia de SFB, conservando sus características morfológicas celulares.

Abstract Introduction. Gingival fibroblasts (GF) are cells of gingival connective tissue that have taken promising relevance in recent years due to their probable use in cell therapy, given their multipotencial and self-renewal capabilities. Objective. To know and to describe the impact of the absence of Fetal Bovine Serum (FBS supplementation on the survival of gingival fibroblasts in cultures. Materials and methods. Gingival fibroblasts were isolated from gingival tissue of healthy patients and cultured in DMEM (Dulbecco's Modified of Eagle Medium) culture media in absence and supplemented with 0.2% FBS at 37 ° C in a humid atmosphere with 5% CO2. A morphological evaluation, survival and proliferation of GF were carried out, as well as the identification by the immunofluorescence technique of cellular cytoskeleton markers such as actin and mitochondria. Results. The GF grown in the absence and with supplementation of 0.2% FBS showed a fusiform shape, with oval nuclei and numerous cytoplasmic extensions during the culture time. A slight increase in the proliferation of GF was observed in those cells in contact with the DMEM medium +0.2% FBS compared to the medium where the supplementation was absent. Immunostaining of actin and mitochondria showed that the absence and supplementation to 0.2% of FBS did not affect its location in the evaluated. Conclusion. Gingival fibroblasts survive and proliferate in the absence of FBS, preserving their cellular morphological characteristics.

Collection and culture of human connective tissue cells from gingival explant technique for oral tissue bioengineering / Colección de células de tejido conectivo humano a partir de la técnica de explante gingival para bioingeniería de tejidos orales

SUMMARY: Cell culture is an important tool in medical, odontological and biological research laboratories, supporting cell therapies and tissue bioengineering strategies. Gingival fibroblasts present structural function, being able to modulate their metabolic capacity, which is reflected in the tissue morphology. The possibility of culturing fibroblasts in vitro, in monolayer or on three-dimensional scaffolds, for subsequent transplants in vivo opens important perspectives for the periodontal surgical clinic. The objective of the present article is to present a method of obtaining and cultivating viable human gingival fibroblasts for in vitro research. Explants derived from periodontal surgical discards were used, grown in 25 cm2 bottles to obtain a primary cell culture. After observing the proliferation and growth of the fibroblasts that interconnected and formed a monolayer network, involving the periphery of the explants, it was possible to remove the explants, to make the passage and the new subcultures were obtained in a ratio of 1:1. After 7 days, the amount of viable cells was analyzed in triplicate, using the Neubauer chamber technique, in cell culture bottles of 25 mm2 (T25) and 75 mm2 (T75). Fibroblasts were described and subclassified morphologically. The results showed a growth pattern in both bottles, but with a larger number in bottles of 75 cm2. Cells with fibroblastic morphology were subclassified into reticular and fusiform, being predominant those with fusiform morphology. In conclusion, culture of explant of human gingival connective tissue is a viable method for obtaining gingival connective tissue cells suitable for laboratory tests in cell culture, aiming at obtaining constructs for gingival tissue engineering.

RESUMEN: El cultivo celular es una herramienta importante en los laboratorios de investigación médica, odontológica y biológica, que apoyan las terapias celulares y las estrategias de bioingeniería de tejidos. Los fibroblastos gingivales presentan una función estructural, pudiendo modular su capacidad metabólica, que se refleja en la morfología tisular. La posibilidad de cultivar fibroblastos in vitro, en monocapa o en andamios tridimensionales, para trasplantes posteriores in vivo abre perspectivas importantes para la clínica de cirugía periodontal. El objetivo del presente artículo es presentar un método para obtener y cultivar fibroblastos gingivales humanos viables para investigación in vitro. Se utilizaron explantes derivados de los descartes quirúrgicos periodontales, crecidos en frascos de 25 cm2 para obtener un cultivo de células primarias. Después de observar la proliferación y el crecimiento de los fibroblastos que se interconectaron y formaron una red de monocapa, que involucraba la periferia de los explantes, fue posible eliminar los explantes, hacer el pasaje y los nuevos subcultivos se obtuvieron en una proporción de 1:1. Después de 7 días, la cantidad de células viables se analizó por triplicado, utilizando la técnica de cámara de Neubauer, en botellas de cultivo celular de 25 mm2 (T25) y 75 mm2 (T75). Los fibroblastos fueron descritos y sub-clasificados morfológicamente. Los resultados mostraron un patrón de crecimiento en ambas botellas, pero con un número mayor en botellas de 75 cm2. Las células con morfología fibroblástica se subclasificaron en reticulares y fusiformes, predominando aquellas con morfología fusiforme. En conclusión, el cultivo de explante de tejido conectivo gingival humano es un método viable para obtener células de tejido conectivo gingival adecuadas para pruebas de laboratorio en cultivos celulares, con el objetivo de obtener construcciones para la ingeniería del tejido gingival.

Histometry Analysis of Osteoclasts, Osteoblasts and Fibroblasts in Immediate Placement Implants with Addition of Aloe Vera Extract

Objective: To analyze the effect of immediate placement of implants with extract from the new bone formation histometically. Material and Methods: In this true-experimental design with randomized post test control group, 9 mongrel dogs weighing 10 to 12 kg were used, which were divided into 3 groups, based on observation time of 14 days, 28 days and 56 days. On the installation of implants (∅3.5x10 mm) sequentially, the former socket extraction of the lower jaw's right second premolar tooth in the study sample injected 10% Aloe vera gel extract and left second left premolar tooth without injection of 10% Aloe vera extract. To compare independent groups use the Mann-Whitney test. All analysis were carried out using SPSS version 20. Results: There was an increase in the number of osteoblast cells in both treatment and control groups, but the value of the treatment group was greater. There were significant differences in the number of osteoblast cells between the treatment and control groups 14 days (p=0.019), 28 days: (p=0.018), and 56 days (p=0.009). There were no significant differences in the number of fibroblast cells between the treatment and control groups (p>0.05). But at observations 28 and 56 days, it was showed a significant difference in the number of fibroblast cells between the treatment and control groups (p=0.353 and p=0.024, respectively). Conclusion: Immediate placement of implants with 10% Aloe vera extract gel on extracted socket increases the number of osteoblasts and suppresses the number of osteoclasts and fibroblasts.

Morphology of the interstitial tissue of active and resting testis of the Guinea fowl / Morfología del tejido intersticial del testículo activo y en reposo de la gallina de Guinea

SUMMARY: The morphology of the interstitial tissue of sexually active and resting testis of the guinea fowl were studied. Six adult health birds of active and resting phases of reproductive cycle were used for this study. The interstitial tissue consisted of loose connective tissue, interstitial cells (Leydig cells), few connective cells, blood vessels and adrenergic nerve fibres in the present study in both active and resting testes. The interstitial tissue was compact in sexually active testis whereas, the volume of the tissue was found to be increased in resting testis. The loose connective tissue of the interstitial tissue composed of mainly of collagen fibres and few reticular fibres whereas elastic fibres were absent in both groups studied. The interstitial cells appeared in clusters of a few cells and were relatively less in the active testis than the resting testis. The interstitial cells were pale staining or polygonal cells with euchromatic nuclei with few large lipid droplets in the active testis whereas the cells were flat and highly heterochromatic with numerous small lipid droplets in resting testis. Few macrophages were found only in resting testis. Interstitial cells showed negative reaction to alkaline, acid phosphatases and PAS in both groups studied but positive for lipids. The interstitial tissue was well vascularised with centrally located blood vessels in the active testis whereas the blood vessels were small and inconspicuous in the resting testis. The lymphatic vessels were not identified in both groups studied.

RESUMEN: Se estudió la morfología del tejido conectivo intersticial en testículos sexualmente activos y en reposo de la gallina de Guinea (Numida meleagris). Se utilizaron seis gallinas de Guinea machos adultos sanos, en fase activa y de reposo del ciclo reproductivo. El tejido conectivo intersticial estaba formado por tejido conectivo laxo, células endocrinas intersticiales, pocas células conectivas, vasos sanguíneos y fibras nerviosas adrenérgicas, tanto en testículos activos como en reposo. El espacio intertubular en los testículos sexualmente activos era menor en comparación a los del testículos en reposo. El tejido conectivo laxo estaba compuesto principalmente de fibras colágenas y en menor cantidad de fibras reticulares. Las fibras elásticas estaban ausentes en ambos grupos. Las células endocrinas intersticiales se organizaban en racimos de pocas células y se observaban con menor frecuencia en los testículos sexualmente activos. Las células endocrinas intersticiales de los testículos activos presentaban forma poligonal, citoplasma levemente eosinofílico con algunas gotas lipídicas de gran tamaño en su interior y nucleos redondos con cromatina laxa. Las células intersticiales de los testículos en reposo eran planas y altamente heterocromáticas, con numerosas gotas lipídicas pequeñas en su citoplasma. Las células intersticiales mostraron una reacción negativa a las fosfatasas ácidas, alcalinas y PAS en ambos grupos, Sin embargo presentaron reacción positivas para los lípidos. El tejido conectivo intersticial estaba bien vascularizado con vasos sanguíneos situados centralmente en el testículo activo y vasos sanguíneos pequeños y discretos en el testículo en reposo. Los vasos linfáticos no fueron identificados en los dos grupos estudiados.Los macrófagos fueron observados solo en los testículos en reposo, aunque en escasa cantidad.

Interacciones epitelio-mesenquimáticas en el desarrollo testicular / Epithelial-mesenchymal transitions in the development of testis

RESUMEN: La espermatogénesis es un proceso continuo que se inicia durante el desarrollo embriofetal. Las relaciones auto, para y yuxtacrinas indican la interdependencia de las células intersticiales (de Leydig) con las células peritubulares (lamina propia) y células sustentaculares (de Sertoli). Ciertos morfógenos son fundamentales en este proceso. Las células sustentaculares son capaces de regular la diferenciación y función de las células peritubulares e intersticiales a través de la producción de IGF1, TGFA, TGFB y DHH. Las células peritubulares son capaces de producir P-Mod-S, regulando la diferenciación de las células sustentaculares, y a través de FGF2 y FGF9 modulan las transiciones epitelio-mesenquimática entre células sustentaculares y mesonefros. También remodelan la membrana basal del condón testicular y regulan la diferenciación y función de las células intersticiales por medio de IGF1, TGFA y TGFB. Las células intersticiales son las reponsables de la producción de testosterona e INSL3, influyendo en la diferenciación sexual masculina. Se plantea que provienen de células mesenquimales del epitelio celómico y mesonefros. Sin embargo, otros autores proponen su origen a partir de células de la cresta neural. Estas influyen a través de mecanismos paracrinos en la proliferación de las células sutentaculares por medio de activina A, teniendo como resultado la expansión del cordón testicular. Las interacciones entre las distintas poblaciones celulares a través de morfógenos inducen una transición epitelio-mesénquima fundamental en la formación y diferenciación de la gónada masculina.

SUMMARY: Spermatogenesis is a continuous process which starts during the embryo-fetal development. Auto, para and juxtacrine relations indicate the interdependence of the interstitial cells (Leydig) with the peritubular cells (lamina propria) and sustentacular cells (Sertoli). Certain morphogens are fundamental in this process. Sustentacular cells are able to regulate differentiation and function and peritubular interstitial cells through production of IGF1, TGFA, TGFB and DHH. Peritubular cells are able to produce P-Mod-S regulating differentiation sustentacular cells and through FGF2 and FGF9 modulate epithelial-mesenchymal transitions between sustentacular cells and mesonephros. They also remodel the basal membrane of the testicular condom and regulate the differentiation and function of the interstitial cells by means of IGF1, TGFA and TGFB. Interstitial cells are responsible for the production of testosterone and INSL3, influencing male sexual differentiation. It is suggested that they come from mesenchymal cells of the coelomic epithelium and mesonephros. However, other authors propose their origin from cells of the neural crest. These influence through paracrine mechanisms proliferation sutentaculares cells by activin A, resulting in the expansion of cord testicular. The interactions between the different cell populations through morphogens induce a fundamental epithelial-mesenchymal transition in the formation and differentiation of the male gonad.

Morphological evidence of telocytes in mice aorta / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Telocytes (TCs) are a novel type of interstitial cells, which have been recently described in a large variety of cavitary and noncavitary organs. TCs have small cell bodies, and remarkably thin, long, and moniliform prolongations called telopodes (Tps). Until now, TCs have been found in various loose connective tissues surrounding the arterioles, venules, and capillaries, but as a histological cellular component, whether TCs exist in large arteries remains unexplored.</p><p><b>METHODS</b>TCs were identified by transmission electron microscope in the aortic arch of male C57BL/6 mice.</p><p><b>RESULTS</b>TCs in aortic arch had small cell bodies (length: 6.06-13.02 μm; width: 1.05-4.25 μm) with characteristics of specific long (7.74-39.05 μm), thin, and moniliform Tps; TCs distributed in the whole connective tissue layer of tunica adventitia: TCs in the innermost layer of tunica adventitia, located at the juncture between media and adventitia, with their long axes oriented parallel to the outer elastic membrane; and TCs in outer layers of tunica adventitia, were embedded among transverse and longitudinal oriented collagen fibers, forming a highly complex three-dimensional meshwork. Moreover, desmosomes were observed, serving as pathways connecting neighboring Tps. In addition, vesicles shed from the surface of TCs into the extracellular matrix, participating in some biological processes.</p><p><b>CONCLUSIONS</b>TCs in aorta arch are a newly recognized complement distinct from other interstitial cells in large arteries, such as fibroblasts. And further biologically functional correlations need to be elucidated.</p>

Coexistencia gist con cáncer gástrico / The gist coexistence with gastric cancer

Los tumores estromales gastrointestinales (GIST) conjuntamente con los adenocarci-nomas, son raros y constituyen la mayoría de neoplasias no epiteliales del estómago. Pueden comprometer el intestino delgado, mesenterio y retroperitoneo. Se origina de las células intersticiales de Cajal, de la pared del tracto gastrointestinal y se puede distinguir de otras neoplasias por técnicas de inmunohistoquímica. La patogénesis del cáncer gástrico se halla relacionada con factores ambientales. Su incidencia ha disminuido en los países como Estados Unidos e Inglaterra, pero permanece alta en otros como Japón, Chile e Italia. Existen evidencias que la predisposición genética juega un rol importante en un subconjunto de pacientes. Los carcinomas gástricos se originan en las células basales de las foveolares en un fondo de gastritis crónica atrófica con displasia progresiva. Presentamos un caso registrado en Cuenca, de adenocarcinoma gástrico y GIST concomitante, que corresponde a una paciente de 75 años de edad, intervenido por adenocarcinoma bien diferenciado intramucoso gástrico; además en el acto quirúrgico se nota la presencia de un tumor a nivel de yeyuno cuyo estudio ana-tomopatológico reveló la presencia de un GIST, confirmado con técnicas de inmuno-histoquímica. Adicionalmente se revisa la literatura internacional disponible de esta infrecuente asociación.

Gastrointestinal stromal tumors (GIST) in con-junction with adenocarcinoma are rare, these constitute the majority of non-epithe-lial neoplasms of the stomach. They can compromise the small intestine, mesentery and retroperitoneum. Originates from the interstitial cells of Cajal, wall of the gastroin-testinal tract and can be distinguished from other tumors by immunohistochemistry.Pathogenesis of gastric cancer is related to environmental factors. Its incidence has declined in countries like the US and Britain, but remains high in others such as Japan, Chile and Italy. There is evidence that ge-netic predisposition plays an important role in a subset of patients. Gastric carcinomas originate from the basal cells of the foveal on a background of chronic atrophic gas-tritis with progressive dysplasia.Meet one recorded case in Cuenca con-comitant gastric adenocarcinoma and GIST, which corresponds to a 75-year-old spoke gastric intramucosal well differentia-ted adenocarcinoma, also during surgery the presence of a tumor in the jejunum is noted for study revealed the presence of pathological GIST confirmed with immuno-histochemistry techniques. Also available international literature of this uncommon association is reviewed.

Trastornos hereditarios de la fibra colágena / Inherited disorders of collagen fiber

A propósito de la publicación en esta revista del artículo titulado Evaluación de la hemostasia en niños con síndrome de Ehlers-Danlos tipo III de los autores Mirta C. Campo Díaz y col1, tenemos a bien comentar algunos de los tópicos que en este se abordan.Los trastornos hereditarios del tejido conectivo incluyen al síndrome Ehlers Danlos como una de las entidades con signos clínicos de hipermovilidad articular, que no es privativa de este síndrome genético, pero como condición genética constituye un espectro con sobrelapamiento clínico y molecular difícil de diferenciar...

Evaluation of quantitative parameters of Leydig cell in diabetic adults rats / Avaliação de parâmetros quantitativos de células de Leydig em ratos diabéticos adultos

The goal of present study was to evaluate the effects of diabetes on quantitative parameters of Leydig cells. Twelve adults Wistar rats were divided in: 1) Diabetic Group (DG), which was induced by a single intraperitoneal injection of streptozotocin (60 mg kg-1 of body weight); and 2) Control Group (CG), which received citrate buffer intraperitoneal. After eight weeks of diabetic induction, the animals were weighted, anesthetized and testicles were removed and routinely processed to paraffin embedded. Body weight (40%) and testicular weight (18%) of diabetic rats were significantly lower than control group. Diabetic rats showed an increase in interstitial compartment but the tubular compartment did not differ. The individual volume of Leydig cells and nuclear diameter were lower in DG. However, the population of these cells was increased. In conclusion, diabetes induced by streptozotocin in adult rats promoted alterations in testicular compartments and changes on volume, nuclear diameter and population of Leydig cells, compromising the testicular function.

O objetivo do presente estudo foi avaliar os efeitos do 'diabetes' nos parâmetros quantitativos de células de Leydig. Doze ratos machos adultos foram divididos em: 1) Grupo Diabético (GD) induzidos por injeção intraperitoneal única de estreptozotocina (60 mg kg-1 de peso corporal); e 2) Grupo Controle (GC) receberam tampão citrato, via intraperitoneal. Após oito semanas da indução, os animais foram pesados, anestesiados, os testículos foram removidos e processados rotineiramente em parafina. O peso corporal (40%) e testicular (18%) dos ratos diabéticos reduziu significativamente em relação ao grupo controle. Ratos diabéticos mostraram aumento no compartimento intersticial, mas o compartimento tubular não apresentou diferença significativa. O volume individual e o diâmetro nuclear de células de Leydig reduziram em GD. No entanto, a população dessas células aumentou. Em conclusão, o 'diabetes' induzido por estreptozotocina, em ratos adultos, promoveu alterações nos compartimentos testiculares e mudanças no volume, diâmetro nuclear e população das células de Leydig, comprometendo a função testicular.

Biocompatibilidad de células madres mesenquimales de tejido gingival humano en cultivoconun andamiaje de polímero sintético de ácido poliláctico (OPLA) / Biocompatibility of mesenchymal stem cells derived from human gingival tissue in culture with a scaffold of a synthetic polymer open porosity polylactic acid polymer (OPLA)

La ingeniería tisular se plantea como tratamiento ideal para la regeneración de tejidos con la utilización de andamiajes, células madres y factores de crecimiento. Las células madres de origen gingival plantean ventajas de obtención, mientras que el OPLA 3D permite cultivos de alta densidad celular. El objetivo de esta investigación fue evaluar la biocompatibilidad de células madres de origen gingival en OPLA. Las células se obtuvieron de tejido gingival y fueron caracterizadas fenotípica y funcionalmente. La biocompatibilidad se evaluó mediante la proliferación celular, prueba de viabilidad con azul tripán y diferenciación celular a linaje condrogénico y osteogénico. El recultivo del constructo se utilizó para evaluar la capacidad de transporte. Las células al interior del OPLA se visualizaron mediante cortes teñidos con H-E. Las células madres mesenquimales en OPLA proliferaron, 80% de confluencia a la cuarta semana. La viabilidad celular en OPLA fue de 83,32%. En el recultivo, las células comienzan a proliferar a la semana. El OPLA permite la diferenciación celular a linaje condrogénico y osteogénico. Se observan células al interior del OPLA, permite la proliferación, viabilidad y diferenciación celular. El OPLA podría ser utilizado como andamiaje celular para la ingeniería de tejidos.

Tissue engineering arises as the ideal treatment for tissue regeneration with the use of scaffolds, stem cells and growth factors. Stem cells derived from gingival tissue present benefits in its objection. 3D OPLA allow high cell density cultures. The objective of this study was to evaluate the biocompatibility of gingival stem cells in OPLA. Cells were obtained from gingival tissue and were characterized phenotypically and functionally. The biocompatibility was evaluated through cell proliferation, viability test with trypan blue and cell differentiation to chondrogenic and osteogenic lineage. Recultivation of the construct was used to evaluate transportability. Cells inside OPLA were visualized by stained sections with H&E. Mesenchymal stem cells proliferated in OPLA, 80% confluence at the fourth week. Cell viability in OPLA was 83.32%. In recultivation, cells start proliferating in a week. OPLA allows cell differentiation to chondrogenic and osteogenic lineage. Cells were observed within OPLA. In conclusion OPLA allows proliferation, viability and cell differentiation. OPLA could be used as scaffolds for cells in tissue engineering.

Análise dos efeitos da eletrolipólise no tratamento do fibro edema gelóide por meio da biofotogrametria computadorizada / Analysis of the effects of electrolipolysis in the treatment of fiber edema geloid by means of computer-aided biophotogrammetry

O Fibro Edema Gelóide (FEG) é uma infiltração edematosa do tecido conjuntivo subcutâneo e considerada uma patologia multifatorial, classificando-se em quatro graus: Grau I; Grau II, Grau IIIe Grau IV. Dentre as inúmeras técnicas terapêuticas a eletrolipólise apresenta-se como um método novo e promissor, pois o estímulo circulatório produzido pela corrente elétrica tem grande importância na drenagem da área tratada. Esse efeito, particularmente, é um dos que justifica o uso da eletrolipólise no tratamento do FEG. Como forma de avaliação quantitativa para verificar a evolução do tratamento,foi utilizada a Biofotogrametria Computadorizada, através da qual foi realizada a avaliação da área e do perímetro das lesões no prée no pós-tratamento. Assim, este estudo teve como objetivo analisar os efeitos da eletrolipólise no tratamento do FEG grau III por meio da biofotogrametria computadorizada. Realizou-se 10 sessões coma eletrolipólise na região glútea. A variável área apresentou maior redução percentual média, se comparada à variável perímetro, tanto para as condições de com contração (CC) como sem contração (SC). Após a análise dos resultados, concluiu-se que a eletrolipólise pode ser utilizada com eficácia em pacientes que apresentem FEG grau III, tendo em vista o biotipo tratado.

The Fiber Edema Geloid (FEG) is an infiltration of the edematous subcutaneous tissue and is considered a multifactorial disease, classified into four grades: Grade I, Grade II, Grade III and Grade IV. Among the many therapeutic techniques, electrolipolysis appears to be a new and promising method, since the circulatory stimulation generated by electrical current is of great importancein the drainage of the treated area. This effect, in particular, is one that justifies the use of eletrolipolysis in the treatment of FEG. In order to check treatment progress as a quantitative assessment, we used a computerized biophotogrammetry. The lesion perimeter and surface area in the pre and post-treatment were evaluated. This study aimed at analyzing the effects of electrolipolysis in the treatment of FEG grade III by means of computerized photogrammetry. We conducted 10 sessions with electrolipolysis in the gluteal region. The variable area showed greater mean percent reduction when compared to the perimeter variable for both conditions with contraction and without contraction. After analyzing the results, we concluded thatthe electrolipolysis can be used successfully in patients with gradeIII FEG, in view of the biotype treated.

Immunohistochemical evaluation of CD25+ cell expression in the progression of periodontal disease

It was assessed the immunohistochemical profile of CD25+ cells in cases of chronic gingivitis (CG) and chronic periodontitis (CP). Immunohistochemistry was carried out using streptoavidin-biotin complex and anti-CD25 antibody in 17 cases of CG and 25 cases of CP. Sixteen cases (94.1%) of CG were immunopositive. CD25 was focally expressed in 50% of the sample and diffusely expressed in 25%. The stained cells were localized not only beneath the epithelium, but also far from it. In relation to the cellular density quantification of CD25+ cells, score ++ was the most common. Concerning CP, all cases were immunopositive. CD25+ cells were expressed in focal or diffuse pattern either close or far from the epithelium. Diffuse distribution of positive cells throughout the connective tissue was seen in 60% of the cases and 32% showed focal or diffuse cellular pattern. Sixteen cases (64%) received score +++. It was identified that CD25+ cells are present in either a focal or a diffuse pattern in connective tissue. Significant differences in the density of cellular immunostaining between CG and CP were found. The greatest density was observed in CP cases, which suggests that the infiltrate of lymphocytes show a higher degree of cellular activation in periodontitis compared with gingivitis.

Foi avaliado o perfil imunohistoquímico das células CD25+ em casos de gengivite (CG) e periodontite crônica (CP). A imunohistoquímica foi realizada utilizando o complexo de streptoavidina-biotina e o anticorpo anti-CD25 em 17 casos de CG e 25 casos de CP. 16 casos (94.1%) de CG foram imunopositivos. O CD25 foi expresso focalmente em 50% da amostra e difusamente em 25% dos casos. As células imunomarcadas estavam localizadas não apenas no epitélio, mas também por todo o tecido conjuntivo. Em relação à quantificação da densidade celular de células CD25+, o escore ++ foi o mais comum. Em relação a CP, todos os casos foram imunopositivos. As células CD25+ foram expressas em padrão ora focal ora difuso, tanto no epitélio como no conjuntivo. A distribuição difusa das células positivas apenas no tecido conjuntivo foi observada em 60% dos casos, e 32% dos casos exibiram padrão celular ora focal ora difuso. 16 casos (64%) foram considerados como escore +++. Identificamos que as células CD25+ estão presentes em padrão ora focal ora difuso no tecido conjuntivo. Diferenças significantes na densidade da imunomarcação celular entre CG and CP foram encontradas. A maior densidade celular foi observada na periodontite, sugerindo que o infiltrado de linfócitos mostrou um maior grau de ativação celular na periodontite comparada à gengivite.

Enxerto ósseo em crânio: princípios básicos para a integração óssea / Bone graft in skull: basic principles for osteointegration

Introdução: A reparação das perdas ósseas evoluiu de maneira significativa a partir do século 20, principalmente na segunda metade, quando a Medicina teve uma evolução importante para resolver os casos de reconstrução de perdas de tecidos. Inicialmente os materiais de inclusão, citando os metais maleáveis, silicone, acrílico de rápida polimerização, apareceram juntamente com o uso de enxerto ósseo autólogo. Os enxertos ósseos autólogos avançaram com muita propriedade, para se tornarem o material de eleição na reparação destas perdas ósseas. Os locais mais importantes para obtenção do enxerto são: crânio, osso ilíaco, costela e tíbia. Nos últimos 30 anos, com o refinamento das técnicas e o uso de material cirúrgico preciso, tanto para retirada do osso para o enxerto, como a colocação do mesmo na área receptora, estes procedimentos se firmaram definitivamente no estudo da reconstrução. Aliado aos recursos citados, salientamos o conhecimento da origem embriológica do osso que é subdividido em ossos cartilaginosos (tíbia, osso ilíaco e costela) e ossos membranosos, portanto, os “ossos parietais” são priorizados para a reparação de ossos cranianos e faciais. Além da origem embriológica, devemos citar também o conhecimento histológico do osso compacto e do osso esponjoso. Citamos, ainda, as células precursoras do tecido ósseo, osteoblastos, osteócitos e osteoclastos.

Background: The repair of bone loss had a great improvement starting in the middle ofthe last century, when the Medicine had a big evolution in solving the problems of tissue loss using initially soft metals, silicone, acrylic of rapid polymerization, came up at the same time of the use of bone graft autologous. The bone graft autologous developed in the way that became the first choice for reconstruction of bone loss. The common area to obtain the bones grafts are: cranial, iliac bones, ribs and tibia. In the last 30 years, with the improvement of techniques and the use of precisely surgical materials, for extraction of the bones for grafts and also for insertion of the bone at the receptor area, these processes established the way to do reconstruction surgeries. Besides the processes listed above we also take in consideration that we had about embryologic origin of the bones with are divided in: cartilaginous bones (tibia, iliac bones and ribs) and membranous bones, therefore the “parietals bones”, are prioritized for the repair of cranial and facial bones. Besides the embryologic origin of the bones we should also emphasize the knowledge of the compact bone and spongy bone history. We would like also to mention the precursor cells of the bone tissues, osteoblasts, osteocytes and osteoclasts.

Tumores estromales gastrointestinales: parámetros morfologicos e histogenéticos de valor pronostico / Tumors estromal gastrointestinal: morphologic parameters and histogenéticos of value I predict

INTRODUCCION: Los tumores estromales gastrintestinales (GISTs) constituyen la categoría más amplia de neoplasias no epiteliales primarias del tracto gastrointestinal. Son más frecuentes en estómago y en intestino delgado, pero también pueden comprometer esófago, colon, recto y ano. Han sido encontrados, además, en mesenterio, retroperitoneo, omento y tejidos blandos (EGIST-Tumor estromal extragastrointestinal). Martín los reconoció como entidad clínico -patológica y Stout los denominó Leinomioblastomas. Mazur y Clark acuñaron el término tumor estromal gastrointesitnal y sugierieron que podían originarse del sistema nervioso mioentérico. Hubo, además, algunos casos con evidencias de diferenciacion neural, y fue introducido el término "Tumor autonómico gastrointestinal" (GANT). Kindblom y col., sugirieron que esta neoplasias presentan un inmunofenotipo similar a células intersticiales de Cajal (Células marcapsos del tracto gastrointestinal). OBJETIVOS: -Correlacionar tamaño tumoral, patrón microscopico e inmunofenotipo con el comportamiento biológico. - Determinar formas macro y microscopicas y analizar la forma de preesentación clínica. - Relacionar topografia, tamaño y grado con la agresividad tumoral. - Confirmar el valor del protooncogen (CD-117) como marcador de CIC y de GISTs.

Histological and functional organization in human testicle: expression of receptors c-kit and androgens / Organización histológica y funcional en el testículo humano: expresión de los receptores c-kit y andrógenos

The objective of this work was identify the presence of interstitial cells of Cajal, muscle cells, nerves and androgen receptor positive cells in adult human testicle, using immunohistochemical detection for c-kit/CD-117, actin smooth muscle specific (ASMS), neurofilament (N) and androgen receptor (AR), respectively. The samples were obtained from patients (n= 10) with diagnosis of prostate cancer, with surgery of orchiectomy. Subsequently were processed by histology and for immunohistochemistry using specific antibodies. It showed the presence of cells c-kit/CD-117, with diverse degrees of positivity, distributed mainly in the interstitial peritubular area of the human testicle. The peritubular myoides cells were positive to the presence of the actin smooth muscle and androgen receptor. The neurofilaments elements (+) only were observed in the vascular tunic. The specific immunohistochemistry describe the presence of the interstitial cells of Cajal in human testicular interstitium, opening a new perspective for the functional interpretation of the testicular cellularity and tubular motility. Possibly associated functionally to peribubulars cells of smooth muscle to regulate the mobility of the seminiferous tabules, whose integration and function would be androgen dependent. The cells that express the c-kit receptor, were found exclusively in the interstitial compartment. This cellular type in addition of the muscular cells of peritubules and the absence of nervous fibers to the interior of the testicle, could be responsible for the regulation of tubular mobility, as it happens in the gastrointestinal apparatus.

El objetivo de este trabajo fue identificar la presencia de células interticiales de Cajal, células musculares lisas, células nerviosas y células que expresan receptores de andrógeno en el testículo de humano adulto, usando inmunohistoquímica específica para: c-kit/CD-117, músculo liso actina específico (ASMS), neurofilamentos (N) y para receptores de andrógenos (AR). Las muestras fueron obtenidas de pacientes (n=10) con diagnóstico de cáncer prostático sometidos a cirugía de orquiectomía. Las biopsias se procesaron para histología e inmunohistoquímica usando anticuerpos específicos. Se muestra la presencia de células c-kit/CD-117, con diversos grados de positividad y distribuidas en el compartimento interticial del testículo. Las células peritubulares mioides fueron positivas para la presencia de músculo liso actina específico y para receptor de andrógenos. La marcación de neurofilamentos positivos, sólo fueron observados en la túnica vascular. Conclusiones: La inmunohistoquímica específica describe la presencia de células interticiales de Cajal en los interticios testiculares humanos, abriendo una nueva visión en la interpretación funcional de la celularidad testicular y la motilidad tubular. Lo anterior asociado a la funcionalidad de las células peritubulares (músculo liso) regularían la motilidad de los túbulos seminíferos. Este proceso posiblemente es andrógeno dependiente. Las células que expresan receptores c-kit se encuentran exclusivamente en los compartimentos interticiales, estas células en conjunto con las células musculares peritubulares agregado a la ausencia de fibras nerviosas al interior del testículo, podrían ser los responsables de la regulación de la motilidad tubular, similar a como se informa para el tracto gastrointestinal.

A Comparative Histomorphometric Study of the Stomach of Rat (Rattus norvegicus), Bat (Eidolon helvum) and Pangolin (Manis tricuspis) in Relation to Diet / Estudio Comparativo Histomorfométrico del Estómago de Rata (Rattus norvegicus), Murciélago (Eidolon helvum) y Pangolin (Manis tricuspis) en Relación con la Dieta

This study verified the comparative histomorphometric adaptations in the stomach of rat, bat and pangolin in relation to diet. Ten rats, ten bats and ten pangolins of both sexes were used for this investigation. The animals were sacrificed after slight anesthesia under chloroform inhalation. The stomach were excised, fixed in 10 percent formol saline and processed for light microscopic study. Stained slides were also subjected to morphometric analysis at a magnification of 400x. The results revealed that the cellular diameter/ density of parietal and zymogenic cells are significantly different in the three mammals (p<0.05) with the exception of the diameter of the zymogenic cells in pangolin which was not statistically significant (p>0.05) when compared with that of rat. Also, histological analysis revealed slight differences in the pattern of organization and distribution of connective tissue fibers. All these observations were reflections of the different pattern the stomachs of the three mammals have adopted to cope with their respective diets.

En este estudio se verificaron las adaptaciones histomorfométricas comparativas en el estómago de ratas, murciélagos y pangolines en relación a la dieta. Se utilizaron para esta investigación 10 ejemplares de cada especie, de ambos sexos. Los animales fueron sacrificados después de anestesia bajo inhalación de cloroformo. Los estómagos fueron extirpados, fijados en formol al 10 por ciento de solución salina y procesados para su estudio microscópico de luz. Los cortes teñidos fueron también objeto de análisis morfométrico con un aumento de X 400. Los resultados revelaron que el diámetro/densidad celular de parietal y las células cimógenas son significativamente diferentes en los tres mamíferos (p <0,05), con la excepción del diámetro de la células cimógenas de pangolines que no era estadísticamente significativa (p> 0,05) en comparación con la de rata. Por otra parte, el análisis histológico reveló ligeras diferencias en las características de organización y distribución de las fibras del tejido conjuntivo. Todas estas observaciones son un reflejo del patrón de los diferentes estómagos de los tres mamíferos, que han adoptado para hacer frente a sus respectivas dietas.

Tumor desmoide: reporte de un caso / Desmoide tumor: case report

El tumor desmoide es una neoplasia rara de tejidos blandos que se desarrolla a partir de músculo, tejido conectivo, fascia y aponeurois. Se presenta esporádicamente y más frecuentemente en mujeres. El caso que se presenta concierne a paciente femenina de 27 años que fue sometida a escisión radical de una gran masa que infiltraba músculo recto abdominal, diagnosticada como tumor desmoide. Aunque tiene características benignas, es de naturaleza infiltrativa y se comporta como una masa localmente agresiva, la cual puede invadir estructuras adyacentes haciendo que la resección quirúrgica sea difícil. El único tratamiento viable es la cirugía amplia dejando bordes sanos, esto causa gran defecto de pared y por tanto serios problemas en la reconstrucción. Además, la tasa de recurrencias locales varía y depende de la edad del paciente, localización y los márgenes de resección.

Desmoid tumor is quite rare soft tissues neoplasm that develops from muscle connective tissue, fasciae and aponeuroses. This neoplasm occurs in sporadic and more frequent in women than men. The presented case report refers to young female (27 years old), who underwent the radical excision of a large desmoid tumour infiltrating the right rectus muscle of the abdomen. Although desmoid is classified pathologically as a benign tumour, its infiltrative nature leads to a locally aggressive mass, which can invade surrounding structures and organs making surgical resection difficult. The only radical treatment for her was the surgical resection carried out far from the tumour borders into the healthy tissues. This resection causes wide muscle-fascial defects determining serious reconstructive problems. Futhermore, overall local recurrence rates vary and depend on patient’s age, tumour location and margins at resection.

Transforming growth factor-beta Does Not Induce Endothelin-1 Secretion in Primary Cultured Human Tenon's Fibroblasts

No abstract available.

DNA microarray analysis of gene expression of MC3T3-E1 osteoblast cell cultured on anodized- or machined titanium surface / 대한치주과학회지

PURPOSE: The aim of this study was to evaluate adhesion and gene expression of the MC3T3-E1 cells cultured on machined titanium surface (MS) and anodized titanium surface (AS) using MTT test, Scanning electron micrograph and cDNA microarray. MATERIALS AND METHODS: The MTT test assay was used for examining the proliferation of MC3T3-E1 cells, osteoblast like cells from Rat calvaria, on MS and AS for 24 hours and 48 hours. Cell cultures were incubated for 24 hours to evaluate the influence of the substrate geometry on both surfaces using a Scanning Electron Micrograph (SEM). The cDNA microarray Agilent Rat 22K chip was used to monitor expressions of genes. RESULTS: After 24 hours of adhesion, the cell density on AS was higher than MS (p0.05). AS had the irregular, rough and porous surface texture. After 48 hours incubation of the MC3T3-E1 cells, connective tissue growth factor (CTGF) was up-regulated on AS than MS (more than 2 fold) and the insulin-like growth factor 1 receptor was down-regulated (more than 2 fold) on AS than MS. CONCLUSION: Microarray assay at 48 hours after culturing the cells on both surfaces revealed that osteoinductive molecules appeared more prominent on AS, whereas the adhesion molecules on the biomaterial were higher on MS than AS, which will affect the phenotype of the plated cells depending on the surface morphology.

Changes in Corpus Cavernosum after Partial Bladder Outlet Obstruction in Rat / 대한비뇨기과학회지

PURPOSE: Abnormalities of the relaxation and contraction of the corpus cavernosum can lead to erectile dysfunction. Therefore, we induced a partial bladder outlet obstruction(PBOO) in male rats, and investigated the mechanisms of penile dysfunction with endothelial nitric oxide synthase(eNOS), vascular endothelial growth factor(VEGF), endothelin-1(ET-1), and apoptosis of peri-vascular smooth muscle and connective tissue cells in the corpus cavernosum. MATERIALS AND METHODS: PBOO was induced in 13 Sprague-Dawley rats by placing a 25 gauge needle sheath around the urethra, then ligating the bladder neck with a 3-0 suture. Three week after surgery, distal penile tissues were dissected for immunohistochemical staining, immunoblotting, and TUNEL staining. RESULTS: The expression of eNOS and VEGF were significantly decreased, whereas the expression of ET-1 and apoptosis of perivascular smooth muscle and connective tissue cells were significantly increased in the corpus cavernosum. CONCLUSIONS: The significant increase of ET-1 and apoptosis along with decreased eNOS and VEGF could mediate erectile dysfunction.