Immature Dental Pulp Stem Cells Showed Renotropic and Pericyte-Like Properties in Acute Renal Failure in Rats.

Acute renal failure (ARF) is a common renal disease that can lead to high mortality. Recovery from ARF occurs with the replacement of necrotic tubular cells by functional tubular epithelial cells and the normalization of microvascular endothelial cell function in the peritubular capillaries. Conventional therapeutic techniques are often ineffective against ARF. Hence, stem cell therapies, which act through multiple trophic and regenerative mechanisms, are encouraging. We investigated the homing of human immature dental pulp stem cells (IDPSCs) after endovenous (EV) or intraperitoneal (IP) injection, in immunocompetent Wistar rats with ARF induced by intramuscular injection of glycerol, without the use of immunosuppression. The cells, which had been cryopreserved for 6 years, were CD105(+), CD73(+), CD44(+), and partly, STRO-1(+) and CD146(+), and presented unaltered mesoderm differentiation potential. The presence of these cells in the tubular region of the kidney and in the peritubular capillaries was demonstrated. These cells accelerate tubular epithelial cell regeneration through significant increase of Ki-67-immunoreactive cells in damaged kidney. Flow cytometry analysis confirmed that IDPSCs home to the kidneys (EV 34.10% and IP 33.25%); a lower percentage of cells was found in the liver (EV 19.05% and IP 9.10%), in the muscles (EV 6.30% and IP 1.35%), and in the lungs (EV 2.0% and IP 1.85%). After infusion into rat, these cells express pericyte markers, such as CD146(+), STRO-1(+), and vascular endothelial growth factor (VEGF(+)). We found that IDPSCs demonstrate renotropic and pericyte-like properties and contributed to restore renal tubule structure in an experimental rat ARF model.

Successful transplant of mesenchymal stem cells in induced osteonecrosis of the ovine femoral head: preliminary results / Sucesso no transplante de células tronco mesenquimais em ovinos com osteonecrose induzida da cabeça do fêmur: resultados preliminares

PURPOSE: Evaluate the bone tissue recovery following transplantation of ovine mesenchymal stem cells (MSC) from bone marrow and human immature dental-pulp stem cells (hIDPSC) in ovine model of induced osteonecrosis of femoral head (ONFH). METHODS: Eight sheep were divided in three experimental groups. First group was composed by four animals with ONFH induced by ethanol through central decompression (CD), for control group without any treatment. The second and third group were compose by two animals, six weeks after ONFH induction received transplantation of heterologous ovine MSC (CD + oMSC), and hIDPSC (CD + hIDPSC), respectively. In both experiments the cells were transplanted without application of any type of immunosupression protocol. RESULTS: Our data indicate that both cell types used in experiments were able to proliferate within injured site providing bone tissue recovery. The histological results obtained from CD+hIDPSC suggested that the bone regeneration in such animals was better than that observed in CD animals. CONCLUSION: Mesenchymal stem cell transplant in induced ovine osteonecrosis of femoral head by central decompression technique is safe, and apparently favors bone regeneration of damaged tissues.

OBJETIVO: Verificar os efeitos das células-tronco mesenquimais da medula óssea de ovinos e da polpa dentária imatura humana em ovinos com osteonecrose induzida, da cabeça do fêmur. MÉTODOS: Oito ovelhas foram distribuídas em três grupos experimentais. O primeiro grupo foi composto por quatro animais com osteonecrose da cabeça do fêmur induzida por etanol através da descompressão central, que não receberam nenhum tratamento. O segundo e o terceiro grupo, cada um composto por dois animais, receberam transplante heterólogo de células tronco mesenquimais de ovinos e polpa dentária imatura humana seis semanas após a indução da osteonecrose da cabeça do fêmur, respectivamente. Em ambos os grupos experimentais as células foram transplantadas sem o uso de drogas imunossupressoras. RESULTADOS: Os achados demonstram que as células-tronco mesenquimais injetadas na cabeça do fêmur se encontravam viáveis após o transplante no novo sítio e proliferaram em pouco tempo. Os dados histológicos sugerem que a regeneração óssea nos animais transplantados com polpa dentária imatura humana foi mais rápida do que nos animais submetidos somente a descompressão central. CONCLUSÃO: O transplante de células tronco mesenquimais na osteonecrose da cabeça do fêmur induzida em ovinos através da técnica de descompressão central é um procedimento seguro, e aparentemente favorece a regeneração óssea de tecidos lesados.

Successful transplant of mesenchymal stem cells in induced osteonecrosis of the ovine femoral head: preliminary results.

PURPOSE: Evaluate the bone tissue recovery following transplantation of ovine mesenchymal stem cells (MSC) from bone marrow and human immature dental-pulp stem cells (hIDPSC) in ovine model of induced osteonecrosis of femoral head (ONFH). METHODS: Eight sheep were divided in three experimental groups. First group was composed by four animals with ONFH induced by ethanol through central decompression (CD), for control group without any treatment. The second and third group were compose by two animals, six weeks after ONFH induction received transplantation of heterologous ovine MSC (CD + oMSC), and hIDPSC (CD + hIDPSC), respectively. In both experiments the cells were transplanted without application of any type of immunosupression protocol. RESULTS: Our data indicate that both cell types used in experiments were able to proliferate within injured site providing bone tissue recovery. The histological results obtained from CD+hIDPSC suggested that the bone regeneration in such animals was better than that observed in CD animals. CONCLUSION: Mesenchymal stem cell transplant in induced ovine osteonecrosis of femoral head by central decompression technique is safe, and apparently favors bone regeneration of damaged tissues.

Morfologia da glândula pineal de gambás (Didelphis sp) / Morphology of the pineal gland in opossum (Didelphis sp)

A glândula pineal deve ser analisada e estudada em animais da fauna brasileira, para que dados da pesquisa básica possam ser aplicados em novas técnicas de manejo reprodutivo destes animais, inclusive em cativeiro, face à íntima relação deste órgão fotorreceptor com o ciclo reprodutivo. Para este estudo, foram utilizados 10 gambás (Didelphis sp), provenientes do Departamento de Anatomia da USP e da UNIFEOB, já mortos e fixados. Nenhum animal foi submetido a situações de dor/sofrimento e ao sacrifício de sua vida. A glândula pineal foi encontrada em todos animais estudados e apresentou-se com diminutas dimensões, não sendo possível, portanto descrever-lhe características macroscópicas. Através da análise microscópica pudemos localizar a glândula no espaço correspondente ao plano mediano, em relação ao encéfalo, rostral e dorsalmente aos colículos rostrais, ventralmente aos hemisférios cerebrais e caudalmente à comissura habenular. Consiste de uma evaginação do teto do diencéfalo e mostra-se em forma de “U” invertido. Comparativamente a características de glândulas pineais de outras espécies animais, a do Didelphis genus, que estudamos, revela peculiaridades tanto em relação ao seu tamanho, apenas perceptível microscopicamente, quanto ao fato de apresentar células semelhantes às secretoras, dispersas também em áreas vizinhas. Tais peculiaridades motivam reflexões sobre o papel funcional da glândula, na espécie considerada.

The pineal gland must to be analyzed and studied in animals of the Brazilian fauna, to apply the data obtained in the basic research of new techniques at reproductive handling of these animals, including in captivity, in view of the close relation between this photoreceptor organ with the circadian and reproductive cycle. For this study, 10 opossums (Didelphis sp), had been used, already died and fixed, proceeding from the Department of Anatomy of USP and UNIFEOB. None animals were submitted to pain/suffering situations and their no life sacrifice. The pineal gland was found in all studied animals with and smaller dimention, not possessing, therefore goss features. By microscopy analysis we could found the gland in the correspondent space to median plan in relation to the encephalon, rostral and dorsally to the rostral coliculli, ventrally to the brain hemispheres and caudally to the habenular comissure. That consistes like an evagination of the diencephalons tectum showing the “U” shape. Considering other pineal glands and its features in different species, we note the gland is extremely small for it specie, possessing dispersed secretory cells in the nervous parenchyma whose form, sufficiently irregular, suggests a small hormonal performance to them in the Didelphis genus. Comparativelly of the pineal gland feactures in different animals, the Didelphis genus, that was our aim, shows pecualirity as in size relation, only microscopically visible, than the fact to prossessing similar secretory cells also dispased in neighbor areas. All pecualiarites suggest refletion about the function action of the gland at the studied specie.

Influência do glicerol e etilenoglicol e da criopreservação sobre o complexo DNA-Proteina de espermatozóides em garanhões / Glycerol, ethyleneglycol and cryopreservation influences on semen an sperm DNA-Protein complex in stallion

O processo de criopreservação causa estresse físico e químico aos espermatozóides, acarretando alterações bioquímicas, diminuição irreverssível da motilidade espermática, aumento da degeneração do DNA e liberação intracelular de enzimas e lipídeos. No presente estudo, foram estudadas a influência das estações não reprodutiva e reprodutiva, dos crioprotetores glicerol e etilenoglicol e do processo de congelação e descongelação sobre o complexo DNA-proteína de espermatozóides em garanhões. Foi comparados o sêmen puro, o sêmen puro e congelado sem crioprotetores, o sêmen diluído e exposto aos crioprotetores sem congelação e o sêmen diluído e congelado com crioprotetores. Foram utilizados seis garanhões, colhendo 12 ejaculados cada. A patologia do complexo DNA-Proteína foi avaliada em espermatozóides fixados com etanol-ácido-acético glacial 3: 1 (v/ v), tratados com HCL 4N a 25°C e corados com azul de toluidina a 0,025% em tampão McIlvaine, empregando microscopia óptica com aumento de 1000x. Os resultados mostraram que a anomalia do complexo DNA-Proteína dos espermatozóides diferem entre os grupos congelados e não congelados (P<0,05). O sêmen congelado sem crioprotetor não apresentou aumento significativo de patologia do complexo DNA-Proteína em relação ao sêmen congelado com crioprotetores, mas ambos mostraram aumento em relação ao sêmen puro ou diluído e exposto aos crioprotetores. A influência da estação reprodutiva mostrou diferença significativa (P<0,05) somente no sêmen puro e no sêmen puro e congelado sem crioprotetor. Conclui-se que o processo de congelação exerce influência negativa sobre o complexo DNA-Proteína de espermatozóides em garanhões.

The cryopreservation process cause stress physical and chemical to the spermatozoa, causing biochemistry alteration, irreversible reduction of the spermatic motility, increase of the DNA degeneration and intracellular enzyme and lipids release. The aim of this study was to evaluate the influence of non-breeding and breeding seasons, glycerol and ethylene glycol, cryopreservation and thawing processes on stallion spermatozoa DN A -protein complexo It was compared fresh semen, diluted semen frozen without cryoprotectants, diluted semen exposured to cryoprotectants but not frozen and d) diluted semen frozen with cryoprotectants. Six stallions had 12 semen collections each. DNA-protein complex pathology was assessed by optical microscopy (1000x) using spermatozoa treated with ethanol-acetic acid 3:1 (v/v), HCl 4N at room temperature and toluidin blue 0,025% in McIlavaine buffer. Results showed that DNA-protein complex were different between frozen and not frozen spermatozoa groups (P<0,05). Frozen semen without cryoprotectants had no increasing of DNA-protein complex pathology compared to semen cryopreserved with cryoprotectant, but both showed increasing in relation to fresh and diluted semen exposured to cryoprotectants. The influence of non breeding and breeding season showed significant difference (P<0,05) in the fresh semen and fresh semen frozen without cryoprotectants. Cryopreservation process had negative influence on spermatozoa DNA-protein complex.