Allogenic mesenchymal stem cell intravenous infusion in reparation of mild intestinal ischemia/reperfusion injury in New Zealand rabbits / Infusão intravenosa de células tronco mesenquimais alógenas na reparação da lesão branda de isquemia/reperfusão intestinal em coelhos Nova Zelândia

The present study aimed to evaluate the efficacy of mesenchymal stem cell (MSC) infusion, derived from adipose tissue, on reduction of local and remote tissue damage caused by the event of experimental intestinal I/R in New Zealand breed rabbits. For obtaining, characterization, and cultivation of MSC derived from adipose tissue (MSC-Adp), 3 juvenile animals (four months old) were used. The cells were considered to be viable for therapy after the fourth passage (in vitro phase). For the in vivo stage, 24 young adult animals (six months old) were used, weighing approximately 3.5 kg, in which were randomly divided into two groups, called: IR treated with MSC (I2H/R5H MSC 3D; I2H/R5H MSC 7D); IR treated with PBS (I2H/R5H PBS 3D; I2H/R5H PBS 7D). The animals were anesthetized and submitted to pre-retro-umbilical midline celiotomy. The extramural peri-intestinal marginal artery was located and clamped (predetermined and standardized region) with the aid of a vascular clip, promoting a 2 hour blood flow interruption. After this period, blood flow was reestablished, inhalatory anesthesia was suspended, and the animals awaken. After 5 hours of reperfusion, the treatments were performed by intravenous infusion according to the experimental groups. The animals were evaluated 72 hours and seven days after the treatment as for the macroscopic appearance (color and peristaltism) of the jejunal segment, and by histological evaluation of the ischemic segment for the presence or absence of destruction of the intestinal mucosa, edema, bleeding, dilation of lymph vessels, and presence of polymorphonuclear inflammatory cells, both in the mucosa and submucosa. The observed results revealed that the groups treated with MSC-Adp obtained smaller mucosal and submucosal lesions when compared to the groups treated with PBS. Also, MSC-Adp treated groups obtained controlled inflammatory response and higher mitotic rate, outcomes related to the therapeutic potential of MSC. Infusion of stem cells attenuated the lesions caused by intestinal I/R in both MSC groups when compared to the group treated with PBS.(AU)

O presente estudo teve como principal objetivo avaliar a eficácia da infusão células tronco mesenquimais (CTM) derivada de tecido adiposo sobre diminuição das lesões teciduais locais e remotas, causadas pelo evento de I/R intestinal experimental, em coelhos da raça Nova Zelândia. Para obtenção, cultivo e caracterização das CTM provenientes de tecido adiposo (ADCTM) foram utilizados 3 animais jovens. As células foram consideradas viáveis para terapia a partir da quarta passagem (fase in vitro). Para etapa in vivo foram utilizados 24 animais, adulto-jovens, pesando aproximadamente 3,5kg, divididos aleatoriamente em dois grupos experimentais, denominados IR Tratado com CTM (I2H/R5H CTM 3D; I2H/R5H CTM 7D); IR Tratado PBS (I2H/R5H PBS 3D; I2H/R5H PBS 7D). Os animais foram anestesiados e submetidos à celiotomia mediana pré-retroumbilical. A artéria marginal peri-intestinal extramural foi localizada e clampeada (região predeterminada e padronizada) com auxílio de um clipe vascular, promovendo uma interrupção do fluxo sanguíneo durante 2 horas. Após esse período, o fluxo sanguíneo foi restabelecido, a anestesia inalatória suspendida e os animais despertados. Após 5 horas de reperfusão realizou-se os tratamentos por infusão endovenosa, conforme grupos experimentais. Os animais foram avaliados 72 horas e sete dias após o tratamento quanto ao aspecto macroscópico (coloração e peristaltismo) do segmento jejunal e por meio de avaliação histológica do segmento isquemiado quanto à presença ou ausência de destruição de mucosa intestinal, edema, hemorragia, dilatação de vasos linfáticos e presença de células inflamatórias polimorfornucleares, tanto em mucosa quanto submucosa. Os resultados observados revelaram que os grupos tratados com ADCTM obtiveram menores lesões em mucosa e submucosa quando comprados aos grupos tratados com PBS. Ainda os grupos tratados com ADCTM obtiveram resposta inflamatória controlada e maior taxa mitótica, resultados relacionados ao potencial terapêutico das CTM.(AU)

Valoración de la tasa de cicatrización de una úlcera aguda en ratas Sprague-Dawley, mediante la aplicación de células madre mesenquimatosas autólogas, obtenidas del tejido adiposo / (Healing rate assessment of an acute ulcer in Sprague-Dawley rats by applying autologous mesenchymal stem cells drawn from adipose tissue)

Resumen Objetivo: se diseñó un estudio que evaluara la tasa de cicatrización cutánea en úlceras agudas en ratas Sprague-Dawley después de recibir la administración de células madre mesenquimatosas derivadas de tejido adiposo. Métodos: al primer grupo, denominado células madre (CM), se le administró células mesenquimatosas derivadas de tejido adiposo inyectadas, tanto periulcerosa como intraulcerosa. El segundo grupo, denominado neobol (N), recibió tratamiento con neobolR tópico, cuyo principio activo es el clostebol. El tercer grupo, denominado control (C), fue sometido a la misma manipulación quirúrgica que los dos grupos anteriores, pero no recibió ningún tipo de tratamiento. Después de realizar una úlcera aguda en el dorso de las ratas y recibir el tratamiento respectivo, se evaluó la tasa de cicatrización (día 1 de la úlcera - día X úlcera) / día 1 de la úlcera en todos los grupos. Resultados: se extrajo un promedio de 1,22 ± 0,46 g de muestra y se aislaron 3,5 x 105 células, con un inóculo promedio de 2,4 x 104 y una viabilidad del 95,5%. La positividad para el antígeno CD29 mediante citometría de flujo fue del 96,5%. El análisis histológico realizado a los 7 días posteriores a la cicatrización clínica, demostró que el grupo de CM presentó la combinación de mayor vascularización y formación de epitelio, así como mayor porcentaje de cicatrización en relación con el grupo N (H(1)=5,61; p < 0,01) y C (H(1)=10,47; p < 0,001). Conclusión: el estudio sugiere que las células madre derivadas del tejido adiposo aumentan la tasa de cicatrización.

Abstract Objective: To evaluate the rate of cutaneous cicatrization of acute ulcers in Sprague-Dawley rats, after receiving the administration of mesenchymal stem cells derived from adipose tissue. Methods: There were three experimental groups. After an acute ulcer was performed in the backs of the rats they received either stem cells (SC), Clostebol, (NeobolR group, N) or no treatment (control group, C), the ulcer cutaneous healing rate was assessed as follows: (ulcer day 1 - ulcer day X)/in all groups day 1. Stem cells were extracted from adipose tissue in the inguinal pad and then injected in the Stem cells group. Results: An average of 1.22 ± 0,46 grams (g) of adipose tissue was extracted and 3.5 x 105 cells were isolated with an average 2.4 x 104 inoculant, 95,5% cell viability. The CD29 antigen positivity on the stem cells assessed by flow cytometry was 96,5%. The histological analysis performed 7 days after the clinical healing showed that the SC group showed the highest vascularization and epithelial tissue formation. When comparing the average healing percentage among groups, only the SC group showed significant differences in contrast with the N group (H (1) = 5.61; p < 0,01) and C (H (1) = 10.47; p< 0.001). Conclusion: This study suggests that mesenchymal cells derived from adipose tissue increase the healing rate.