Clinical evaluation of allogeneic adipose tissue-derived stem cells for the treatment of osteoarthritis secondary to hip dysplasia in dogs / Avaliação da eficácia clínica do uso de células-tronco mesenquimais alógenas derivadas de tecido adiposo no tratamento da osteoartrite secundária a displasia coxofemoral em cães

This study assessed the efficacy of an intra-articular injection of allogeneic adipose tissue-derived from mesenchymal stem cells (AD-MSCs) for the treatment of hip dysplasia in dogs. The study group included 12 otherwise healthy dogs of different breeds, ages, weights, and degrees of hip dysplasia diagnosed using radiography. An orthopedic assessment was performed on all dogs before and at 30, 60, and 90 days after infusion of AD-MSCs(2 × 106cells). On the same days, each dog's owner answered a questionnaire based on theHelsinkiChronic Pain Index. The data were converted to ordinal data based on the score for each variable, and the Friedman test for multiple comparisons was used to verify the results. Compared with the corresponding values on day 0, orthopedic and gait assessments and owners' reported pain indexes improved over the 90-day observation period. These results suggested that treatment with allogeneic AD-MSCs significantly reduced the clinical signs associated with hip dysplasia during the study period. However, long-term studies are needed to determine the optimal therapeutic protocol for routine clinical use of AD-MSCs in hip dysplasia.

O objetivo deste estudo foi avaliar a eficácia clínica da aplicação intra-articular de células-tronco mesenquimais alógenas derivadas do tecido adiposo (AD-CTM) no tratamento de cães portadores de osteoartrite secundária a displasia do coxofemoral (DCF). Doze cães de ambos os sexos, diferentes raças, idades e peso, portadores de graus variados de DCF comprovada em radiografia e livres de quaisquer outras alterações clínicas ou ortopédicas, foram utilizados no estudo. Todos os cães foram submetidos a avaliação ortopédica nos dias 0, 30, 60 e 90 após aplicação de AD-CTM na dose de 2 x 106. Além disso, os tutores preencheram a um questionário, baseado no Índice de dor crônica de Helsinque (IDCH) nos mesmos intervalos. Em comparação com o dia 0, observou-se melhora significativa na avalição em locomoção e físico-ortopédica assim como na avaliação dos tutores pelo IDCH ao longo dos 90 dias. Os resultados permitem inferir que as AD-CTM alógenas contribuíram significativamente para a redução dos sinais clínicos comumente associados a DCF durante o período de estudo. Entretanto, há necessidade de estudos de longo prazo para melhor determinação de protocolos terapêuticos baseados no uso de AD-CTM na rotina clínica.

Para além dos limites da saúde: cuidado em perspectiva interdisciplinar / Beyond the limits of health: care from an interdisciplinary perspectivea / Más allá de los límites de la salud: cuidado desde una perspectiva interdisciplinaria

Este relato apresenta reflexões sobre uma experiência interdisciplinar que envolveu idealização, planejamento, organização, realização e divulgação de um evento científico virtual como requisito da disciplina Seminários Avançados de Pesquisa 1, oferecida pelo Programa de Pós-graduação em Informação e Comunicação em Saúde desenvolvido pelo Instituto de Comunicação e Informação Científica e Tecnológica em Saúde, uma das unidades técnico-científicas da Fundação Oswaldo Cruz. Os doutorandos da turma de 2020 promoveram o webinário "Para além dos limites da saúde: cuidado em perspectiva interdisciplinar", como parte da formação acadêmica, e este relato é produto científico daquele evento. Além do desenvolvimento de habilidades e atitudes, essa experiência representou um aprendizado pessoal, intangível e emocional do cuidado para além dos limites da saúde, e também ético sobre as "coisas negligenciadas" e acerca da força dos consensos diante de um cenário complexo marcado por uma pandemia causada pelo vírus SARS-CoV-2, a pandemia de covid-19

This report presents reflections about an interdisciplinary experience that involved the idealization, planning, organization, implementation and dissemination of a virtual scientific event as requisite for the Seminários Avançados de Pesquisa 1 (Advanced Research Seminars 1), one of the disciplines offered by the Programa de Pós-graduação em Informação e Comunicação em Saúde (Postgraduate Program in Health Information and Communication) developed by the Instituto de Comunicação e Informação Científica e Tecnológica em Saúde (Institute of Communication and Scientific and Technological Information in Health), one of the technical-scientific units of the Fundação Oswaldo Cruz (Oswaldo Cruz Foundation). The students who began in 2020 their classes to obtain the PhD Degree promoted the webinar "Beyond the limits of health: care from an interdisciplinary perspective" as part of the academic education, and this report is a scientific product of that event. In addition to the development of skills and attitudes, that experience represented a personal, intangible and emotional learning of the care beyond the limits of health as well as an important ethical learning about the "neglected things" and the strength of consensus in the face of a complex scenario marked by a pandemic caused by the SARS-CoV-2, the Covid-19 pandemic

Este relato presenta reflexiones sobre una experiencia interdisciplinar que implicó la idealización, planificación, organización, realización y difusión de un evento científico virtual como requisito de la disciplina Seminários Avançados de Pesquisa 1 (Seminarios Avanzados de Investigación 1), ofrecida por el Programa de Pós-graduação em Informação e Comunicação em Saúde (Programa de Posgrado en Información y Comunicación en Salud) desarrollado por el Instituto de Comunicação e Informação Científica e Tecnológica em Saúde (Instituto de Comunicación y Información Científica y Tecnológica en Salud), una de las unidades técnicas y científicas de la Fundação Oswaldo Cruz (Fundación Oswaldo Cruz). Los doctorandos del curso de 2020 promovieron el webinario "Más allá de los límites de la salud: cuidado desde una perspectiva interdisciplinaria" como parte de la formación académica, y este relatoes producto científico de aquel evento. Además del desarrollo de habilidades y actitudes, esa experiencia representó un aprendizaje personal, intangible y emocional del cuidado más allá de los límites de la salud, y también un importante aprendizaje ético sobre las "cosas tratadas con negligencia" y aún acerca de la fuerza de los consensos ante un escenario complejo marcado por una pandemia provocada por el virus SARS-CoV-2, la pandemia de Covid-19

Subconjunctival use of allogeneic mesenchymal stem cells to treat chronic superficial keratitis in German shepherd dogs: Pilot study.

Background: Chronic superficial keratitis (CSK) is an ocular condition in dogs characterized by corneal opacification leading to visual function impairment. Control of this chronic condition requires the use of topical immunomodulators or corticosteroids daily. Regenerative medicine has shown promising results in several fields of medicine. Aim: The aim of this study was to evaluate the clinical effect of allogeneic mesenchymal stem cells (MSCs) of adipose tissue applied via subconjunctival in dogs with CSK. Methods: A series of cases of eight dogs diagnosed with CSK were divided into two groups, four dogs each; the conventional treatment group received prednisolone 1% as topical eye drops and the experimental group (EG) received allogeneic MSCs transplantation. The dogs had not previously been treated for CSK. Systemic and ophthalmologic examinations were performed to exclude other abnormalities. An administered amount of MSC (1 × 106 cells each time) was injected via subconjunctival in the peri-limbal region at 0 and 30 days. The animals were followed for 110 days for clinical evaluation, and, at the same time, the images of the corneal abnormalities were obtained and analyzed in the ImageJ software. The statistical analysis was performed in the GrandPrism 7.0 software. Results: Initial and final images revealed that areas with neovascularization, inflammatory infiltrate, and opacity regressed in most eyes in both groups (7/8 eyes in each group) at the end of the 110 days, p = 0.0391 and p = 0.0078 respectively, but this response was minor in the EG comparing to conventional group (CG) (p = 0.026). No local or systemic side effects were observed. Conclusions: Despite the small melioration, MSCs treatment suggests clinical improvement in patients with CSK after 110 days without any local or systemic side effects. However, the improvement achieved was significantly less than the observed within CG. Further studies still are needed to evaluate the use and benefits of stem cells as an adjunct treatment for CSK.

AeMOPE-1, a Novel Salivary Peptide From Aedes aegypti, Selectively Modulates Activation of Murine Macrophages and Ameliorates Experimental Colitis.

The sialotranscriptomes of Aedes aegypti revealed a transcript overexpressed in female salivary glands that codes a mature 7.8 kDa peptide. The peptide, specific to the Aedes genus, has a unique sequence, presents a putative secretory nature and its function is unknown. Here, we confirmed that the peptide is highly expressed in the salivary glands of female mosquitoes when compared to the salivary glands of males, and its secretion in mosquito saliva is able to sensitize the vertebrate host by inducing the production of specific antibodies. The synthetic version of the peptide downmodulated nitric oxide production by activated peritoneal murine macrophages. The fractionation of a Ae. aegypti salivary preparation revealed that the fractions containing the naturally secreted peptide reproduced the nitric oxide downmodulation. The synthetic peptide also selectively interfered with cytokine production by murine macrophages, inhibiting the production of IL-6, IL-12p40 and CCL2 without affecting TNF-α or IL-10 production. Likewise, intracellular proteins associated with macrophage activation were also distinctively modulated: while iNOS and NF-κB p65 expression were diminished, IκBα and p38 MAPK expression did not change in the presence of the peptide. The anti-inflammatory properties of the synthetic peptide were tested in vivo on a dextran sulfate sodium-induced colitis model. The therapeutic administration of the Ae. aegypti peptide reduced the leukocytosis, macrophage activity and nitric oxide levels in the gut, as well as the expression of cytokines associated with the disease, resulting in amelioration of its clinical signs. Given its biological properties in vitro and in vivo, the molecule was termed Aedes-specific MOdulatory PEptide (AeMOPE-1). Thus, AeMOPE-1 is a novel mosquito-derived immunobiologic with potential to treat immune-mediated disorders.

AeMOPE-1: a novel salivary peptide from Aedes aegypti, selectively modulates activation of murine macrophages and ameliorates experimental Colitis

The sialotranscriptomes of Aedes aegypti revealed a transcript overexpressed in female salivary glands that codes a mature 7.8 kDa peptide. The peptide, specific to the Aedes genus, has a unique sequence, presents a putative secretory nature and its function is unknown. Here, we confirmed that the peptide is highly expressed in the salivary glands of female mosquitoes when compared to the salivary glands of males, and its secretion in mosquito saliva is able to sensitize the vertebrate host by inducing the production of specific antibodies. The synthetic version of the peptide downmodulated nitric oxide production by activated peritoneal murine macrophages. The fractionation of a Ae. aegypti salivary preparation revealed that the fractions containing the naturally secreted peptide reproduced the nitric oxide downmodulation. The synthetic peptide also selectively interfered with cytokine production by murine macrophages, inhibiting the production of IL-6, IL-12p40 and CCL2 without affecting TNF-α or IL-10 production. Likewise, intracellular proteins associated with macrophage activation were also distinctively modulated: while iNOS and NF-κB p65 expression were diminished, IκBα and p38 MAPK expression did not change in the presence of the peptide. The anti-inflammatory properties of the synthetic peptide were tested in vivo on a dextran sulfate sodium-induced colitis model. The therapeutic administration of the Ae. aegypti peptide reduced the leukocytosis, macrophage activity and nitric oxide levels in the gut, as well as the expression of cytokines associated with the disease, resulting in amelioration of its clinical signs. Given its biological properties in vitro and in vivo, the molecule was termed Aedes-specific MOdulatory PEptide (AeMOPE-1). Thus, AeMOPE-1 is a novel mosquito-derived immunobiologic with potential to treat immune-mediated disorders.

Effects of canine adipose-derived mesenchymal stem cells on the epithelialization of rabbits' skin autograft (Oryctolagus cuniculus) / Efeitos das células-tronco mesenquimais derivadas do tecido adiposo de cães na epitelização de autoenxertos de pele em coelhos (Oryctolagus cuniculus)

The present study aimed to evaluate the effects of mesenchymal stem cells derived from canine adipose tissue in the healing process of full-thickness mesh skin grafts in rabbits. The stem cells were collected from young dogs; and, after characterization, remained in cryopreservation, in independent doses containing 2 x 106 cells. The mesh distal limb graft technique was performed in 60 rabbits, divided into three groups, CG (Control Group), GT1 (Intralesional Stem Cell Treated Group), and GT2 (Intravenous Stem Cell Treated Group), containing 20 animals each. After grafting, each group was randomly divided into four subgroups according to euthanasia time 3, 7, 14, and 30 days, containing five animals in each group. Animals of GT1_14, GT1_30, and GT2_14, GT2_30 subgroups received a second dose of xenogeneic cells on the seventh day. Meanwhile, animals from GT1_30 and GT2_30 received the third dose of xenogeneic cells on day 14. The groups treated with xenogeneic stem cells positively affected type III collagen re-epithelialization and deposition, and possibly GT1 had a controlled inflammatory response. However, no effect on angiogenesis. Thus, it was possible to demonstrate tolerance and therapeutic action of mesenchymal stem cells from canine adipose tissue in skin grafts in rabbits.(AU)

O presente estudo teve como principal objetivo avaliar os efeitos das células-tronco mesenquimais derivadas do tecido adiposo de cães no processo de cicatrização de autoenxertos de pele de espessura total em malha em coelhos. As células-tronco foram coletadas de cães jovens, após a caracterização estas permaneceram em criopreservação, em doses individuais contendo 2 x 106 células. A técnica de enxerto em malha na região distal do membro foi realizada em 60 coelhos, divididos em três grupos, GC (Grupo Controle), GT1 (Grupo tratado com células-tronco intralesional) e GT2 (Grupo tratado com células-tronco via endovenosa), contendo 20 animais cada. Imediatamente após a enxertia, cada grupo foi dividido aleatoriamente em quatro subgrupos, de acordo com o tempo de eutanásia 3, 7, 14 e 30 dias contendo cinco animais cada. Animais dos subgrupos GT1_14, GT1_30 e GT2_14, GT2_30 receberam uma segunda dose de células xenógenas no sétimo dia. Ademais, animais do GT1_30 e do GT2_30 receberam a terceira dose de células xenógenas no dia 14. Os grupos tratados com células-tronco xenógenas tiveram um efeito positivo na reepitelização e deposição de colágeno tipo III, e possivelmente, o GT1 teve uma resposta inflamatória controlada, entretanto o efeito na angiogênese não foi observado. Dessa forma, foi possível demonstrar que houve tolerância e ação terapêutica das células-tronco mesenquimais derivadas do tecido adiposo de cães em enxertos de pele em coelhos.(AU)

Effects of canine adipose-derived mesenchymal stem cells on the epithelialization of rabbits' skin autograft (Oryctolagus cuniculus) / Efeitos das células-tronco mesenquimais derivadas do tecido adiposo de cães na epitelização de autoenxertos de pele em coelhos (Oryctolagus cuniculus)

The present study aimed to evaluate the effects of mesenchymal stem cells derived from canine adipose tissue in the healing process of full-thickness mesh skin grafts in rabbits. The stem cells were collected from young dogs; and, after characterization, remained in cryopreservation, in independent doses containing 2 x 106 cells. The mesh distal limb graft technique was performed in 60 rabbits, divided into three groups, CG (Control Group), GT1 (Intralesional Stem Cell Treated Group), and GT2 (Intravenous Stem Cell Treated Group), containing 20 animals each. After grafting, each group was randomly divided into four subgroups according to euthanasia time 3, 7, 14, and 30 days, containing five animals in each group. Animals of GT1_14, GT1_30, and GT2_14, GT2_30 subgroups received a second dose of xenogeneic cells on the seventh day. Meanwhile, animals from GT1_30 and GT2_30 received the third dose of xenogeneic cells on day 14. The groups treated with xenogeneic stem cells positively affected type III collagen re-epithelialization and deposition, and possibly GT1 had a controlled inflammatory response. However, no effect on angiogenesis. Thus, it was possible to demonstrate tolerance and therapeutic action of mesenchymal stem cells from canine adipose tissue in skin grafts in rabbits.(AU)

O presente estudo teve como principal objetivo avaliar os efeitos das células-tronco mesenquimais derivadas do tecido adiposo de cães no processo de cicatrização de autoenxertos de pele de espessura total em malha em coelhos. As células-tronco foram coletadas de cães jovens, após a caracterização estas permaneceram em criopreservação, em doses individuais contendo 2 x 106 células. A técnica de enxerto em malha na região distal do membro foi realizada em 60 coelhos, divididos em três grupos, GC (Grupo Controle), GT1 (Grupo tratado com células-tronco intralesional) e GT2 (Grupo tratado com células-tronco via endovenosa), contendo 20 animais cada. Imediatamente após a enxertia, cada grupo foi dividido aleatoriamente em quatro subgrupos, de acordo com o tempo de eutanásia 3, 7, 14 e 30 dias contendo cinco animais cada. Animais dos subgrupos GT1_14, GT1_30 e GT2_14, GT2_30 receberam uma segunda dose de células xenógenas no sétimo dia. Ademais, animais do GT1_30 e do GT2_30 receberam a terceira dose de células xenógenas no dia 14. Os grupos tratados com células-tronco xenógenas tiveram um efeito positivo na reepitelização e deposição de colágeno tipo III, e possivelmente, o GT1 teve uma resposta inflamatória controlada, entretanto o efeito na angiogênese não foi observado. Dessa forma, foi possível demonstrar que houve tolerância e ação terapêutica das células-tronco mesenquimais derivadas do tecido adiposo de cães em enxertos de pele em coelhos.(AU)

Reduction in the inflammatory markers CD4, IL-1, IL-6 and TNFα in dogs with keratoconjunctivitis sicca treated topically with mesenchymal stem cells.

Keratoconjunctivitis sicca (KCS) is of predominantly immune-mediated origin. Dogs are an excellent model for understanding this disease, as the origin of KCS in dogs is like that in humans. The objective of this study was to localize and quantify immunological markers, such as CD4 lymphocytes, interleukin (IL)-1, IL-6 and tumor necrosis factor alpha (TNFα), before and after topical treatment with mesenchymal stem cells (MSCs). Twenty-two dogs positive for KCS were topically treated with 50 µL (1 × 106 MSCs) in the conjunctival sac and were evaluated for 6 months. The levels of the markers CD4, IL-6, IL-1 and TNFα were analyzed in conjunctival biopsy and cytology of the third eyelid gland by immunohistochemistry and immunocytochemistry. The results showed that before treatment, there was marked expression of all the markers (CD4, IL-6, IL-1 and TNFα), and after 6 months, there were significant (p < .05) reductions in the expression levels of all the markers. These results demonstrated that topical MSC treatment promotes a significant decrease in the expression levels of these inflammatory markers and could be used as adjuvant therapy in the treatment of KCS in dogs and humans. In addition, these markers can be excellent tools for diagnosing and analyzing the progression of KCS.

Evaluation of inflammatory skin infiltrate following Aedes aegypti bites in sensitized and non-sensitized mice reveals saliva-dependent and immune-dependent phenotypes.

During probing and blood feeding, haematophagous mosquitoes inoculate a mixture of salivary molecules into their vertebrate hosts' skin. In addition to the anti-haemostatic and immunomodulatory activities, mosquito saliva also triggers acute inflammatory reactions, especially in sensitized hosts. Here, we characterize the oedema and the cellular infiltrate following Aedes aegypti mosquito bites in the skin of sensitized and non-sensitized BALB/c mice by flow cytometry. Ae. aegypti bites induced an increased oedema in the ears of both non-sensitized and salivary gland extract- (SGE-)sensitized mice, peaking at 6 hr and 24 hr after exposure, respectively. The quantification of the total cell number in the ears revealed that the cellular recruitment was more robust in SGE-sensitized mice than in non-sensitized mice, and the histological evaluation confirmed these findings. The immunophenotyping performed by flow cytometry revealed that mosquito bites were able to produce complex changes in cell populations present in the ears of non-sensitized and SGE-sensitized mice. When compared with steady-state ears, the leucocyte populations significantly recruited to the skin after mosquito bites in non-sensitized and sensitized mice were eosinophils, neutrophils, monocytes, inflammatory monocytes, mast cells, B-cells and CD4+ T-cells, each one with its specific kinetics. The changes in the absolute number of cells suggested two cell recruitment profiles: (i) a saliva-dependent migration; and (ii) a migration dependent on the immune status of the host. These findings suggest that mosquito bites influence the skin microenvironment by inducing differential cell migration, which is dependent on the degree of host sensitization to salivary molecules.

Aedes aegypti saliva impairs M1-associated proinflammatory phenotype without promoting or affecting M2 polarization of murine macrophages.

BACKGROUND: During the feeding process, the mouthparts of hematophagous mosquitoes break the skin barrier and probe the host tissue to find the blood. The saliva inoculated in this microenvironment modulates host hemostasis, inflammation and adaptive immune responses. However, the mechanisms involved in these biological activities remain poorly understood and few studies explored the potential roles of mosquito saliva on the individual cellular components of the immune system. Here, we report the immunomodulatory activities of Aedes aegypti salivary cocktail on murine peritoneal macrophages. RESULTS: The salivary gland extract (SGE) of Ae. aegypti inhibited the production of nitric oxide and inflammatory cytokines such as interleukin-6 (IL-6) and IL-12, as well as the expression of inducible nitric oxide synthase and NF-κB by murine macrophages stimulated by lipopolysaccharide (LPS) plus interferon-γ (IFN-γ). The spare respiratory capacity, the phagocytic and microbicidal activities of these macrophages were also reduced by Ae. aegypti SGE. These phenotypic changes are consistent with SGE suppressing the proinflammatory program of M1 macrophages. On the other hand, Ae. aegypti SGE did not influence M2-associated markers (urea production, arginase-1 and mannose receptor-1 expression), either in macrophages alternatively activated by IL-4 or in those classically activated by LPS plus IFN-γ. In addition, Ae. aegypti SGE did not display any cytokine-binding activity, nor did it affect macrophage viability, thus excluding supposed experimental artifacts. CONCLUSIONS: Given the importance of macrophages in a number of biological processes, our findings help to enlighten how vector saliva modulates vertebrate host immunity.

Mice Selected for Acute Inflammation Present Altered Immune Response during Pristane-Induced Arthritis Progression.

Mouse lines selected for maximal (AIRmax) or minimal acute inflammatory reaction (AIRmin) were used to characterize the immune response and the influence of genetic background during pristane-induced arthritis (PIA). Susceptible AIRmax mice demonstrated exacerbated cellular profiles during PIA, with intense infiltration of lymphocytes, as well as monocytes/macrophages and neutrophils, producing higher levels of IL-1ß, IFN-γ, TNF-α, IL-10, total IgG3, and chemokines. Resistant AIRmin mice controlled cell activation more efficiently than the AIRmax during arthritis progression. The weight alterations of the spleen and thymus in the course of PIA were observed. Our data suggest that selected AIRmax cellular and genetic immune mechanisms contribute to cartilage damage and arthritis severity, evidencing many targets for therapeutic actions.

Mice selected for acute inflammation present altered immune response during Pristane-induced arthritis progression

Mouse lines selected for maximal (AIRmax) or minimal acute inflammatory reaction (AIRmin) were used to characterize the immune response and the influence of genetic background during pristane-induced arthritis (PIA). Susceptible AIRmax mice demonstrated exacerbated cellular profiles during PIA, with intense infiltration of lymphocytes, as well as monocytes/macrophages and neutrophils, producing higher levels of IL-1ß, IFN-γ, TNF-α, IL-10, total IgG3, and chemokines. Resistant AIRmin mice controlled cell activation more efficiently than the AIRmax during arthritis progression. The weight alterations of the spleen and thymus in the course of PIA were observed. Our data suggest that selected AIRmax cellular and genetic immune mechanisms contribute to cartilage damage and arthritis severity, evidencing many targets for therapeutic actions.

Mice selected for acute inflammation present altered immune response during pristane-induced arthritis progression

Mouse lines selected for maximal (AIRmax) or minimal acute inflammatory reaction (AIRmin) were used to characterize the immune response and the influence of genetic background during pristane-induced arthritis (PIA). Susceptible AIRmax mice demonstrated exacerbated cellular profiles during PIA, with intense infiltration of lymphocytes, as well as monocytes/macrophages and neutrophils, producing higher levels of IL-1ß, IFN-?, TNF-a, IL-10, total IgG3, and chemokines. Resistant AIRmin mice controlled cell activation more efficiently than the AIRmax during arthritis progression. The weight alterations of the spleen and thymus in the course of PIA were observed. Our data suggest that selected AIRmax cellular and genetic immune mechanisms contribute to cartilage damage and arthritis severity, evidencing many targets for therapeutic actions.

Células-Tronco na cirurgia reconstrutiva: revisão de literatura / Stem cells in reconstructive surgery: review of literature / Las células madre en cirugía reconstructiva: revisión de la literatura

As células-tronco mesenquimais (MSCs) são excelente alternativa para estimular a cicatrização de feridas agudas e crônicas, mediante uma variedade de funções incluindo suporte trófico, anti-inflamatória,imunomoduladora, revascularização, atividade antiapoptótica, capacidade de diferenciação e quimiotaxia. Assim,as cirurgias reconstrutivas são utilizadas para corrigir defeitos extensos, promovendo melhor resultados estéticos e proporcionando que o paciente retorne com maior rapidez a rotina normal. Uma das modalidades da cirurgia reconstrutiva são os enxertos cutâneos, os quais consistem de um segmento de pele separado por completo da área doadora, além disso são desprovidos de pedículo vascular, implicando em complicações pós-cirúrgicas como isquemia e necrose. Sendo assim, o uso de substâncias que estimulam a angiogênese e favoreçam a cicatrização são alvo de muitos estudos, como no caso das células-troncos. Dessa forma, objetivou-se com esse estudo realizar uma revisão de literatura sobre as células-tronco associadas aos enxertos cutâneos, a fim de avaliar seus efeitos na cicatrização.

Mesenchymal stem cells (MSCs) are an excellent alternative to stimulate the healing of acute and chronic wounds, acting through a variety of functions including trophic support, anti-inflammatory,immunomodulatory, revascularization, antiapoptotic activity, differentiation capacity and chemotaxis. Like this,reconstructive surgeries are used to correct extensive defects, promoting better aesthetic results and allowing the patient to return to normal routine more quickly. One of the modalities of reconstructive surgery are the cutaneous grafts, which involve a segment of skin completely separated from the donor area, besides they lack of vascular pedicle, resulting in postoperative complications such as ischemia and necrosis. Therefore, the use of substances that stimulate angiogenesis and promote wound healing are the target of many studies, as in the case of stem cells. Thus, this study aimed to review the literature on stem cells associated with cutaneous grafts,in order to evaluate their effects on wound healing.

Las células madre mesenquimales (MSCs) constituyen una excelente alternativa para estimular la cicatrización de heridas agudas y crónicas, mediante vários tipos de funciones tales como soporte trófico,antiinflamatório, inmunomodulación, neovascularización, actividad antiapoptosis, capacidad de diferenciación y quimiotaxis. Mientras que, las cirugías reconstructivas son utilizadas para corregir defectos extensos,proporcionando mejores resultados estéticos y permitiendo que el paciente retorne más rápido a la rutina normal.Uno de los métodos de cirugía recosntructiva son los injertos de piel, consisten en un segmento de piel separada por completo de la zona donante y desprovistos de pedículo vascular, reultando en complicaciones posquirúrgicas tales como isquemia y necrosis. Por lo tanto, el uso de substancias que estimulan la angiogénesis y cicatrización de heridas, tales como las células madre, son objeto de numerosos estúdios. Por esta razón, el objetivo de este trabajo es realizar una revisión de literatura sobre las células madres e injertos cutâneos, con el propósito de evaluar los efectos en la cicatrización.

Células-Tronco na cirurgia reconstrutiva: revisão de literatura / Stem cells in reconstructive surgery: review of literature / Las células madre en cirugía reconstructiva: revisión de la literatura

As células-tronco mesenquimais (MSCs) são excelente alternativa para estimular a cicatrização de feridas agudas e crônicas, mediante uma variedade de funções incluindo suporte trófico, anti-inflamatória,imunomoduladora, revascularização, atividade antiapoptótica, capacidade de diferenciação e quimiotaxia. Assim,as cirurgias reconstrutivas são utilizadas para corrigir defeitos extensos, promovendo melhor resultados estéticos e proporcionando que o paciente retorne com maior rapidez a rotina normal. Uma das modalidades da cirurgia reconstrutiva são os enxertos cutâneos, os quais consistem de um segmento de pele separado por completo da área doadora, além disso são desprovidos de pedículo vascular, implicando em complicações pós-cirúrgicas como isquemia e necrose. Sendo assim, o uso de substâncias que estimulam a angiogênese e favoreçam a cicatrização são alvo de muitos estudos, como no caso das células-troncos. Dessa forma, objetivou-se com esse estudo realizar uma revisão de literatura sobre as células-tronco associadas aos enxertos cutâneos, a fim de avaliar seus efeitos na cicatrização.(AU)

Mesenchymal stem cells (MSCs) are an excellent alternative to stimulate the healing of acute and chronic wounds, acting through a variety of functions including trophic support, anti-inflammatory,immunomodulatory, revascularization, antiapoptotic activity, differentiation capacity and chemotaxis. Like this,reconstructive surgeries are used to correct extensive defects, promoting better aesthetic results and allowing the patient to return to normal routine more quickly. One of the modalities of reconstructive surgery are the cutaneous grafts, which involve a segment of skin completely separated from the donor area, besides they lack of vascular pedicle, resulting in postoperative complications such as ischemia and necrosis. Therefore, the use of substances that stimulate angiogenesis and promote wound healing are the target of many studies, as in the case of stem cells. Thus, this study aimed to review the literature on stem cells associated with cutaneous grafts,in order to evaluate their effects on wound healing.(AU)

Las células madre mesenquimales (MSCs) constituyen una excelente alternativa para estimular la cicatrización de heridas agudas y crónicas, mediante vários tipos de funciones tales como soporte trófico,antiinflamatório, inmunomodulación, neovascularización, actividad antiapoptosis, capacidad de diferenciación y quimiotaxis. Mientras que, las cirugías reconstructivas son utilizadas para corregir defectos extensos,proporcionando mejores resultados estéticos y permitiendo que el paciente retorne más rápido a la rutina normal.Uno de los métodos de cirugía recosntructiva son los injertos de piel, consisten en un segmento de piel separada por completo de la zona donante y desprovistos de pedículo vascular, reultando en complicaciones posquirúrgicas tales como isquemia y necrosis. Por lo tanto, el uso de substancias que estimulan la angiogénesis y cicatrización de heridas, tales como las células madre, son objeto de numerosos estúdios. Por esta razón, el objetivo de este trabajo es realizar una revisión de literatura sobre las células madres e injertos cutâneos, con el propósito de evaluar los efectos en la cicatrización.(AU)

Allogeneic Mesenchymal Stem Cell Transplantation in Dogs With Keratoconjunctivitis Sicca.

Keratoconjunctivitis sicca (KCS) is a dysfunction in tear production associated with clinical signs, which include conjunctival hyperemia, ocular discharge, discomfort, pain, and, eventually, corneal vascularization and pigmentation. Immunosuppressive drugs are routinely administrated for long periods to treat KCS but with side effects and limited results. Evaluation of the clinical benefits of intralacrimal transplantation of allogeneic mesenchymal stem cells (MSCs) in dogs with mild-moderate and severe KCS was done. A total of 24 eyes with KCS from 15 dogs of different breeds were enrolled in the present study. A single transplantation of MSCs (1 × 106) directly into lacrimal glands (dorsal and third eyelid) was performed. The Schirmer tear tests (STTs) and ocular surface improvements were used to assess short- and long-term effects of these cells. The STTs were carried out on day 0 (before MSCs transplantation) and on days 7, 14, 21, and 28, as well as 6 and 12 months after MSC transplantation. Our data demonstrate that allogeneic MSC transplantation in KCS dogs is safe since no adverse effects were observed immediately after transplantation and in short- and long-term follow-ups. A statistically significant increase in the STT and ocular surface improvements was found in all eyes studied. In all the eyes with mild-moderate KCS, STT values reverted to those of healthy eyes, while in eyes with severe KCS, although complete reversion was not found, there was improvement in tear production and in other clinical signs. Our study shows that a single dose of a low number of MSCs can be used to treat KCS in dogs. In contrast to immunosuppressive drug use, MSC transplantation has an effect over a long period (up to 12 months), even after a single administration, and does not require daily drug administration.

Exposure to Aedes aegypti Bites Induces a Mixed-Type Allergic Response following Salivary Antigens Challenge in Mice.

Classical studies have shown that Aedes aegypti salivary secretion is responsible for the sensitization to mosquito bites and many of the components present in saliva are immunogenic and capable of inducing an intense immune response. Therefore, we have characterized a murine model of adjuvant-free systemic allergy induced by natural exposure to mosquito bites. BALB/c mice were sensitized by exposure to A. aegypti mosquito bites and intranasally challenged with phosphate-buffered saline only or the mosquito's salivary gland extract (SGE). Blood, bronchoalveolar lavage (BAL) and lung were collected and evaluated for cellularity, histopathological analyses, cytokines and antibody determination. Respiratory pattern was analyzed by Penh measurements and tracheal segments were obtained to study in vitro reactivity to methacholine. BAL recovered from sensitized mice following challenge with SGE showed an increased number of eosinophils and Th2 cytokines such as IL-4, IL-5 and IL-13. Peribronchoalveolar eosinophil infiltration, mucus and collagen were also observed in lung parenchyma of sensitized mice, suggesting the development of a typical Th2 response. However, the antibody profile in serum of these mice evidenced a mixed-type response with presence of both, IgG1/IgE (Th2-related) and IgG2a (Th1-related) isotypes. In addition, changes in breathing pattern and tracheal reactivity to methacholine were not found. Taken together, our results show that A. aegypti bites trigger an atypical allergic reaction, with some classical cellular and soluble Th2 components in the lung, but also systemic Th1 and Th2 antibody isotypes and no change in either the respiratory pattern or the trachea responsiveness to agonist.

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.

Transcriptome sequencing and developmental regulation of gene expression in Anopheles aquasalis.

BACKGROUND: Anopheles aquasalis is a major malaria vector in coastal areas of South and Central America where it breeds preferentially in brackish water. This species is very susceptible to Plasmodium vivax and it has been already incriminated as responsible vector in malaria outbreaks. There has been no high-throughput investigation into the sequencing of An. aquasalis genes, transcripts and proteins despite its epidemiological relevance. Here we describe the sequencing, assembly and annotation of the An. aquasalis transcriptome. METHODOLOGY/PRINCIPAL FINDINGS: A total of 419 thousand cDNA sequence reads, encompassing 164 million nucleotides, were assembled in 7544 contigs of ≥ 2 sequences, and 1999 singletons. The majority of the An. aquasalis transcripts encode proteins with their closest counterparts in another neotropical malaria vector, An. darlingi. Several analyses in different protein databases were used to annotate and predict the putative functions of the deduced An. aquasalis proteins. Larval and adult-specific transcripts were represented by 121 and 424 contig sequences, respectively. Fifty-one transcripts were only detected in blood-fed females. The data also reveal a list of transcripts up- or down-regulated in adult females after a blood meal. Transcripts associated with immunity, signaling networks and blood feeding and digestion are discussed. CONCLUSIONS/SIGNIFICANCE: This study represents the first large-scale effort to sequence the transcriptome of An. aquasalis. It provides valuable information that will facilitate studies on the biology of this species and may lead to novel strategies to reduce malaria transmission on the South American continent. The An. aquasalis transcriptome is accessible at http://exon.niaid.nih.gov/transcriptome/An_aquasalis/Anaquexcel.xlsx.

Análise comparativa do potencial de migração de células-tronco imaturas de polpa dentária humana em ratos induzidos a necrose tubular aguda utilizando as vias: intraperitoneal e endovenosa / Comparative analysis of the potential for migration of immature stem cells from human dental pulp in rats induced acute tubular necrosis using routes: intraperitoneal and intravenous

Pesquisas com células tronco têm demonstrado que essa alternativa terapêutica tem grande potencial em diversas patologias. Contudo, muito ainda precisa ser elucidado quanto aos mecanismos de ação, dose e melhores vias de aplicação, para que essa alternativa terapêutica seja efetivamente incorporada às outras. Neste contexto, a via de aplicação se torna crucial na efetividade do tratamento. O objetivo do presente estudo foi avaliar no modelo de necrose tubular aguda (NTA) induzido pelo glicerol o potencial de migração das células tronco mesenquimais derivadas de polpa dentária humana, utilizando as vias intraperitoneal e a via intravenosa. Os nossos resultados demonstraram que neste modelo, não há diferença na migração das células para o órgão alvo, o rim. Contudo, a via intravenosa proporcionou uma melhora da função renal mais significativa do que quando comparada a via intraperitoneal.

Stem cell research have demonstrated that this alternative therapy has great potential in various diseases. However, much remains to be elucidated about the mechanisms of action, dosage and best means of implementation, so that alternative therapy may be incorporated other. In this context, the route of application becomes crucial in the effectiveness of treatment. The aim of this study was to evaluate the model of acute tubular necrosis (NTA) induced by glycerol the potential for migration of mesenchymal stem cells derived from human dental pulp, using the intraperitoneal and intravenous route. Our results demonstrate that this model, there is no difference in cell migration to the target organ, the kidney. However, the intravenous route provided an improvement in renal function more significantly than when compared to the intraperitoneal route.