Bone-marrow mononuclear cells and acellular human amniotic membrane improve global cardiac function without inhibition of the NLRP3 Inflammasome in a rat model of heart failure.

Recent studies have suggested that therapies with stem cells and amniotic membrane can modulate the inflammation following an ischemic injury in the heart. This study evaluated the effects of bone-marrow mononuclear cells (BMMC) and acellular human amniotic membrane (AHAM) on cardiac function and NLRP3 complex in a rat model of heart failure.On the 30th day,the echocardiographic showed improvements on ejection fraction and decreased pathological ventricular remodeling on BMMC and AHAM groups.Oxidative stress analysis was similar between the three groups,and the NLRP3 inflammasome activity were not decreased with the therapeutic use of both BMMC and AHAM,in comparison to the control group.

Tracheal regeneration with acellular human amniotic membrane and 15-deoxy-∆12,14prostaglandinj2 nanoparticles in a rabbit model.

The treatment of tracheal pathologies remains challenging.Nanotechnology allows adding substances to decellularized human amniotic membrane (DHAM), such as 15-Deoxy-∆12,14ProstaglandinJ2 nanoparticles (15D-PGJ2-NC).This study performed a tracheotomy in rabbits randomized into three groups.The tissue repair process was evaluated when treated with DHAM associated or not with 15D-PGJ2-NC.The average of the area in the control group was 54.76% smaller than DHAM group and 41.98% smaller than DHAM + 15D-PGJ2-NC group (p=0.004 for both).The DHAM + 15D-PGJ2-NC group had significantly more immature cartilage (p=0.015).DHAM impregnated with 15D-PGJ2-NC could provide support for the healing of the tracheal defect and may prevent reduction of its lumen.

Histological, Laboratorial, and Clinical Benefits of an Optimized Maintenance Strategy of a Potential Organ Donor-A Rabbit Experimental Model.

INTRODUCTION: Most transplanted organs are obtained from brain-dead donors. Inflammation results in a higher rate of rejection. Objectives: The objective of this animal model of brain death (BD) was to evaluate the effect of the progressive institution of volume expansion, norepinephrine, and combined hormone therapy on clinical, laboratory, and histological aspects. Methods: Twenty rabbits were divided: A (control), B (induction of BD + infusion of crystalloid), C (BD + infusion of crystalloid and noradrenaline (NA)), and D (BD + infusion of crystalloid + vasopressin + levothyroxine + methylprednisolone + NA). The animals were monitored for four hours with consecutives analysis of vital signs and blood samples. The organs were evaluated by a pathologist. Results: In Group D, we observed fewer number and lesser volume of infusions (p = 0.032/0.014) when compared with groups B and C. Mean arterial pressure levels were higher in group D when compared with group B (p = 0.008). Group D had better glycemic control when compared with group C (p = 0.016). Sodium values were elevated in group B in relation to groups C and D (p = 0.021). In Group D, the organ perfusion was better. Conclusion: The optimized strategy of management of BD animals is associated with better hemodynamic, glycemic, and natremia control, besides reducing early signs of ischemia.

Acellular Biomaterials Associated with Autologous Bone Marrow-Derived Mononuclear Stem Cells Improve Wound Healing through Paracrine Effects.

Wound healing is a complex process of repair that involves the interaction between different cell types and involves coordinated interactions between intracellular and extracellular signaling. Bone Marrow Mesenchymal Stem Cells (BMSCs) based and acellular amniotic membrane (AM) therapeutic strategies with the potential for treatment and regeneration of tissue. We aimed to evaluate the involvement of paracrine effects in tissue repair after the flap skin lesion rat model. In the full-thickness flap skin experiment of forty Wistar rats: A total of 40 male Wistar rats were randomized into four groups: group I: control (C; n = 10), with full-thickness lesions on the back, without (BMSCs) or AM (n = 10); group II: injected (BMSCs; n = 10); group III: covered by AM; group IV-injected (AM + BMSCs; n = 10). Cytokine levels, IL-1, and IL-10 assay kits, superoxide dismutase (SOD), glutathione reductase (GRs) and carbonyl activity levels were measured by ELISA 28th day, and TGF-ß was evaluated by immunohistochemical, the expression collagen expression was evaluated by Picrosirius staining. Our results showed that the IL-1 interleukin was higher in the control group, and the IL-10 presented a higher mean when compared to the control group. The groups with BMSCs and AM showed the lowest expression levels of TGF-ß. SOD, GRs, and carbonyl activity analysis showed a predominance in groups that received treatment from 80%. The collagen fiber type I was predominant in all groups; however, the AM + BMSCs group obtained a higher average when compared to the control group. Our findings suggest that the AM+ BMSCs promote skin wound healing, probably owing to their paracrine effect attributed to the promotion of new collagen for tissue repair.

Decellularized Wharton Jelly Implants Do Not Trigger Collagen and Cartilaginous Tissue Production in Tracheal Injury in Rabbits.

BACKGROUND: Tracheal lesions are pathologies derived from the most diverse insults that can result in a fatal outcome. Despite the number of techniques designed for the treatment, a limiting factor is the extent of the extraction. Therefore, strategies with biomaterials can restructure tissues and maintain the organ's functionality, like decellularized Wharton's jelly (WJ) as a scaffold. The aim is to analyze the capacity of tracheal tissue regeneration after the implantation of decellularized WJ in rabbits submitted to a tracheal defect. METHODS: An in vivo experimental study was undertaken using twenty rabbits separated into two groups (n = 10). Group 1 submitted to a tracheal defect, group 2 tracheal defect, and implantation of decellularized WJ. The analyses were performed 30 days after surgery through immunohistochemistry. RESULTS: Inner tracheal area diameter (p = 0.643) didn't show significance. Collagen type I, III, and Aggrecan highlighted no significant difference between the groups (both collagens with p = 0.445 and the Aggrecan p = 0.4). CONCLUSION: The scaffold appears to fit as a heterologous implant and did not trigger reactions such as rejection or extrusion of the material into the recipient. However, these results suggested that although the WJ matrix presents several characteristics as a biomaterial for tissue regeneration, it did not display histopathological benefits in trachea tissue regeneration.

Tracheal Repair with Human Umbilical Cord Mesenchymal Stem Cells Differentiated in Chondrocytes Grown on an Acellular Amniotic Membrane: A Pre-Clinical Approach.

Acellular amniotic membrane (AM) has been studied, with promising results on the reconstruction of lesioned tissues, and has become an attractive approach for tracheal repair. This study aimed to evaluate the repair of the trachea with human umbilical cord mesenchymal stem cells (hucMSCs) differentiated in chondrocytes, grown on an experimental model. Tracheal defects were induced by surgical tracheostomy in 30 New Zealand rabbits, and the acellular amniotic membrane, with or without cells, was covering the defect. The hucMSCs were isolated and cultivated with chondrogenic differentiation over the culture of 14 days, and then grown on the AM. In this study, the AM was biocompatible and hucMSCs differentiated into chondrocytes. Our results demonstrated an important role for AM with cultured cells in the promotion of immature collagen, known to produce tissue regeneration. In addition, cartilaginous tissue was found at the tracheal defects, demonstrated by immunohistology results. This study suggests that this biomaterial implantation can be an effective future therapeutic alternative for patients with tracheal injury.

Bone-Marrow Stem Cells and Acellular Human Amniotic Membrane in a Rat Model of Heart Failure.

Myocardial infarction (MI) remains the leading cause of cardiovascular death worldwide and a major cause of heart failure. Recent studies have suggested that cell-based therapies with bone marrow stem cells (BMSC) and human amniotic membrane (hAM) would recover the ventricular function after MI; however, the mechanisms underlying these effects are still controversial. Herein, we aimed to compare the effects of BMSC and hAM in a rat model of heart failure. MI was induced through coronary occlusion, and animals with an ejection fraction (EF) < 50% were included and randomized into three groups: control, BMSC, and hAM. The BMSC and hAM groups were implanted on the anterior ventricular wall seven days after MI, and a new echocardiographic analysis was performed on the 30th day, followed by euthanasia. The echocardiographic results after 30 days showed significant improvements on EF and left-ventricular end-sistolic and end-diastolic volumes in both BMSC and hAM groups, without significant benefits in the control group. New blood vessels, desmine-positive cells and connexin-43 expression were also elevated in both BMSC and hAM groups. These results suggest a recovery of global cardiac function with the therapeutic use of both BMSC and hAM, associated with angiogenesis and cardiomyocyte regeneration after 30 days.

Tissue-engineered amniotic membrane in the treatment of myocardial infarction: a systematic review of experimental studies.

OBJECTIVE: myocardial infarction (MI) remains the leading cause of death worldwide. Cell-based therapies have become potential therapeutic approaches, attempting to recover the contractility of necrotic cardiomyocytes. In the present study, we aimed to systematically evaluate experimental studies on the use of tissue-engineered amniotic membrane (hAMC) in MI treatment. METHODS: a systematic review of literature published in PubMed, Embase and CENTRAL databases was conducted, until March 31, 2020, for experimental studies reporting on hAMC cell-therapy performed on LV function, MI size, paracrine effects, angiogenesis, and cell differentiation. Two reviewers selected the articles that met the inclusion criteria and disagreements were solved through a consensus. RESULTS: a total of 11 studies were included for data extraction. For the acute scenario, therapeutic use of hAMC after MI was capable of improving LV function in rats, mainly due to its paracrine effects (anti-apoptotic and anti-inflammatory) and associated with cardiomyocyte differentiation, MI size reduction and neo-angiogenesis. CONCLUSION: tissue engineered hAMC following MI provided clinically relevant benefits on cardiac function and ventricular remodeling.

Combined Biomaterials: Amniotic Membrane and Adipose Tissue to Restore Injured Bone as Promoter of Calcification in Bone Regeneration: Preclinical Model.

Discarded tissues, like human amniotic membranes and adipose tissue, were investigated for the application of Decellularized Human Amniotic Membrane (DAM) as a viable scaffold for transplantation of Adipose-derived stromal cells (ASCs) in bone regeneration of non-healing calvarial defects in rats. Amniotic membrane was decellularized to provide a scaffold for male Wistar rats ASCs expansion and transplantation. ASCs osteoinduction in vitro promoted the deposition of a mineralized bone-like matrix by ASCs, as calcified globular accretions associated with the cells on the DAM surface and inside the collagenous matrix. Non-healing calvarial defects on male Wistar rats were randomly divided in control without treatment, treatment with four layers of DAM, or four layers of DAM associated with ASCs. After 12 weeks, tissue blocks were examined by micro-computed tomography and histology. DAM promoted osteoconduction by increasing the collagenous matrix on both DAM treatments. DAM with ASCs stimulated bone deposition, demonstrated by a higher percentage of bone volume and trabecular bone number, compared to control. Besides the osteogenic capacity in vitro, ASCs stimulated the healing of calvarial defects with significant DAM graft incorporation concomitant with higher host bone deposition. The enhanced in vivo bone regeneration by undifferentiated ASCs loaded onto DAM confirmed the potential of an easily collected autologous cell source associated with a broadly available collagenous matrix in tissue engineering.

Acellular Human Amniotic Membrane Scaffold with 15d-PGJ2 Nanoparticles in Postinfarct Rat Model.

The difficulty in the regeneration of cardiomyocytes after myocardial infarction is a major cause of heart failure. Together, the amniotic membrane and 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) can help in the recovery of cardiomyocyte, as they present many growth factors and anti-inflammatory effect, respectively. The objective of this study is to compare the efficacy of Human Decellularized Amniotic Membrane Scaffold (AHAS) loaded with 15d-PGJ2 in improving ventricular function in a rat model of postinfarct ventricular dysfunction. Myocardial infarction was induced in 24 rats by left coronary occlusion. After a week, the animals were subjected to echocardiography for evaluation of left ventricle ejection fraction (LVEF), left ventricle end diastolic volume (LVEDV), and left ventricle end systolic volume (LVESV). Animals with ejection fraction <40% were included in the study and were randomized into three groups: control (n = 8), AHAS (n = 8) and AHAS +15d-PGJ2 (n = 8). In the AHAS group only the membrane was implanted, whereas in the AHAS +15d-PGJ2 the membrane +15d-PGJ2 was implanted on myocardial infarction. Echocardiographic evaluation was performed after 1 month. For histological analysis, heart tissue was stained with Gomori trichome, Sirius Red, the antibody against CD31 and connexin 43 (Cx43). There were no significant differences in the baseline LVEF, LVEDV, and LVESV in all groups. After 1 month, ejection fraction decreased in the control group but increased in the AHAS group and in the AHAS +15d-PGJ2 group in comparison with the control group. The LVEDV and LVESV in the AHAS and AHAS +15d-PGJ2 groups decreased compared with the control group, featuring a ventricular antiremodeling effect. Histopathology of the infarcted area identified the reduction of infarct size and collagen type 1 in the AHAS and AHAS +15d-PGJ2 groups. New blood vessels and cardiomyocytes have been identified in an infarcted area by CD31 and Cx43. AHAS +15d-PGJ2 provided an increase in the ejection fraction and prevented ventricular dilation in this postinfarction ventricular dysfunction model. Impact Statement Our study demonstrated reduction of myocardial fibrosis, proliferation of cardiomyocytes and increase in ejection fraction in rats after experimental acellular amniotic membrane scaffold (AHAS) carrying nanoparticles of 15d-PGJ2 scaffold engraftment in infarcted myocardium. AHAS grafts facilitated colonization of fibrotic myocardium regions with new contractile cells, in addition to preventing reduction of left ventricle wall thickness. This contribution is theoretically and practically relevant as current literature describes experimental studies performed on cardiac ischemic models which present conflicting results concerning cell types used in a research model.

Stem Cells and Platelet-Rich Plasma Enhance the Healing Process of Tendinitis in Mice.

OBJECTIVE: Achilles tendon pathologies occur frequently and have a significant socioeconomic impact. Currently, there is no evidence on the best treatment for these pathologies. Cell therapy has been studied in several animal models, and encouraging results have been observed with respect to tissue regeneration. This study is aimed at evaluating the functional and histological effects of bone marrow stem cell or platelet-rich plasma implantation compared to eccentric training in the treatment of Achilles tendinopathy in rats. METHODS: Fourty-one male Wistar rats received collagenase injections into their bilateral Achilles tendons (collagenase-induced tendinopathy model). The rats were randomly divided into four groups: stem cells (SC), platelet-rich plasma (PRP), stem cells+platelet-rich plasma (SC+PRP), and control (eccentric training (ET)). After 4 weeks, the Achilles tendons were excised and subjected to biomechanical and histological analyses (Sirius red and hematoxylin-eosin staining). RESULTS: Biomechanical assessments revealed no differences among the groups in ultimate tensile strength or yield strength of the tendons (p = 0.157), but there were significant differences in the elastic modulus (MPa; p = 0.044) and maximum tensile deformation (p = 0.005). The PRP group showed the greatest maximum deformation, and the SC group showed the highest Young's modulus (elasticity) measurement. In histological analysis (hematoxylin-eosin and Sirius red staining), there were no differences among the groups. CONCLUSION: PRP and SC+PRP yielded better biomechanical results than eccentric training, showing that these treatments offer better tend function outcomes. This theoretical rationale for the belief that cell therapies can serve as viable alternatives to current treatments chronic fibrotic opens the door for opportunities to continue this research.

Fluorescence properties of curcumin-loaded nanoparticles for cell tracking.

BACKGROUND: Posttransplant cell tracking, via stem cell labeling, is a crucial strategy for monitoring and maximizing benefits of cell-based therapies. The structures and functionalities of polysaccharides, proteins, and lipids allow their utilization in nanotechnology systems. MATERIALS AND METHODS: In the present study, we analyzed the potential benefit of curcumin-loaded nanoparticles (NPC) using Vero cells (in vitro) and NPC-labeled adipose-derived mesenchymal stem cells (NPC-ADMSCs) (in vivo) in myocardial infarction and sciatic nerve crush preclinical models. Thereafter, transplantation, histological examination, real time imaging, and assessment of tissue regeneration were done. RESULTS: Transplanted NPC-ADMSCs were clearly identified and revealed potential benefit when used in cell tracking. CONCLUSION: This approach may have broad applications in modeling labeled transplanted cells and in developing improved stem cell therapeutic strategies.

Impact of a High-Intensity Training on Ventricular Function in Rats After Acute Myocardial Infarction / Repercussão do Treinamento de Alta Intensidade sobre a Função Ventricular de Ratos após Infarto Agudo do Miocárdio

Abstract Background: Physical exercise should be part of the treatment of post-acute myocardial infarction (AMI) patients. Objective: To evaluate the effects of two training prescription models (continuous x interval) and its impact on ventricular function in rats after AMI with normal ventricular function. Methods: Forty Wistar rats were evaluated by echocardiography 21 days after the AMI. Those with LVEF = 50% (n = 29) were included in the study and randomized to control group (CG n = 10), continuous training group (CTG n = 9) or interval training group (ITG, n = 10). Then, a swimming test with control of lactate production was performed. Based on its result, the lactate threshold (LT) was established to define the training intensities. After six weeks, the animals were reassessed by echocardiography and lactate production. Outcome measures were end-diastolic diameter (EDD), end-systolic diameter (ESD), left ventricular ejection fraction (LVEF, %) lactate at rest, lactate without overload, and lactate with 12g and 13.5g of additional load. Group comparisons of quantitative variables of the study were performed by one-factor analysis of variance (ANOVA). The Newman-Keuls test was used for multiple comparisons of the groups. Within-group comparisons of dependent variables between the two training protocols were performed by Student's t-test. Normality of the variables was tested by the Shapiro-Wilks test. Values of p < 0.05 indicated statistical significance. Results: EDD, ESD, and LVEF before and after the training period were similar in within-group comparisons. However, EDD was significantly different (p=0.008) in the CG. Significant differences were found for L12g (p=0.002) and L13.5g (p = 0.032) in the ITG, and for L12g (p = 0.014) in the CG. No differences were found in the echocardiographic parameters between the groups. Significant differences were found in lactate without overload (p = 0.016) and L12 (p = 0.031) in the second assessment compared with the first, and between the groups - ITG vs. CG (p = 0.019) and CTG vs. CG (p = 0.035). Conclusion: Both methods produced a training effect without altering ventricular function.

Resumo Fundamento: O exercício físico deve fazer parte do tratamento de pacientes pós-infarto agudo do miocárdio (IAM). Objetivo: Avaliar os efeitos de treinamento produzidos por dois modelos distintos (contínuo x intervalado) e sua repercussão sobre a função ventricular de ratos pós-IAM com função ventricular normal. Métodos: Quarenta ratos Wistar pós-IAM foram avaliados ecocardiograficamente 21 dias após o evento. Aqueles com FEVE = 50% (n = 29) foram incluídos e randomizados: controle (GC n = 10), treinamento contínuo (GTC n = 9) e treinamento intervalado (GTI n = 10). Após, foi realizado um teste de natação com controle de lactato. A partir do resultado foi definido o limiar de lactato (LL) para determinar as intensidades do treinamento. Após seis semanas, foram reavaliados com ecocardiografia e controle de lactato. Como desfecho, foram avaliados: diâmetros diastólico e sistólico final (DDF, DSF, mL), fração de ejeção do ventrículo esquerdo (FEVE, %), lactato de repouso, livre de carga (LC), lactato com 12 g e 13,5 g de carga adicional. Para a comparação dos grupos em relação às variáveis quantitativas do estudo, foi considerado o modelo de análise da variância com um fator (ANOVA). Nas comparações múltiplas dos grupos foi usado o teste de Newman-Keuls. Na comparação entre as duas avaliações, dentro de cada grupo, foi usado o teste t de Student para amostras dependentes. A condição de normalidade das variáveis foi avaliada pelo teste de Shapiro-Wilks. Valores de p < 0,05 indicaram significância estatística. Resultados: Com relação à análise intragrupos, entre o período pré- e pós-treinamento foi identificado semelhança para DDF, DSF, FEVE, porém o GC apresentou diferença significativa para a variável DDF (p = 0,008). Houve diferença do GTI para L12g (p = 0,002) e L13,5g (p = 0,032) e para o GTC na variável L12g (p = 0,014). Não houve diferença para as variáveis ecocardiográficas entre os grupos. Houve diferença nas variáveis LC e L12g na segunda avaliação (p = 0,016 e p = 0,031, respectivamente) e entre os grupos: GTI vs. GC (p = 0,019) e GTC vs. GC (p = 0,035). Conclusão: Os dois métodos produziram efeito de treinamento sem alterar a função ventricular.

Impact of a High-Intensity Training on Ventricular Function in Rats After Acute Myocardial Infarction.

BACKGROUND: Physical exercise should be part of the treatment of post-acute myocardial infarction (AMI) patients. OBJECTIVE: To evaluate the effects of two training prescription models (continuous x interval) and its impact on ventricular function in rats after AMI with normal ventricular function. METHODS: Forty Wistar rats were evaluated by echocardiography 21 days after the AMI. Those with LVEF = 50% (n = 29) were included in the study and randomized to control group (CG n = 10), continuous training group (CTG n = 9) or interval training group (ITG, n = 10). Then, a swimming test with control of lactate production was performed. Based on its result, the lactate threshold (LT) was established to define the training intensities. After six weeks, the animals were reassessed by echocardiography and lactate production. Outcome measures were end-diastolic diameter (EDD), end-systolic diameter (ESD), left ventricular ejection fraction (LVEF, %) lactate at rest, lactate without overload, and lactate with 12g and 13.5g of additional load. Group comparisons of quantitative variables of the study were performed by one-factor analysis of variance (ANOVA). The Newman-Keuls test was used for multiple comparisons of the groups. Within-group comparisons of dependent variables between the two training protocols were performed by Student's t-test. Normality of the variables was tested by the Shapiro-Wilks test. Values of p < 0.05 indicated statistical significance. RESULTS: EDD, ESD, and LVEF before and after the training period were similar in within-group comparisons. However, EDD was significantly different (p=0.008) in the CG. Significant differences were found for L12g (p=0.002) and L13.5g (p = 0.032) in the ITG, and for L12g (p = 0.014) in the CG. No differences were found in the echocardiographic parameters between the groups. Significant differences were found in lactate without overload (p = 0.016) and L12 (p = 0.031) in the second assessment compared with the first, and between the groups - ITG vs. CG (p = 0.019) and CTG vs. CG (p = 0.035). CONCLUSION: Both methods produced a training effect without altering ventricular function.

Functional Effects of Hyperthyroidism on Cardiac Papillary Muscle in Rats / Efeitos Funcionais do Hipertireoidismo no Músculo Papilar Cardíaco de Ratos

Abstract Background: Hyperthyroidism is currently recognized to affect the cardiovascular system, leading to a series of molecular and functional changes. However, little is known about the functional influence of hyperthyroidism in the regulation of cytoplasmic calcium and on the sodium/calcium exchanger (NCX) in the cardiac muscle. Objectives: To evaluate the functional changes in papillary muscles isolated from animals with induced hyperthyroidism. Methods: We divided 36 Wistar rats into a group of controls and another of animals with hyperthyroidism induced by intraperitoneal T3 injection. We measured in the animals' papillary muscles the maximum contraction force, speed of contraction (+df/dt) and relaxation (-df/dt), contraction and relaxation time, contraction force at different concentrations of extracellular sodium, post-rest potentiation (PRP), and contraction force induced by caffeine. Results: In hyperthyroid animals, we observed decreased PRP at all rest times (p < 0.05), increased +df/dt and -df/dt (p < 0.001), low positive inotropic response to decreased concentration of extracellular sodium (p < 0.001), reduction of the maximum force in caffeine-induced contraction (p < 0.003), and decreased total contraction time (p < 0.001). The maximal contraction force did not differ significantly between groups (p = 0.973). Conclusion: We hypothesize that the changes observed are likely due to a decrease in calcium content in the sarcoplasmic reticulum, caused by calcium leakage, decreased expression of NCX, and increased expression of a-MHC and SERCA2.

Resumo Fundamento: Sabe-se atualmente que o hipertireoidismo afeta o sistema cardiovascular, ocasionando uma série de alterações funcionais e moleculares. No entanto, pouco se sabe sobre a influência funcional do hipertireoidismo na regulação do cálcio citoplasmático e no trocador de sódio/cálcio (NCX) no músculo cardíaco. Objetivos: Avaliar as alterações funcionais de músculos papilares isolados de animais com hipertireoidismo induzido. Métodos: Ao todo, 36 ratos Wistar foram distribuídos em um grupo controle e outro grupo com hipertireoidismo induzido por injeção intraperitoneal de T3. Nos músculos papilares isolados dos animais foram medidos a força máxima de contração, a velocidade de contração (+df/dt) e relaxamento (-df/dt), o tempo de contração e relaxamento, a força de contração em diferentes concentrações de sódio extracelular, o potenciação pós-pausa (PPP) e a força de contração induzida por cafeína. Resultados: Em animais com hipertireoidismo, observamos uma diminuição da PPP em todos os períodos de repouso (p < 0,05), aumento do +df/dt e -df/dt (p < 0,001), baixa resposta inotrópica positiva à concentração reduzida de sódio extracelular (p < 0,001), diminuição da força máxima de contração induzida por cafeína (p < 0,003) e diminuição do tempo total de contração (p < 0,001). A força máxima de contração não diferiu significativamente entre os grupos (p = 0,973). Conclusões: Nossa hipótese é de que as alterações observadas são provavelmente resultantes de uma diminuição do conteúdo de cálcio do retículo sarcoplasmático causada por vazamento de cálcio, redução da expressão do NCX e aumento da expressão de a-MHC e SERCA2.

Human Adipose-Derived Mesenchymal Stem Cells Cryopreservation and Thawing Decrease α4-Integrin Expression.

Aim. The effects of cryopreservation on adipose tissue-derived mesenchymal stem cells are not clearly documented, as there is a growing body of evidence about the importance of adipose-derived mesenchymal stem cells for regenerative therapies. The aim of this study was to analyze human adipose tissue-derived mesenchymal stem cells phenotypic expression (CD34, CD45, CD73, CD90, CD105, and CD49d), colony forming unit ability, viability, and differentiation potential before and after cryopreservation. Materials and Methods. 12 samples of the adipose tissue were collected from a healthy donor using the liposuction technique. The cell isolation was performed by enzymatic digestion and then the cells were cultured up to passage 2. Before and after cryopreservation the immunophenotype, cellular viability analysis by flow cytometer, colony forming units ability, differentiation potential into adipocytes and osteoblasts as demonstrated by Oil Red O and Alizarin Red staining, respectively. Results. The immunophenotypic markers expression was largely preserved, and their multipotency was maintained. However, after cryopreservation, the cells decreased α4-integrin expression (CD49d), cell viability, and number of colony forming units. Conclusions. These findings suggest that ADMSC transplanted after cryopreservation might compromise the retention of transplanted cells in the host tissue. Therefore, further studies are warranted to standardize protocols related to cryopreservation to attain full benefits of stem cell therapy.

Functional Effects of Hyperthyroidism on Cardiac Papillary Muscle in Rats.

BACKGROUND: Hyperthyroidism is currently recognized to affect the cardiovascular system, leading to a series of molecular and functional changes. However, little is known about the functional influence of hyperthyroidism in the regulation of cytoplasmic calcium and on the sodium/calcium exchanger (NCX) in the cardiac muscle. OBJECTIVES: To evaluate the functional changes in papillary muscles isolated from animals with induced hyperthyroidism. METHODS: We divided 36 Wistar rats into a group of controls and another of animals with hyperthyroidism induced by intraperitoneal T3 injection. We measured in the animals' papillary muscles the maximum contraction force, speed of contraction (+df/dt) and relaxation (-df/dt), contraction and relaxation time, contraction force at different concentrations of extracellular sodium, post-rest potentiation (PRP), and contraction force induced by caffeine. RESULTS: In hyperthyroid animals, we observed decreased PRP at all rest times (p < 0.05), increased +df/dt and -df/dt (p < 0.001), low positive inotropic response to decreased concentration of extracellular sodium (p < 0.001), reduction of the maximum force in caffeine-induced contraction (p < 0.003), and decreased total contraction time (p < 0.001). The maximal contraction force did not differ significantly between groups (p = 0.973). CONCLUSION: We hypothesize that the changes observed are likely due to a decrease in calcium content in the sarcoplasmic reticulum, caused by calcium leakage, decreased expression of NCX, and increased expression of a-MHC and SERCA2.

Evaluation of the prevalence of stress and its phases in acute myocardial infarction in patients active in the labor market.

INTRODUCTION: Acute myocardial infarction is a social health problem of epidemiological relevance, with high levels of morbidity and mortality. Stress is one of the modifiable risk factors that triggers acute myocardial infarction. Stress is a result of a set of physiological reactions, which when exaggerated in intensity or duration can lead to imbalances in one's organism, resulting in vulnerability to diseases. OBJECTIVE: To identify the presence of stress and its phases in hospitalized and active labor market patients with unstable myocardial infarction and observe its correlation with the life of this population with stress. METHODS: The methodology used was a quantitative, descriptive and transversal research approach conducted with a total of 43 patients, who were still active in the labor market, presenting or not morbidities. Data collection occurred on the fourth day of their hospitalization and patients responded to Lipp's Stress Symptom Inventory for adults. RESULTS: Thirty-one patients (72.1%) presented stress and twelve (27.8%) did not. In patients with stress, the identified phases were: alert - one patient (3.2%); resistance -twenty-two patients (71.0%); quasi-exhaustion - six patients (19.4%) and exhaustion - two patients (6.5%). All women researched presented stress. CONCLUSION: The results suggest a high level of stress, especially in the resistance phase, in the male infarcted population, hospitalized and active in the labor market.