LUIZ FELIPE PINHO MOREIRA
Projetos de Pesquisa
Unidades Organizacionais
Departamento de Cardio-Pneumologia, Faculdade de Medicina - Docente
Instituto do Coração, Hospital das Clínicas, Faculdade de Medicina
LIM/11 - Laboratório de Cirurgia Cardiovascular e Fisiopatologia da Circulação, Hospital das Clínicas, Faculdade de Medicina - Líder
Instituto do Coração, Hospital das Clínicas, Faculdade de Medicina
LIM/11 - Laboratório de Cirurgia Cardiovascular e Fisiopatologia da Circulação, Hospital das Clínicas, Faculdade de Medicina - Líder
13 resultados
Resultados de Busca
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- Molecular and Biomechanical Clues From Cardiac Tissue Decellularized Extracellular Matrix Drive Stromal Cell Plasticity(2020) LIGUORI, Gabriel Romero; LIGUORI, Tacia Tavares Aquinas; MORAES, Sergio Rodrigues de; SINKUNAS, Viktor; TERLIZZI, Vincenzo; DONGEN, Joris A. van; SHARMA, Prashant K.; MOREIRA, Luiz Felipe Pinho; HARMSEN, Martin ConradDecellularized-organ-derived extracellular matrix (dECM) has been used for many years in tissue engineering and regenerative medicine. The manufacturing of hydrogels from dECM allows to make use of the pro-regenerative properties of the ECM and, simultaneously, to shape the material in any necessary way. The objective of the present project was to investigate differences between cardiovascular tissues (left ventricle, mitral valve, and aorta) with respect to generating dECM hydrogels and their interaction with cells in 2D and 3D. The left ventricle, mitral valve, and aorta of porcine hearts were decellularized using a series of detergent treatments (SDS, Triton-X 100 and deoxycholate). Mass spectrometry-based proteomics yielded the ECM proteins composition of the dECM. The dECM was digested with pepsin and resuspended in PBS (pH 7.4). Upon warming to 37 degrees C, the suspension turns into a gel. Hydrogel stiffness was determined for samples with a dECM concentration of 20 mg/mL. Adipose tissue-derived stromal cells (ASC) and a combination of ASC with human pulmonary microvascular endothelial cells (HPMVEC) were cultured, respectively, on and in hydrogels to analyze cellular plasticity in 2D and vascular network formation in 3D. Differentiation of ASC was induced with 10 ng/mL of TGF-beta 1 and SM22 alpha used as differentiation marker. 3D vascular network formation was evaluated with confocal microscopy after immunofluorescent staining of PECAM-1. In dECM, the most abundant protein was collagen VI for the left ventricle and mitral valve and elastin for the aorta. The stiffness of the hydrogel derived from the aorta (6,998 +/- 895 Pa) was significantly higher than those derived from the left ventricle (3,384 +/- 698 Pa) and the mitral valve (3,233 +/- 323 Pa) (One-way ANOVA,p= 0.0008). Aorta-derived dECM hydrogel drove non-induced (without TGF-beta 1) differentiation, while hydrogels derived from the left ventricle and mitral valve inhibited TGF-beta 1-induced differentiation. All hydrogels supported vascular network formation within 7 days of culture, but ventricular dECM hydrogel demonstrated more robust vascular networks, with thicker and longer vascular structures. All the three main cardiovascular tissues, myocardium, valves, and large arteries, could be used to fabricate hydrogels from dECM, and these showed an origin-dependent influence on ASC differentiation and vascular network formation.
- Comparison of acute kidney injury following brain death between male and female rats(2023) ARMSTRONG JR., Roberto; RICARDO-DA-SILVA, Fernanda Yamamoto; VIDAL-DOS-SANTOS, Marina; ANUNCIACAO, Lucas Ferreira da; OTTENS, Petra J.; CORREIA, Cristiano Jesus; MOREIRA, Luiz Felipe Pinho; LEUVENINK, Hendrik Gerrit Derk; BREITHAUPT-FALOPPA, Ana CristinaBackground: Clinical reports associate kidneys from female donors with worse prognostic in male recipients. Brain Death (BD) produces immunological and hemodynamic disorders that affect organ viability. Following BD, female rats are associated with increased renal inflammation interrelated with female sex hormone reduction. Here, the aim was to investigate the effects of sex on BD-induced Acute Kidney Injury (AKI) using an Isolated Perfused rat Kidney (IPK) model.Methods: Wistar rats, females, and males (8 weeks old), were maintained for 4h after BD. A left nephrectomy was performed and the kidney was preserved in a cold saline solution (30 min). IPK was performed under normothermic temperature (37 & DEG;C) for 90 min using WME as perfusion solution. AKI was assessed by morphological analyses, staining of complement system components and inflammatory cell markers, perfusion flow, and creatinine clearance. Results: BD-male kidneys had decreased perfusion flow on IPK, a phenomenon that was not observed in the kidneys of BD-females (p < 0.0001). BD-male kidneys presented greater proximal (p = 0.0311) and distal tubule (p = 0.0029) necrosis. However, BD-female kidneys presented higher expression of eNOS (p = 0.0060) and greater upregulation of inflammatory mediators, iNOS (p = 0.0051), and Caspase-3 (p = 0.0099). In addition, both sexes had increased complement system formation (C5b-9) (p=0.0005), glomerular edema (p = 0.0003), and nNOS (p = 0.0051).Conclusion: The present data revealed an important sex difference in renal perfusion in the IPK model, evidenced by a pronounced reduction in perfusate flow and low eNOS expression in the BD-male group. Nonetheless, the upregulation of genes related to the proinflammatory cascade suggests a progressive inflammatory process in BDfemale kidneys.
- Male versus female inflammatory response after brain death model followed by ex vivo lung perfusion(2024) RICARDO-DA-SILVA, Fernanda Yamamoto; ARMSTRONG-JR, Roberto; RAMOS, Mayara Munhoz de Assis; VIDAL-DOS-SANTOS, Marina; CORREIA, Cristiano Jesus; OTTENS, Petra J.; MOREIRA, Luiz Felipe Pinho; LEUVENINK, Henri G. D.; BREITHAUPT-FALOPPA, Ana CristinaBackgroundEx vivo lung perfusion (EVLP) is a useful tool for assessing lung grafts quality before transplantation. Studies indicate that donor sex is as an important factor for transplant outcome, as females present higher inflammatory response to brain death (BD) than males. Here, we investigated sex differences in the lungs of rats subjected to BD followed by EVLP.MethodsMale and female Wistar rats were subjected to BD, and as controls sham animals. Arterial blood was sampled for gas analysis. Heart-lung blocks were kept in cold storage (1 h) and normothermic EVLP carried out (4 h), meanwhile ventilation parameters were recorded. Perfusate was sampled for gas analysis and IL-1 beta levels. Leukocyte infiltration, myeloperoxidase presence, IL-1 beta gene expression, and long-term release in lung culture (explant) were evaluated.ResultsBrain dead females presented a low lung function after BD, compared to BD-males; however, at the end of the EVLP period oxygenation capacity decreased in all BD groups. Overall, ventilation parameters were maintained in all groups. After EVLP lung infiltrate was higher in brain dead females, with higher neutrophil content, and accompanied by high IL-1 beta levels, with increased gene expression and concentration in the culture medium (explant) 24 h after EVLP. Female rats presented higher lung inflammation after BD than male rats. Despite maintaining lung function and ventilation mechanics parameters for 4 h, EVLP was not able to alter this profile.ConclusionIn this context, further studies should focus on therapeutic measures to control inflammation in donor or during EVLP to increase lung quality. Ex vivo lung perfusion maintains lung function in lung grafts from brain dead rats, independently of sex;Inflammation is greater in female's lung grafts even after ex vivo perfusion when compared to males. As there is a shortage of viable lungs for transplantation, methods of lung preservation, such as ex vivo perfusion, are important. This method is a good alternative, as it will not only preserve the lungs, but also enable lung function assessment and treatment of the organs. Studies have showed that lungs from donors of the female sex have greater risk of being rejected, when transplanted to male receptors. However, it's not certain if sex differences in anatomy, physiology and specially in immune response could interfere with the transplant result. Females do present a greater and more efficient immune response to any hazard, however after brain death this control is lost, producing a great inflammatory response as a result. Therefore, in this study we have investigated in more detail the influence of sex on the effects of brain death followed by the preservation method. Thus, we performed a brain death model in males and females rats and placed their lungs in an ex vivo lung perfusion machine. At the end of the experiment, we analyzed lung ventilation, gas exchange, and inflammatory parameters. The obtained data indicated that overall the lung ventilation and gas exchange is maintained by the ex vivo perfusion machine. Also, that lung inflammation is influenced by the sex of the donor; where the lungs from females present greater inflammation compared to the lungs from males.
- Protective role of 17 beta-estradiol treatment in renal injury on female rats submitted to brain death(2021) ARMSTRONG-JR, Roberto; RICARDO-DA-SILVA, Fernanda Yamamoto; VIDAL-DOS-SANTOS, Marina; CORREIA, Cristiano de Jesus; ANUNCIACAO, Lucas Ferreira; SILVA, Raphael dos Santos Coutinho e; MOREIRA, Luiz Felipe Pinho; LEUVENINK, Henri Gerrit Derk; BREITHAUPT-FALOPPA, Ana CristinaBackground: Clinical and experimental data highlight the consequences of brain death on the quality of organs and demonstrate the importance of donor state to the results of transplantation. Female rats show higher cardio-pulmonary injury linked to decreased concentrations of female sex hormones after brain-dead (BD). This study evaluated the effect of 17 beta-estradiol on brain death induced renal injury in female rats. Methods: Female Wistar rats were randomically allocated into 4 groups: false-operation (Sham), BD, treatment with 17 beta-estradiol (50 mu g/mL, 2 mL/h) 3 h after brain death (E2-T3), or immediately after brain death confirmation (E2-T0). Creatinine, urea, cytokines, and complement system components were quantified. Renal injury markers, such as KIM-1, Caspase-3, BCL-2 and MMP2/9 were evaluated. Results: Brain death leads to increased kidney KIM-1 expression and longer 17 beta-estradiol treatment resulted in downregulation (P<0.0001). There was increase of neutrophil numbers in kidney from BD rats and E2 treatment was able to reduce it (P=0.018). Regarding complement elements, E2-T3 group evidenced E2 therapeutic effects, reducing C5b-9 (P=0.0004), C3aR (P=0.054) and C5aR (P=0.019). In parallel, there were 17 beta-estradiol effects in reducing MMP2 (P=0.0043), MMP9 (P=0.011), and IL-6 (P=0.024). Moreover, E2-T3 group improved renal function in comparison to BD group (P=0.0938). Conclusions: 17 beta-estradiol treatment was able to reduce acute kidney damage in BD female rats owing to its ability to prevent tissue damage, formation of C5b-9, and local synthesis of inflammatory mediators.
- Evaluation of the therapeutic effects of oestradiol on the systemic inflammatory response and on lung injury caused by the occlusion of the proximal descending aorta in male rats(2023) SOUSA, Marcelo Nunes de; ANUNCIACAO, Lucas Ferreira da; FREITAS, Pedro Luiz Zonta de; RICARDO-DA-SILVA, Fernanda Yamamoto; MOREIRA, Luiz Felipe Pinho; CORREIA, Cristiano Jesus; BREITHAUPT-FALOPPA, Ana CristinaOBJECTIVES: Ischaemia and reperfusion-induced microvascular dysfunction is a serious problem encountered during a variety surgical procedures, leading to systemic inflammation and affecting remote organs, specially the lungs. 17 beta-Oestradiol reduces pulmonary repercussions from various acute lung injury forms. Here, we focused on the 17 beta-oestradiol therapeutic effects after aortic ischaemia and reperfusion (I/R) by evaluating lung inflammation. METHODS: Twenty-four Wistar rats were submitted to I/R by insufflation of a 2-F catheter in thoracic aorta for 20 min. Reperfusion took 4 h and 17 beta-oestradiol (280 mg/kg, i.v.) was administered after 1 h of reperfusion. Sham-operated rats were controls. Bronchoalveolar lavage was performed and lung samples were prepared for histopathological analysis and tissue culture (explant). Interleukin (IL)-1 beta, IL-10 and tumour necrosis factor-a were quantified. RESULTS: After I/R, higher number of leukocytes in bronchoalveolar lavage were reduced by 17 beta-oestradiol. The treatment also decreased leukocytes in lung tissue. I/R increased lung myeloperoxidase expression, with reduction by 17 beta-oestradiol. Serum cytokine-induced neutrophil chemoattractant 1 and IL-1 beta increased after I/R and 17 beta-oestradiol decreased cytokine-induced neutrophil chemoattractant 1. I/R increased IL-1 beta and IL-10 in lung explants, reduced by 17 beta-oestradiol. CONCLUSIONS: Our results showed that 17 beta-oestradiol treatment performed in the period of reperfusion, modulated the systemic response and the lung repercussions of I/R by thoracic aortic occlusion. Thus, we can suggest that 17 beta-oestradiol might be a supplementary approach leading the lung deterioration after aortic clamping in surgical procedures.
- MiRNA-30d and miR-770-5p as potential clinical risk predictors of Vasoplegic Syndrome in Patients undergoing on-pump coronary artery bypass grafting(2023) MEJIA, Omar Asdrubal Vilca; SOUZA, Renato Cesar de; SANTOS, Aritania S.; MENEGHINI, Bianca; SILVA, Ana Carolina Carvalho; BRASIL, Guilherme Visconde; RIGAUD, Vagner Oliveira Carvalho; DALLAN, Luis Roberto Palma; MOREIRA, Luiz Felipe Pinho; LISBOA, Luiz Augusto Ferreira; DALLAN, Luis Alberto Oliveira; KALIL, Jorge; CUNHA-NETO, Edecio; FERREIRA, Ludmila Rodrigues Pinto; JATENE, Fabio BiscegliThe aims of this study were to perform pre-surgery miRNA profiling of patients who develop Vasoplegic syndrome (VS) after coronary artery bypass grafting (CABG) and identify those miRNAs that could be used as VS prognostic tools and biomarkers. The levels of 754 microRNAs (miRNAs) were measured in whole blood samples from a cohort of patients collected right before the coronary artery bypass grafting (CABG) surgery. We compared the miRNA levels of those who developed VS (VASO group) with those who did not (NONVASO group) after surgery. Six miRNAs (hsa-miR-548c-3p, -199b-5p, -383-5p -571 -183-3p, -30d-5p) were increased and two (hsa-1236-3p, and hsa-miR770-5p) were decreased in blood of VASO compared to NONVASO groups. Receiver Operating Characteristic (ROC) curve analysis revealed that a combination of the miRNAs, hsa-miR-30d-5p and hsa-miR-770-5p can be used as VS predictors (AUC = 0.9615, p < 0.0001). The computational and functional analyses were performed to gain insights into the potential role of these dysregulated miRNAs in VS and have identified the ""Apelin Liver Signaling Pathway"" as the canonical pathway containing the most target genes regulated by these miRNAs. The expression of the combined miRNAs hsa-miR-30d and hsa-miR-770-5p allowed the ability to distinguish between patients who could and could not develop VS, representing a potential predictive biomarker of VS.
conferenceObject Kidney Release of Inflammatory Mediators Is Modulated by 17BETA-Estradiol Associated With Methylprednisolone After Brain Death in Female Rats(2022) SANTOS, Marina Vidal dos; ANUNCIACAO, Lucas Ferreira da; ARMSTRONG JR., Roberto; SILVA, Fernanda Yamamoto Ricardo da; RAMOS, Mayara Munhoz de Assis; CORREIA, Cristiano de Jesus; MOREIRA, Luiz Felipe Pinho; LEUVENINK, Henri; FALOPPA, Ana Cristina Breithaupt- Regeneration of Tracheal Tissue in Partial Defects Using Porcine Small Intestinal Submucosa(2018) NETO, Nelson Bergonse; JORGE, Lianna Ferrari; FRANCISCO, Julio C.; ERBANO, Bruna Olandoski; BARBOZA, Barbara Evelin Goncalves; SILVA, Larissa Luvison Gomes da; OLANDOSKI, Marcia; CARVALHO, Katherine Athayde Teixeira de; MOREIRA, Luiz Felipe Pinho; FARIA NETO, Jose Rocha; ABDELWAHID, Eltyeb; GUARITA-SOUZA, Luiz CesarBackground. Surgical correction of tracheal defects is a complex procedure when the gold standard treatment with primary end-to-end anastomosis is not possible. An alternative treatment may be the use of porcine small intestinal submucosa (SIS). It has been used as graft material for bioengineering applications and to promote tissue regeneration. The aim of this study was to evaluate whether SIS grafts improved tracheal tissue regeneration in a rabbit model of experimental tracheostomy. Methods. Sixteen rabbits were randomized into two groups. Animals in the control group underwent only surgical tracheostomy, while animals in the SIS group underwent surgical tracheostomy with an SIS graft covering the defect. We examined tissues at the site of tracheostomy 60 days after surgery using histological analysis with hematoxylin and eosin (H&E) staining and analyzed the perimeter and area of the defect with Image-Pro (R) PLUS 4.5 (Media Cybernetics). Results. The average perimeter and area of the defects were smaller by 15.3% (p = 0 034) and 21.8% (p = 0 151), respectively, in the SIS group than in the control group. Histological analysis revealed immature cartilage, pseudostratified ciliated epithelium, and connective tissue in 54.5% (p = 0 018) of the SIS group, while no cartilaginous regeneration was observed in the control group. Conclusions. Although tracheal SIS engraftment could not prevent stenosis in a rabbit model of tracheal injury, it produced some remarkable changes, efficiently facilitating neovascularization, reepithelialization, and neoformation of immature cartilage.
- Directional Topography Influences Adipose Mesenchymal Stromal Cell Plasticity: Prospects for Tissue Engineering and Fibrosis(2019) LIGUORI, Gabriel Romero; ZHOU, Qihui; LIGUORI, Tacia Tavares Aquinas; BARROS, Guilherme Garcia; KUHN, Philipp Till; MOREIRA, Luiz Felipe Pinho; RIJN, Patrick van; HARMSEN, Martin C.Introduction. Progenitor cells cultured on biomaterials with optimal physical-topographical properties respond with alignment and differentiation. Stromal cells from connective tissue can adversely differentiate to profibrotic myofibroblasts or favorably to smooth muscle cells (SMC). We hypothesized that myogenic differentiation of adipose tissue-derived stromal cells (ASC) depends on gradient directional topographic features. Methods. Polydimethylsiloxane (PDMS) samples with nanometer and micrometer directional topography gradients (wavelength49-3, 425nm) were fabricated. ASC were cultured on patterned PDMS and stimulated with TGF-1 to induce myogenic differentiation. Cellular alignment and adhesion were assessed by immunofluorescence microscopy after 24h. After seven days, myogenic differentiation was examined by immunofluorescence microscopy, gene expression, and immunoblotting. Results. Cell alignment occurred on topographies larger than w=1758nm/a=630nm. The number and total area of focal adhesions per cell were reduced on topographies from w=562nm/a=96nm to w=3919nm/a=1430nm. Focal adhesion alignment was increased on topographies larger than w=731nm/a=146nm. Less myogenic differentiation of ASC occurred on topographies smaller than w=784nm/a=209nm. Conclusion. ASC adherence, alignment, and differentiation are directed by topographical cues. Our evidence highlights a minimal topographic environment required to facilitate the development of aligned and differentiated cell layers from ASC. These data suggest that nanotopography may be a novel tool for inhibiting fibrosis.
conferenceObject Sex Differences in Acute Kidney Injury Following BD - An Isolated Rat Organ Perfusion Model(2022) ARMSTRONG JUNIOR, Roberto; RICARDO-DA-SILVA, Fernanda F. Yamamoto; VIDAL-DOS-SANTOS, Marina M.; OTTENS, Petra; CORREIA, Cristiano C. Jesus; MOREIRA, Luiz Felipe Pinho; LEUVENINK, Henri H.; BREITHAUPTO-FALOPPA, Ana Cristina A. C.