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
9 resultados
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- 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.
- 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 BreithauptconferenceObject 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.conferenceObject 17beta-Estradiol and Methylprednisolone Associated Treatment Modulates Early and Late Cytokine Release in Lungs From Female Rats Submitted to Brain Death(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- Limited Efficacy of Adipose Stromal Cell Secretome-Loaded Skin-Derived Hydrogels to Augment Skin Flap Regeneration in Rats(2022) VRIEND, Linda; DONGEN, Joris A. van; SINKUNAS, Viktor; BROUWER, Linda A.; BUIKEMA, Henk J.; MOREIRA, Luiz F.; GEMPERLI, Rolf; BONGIOVANNI, Laura; BRUIN, Alain de; LEI, Berend van der; CAMARGO, Cristina P.; HARMSEN, Martin C.Insufficient vascularization is a recurring cause of impaired pedicled skin flap healing. The administration of adipose tissue-derived stromal cells' (ASCs') secretome is a novel approach to augment vascularization. Yet, the secretome comprised of soluble factors that require a sustained-release vehicle to increase residence time. We hypothesized that administration of a hydrogel derived from decellularized extracellular matrix (ECM) of porcine skin with bound trophic factors from ASCs enhances skin flap viability and wound repair in a rat model. Porcine skin was decellularized and pepsin-digested to form a hydrogel at 37 degrees C. Conditioned medium (CMe) of human ASC was collected, concentrated 20-fold, and mixed with the hydrogel. Sixty Wistar rats were included. A dorsal skin flap (caudal based) of 3 x 10 cm was elevated for topical application of DMEM (group I), a prehydrogel with or without ASC CMe (groups II and III), or ASC CMe (group IV). After 7, 14, and 28 days, perfusion was measured, and skin flaps were harvested for wound healing assessment and immunohistochemical analysis. Decellularized skin ECM hydrogel contained negligible amounts of DNA (11.6 +/- 0.6 ng/mg), was noncytotoxic and well tolerated by rats. Irrespective of ASC secretome, ECM hydrogel application resulted macroscopically and microscopically in similar dermal wound healing in terms of proliferation, immune response, and matrix remodeling as the control group. However, ASC CMe alone increased vessel density after 7 days. Porcine skin-derived ECM hydrogels loaded with ASC secretome are noncytotoxic but demand optimization to significantly augment wound healing of skin flaps.
- A modified hydrogel production protocol to decrease cellular content(2022) BRAGA, Gabriela Catao Diniz; CAMARGO, Cristina Pires; HARMSEN, Martin Conrad; CORREIA, Aristides Tadeu; SOUZA, Sonia; SEELAENDER, Marilia; NUNES, Viviane Araujo; SANTOS, Jeniffer Farias dos; NERI, Elida Adalgisa; VALADAO, Iuri Cordeiro; MOREIRA, Luiz Felipe Pinho; GEMPERLI, RolfPurpose: To analyze the cytotoxicity and cell in porcine-derived decellularized skin matrix. Methods: We analyzed the effect of multiple decellularization processes by histological analysis, DNA quantification, and flow cytometry. Subsequently, we analyzed the most appropriate hydrogel concentration to minimize cytotoxicity on fibroblast culture and to maximize cell proliferation. Results: After the fourth decellularization, the DNA quantification showed the lowest DNA concentration (< 50 ng/mg). Histological analysis showed no cell components in the hydrogel. Moreover, hematoxylin and eosin showed a heterogeneous structure of collagen fibers. The best hydrogel concentration ranged from 3 to 25%, and there was no significant difference between the 24 hours and seven days. Conclusion: The process of hydrogel production was effective for removing cells and DNA elements. The best hydrogel concentration ranged from 3 to 25%.