The effect of bone marrow-derived stem cells associated with platelet-rich plasma on the osseointegration of immediately placed implants

Stem cells associated with growth factors have been shown to improve bone healing and the osseointegration of dental implants. A Brazilian miniature pig model was used to evaluate the effect of autologous bone marrow-derived mesenchymal stem cells (BM-MS

Brazilian minipig as a large-animal model for basic research and stem cell-based tissue engineering. Characterization and in vitro differentiation of bone marrow-derived mesenchymal stem cells

Stem cell-based regenerative medicine is one of the most intensively researched medical issues. Pre-clinical studies in a large-animal model, especially in swine or miniature pigs, are highly relevant to human applications. Mesenchymal stem cells (MSCs) have been isolated and expanded from different sources. Objective: This study aimed at isolating and characterizing, for the first time, bone marrow-derived MSCs (BM-MSCs) from a Brazilian minipig (BR1). Also, this aimed to validate a new large-animal model for stem cell-based tissue engineering. Material and Methods: Bone marrow (BM) was aspirated from the posterior iliac crest of twelve adult male BR1 under general anesthesia. MSCs were selected by plastic-adherence as originally described by Friedenstein. Cell morphology, surface marker expression, and cellular differentiation were examined. The immunophenotypic profile was determined by flow cytometry. The differentiation potential was assessed by cytological staining and by RT-PCR. Results: MSCs were present in all minipig BM samples. These cells showed fibroblastic morphology and were positive for the surface markers CD90 (88.6%), CD29 (89.8%), CD44 (86.9%) and negative for CD34 (1.61%), CD45 (1.83%), CD14 (1.77%) and MHC-II (2.69%). MSCs were differentiated into adipocytes, osteoblasts, and chondroblasts as demonstrated by the presence of lipidic-rich vacuoles, the mineralized extracellular matrix, and the great presence of glycosaminoglycans, respectively. The higher gene expression of adipocyte fatty-acid binding protein (AP2), alkaline phosphatase (ALP) and collagen type 2 (COLII) also confirmed the trilineage differentiation (

IFI27 transcription is an early predictor for COVID-19 outcomes, a multi-cohort observational study

PurposeRobust biomarkers that predict disease outcomes amongst COVID-19 patients are necessary for both patient triage and resource prioritisation. Numerous candidate biomarkers have been proposed for COVID-19. However, at present, there is no consensus on the best diagnostic approach to predict outcomes in infected patients. Moreover, it is not clear whether such tools would apply to other potentially pandemic pathogens and therefore of use as stockpile for future pandemic preparedness.MethodsWe conducted a multi-cohort observational study to investigate the biology and the prognostic role of interferon alpha-inducible protein 27 (IFI27) in COVID-19 patients.ResultsWe show that IFI27 is expressed in the respiratory tract of COVID-19 patients and elevated IFI27 expression in the lower respiratory tract is associated with the presence of a high viral load. We further demonstrate that the systemic host response, as measured by blood IFI27 expression, is associated with COVID-19 infection. For clinical outcome prediction (e.g., respiratory failure), IFI27 expression displays a high sensitivity (0.95) and specificity (0.83), outperforming other known predictors of COVID-19 outcomes. Furthermore, IFI27 is upregulated in the blood of infected patients in response to other respiratory viruses. For example, in the pandemic H1N1/09 influenza virus infection, IFI27-like genes were highly upregulated in the blood samples of severely infected patients.ConclusionThese data suggest that prognostic biomarkers targeting the family of IFI27 genes could potentially supplement conventional diagnostic tools in future virus pandemics, independent of whether such pandemics are caused by a coronavirus, an influenza virus or another as yet-to-be discovered respiratory virus

Mesenchymal stromal cells as a choice for spinal cord injury treatment

Spinal cord injury (SCI) is a serious clinical problem that affects approximately 17,500 new patients per year in the United States. The main causes of SCI are vehicle collisions, falls, violence (mainly gunshot wounds), and sports/recreational activities. The final severity of the damage results from primary and secondary mechanisms that begin at the time of injury and last for months after trauma. To reduce the extent of damage, several treatments have been proposed. This review summarizes results from several studies that have pointed to cell therapy as the main form of neuroregenerative treatment. Mesenchymal stromal cells (MSCs) are important candidates for tissue regeneration due to the release of bioactive factors, as well as antiapoptotic effects, scar inhibitors, and angiogenic effects. Studies have shown that MSCs act in various ways on injured tissue, such as immunomodulation of the inflamed environment, release of bioactive factors, restoration of axon myelin, prevention of neuronal apoptosis, and neuroregeneration. Current research using MSCs aims to prevent secondary injury, promote regeneration, and replace destroyed spinal cord tissue. This review presents information about the damage from primary and secondary events after SCI, treatments usually used, and pre-clinical and clinical results aiming at the cell therapy using MSCs as a tissue regeneration strategy

Inhibition of Hedgehog signaling pathway affects the expression of miR-20a and miR-3

Submitted by Luciane Willcox ([email protected]) on 2016-09-19T16:28:28Z No. of bitstreams: 1 Inibição da via de sinalização Hedgehog.pdf: 1093460 bytes, checksum: 4a5d3de417f100e17c84b5b6a4f0e2cf (MD5)Approved for entry into archive by Luciane Willcox ([email protected]) on 2016-09-19T16:39:21Z (GMT) No. of bitstreams: 1 Inibição da via de sinalização Hedgehog.pdf: 1093460 bytes, checksum: 4a5d3de417f100e17c84b5b6a4f0e2cf (MD5)Made available in DSpace on 2016-09-19T16:39:21Z (GMT). No. of bitstreams: 1 Inibição da via de sinalização Hedgehog.pdf: 1093460 bytes, checksum: 4a5d3de417f100e17c84b5b6a4f0e2cf (MD5) Previous issue date: 2013-11Fundação Oswaldo Cruz (FIOCRUZ-PR), Ministério da Saúde, Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) e Fundação Araucária.Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-Tronco. Curitiba, PR, Brasil.Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-Tronco. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células-Tronco. Curitiba, PR, Brasil.MicroRNAs são pequenas moléculas de RNA (~ 22 nucleotídeos), que atuam como reguladores pós-transcricional da expressão de genes, inibindo a tradução do RNA mensageiro. MicroRNAs estão envolvidos na regulação de muitos processos biológicos e patológicos, assim como a via de sinalização Hedgehog (Hh). Nós avaliamos a expressão de quatro microRNAs (miR-20a, miR-31, miR-17 e miR-199b5p) em células-tronco derivadas de tecido adiposo humano sob ativação e bloqueio da via de sinalização Hh. As células foram tratadas com purmorfamina (ativador da via Hh) ou ciclopamina (bloqueador da via Hh) durante 24 horas. Extração de RNA e cDNA foram realizadas para fazer a analise da expressão dos miRNAs por RT-PCR quantitativa. O nível de expressão do miR-20a e miR-31 aumentou após o bloqueio da via de sinalização Hh, sendo assim, esses miRNAs podem estar envolvidos no controle da proliferação de células-tronco e câncer. O miR-20a e o miR-31 são fortes candidatos para biomarcadores da via Hh.MicroRNAs are small RNA molecules (~ 22 nucleotides) that act as post-transcriptional regulators of gene expression by inhibiting translation of messenger RNAs. MicroRNAs are involved in regulating many biological and pathological processes, as well as the Hedgehog (Hh) signaling pathway. We evaluate the expression of four miRNAs (miR-20a, miR-31, miR-17 and miR-199b5p) in human adipose-derived stem cells under activation and blockade of Hh signaling pathway. Cells were treated with purmorphamine (Hh activator) or cyclopamine (Hh blocker) for 24 hours. RNA extraction and cDNA were performed to analyze of miRNAs expression by quantitative RT-PCR. The expression level of miR-20a and miR-31 increased after blocking Hh signaling pathway, thus these miRNAs may be involved in control of proliferation in stem and cancer cells. MiR-20a and miR-31 are strong candidates for biomarkers of Hh pathway

Proliferation and differentiation of stem cells in contact with eluate from fibrin-rich plasma membrane

Submitted by Manoel Barata ([email protected]) on 2018-02-09T12:37:17Z No. of bitstreams: 1 AguiarProliferat.pdf: 2378251 bytes, checksum: a4efca8744d7c251c7205603761e7fa0 (MD5)Approved for entry into archive by Manoel Barata ([email protected]) on 2018-02-28T12:24:31Z (GMT) No. of bitstreams: 1 AguiarProliferat.pdf: 2378251 bytes, checksum: a4efca8744d7c251c7205603761e7fa0 (MD5)Made available in DSpace on 2018-02-28T12:24:31Z (GMT). No. of bitstreams: 1 AguiarProliferat.pdf: 2378251 bytes, checksum: a4efca8744d7c251c7205603761e7fa0 (MD5) Previous issue date: 2018Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.To evaluate the ability of the eluate from fibrin-rich plasma (FRP) membrane to induce proliferation and differentiation of isolated human adipose-derived stem cells (ASCs) into chondrocytes. How method: FRP membranes were obtained by centrifugation of peripheral blood from two healthy donors, cut, and maintained in culture plate wells for 48h to prepare the fibrin eluate. The SCATh were isolated from adipose tissue by collagenase digestion solution, and expanded in vitro. Cells were expanded and treated with DMEM-F12 culture, a commercial media for chondrogenic differentiation, and eluate from FRP membrane for three days, and labeled with BrdU for quantitative assessment of cell proliferation using the HighContent Operetta® imaging system. For the chondrogenic differentiation assay, the SCATh were grown in micromass for 21 days and stained with toluidine blue and aggrecan for qualitative evaluation by light microscopy. The statistical analysis was performed using ANOVA and Tukey’s test. The results were that there was a greater proliferation of cells treated with the eluate from FRP membrane compared to the other two treatments, where the ANOVA test showed significance (p < 0.001). The differentiation into chondrocytes was visualized by the presence of mucopolysaccharide in the matrix of the cells marked in blue toluidine and aggrecan. This way concluded in by treatment with eluate from FRP membrane stimulated cell proliferation and induced differentiation of the stem cells into chondrocytes, suggesting a potential application of FRP membranes in hyaline cartilage regeneration therapies

Formação in vitro de túbulos capilares a partir de células de sangue de cordão umbilical humano com perspectivas para aplicação terapêutica

OBJETIVO: As células progenitoras endoteliais (CPE), caracterizadas pelo marcador CD133+, contribuem para a neovascularização, e o aumento no número dessas células pode ser uma ferramenta terapêutica promissora. O sangue de cordão umbilical humano contém um número significante de CPE, sugerindo a possibilidade do uso destas células para a revascularização de tecidos isquêmicos. O objetivo desse trabalho foi analisar a funcionalidade das células CD133+ diferenciadas in vitro. MÉTODOS: As células diferenciadas foram caracterizadas por citometria de fluxo; a expressão do mRNA de VEGF foi avaliada por RT-PCR e a funcionalidade, por meio de ensaios de formação de túbulos capilares. RESULTADOS: As células diferenciadas perderam os marcadores de CPE, mantiveram em níveis baixos os marcadores das linhagens hematopoética e monocíticas e aumentaram a expressão dos marcadores de células endoteliais adultas. As células diferenciadas apresentaram transcritos no mRNA de VEGF e mostraram-se capazes de formar túbulos capilares in vitro. CONCLUSÃO: As células CD133+ diferenciadas in vitro em células endoteliais demonstraram serem funcionalmente ativas, abrindo perspectiva para seu uso futuro em aplicações terapêuticas

Cell cycle genes are downregulated after adipogenic triggering in human adipose tissue-derived stem cells by regulation of mRNA abundance

Submitted by Manoel Barata ([email protected]) on 2019-04-30T15:31:00Z No. of bitstreams: 1 s41598-019-42005-3.pdf: 2386137 bytes, checksum: dccd189dd711688e0f6fcd0b24ad0c8c (MD5)Approved for entry into archive by Manoel Barata ([email protected]) on 2019-04-30T18:19:40Z (GMT) No. of bitstreams: 1 s41598-019-42005-3.pdf: 2386137 bytes, checksum: dccd189dd711688e0f6fcd0b24ad0c8c (MD5)Made available in DSpace on 2019-04-30T18:19:40Z (GMT). No. of bitstreams: 1 s41598-019-42005-3.pdf: 2386137 bytes, checksum: dccd189dd711688e0f6fcd0b24ad0c8c (MD5) Previous issue date: 2019Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Instituto Pasteur. Unidad de Bioinformática. Montevideo, Uruguay.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Instituto Pasteur. Unidad de Bioinformática. Montevideo, Uruguay.Pontifícia Universidade Católica do Paraná. Núcleo de Tecnologia Celular. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.The adipogenic process is characterized by the expression of adipocyte differentiation markers that lead to changes in cell metabolism and to the accumulation of lipid droplets. Moreover, during early adipogenesis, cells undergo a strong downregulation of translational activity with a decrease in cell size, proliferation and migration. In the present study, we identified that after 24 hours of adipogenic induction, human adipose tissue-derived stem cells (hASCs) undergo a G1-cell cycle arrest consistent with reduced proliferation, and this effect was correlated with a shift in polysome profile with an enrichment of the monosomal fraction and a reduction of the polysomal fraction. Polysome profiling analysis also revealed that this change in the monosomal/polysomal ratio was related to a strong downregulation of cell cycle and proliferation genes, such as cyclins and cyclin-dependent kinases (CDKs). Comparing total and polysome-associated mRNA sequencing, we also observed that this downregulation was mostly due to a reduction of cell cycle and proliferation transcripts via control of total mRNA abundance, rather than by translational control