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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

The study of adhesive interactions between haemopoietic progenitor cells and bone marrow sinusoidal endothelial cells

Masek, Lisa Christina January 1997 (has links)
No description available.
2

Articular cartilage tissue engineering using chondrogenic progenitor cell homing and 3D bioprinting

Yu, Yin 01 May 2015 (has links)
Articular cartilage damage associated with joint trauma seldom heals and often leads to osteoarthritis (OA). Current treatment often fails to regenerated functional cartilage close to native tissue. We previously identified a migratory chondrogenic progenitor cell (CPC) population that responded chemotactically to cell death and rapidly repopulated the injured cartilage matrix, which suggested their potential for cartilage repair. To test that potential we filled experimental full thickness chondral defects with an acellular hydrogel containing SDF-1α. We expect that SDF-1α can increase the recruitment of CPCs, and then promote the formation of a functional cartilage matrix with chondrogenic factors. Full-thickness bovine chondral defects were filled with hydrogel comprised of fibrin and hyaluronic acid and containing SDF-1α. Cell migration was monitored, followed by chondrogenic induction. Regenerated tissue was evaluated by histology, immunohistochemistry, and scanning electron microscopy. Push-out tests were performed to assess the strength of integration between regenerated tissue and host cartilage. Significant numbers of progenitor cells were recruited by SDF-1α within 12 days. By 5 weeks chondrogenesis, repair tissue cell morphology, proteoglycan density and surface ultrastructure were similar to native cartilage. SDF-1α treated defects had significantly greater interfacial strength than untreated controls. However, regenerated neocartilage had relatively inferior mechanical properties compared with native cartilage. In addition to that, we developed a 3D bioprinting platform, which can directly print chondrocytes as well as CPCs to fabricated articular cartilage tissue in vitro. We successfully implanted the printed tissue into an osteochondral defect, and observed tissue repair after implantation. The regerated tissue has biochemical and mechanical properties within the physiological range of native articular cartilage. This study showed that, when CPC chemotaxis and chondrogenesis are stimulated sequentially, in situ full thickness cartilage regeneration and bonding of repair tissue to surrounding cartilage could occur without the need for cell transplantation from exogenous sources. This study also demonstrated the potential of using 3D bioprinting to engineer articular cartilage implants for repairing cartilage defect.
3

Homing and Differentiation of Mesenchymal Stem Cells in 3D In Vitro Models

Popielarczyk, Tracee 31 August 2017 (has links)
Mesenchymal stem cells (MSCs) have great potential to improve clinical outcomes for many inflammatory and degenerative diseases through delivery of exogenous MSCs via injection or cell-laden scaffolds and through mobilization and migration of endogenous MSCs to injury sites. MSC fate and function is determined by microenvironmental cues, specifically dimensionality, topography, and cell-cell interactions. MSC responses of migration and differentiation are the focus of this dissertation. Cell migration occurs in several physiological and pathological processes; migration mode and cell signaling are determined by the environment and type of confinement in three-dimensional (3D) models. Tendon injury is a common musculoskeletal disorder that occurs through cumulative damage to the extracellular matrix (ECM). Studies combining nanofibrous scaffolds and MSCs to determine an optimal topographical environment have promoted tenogenic differentiation under various conditions. We investigated cellular response of MSCs on specifically designed nanofiber matrices fabricated using a novel spinneret-based tunable engineered parameters production method (STEP). We designed suspended and aligned nanofiber scaffolds to study cellular morphology, tendon marker gene expression, and matrix deposition as determinants for tendon differentiation. The delivery and maintenance of MSCs at sites of inflammation or injury are major challenges in stem cell therapies. Enhancing stem cell homing could improve their therapeutic effects. Homing is a process that involves cell migration through the vasculature to target organs. This process is defined in leukocyte transendothelial migration (TEM); however, far less is known about MSC homing. We investigated two population subsets of MSCs in a Transwell system mimicking the vasculature; migrated cells that initiated transmigration on the endothelium and nonmigrated cells in the apical chamber that failed to transmigrate. Gene and protein expression changes were observed between these subsets and evidence suggests that multiple signaling pathways regulate TEM. The results of these experiments have demonstrated that microenvironmental cues are critical to understanding the cellular and molecular mechanisms of MSC response, specifically in homing and differentiation. This knowledge has identified scaffold parameters required to stimulate tenogenesis and signaling pathways controlling MSC homing. These findings will allow us to target key regulatory molecules and cell signaling pathways involved in MSC response towards development of regenerative therapies. / Ph. D.
4

ROLE OF OXIDATIVE STRESS AND T CELL HOMING IN THE DEVELOPMENT OF MURINE SYNGENEIC GRAFT-VERSUS-HOST DISEASE

Perez-Rodriguez, Jacqueline 01 January 2009 (has links)
Syngeneic graft-versus-host disease (SGVHD) is induced by reconstituting lethally irradiated mice with syngeneic BM cells followed by a 21 day treatment with the immunosuppressive agent cyclosporine A (CsA). Clinical symptoms of the disease appear 2-3 weeks following cessation of CsA therapy and disease-associated inflammation occurs primarily in the colon and liver. The development of SGVHD is a complex process resulting from the cooperative interaction of multiple effector cell populations including NK cells, T cells and macrophages. TH1 cytokines (IL-12, TNF-α, IFN- γ), produced by these effector cells, serve as inflammatory mediators contributing to the pathogenesis of SGVHD. The SGVHD conditioning agents, irradiation and CsA, are both required for the development of disease and contribute to the production of oxidative stress. Time course studies revealed increased reactive oxygen and nitrogen species (ROS/RNS), as well as, increased colon mRNA levels for TNF-α and iNOS in CsA-treated versus control BMT animals. Since ROS/RNS are known to mediate CsA toxicity, studies were undertaken to determine the effect of oxidative stress on the induction of SGVHD. In vivo treatment with the antioxidant MnTBAP caused a reduction in colon mRNA levels for iNOS and TNF-α, as well as delayed disease development, suggesting a role for oxidative stress in the development of SGVHD. In addition, CD4+ T cells have been shown to play an important role in the inflammatory response observed in the gut of SGVHD mice. Time course studies revealed significant increases in the migration of CD4+ T cells as early as day 14 post- BMT into the colon of CsA mice as well as significant elevated mRNA levels of cell adhesion molecules. Homing studies revealed that a labeled CD4+ T cell line, generated from SGVHD mice, migrated in larger numbers into the gut of CsA-treated mice compared to control animals. This study demonstrated that CD4+ T cells responsible for the pathogenesis observed in murine SGVHD are present early after BMT in colons of CsA-treated mice, suggesting that during the 21 days of immunosuppression therapy functional mechanisms are in place that result in increased homing of effector cells to colons of CsA-treated mice.
5

Estudo da biodistribuição de células tronco de polpa de dente decíduo humana (CTPDDh) após o transplante intra-uterino no modelo canino (Canis lupus familiares) / Biodistribution of human immature dental pulp stem cells following in utero transplantation in canine model (Canis lupus familiaris)

Reginato, Ana Luísa 19 June 2012 (has links)
O transplante intrauterino de células-tronco (TIUCT) é um método de tratamento de doenças genéticas, congênitas, hematológicas e imunológicas em um feto durante a gestação. Em pesquisa básica este modelo permite o estudo da dinâmica de migração, enxertia e estado funcional de diferentes tipos de células-tronco (CT). Estas células podem ser transplantadas em diferentes momentos do período gestacional, que pode ser dividido em três momentos do desenvolvimento fetal, sendo estes, diferentes funcionalmente. A escolha deste momento para o transplante influenciará tanto no comportamento celular quanto no resultado. Para o TIUCT são utilizadas as CT mesenquimais derivadas da medula óssea ou fetais ou hematopoiéticas. Para esta pesquisa utilizamos células-tronco derivadas da polpa dentária imatura humana (CTIPDh) as quais apresentam potencial pluripotente e propriedades imunomodulatórias. Nosso principal objetivo foi avaliar a capacidade migratória, bem como de proliferação e endereçamento (homing) das CTIPDh durante o terceiro período gestacional do desenvolvimento fetal no modelo canino. Todos os procedimentos experimentais foram elaborados sob protocolo anestésico apropriado e aprovados pelo comitê de ética da FMVZ da USP. Foram transplantadas via intraperitoneal (IP) 1x106 CTIPDh GFP+ em cada feto, durante procedimento cirúrgico de laparotomia exploratória com ultrassonografia guiada intraoperatóriamente em quatro fetos com idade gestacional aproximada de 45 dias, e outros dois fetos os quais não receberam o transplante, utilizados como controle. Avaliamos os fetos pré e pós-transplante através do ultrasson. Após sete dias, realizamos a ovário-salpingo-histerectomia (OSH) para a colheita dos fetos. Em seguida coletamos seus órgãos e tecidos os quais foram fixados em paraformoldeído a 4% e criopreservados a temperatura de -80oC. Analisamos a biodistribuição das CTIPDh dentro dos órgãos e tecidos em criocortes de 5µm sob microscopia Confocal. Constatamos o homing das CTIPDh nos órgãos derivados das linhas germinativas endodermais, ectodermais e mesodermais. No estômago e intestinos as CTIPD/GFP+ foram identificadas tanto no espaço intraglandular, como na camada muscular da mucosa; no fígado no parenquima hepático; no coração especialmente no tecido muscular do miocárdio; no cérebro nos vasos da substância branca, e cerebelo entre células de Purkinje. Na placenta estas células foram encontradas especialmente junto aos vasos. Quantificamos as CTIPD GFP+ utilizando a citometria de fluxo. Comparativamente dentre os órgãos analisados, obtivemos resultados expressivos do homing celular no miocárdio (~50%), no baço e fígado. Nossos resultados foram confirmados através das análises de imunohistoquímica e imunofluorescência utilizando os anticorpos Anti-núcleo (HuNu), Anti-CTIPD e Anti-GFP humanos. Concluímos que as CTIPDh apresentam grande potencial migratório e proliferativo após o TIUCT em fetos caninos. Estas células indiferenciadas demonstraram homing, especialmente nos tecidos: hematopoiéticos fetais (placenta, fígado e baço), tecido epitelial e glandular de órgãos, bem como de nichos perivasculares de CT. Estes dados sugerem que as CTIPD através do TIU, é uma alternativa viável, segura e promissora para o tratamento de doenças genéticas, congênitas, hematológicas e imunológicas. / Intra-uterine stem cells transplantation (IUSCT) is a method for the treatment of genetic, congenital, hematological, and immunological diseases. In basic research it provides a model for studying the dynamics of migration, graft and functional status of different types of stem cells. The cells can be transplanted in different moments of gestational period, which can be divided into quarters that are not functionally equivalent. The choice of the cells and quarter where the stem cells will be applied can influence cells behavior and results of transplantation. Fetal and adult hematopoietic or bone marrow derived mesenchymal stem cells (MSCs) were mainly used for IUSCT. We previously obtained human immature dental pulp stem cell (IDPSCs), which showed pluripotent potential and immune-compatible properties. The goal of our study was to evaluate migration capacity, proliferation and homing of IDPSCs after IUSCT during the third fetal period in dogs. All experimental procedures were approved by the Ethical Committee of the School of Veterinary Medicine and Animal Science of São Paulo University and were performed under appropriate anesthesia. 1x106 of undifferentiated GFP-positive human IDPSCs were transplanted following laparotomy and intraperitoneal injection under intra-operative ultrasound control into 5 fetuses at the 45 days of gestation. Five fetuses, which did not receive IDPSCs, were used as a control. Ultrasound analyses were performed daily before collection of the fetuses. After 7 days ovarian hysterectomy was performed, fetuses were collected; organs and tissues were isolated and fixed in 4% paraformaldehyde or cryopreserved. Biodistribution of IDPSCs within the organs and tissues were analyzed on cryosections (5µm) under Confocal Microscopy. Homing of IDPSCs was observed in organs derived from three germ lines, endoderm, ectoderm and mesoderm. In stomach and in intestine GFP IDPSCs were found in intraglandular space as well as in muscularis mucosae. In liver they appeared in hepatic parenchyma; in heart in myocardium and in brain in bold vessels, in cerebellum within Purkinje cells. Using Flow cytometry assay GFP IDPSCs graft was quantified. Among the different organs an expressive homing was observed in myocardium of heart (~50%), in spleen and liver. The IDPSCs were also found in canine placenta, especially in blood vessels. These data were confirmed using anti-human nucleus (HuNu), anti-GFP and anti-IDPSCs anti-bodies. Human IDPSCs showed high migration and proliferation potential after IUSCT in dog fetuses. Undifferentiated IDPSCs demonstrated homing in fetal hematopoietic (placenta), epithelial (gastric glands) and perivascular stem cells niches. Our data suggest that IDPSCs is a new promising source for genetic, congenital, hematological, and immunological treatment for those diseases through IUSCT.
6

Estudo da biodistribuição de células tronco de polpa de dente decíduo humana (CTPDDh) após o transplante intra-uterino no modelo canino (Canis lupus familiares) / Biodistribution of human immature dental pulp stem cells following in utero transplantation in canine model (Canis lupus familiaris)

Ana Luísa Reginato 19 June 2012 (has links)
O transplante intrauterino de células-tronco (TIUCT) é um método de tratamento de doenças genéticas, congênitas, hematológicas e imunológicas em um feto durante a gestação. Em pesquisa básica este modelo permite o estudo da dinâmica de migração, enxertia e estado funcional de diferentes tipos de células-tronco (CT). Estas células podem ser transplantadas em diferentes momentos do período gestacional, que pode ser dividido em três momentos do desenvolvimento fetal, sendo estes, diferentes funcionalmente. A escolha deste momento para o transplante influenciará tanto no comportamento celular quanto no resultado. Para o TIUCT são utilizadas as CT mesenquimais derivadas da medula óssea ou fetais ou hematopoiéticas. Para esta pesquisa utilizamos células-tronco derivadas da polpa dentária imatura humana (CTIPDh) as quais apresentam potencial pluripotente e propriedades imunomodulatórias. Nosso principal objetivo foi avaliar a capacidade migratória, bem como de proliferação e endereçamento (homing) das CTIPDh durante o terceiro período gestacional do desenvolvimento fetal no modelo canino. Todos os procedimentos experimentais foram elaborados sob protocolo anestésico apropriado e aprovados pelo comitê de ética da FMVZ da USP. Foram transplantadas via intraperitoneal (IP) 1x106 CTIPDh GFP+ em cada feto, durante procedimento cirúrgico de laparotomia exploratória com ultrassonografia guiada intraoperatóriamente em quatro fetos com idade gestacional aproximada de 45 dias, e outros dois fetos os quais não receberam o transplante, utilizados como controle. Avaliamos os fetos pré e pós-transplante através do ultrasson. Após sete dias, realizamos a ovário-salpingo-histerectomia (OSH) para a colheita dos fetos. Em seguida coletamos seus órgãos e tecidos os quais foram fixados em paraformoldeído a 4% e criopreservados a temperatura de -80oC. Analisamos a biodistribuição das CTIPDh dentro dos órgãos e tecidos em criocortes de 5µm sob microscopia Confocal. Constatamos o homing das CTIPDh nos órgãos derivados das linhas germinativas endodermais, ectodermais e mesodermais. No estômago e intestinos as CTIPD/GFP+ foram identificadas tanto no espaço intraglandular, como na camada muscular da mucosa; no fígado no parenquima hepático; no coração especialmente no tecido muscular do miocárdio; no cérebro nos vasos da substância branca, e cerebelo entre células de Purkinje. Na placenta estas células foram encontradas especialmente junto aos vasos. Quantificamos as CTIPD GFP+ utilizando a citometria de fluxo. Comparativamente dentre os órgãos analisados, obtivemos resultados expressivos do homing celular no miocárdio (~50%), no baço e fígado. Nossos resultados foram confirmados através das análises de imunohistoquímica e imunofluorescência utilizando os anticorpos Anti-núcleo (HuNu), Anti-CTIPD e Anti-GFP humanos. Concluímos que as CTIPDh apresentam grande potencial migratório e proliferativo após o TIUCT em fetos caninos. Estas células indiferenciadas demonstraram homing, especialmente nos tecidos: hematopoiéticos fetais (placenta, fígado e baço), tecido epitelial e glandular de órgãos, bem como de nichos perivasculares de CT. Estes dados sugerem que as CTIPD através do TIU, é uma alternativa viável, segura e promissora para o tratamento de doenças genéticas, congênitas, hematológicas e imunológicas. / Intra-uterine stem cells transplantation (IUSCT) is a method for the treatment of genetic, congenital, hematological, and immunological diseases. In basic research it provides a model for studying the dynamics of migration, graft and functional status of different types of stem cells. The cells can be transplanted in different moments of gestational period, which can be divided into quarters that are not functionally equivalent. The choice of the cells and quarter where the stem cells will be applied can influence cells behavior and results of transplantation. Fetal and adult hematopoietic or bone marrow derived mesenchymal stem cells (MSCs) were mainly used for IUSCT. We previously obtained human immature dental pulp stem cell (IDPSCs), which showed pluripotent potential and immune-compatible properties. The goal of our study was to evaluate migration capacity, proliferation and homing of IDPSCs after IUSCT during the third fetal period in dogs. All experimental procedures were approved by the Ethical Committee of the School of Veterinary Medicine and Animal Science of São Paulo University and were performed under appropriate anesthesia. 1x106 of undifferentiated GFP-positive human IDPSCs were transplanted following laparotomy and intraperitoneal injection under intra-operative ultrasound control into 5 fetuses at the 45 days of gestation. Five fetuses, which did not receive IDPSCs, were used as a control. Ultrasound analyses were performed daily before collection of the fetuses. After 7 days ovarian hysterectomy was performed, fetuses were collected; organs and tissues were isolated and fixed in 4% paraformaldehyde or cryopreserved. Biodistribution of IDPSCs within the organs and tissues were analyzed on cryosections (5µm) under Confocal Microscopy. Homing of IDPSCs was observed in organs derived from three germ lines, endoderm, ectoderm and mesoderm. In stomach and in intestine GFP IDPSCs were found in intraglandular space as well as in muscularis mucosae. In liver they appeared in hepatic parenchyma; in heart in myocardium and in brain in bold vessels, in cerebellum within Purkinje cells. Using Flow cytometry assay GFP IDPSCs graft was quantified. Among the different organs an expressive homing was observed in myocardium of heart (~50%), in spleen and liver. The IDPSCs were also found in canine placenta, especially in blood vessels. These data were confirmed using anti-human nucleus (HuNu), anti-GFP and anti-IDPSCs anti-bodies. Human IDPSCs showed high migration and proliferation potential after IUSCT in dog fetuses. Undifferentiated IDPSCs demonstrated homing in fetal hematopoietic (placenta), epithelial (gastric glands) and perivascular stem cells niches. Our data suggest that IDPSCs is a new promising source for genetic, congenital, hematological, and immunological treatment for those diseases through IUSCT.

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