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1.
Radiographics ; 44(6): e230182, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38781089

RESUMO

Renal transplant is the first-line treatment of end-stage renal disease. The increasing number of transplants performed every year has led to a larger population of transplant patients. Complications may arise during the perioperative and postoperative periods, and imaging plays a key role in this scenario. Contrast-enhanced US (CEUS) is a safe tool that adds additional value to US. Contrast agents are usually administered intravenously, but urinary tract anatomy and complications such as stenosis or leak can be studied using intracavitary administration of contrast agents. Assessment of the graft and iliac vessels with CEUS is particularly helpful in identifying vascular and parenchymal complications, such as arterial or venous thrombosis and stenosis, acute tubular injury, or cortical necrosis, which can lead to graft loss. Furthermore, infectious and malignant graft involvement can be accurately studied with CEUS, which can help in detection of renal abscesses and in the differentiation between benign and malignant disease. CEUS is also useful in interventional procedures, helping to guide percutaneous aspiration of collections with better delimitation of the graft boundaries and to guide renal graft biopsies by avoiding avascular areas. Potential postprocedural vascular complications, such as pseudoaneurysm, arteriovenous fistula, or active bleeding, are identified with CEUS. In addition, newer quantification tools such as CEUS perfusion are promising, but further studies are needed to approve its use for clinical purposes. ©RSNA, 2024 Supplemental material is available for this article.


Assuntos
Meios de Contraste , Transplante de Rim , Complicações Pós-Operatórias , Ultrassonografia , Humanos , Transplante de Rim/efeitos adversos , Complicações Pós-Operatórias/diagnóstico por imagem , Ultrassonografia/métodos , Falência Renal Crônica/diagnóstico por imagem , Falência Renal Crônica/terapia , Falência Renal Crônica/cirurgia
2.
Mar Environ Res ; 196: 106435, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38467089

RESUMO

Cnidarians may dominate benthic communities, as in the case of coral reefs that foster biodiversity and provide important ecosystem services. Polyps may feed by predating mesozooplantkon and large motile prey, but many species further obtain autotrophic nutrients from photosymbiosis. Anthropogenic disturbance, such as the rise of seawater temperature and turbidity, can lead to the loss of symbionts, causing bleaching. Prolonged periods of bleaching can induce mortality events over vast areas. Heterotrophy may allow bleached cnidarians to survive for long periods of time. We tested the reinforcement of heterotrophic feeding of bleached polyps of Exaiptasia diaphana fed with both small zooplantkon and large prey, in order to evaluate if heterotrophy allows this species to compensate the reduction of autotrophy. Conversely to expected, heterotrophy was higher in unbleached polyps (+54% mesozooplankton prey and +11% large prey). The increase of heterotrophic intake may not be always used as a strategy to compensate autotrophic depletion in bleached polyps. Such a resilience strategy might be more species-specific than expected.


Assuntos
Antozoários , Anêmonas-do-Mar , Animais , Ecossistema , Comportamento Predatório , Recifes de Corais , Simbiose
3.
Adv Healthc Mater ; : e2301966, 2024 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-38345178

RESUMO

Neutrophils are the first line of defense of the innate immune system. In response to methicillin-resistant Staphylococcus aureus infection in the skin, hematopoietic stem, and progenitor cells (HSPCs) traffic to wounds and undergo extramedullary granulopoiesis, producing neutrophils necessary to resolve the infection. This prompted the engineering of a gelatin methacrylate (GelMA) hydrogel that encapsulates HSPCs within a matrix amenable to subcutaneous delivery. The authors study the influence of hydrogel mechanical properties to produce an artificial niche for granulocyte-monocyte progenitors (GMPs) to efficiently expand into functional neutrophils that can populate infected tissue. Lin-cKIT+ HSPCs, harvested from fluorescent neutrophil reporter mice, are encapsulated in GelMA hydrogels of varying polymer concentration and UV-crosslinked to produce HSPC-laden gels of specific stiffness and mesh sizes. Softer 5% GelMA gels yield the most viable progenitors and effective cell-matrix interactions. Compared to suspension culture, 5% GelMA results in a twofold expansion of mature neutrophils that retain antimicrobial functions including degranulation, phagocytosis, and ROS production. When implanted dermally in C57BL/6J mice, luciferase-expressing neutrophils expanded in GelMA hydrogels are visualized at the site of implantation for over 5 days. They demonstrate the potential of GelMA hydrogels for delivering HSPCs directly to the site of skin infection to promote local granulopoiesis.

4.
Tissue Eng Part A ; 30(7-8): 259-271, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38126327

RESUMO

Osteoarthritis (OA) is the most prevalent musculoskeletal disorder and a leading cause of disability globally. Although many efforts have been made to treat this condition, current tissue engineering (TE) and regenerative medicine strategies fail to address the inflammatory tissue environment that leads to the rapid progression of the disease and prevents cartilage tissue formation. First, this review addresses in detail the current anti-inflammatory therapies for OA with a special emphasis on pharmacological approaches, gene therapy, and mesenchymal stromal cell (MSC) intra-articular administration, and discusses the reasons behind the limited clinical success of these approaches at enabling cartilage regeneration. Then, we analyze the state-of-the-art TE strategies and how they can be improved by incorporating immunomodulatory capabilities such as the optimization of biomaterial composition, porosity and geometry, and the loading of anti-inflammatory molecules within an engineered structure. Finally, the review discusses the future directions for the new generation of TE strategies for OA treatment, specifically focusing on the spatiotemporal modulation of anti-inflammatory agent presentation to allow for tailored patient-specific therapies. Impact statement Osteoarthritis (OA) is a prevalent and debilitating musculoskeletal disorder affecting millions worldwide. Despite significant advancements in regenerative medicine and tissue engineering (TE), mitigating inflammation while simultaneously promoting cartilage tissue regeneration in OA remains elusive. In this review article, we discuss current anti-inflammatory therapies and explore their potential synergy with cutting-edge cartilage TE strategies, with a special focus on novel spatiotemporal and patient-specific anti-inflammatory strategies.


Assuntos
Cartilagem Articular , Transplante de Células-Tronco Mesenquimais , Osteoartrite , Humanos , Osteoartrite/terapia , Engenharia Tecidual , Anti-Inflamatórios
5.
JMIR Res Protoc ; 12: e48925, 2023 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-37962929

RESUMO

BACKGROUND: Only 5% of the molecules tested in oncology phase 1 trials reach the market after an average of 7.5 years of waiting and at a cost of tens of millions of dollars. To reduce the cost and shorten the time of discovery of new treatments, "drug repurposing" (research with molecules already approved for another indication) and the use of secondary data (not collected for the purpose of research) have been proposed. Due to advances in informatics in clinical care, secondary data can, in some cases, be of equal quality to primary data generated through prospective studies. OBJECTIVE: The objective of this study is to identify drugs currently marketed for other indications that may have an effect on the prognosis of patients with cancer. METHODS: We plan to monitor a cohort of patients with high-lethality cancers treated in the public health system of Catalonia between 2006 and 2012, retrospectively, for survival for 5 years after diagnosis or until death. A control cohort, comprising people without cancer, will also be retrospectively monitored for 5 years. The following study variables will be extracted from different population databases: type of cancer (patients with cancer cohort), date and cause of death, pharmacological treatment, sex, age, and place of residence. During the first stage of statistical analysis of the patients with cancer cohort, the drugs consumed by the long-term survivors (alive at 5 years) will be compared with those consumed by nonsurvivors. In the second stage, the survival associated with the consumption of each relevant drug will be analyzed. For the analyses, groups will be matched for potentially confounding variables, and multivariate analyses will be performed to adjust for residual confounding variables if necessary. The control cohort will be used to verify whether the associations found are exclusive to patients with cancer or whether they also occur in patients without cancer. RESULTS: We anticipate discovering multiple significant associations between commonly used drugs and the survival outcomes of patients with cancer. We expect to publish the initial results in the first half of 2024. CONCLUSIONS: This retrospective study may identify several commonly used drugs as candidates for repurposing in the treatment of various cancers. All analyses are considered exploratory; therefore, the results will have to be confirmed in subsequent clinical trials. However, the results of this study may accelerate drug discovery in oncology. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/48925.

7.
Polymers (Basel) ; 14(19)2022 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-36236029

RESUMO

Urothelial tumour of the upper urinary tract is a rare neoplasm, but unfortunately, it has a high recurrence rate. The reduction of these tumour recurrences could be achieved by the intracavitary instillation of adjuvant chemotherapy after nephron-sparing treatment in selected patients, but current instillation methods are ineffective. Therefore, the aim of this in vitro study is to evaluate the cytotoxic capacity of a new instillation technology through a biodegradable ureteral stent/scaffold coated with a silk fibroin matrix for the controlled release of mitomycin C as an anti-cancer drug. Through a comparative study, we assessed, in urothelial carcinoma cells in a human cancer T24 cell culture for 3 and 6 h, the cytotoxic capacity of mitomycin C by viability assay using the CCK-8 test (Cell counting Kit-8). Cell viability studies in the urothelial carcinoma cell line confirm that mitomycin C embedded in the polymeric matrix does not alter its cytotoxic properties and causes a significant decrease in cell viability at 6 h versus in the control groups. These findings have a clear biomedical application and could be of great use to decrease the recurrence rate in patients with upper tract urothelial carcinomas by increasing the dwell time of anti-cancer drugs.

8.
Knee Surg Sports Traumatol Arthrosc ; 29(6): 1822-1829, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32809118

RESUMO

PURPOSE: In patients with open growth plates, the direction of tunneling that avoids distal femoral physis (DFP) damage in anatomic reconstructions of the medial patellofemoral ligament (MPFL) has been a topic of discussion. The objective of this study was to determine the ideal orientation for anatomic reconstructions of MPFL tunneling that minimized DFP damage while avoiding breaching the intercondylar notch. METHODS: Eighty magnetic resonance images of patients aged 10 through 17 were obtained, randomly sampled from the institutional database. A de novo software was developed to obtain 3D models of the distal femur and DFP. In each model, the anatomical insertion point of the MPFL was determined as defined by Stephen. A 20-mm-depth drilling was simulated, starting from the insertion point at every possible angle within a 90° cone using 5-, 6- and 7-mm drills. Physeal damage for each pair of angles and each drill size was determined. Damage was expressed as a percentage of total physis volume. Statistical analysis was conducted using Student's t test and one-way ANOVA. RESULTS: Maximum physeal damage (5.35% [4.47-6.24]) was obtained with the 7-mm drill when drilling 3° cephalic and 15° posterior from insertion without differences between sexes (n.s.). Minimal physeal damage (0.22% [0.07-0.37]) was obtained using the 5-mm drill aimed 45° distal and 0° anteroposterior, not affected by sex (n.s.). Considering intra-articular drilling avoidance, the safest zone was obtained when aiming 30°-40° distal and 5°-35° anterior, regardless of sex. CONCLUSION: Ideal femoral tunnel orientation, avoiding physeal damage and breaching of the intercondylar notch, was obtained when aiming 30°-40° distal and 5°-35° anterior, regardless of sex. This area is a safe zone that allows anatomic MPFL reconstruction of patients with an open physis.


Assuntos
Lâmina de Crescimento/cirurgia , Ligamentos Articulares/cirurgia , Luxação Patelar/cirurgia , Articulação Patelofemoral/cirurgia , Procedimentos de Cirurgia Plástica/métodos , Adolescente , Criança , Feminino , Fêmur/diagnóstico por imagem , Fêmur/cirurgia , Lâmina de Crescimento/diagnóstico por imagem , Lâmina de Crescimento/patologia , Humanos , Articulação do Joelho/diagnóstico por imagem , Articulação do Joelho/cirurgia , Ligamentos Articulares/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Masculino , Orientação Espacial , Articulação Patelofemoral/diagnóstico por imagem , Software
9.
3D Print Addit Manuf ; 7(3): 139-147, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32855996

RESUMO

Aggregation of cells into spheroids and organoids is a promising tool for regenerative medicine, cancer and cell biology, and drug discovery due to their recapitulation of the cell-cell and cell-matrix interactions found in vivo. Traditional approaches for the production of spheroids, such as the hanging drop method, are limited by the lack of reproducibility and the use of labor-intensive and time-consuming techniques. The need for high-throughput approaches allowing for the quick and reproducible formation of cell aggregates has driven the development of soft lithography techniques based on the patterning of microwells into nonadherent hydrogels. However, these methods are also limited by costly, labor-intensive, and multistep protocols that could impact the sterility of the process and efficiency of spheroid formation. In this study, we describe a one-step method for the fabrication of patterned nonadherent microwells into tissue culture plates using three-dimensional (3D) printed stamps and evaluate the production of cell spheroids of different sizes and cell sources. The generation of bone marrow-derived mesenchymal stromal cell and endothelial cell spheroids by the use of 3D printed stamps was superior in comparison with a widely used multistep mold technique, yielding spheroids of larger sizes and higher DNA content. The 3D stamps produced spheroids of more consistent diameter and DNA content when compared with other commercially available methods. These 3D printed stamps offer a tunable, simple, fast, and cost-effective approach for the production of reproducible spheroids and organoids for a wide range of applications.

10.
Biomaterials ; 245: 119973, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32244091

RESUMO

The native extracellular matrix (ECM) contains a host of matricellular proteins and bioactive factors that regulate cell behavior, and many ECM components have been leveraged to guide cell fate. However, the large size and chemical characteristics of these constituents complicate their incorporation into biomaterials without interfering with material properties, motivating the need for alternative approaches to regulate cellular responses. Mesenchymal stromal cells (MSCs) can promote osseous regeneration in vivo directly or indirectly through multiple means including (1) secretion of proangiogenic and mitogenic factors to initiate formation of a vascular template and recruit host cells into the tissue site or (2) direct differentiation into osteoblasts. As MSC behavior is influenced by the properties of engineered hydrogels, we hypothesized that the biochemical and biophysical properties of alginate could be manipulated to promote the dual contributions of encapsulated MSCs toward bone formation. We functionalized alginate with QK peptide to enhance proangiogenic factor secretion and RGD to promote adhesion, while biomechanical-mediated osteogenic cues were controlled by modulating viscoelastic properties of the alginate substrate. A 1:1 ratio of QK:RGD resulted in the highest levels of both proangiogenic factor secretion and mineralization in vitro. Viscoelastic alginate outperformed purely elastic gels in both categories, and this effect was enhanced by stiffness up to 20 kPa. Furthermore, viscoelastic constructs promoted vessel infiltration and bone regeneration in a rat calvarial defect over 12 weeks. These data suggest that modulating viscoelastic properties of biomaterials, in conjunction with dual peptide functionalization, can simultaneously enhance multiple aspects of MSC regenerative potential and improve neovascularization of engineered tissues.


Assuntos
Hidrogéis , Células-Tronco Mesenquimais , Animais , Diferenciação Celular , Osteogênese , Peptídeos , Ratos , Células Estromais
11.
Mar Environ Res ; 155: 104887, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32072989

RESUMO

The impact of plastic debris, and in particular of microplastics (here referred as particles smaller than 5 mm) on aquatic environments has now become a topic of raising concern. Microplastics are particularly abundant in the Mediterranean Sea, potentially exerting substantial pressures on marine organisms at different levels of organization. Ingestion of microplastics has been observed in a large number of marine species. The aim of this work is to test if microplastics produce a feeding impairment in Astroides calycularis, a shallow water, habitat-forming coral endemic to the Mediterranean Sea. Our findings suggest a lack of any avoidance mechanism allowing the polyps to discern between food items and microplastics when occurring simultaneously. Moreover, polyps spend a considerable amount of time on handling microplastic particles. As a consequence, microplastics impair the feeding efficiency in A. calycularis, since polyps may not be fully able to profit from the drifting plankton aggregations. Therefore, we suggest that microplastics can cause a reduction of fitness in A. calycularis, and presumably also in other species characterized by suspension feeding strategy.


Assuntos
Antozoários , Comportamento Alimentar , Microplásticos/efeitos adversos , Poluentes Químicos da Água/efeitos adversos , Animais , Ecossistema , Monitoramento Ambiental , Mar Mediterrâneo
12.
Biomaterials ; 239: 119833, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32062479

RESUMO

Engineering a pro-regenerative immune response following scaffold implantation is integral to functional tissue regeneration. The immune response to implanted biomaterials is determined by multiple factors, including biophysical cues such as material stiffness, topography and particle size. In this study we developed an immune modulating scaffold for bone defect healing containing bone mimetic nano hydroxyapatite particles (BMnP). We first demonstrate that, in contrast to commercially available micron-sized hydroxyapatite particles, in-house generated BMnP preferentially polarize human macrophages towards an M2 phenotype, activate the transcription factor cMaf and specifically enhance production of the anti-inflammatory cytokine, IL-10. Furthermore, nano-particle treated macrophages enhance mesenchymal stem cell (MSC) osteogenesis in vitro and this occurs in an IL-10 dependent manner, demonstrating a direct pro-osteogenic role for this cytokine. BMnPs were also capable of driving pro-angiogenic responses in human macrophages and HUVECs. Characterization of immune cell subsets following incorporation of functionalized scaffolds into a rat femoral defect model revealed a similar profile, with micron-sized hydroxyapatite functionalized scaffolds eliciting pro-inflammatory responses characterized by infiltrating T cells and elevated expression of M1 macrophages markers compared to BMnP functionalized scaffolds which promoted M2 macrophage polarization, tissue vascularization and increased bone volume. Taken together these results demonstrate that nano-sized Hydroxyapatite has immunomodulatory potential and is capable of directing anti-inflammatory innate immune-mediated responses that are associated with tissue repair and regeneration.


Assuntos
Células-Tronco Mesenquimais , Osteogênese , Animais , Regeneração Óssea , Interleucina-10 , Ativação de Macrófagos , Macrófagos , Ratos , Alicerces Teciduais
13.
Rev. cuba. reumatol ; 22(supl.1): e849, graf
Artigo em Espanhol | LILACS, CUMED | ID: biblio-1280395

RESUMO

La incidencia de la miopatía inflamatoria idiopática es de 4 a 15 casos por cada millón de habitantes y su prevalencia de 60 por cada millón de habitantes. La dermatomiositis idiopática es más frecuente en las mujeres, aunque su asociación a fibrosis pulmonar es muy rara y solo se reporta en un 2 por ciento de los casos. Se describe el caso de un paciente de 50 años de edad, femenina, que presentó debilidad a nivel de la cintura escapular acompañada de fatiga. Tenía lesiones de rascado en diferentes regiones del cuerpo por prurito y lesiones eritematosas en la piel en ambos muslos. Además, se quejaba de dolores articulares generalizados con impotencia funcional y mialgias generalizadas progresivas e hipotrofia muscular de varios grupos musculares. El estudio analítico reveló enzimas musculares elevadas. La biopsia de piel y músculo mostró elementos sugestivos de dermatomiositis. Con la espirometría se detectó trastornos ventilatorios restrictivos de grave intensidad. Mediante la radiografía de tórax se halló infiltrado difuso peribroncovascular asociado a un trayecto fibroso y la tomografía axial computarizada precisó el pulmón con consolidación alveolar y discreto engrosamiento pleural. La paciente fue tratada con prednisona a 1 mg/kg/día asociado con azatioprina 1,5 mg/kg/día. Este tratamiento fue muy eficaz, y se logró una notable recuperación clínica y por estudios de laboratorio. Reportamos el caso de una paciente con dermatomiositis idiopática y fibrosis pulmonar. Esta asociación constituye un hallazgo infrecuente en nuestro medio y más aun con el paciente asintomático(AU)


The incidence of Idiopathic Inflammatory Myopathy is from 4 to 15 cases per million inhabitants and its prevalence of 60 per million inhabitants. Idiopathic dermatomyositis is more frequent in women; Although its association with pulmonary fibrosis is described, it is very infrequent, it is only reported in 2 percent of cases. To describe a diagnosed case of idiopathic dermatomyositis and pulmonary fibrosis. A 50-year-old patient presented weakness at the level of the shoulder girdle accompanied by fatigue. Physical examination: Skin: scratching lesions in different regions of the body due to pruritus, erythematous lesions at the level of the skin on both thighs. Osteomyoarticular system: generalized joint pains with functional impotence and progressive generalized myalgias and muscular hypotrophy of several muscle groups. The analytical study revealed elevated muscle enzymes. The skin and muscle biopsy showed elements suggestive of dermatomyositis. Chest X-ray: diffuse peribronchovascular infiltrate associated with fibrous path. Spirometry: restrictive ventilatory disorders of severe intensity. Computed tomography of the lung with alveolar consolidation and discrete pleural thickening. We report the case of a patient with idiopathic dermatomyositis and pulmonary fibrosis. This association is an uncommon finding in our environment and even more so when the patient is asymptomatic(AU)


Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Espirometria/métodos , Azatioprina/uso terapêutico , Prednisona/uso terapêutico , Dermatomiosite/diagnóstico , Miosite/epidemiologia , Fadiga , Relatório de Pesquisa
14.
Sci Rep ; 9(1): 13469, 2019 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-31530904

RESUMO

Shallow-water marine organisms are among the first to suffer from combined effects of natural and anthropogenic drivers. The orange coral Astroides calycularis is a shallow-water bioconstructor species endemic to the Mediterranean Sea. Although raising conservation interest, also given its special position within the Dendrophylliidae, information about the threats to its health is scant. We investigated the health status of A. calycularis at five locations in northwestern Sicily along a gradient of cumulative human impact and the most probable origin of the threats to this species, including anthropogenic land-based and sea-based threats. Cumulative human impact appeared inversely related to the performance of A. calycularis at population, colony, and polyp levels. Sea-based human impacts appeared among the most likely causes of the variation observed. The reduction in polyp length can limit the reproductive performance of A. calycularis, while the decrease of percent cover and colony area is expected to impair its peculiar feeding behaviour by limiting the exploitable dimensional range of prey and, ultimately, reef functioning. This endangered habitat-forming species appeared susceptible to anthropogenic pressures, suggesting the need to re-assess its vulnerability status. Creating microprotected areas with specific restrictions to sea-based human impacts could be the best practice preserve these bioconstructions.


Assuntos
Antozoários , Ecossistema , Animais , Antozoários/anatomia & histologia , Antozoários/crescimento & desenvolvimento , Conservação dos Recursos Naturais , Humanos , Mar Mediterrâneo , Sicília
15.
Acta Biomater ; 88: 314-324, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30825603

RESUMO

Controlling the phenotype of transplanted stem cells is integral to ensuring their therapeutic efficacy. Hypoxia is a known regulator of stem cell fate, the effects of which can be mimicked using hypoxia-inducible factor (HIF) prolyl hydroxylase inhibitors such as dimethyloxalylglycine (DMOG). By releasing DMOG from mesenchymal stem cell (MSC) laden alginate hydrogels, it is possible to stabilize HIF-1α and enhance its nuclear localization. This correlated with enhanced chondrogenesis and a reduction in the expression of markers associated with chondrocyte hypertrophy, as well as increased SMAD 2/3 nuclear localization in the encapsulated MSCs. In vivo, DMOG delivery significantly reduced mineralisation of the proteoglycan-rich cartilaginous tissue generated by MSCs within alginate hydrogels loaded with TGF-ß3 and BMP-2. Together these findings point to the potential of hypoxia mimicking hydrogels to control the fate of stem cells following their implantation into the body. STATEMENT OF SIGNIFICANCE: There are relatively few examples where in vivo delivery of adult stem cells has demonstrated a true therapeutic benefit. This may be attributed, at least in part, to a failure to control the fate of transplanted stem cells in vivo. In this paper we describe the development of hydrogels that mimic the effects of hypoxia on encapsulated stem cells. In vitro, these hydrogels enhance chondrogenesis of MSCs and suppress markers associated with chondrocyte hypertrophy. In an in vivo environment that otherwise supports progression along an endochondral pathway, we show that these hydrogels will instead direct mesenchymal stem cells (MSCs) to produce a more stable, cartilage-like tissue. In addition, we explore potential molecular mechanisms responsible for these phenotypic changes in MSCs.


Assuntos
Hidrogéis/farmacologia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Alginatos/química , Aminoácidos Dicarboxílicos/farmacologia , Animais , Proteína Morfogenética Óssea 2/farmacologia , Hipóxia Celular/efeitos dos fármacos , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Condrogênese/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Hipertrofia , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Células-Tronco Mesenquimais/efeitos dos fármacos , Camundongos Nus , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Estabilidade Proteica/efeitos dos fármacos , Transporte Proteico/efeitos dos fármacos , Proteínas Smad/metabolismo , Suínos , Fator de Crescimento Transformador beta3/farmacologia
16.
Biomaterials ; 189: 1-10, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30384124

RESUMO

The maintenance and direction of stem cell lineage after implantation remains challenging for clinical translation. Aggregation and encapsulation into instructive biomaterials after preconditioning can bolster retention of differentiated phenotypes. Since these procedures do not depend on cell type or lineage, we hypothesized we could use a common, tunable platform to engineer formulations that retain and enhance multiple lineages from different cell populations. To test this, we varied alginate stiffness and adhesive ligand content, then encapsulated spheroids of varying cellularity. We used Design-of-Experiments to determine the effect of these parameters and their interactions on phenotype retention. The combination of parameters leading to maximal differentiation varied with lineage and cell type, inducing a 2-4-fold increase over non-optimized levels. Phenotype was also retained for 4 weeks in a murine subcutaneous model. This widely applicable approach can facilitate translation of cell-based therapies by instructing phenotype in situ without prolonged induction or costly growth factors.


Assuntos
Alginatos/química , Materiais Biocompatíveis/química , Diferenciação Celular , Hidrogéis/química , Células-Tronco Mesenquimais/citologia , Animais , Células Cultivadas , Feminino , Masculino , Transplante de Células-Tronco Mesenquimais , Camundongos SCID , Esferoides Celulares/citologia
18.
Biomaterials ; 161: 240-255, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29421560

RESUMO

High-density mesenchymal stem cell (MSC) aggregates can be guided to form bone-like tissue via endochondral ossification in vitro when culture media is supplemented with proteins, such as growth factors (GFs), to first guide the formation of a cartilage template, followed by culture with hypertrophic factors. Recent reports have recapitulated these results through the controlled spatiotemporal delivery of chondrogenic transforming growth factor-ß1 (TGF-ß1) and chondrogenic and osteogenic bone morphogenetic protein-2 (BMP-2) from microparticles embedded within human MSC aggregates to avoid diffusion limitations and the lengthy, costly in vitro culture necessary with repeat exogenous supplementation. However, since GFs have limited stability, localized gene delivery is a promising alternative to the use of proteins. Here, mineral-coated hydroxyapatite microparticles (MCM) capable of localized delivery of Lipofectamine-plasmid DNA (pDNA) nanocomplexes encoding for TGF-ß1 (pTGF-ß1) and BMP-2 (pBMP-2) were incorporated, alone or in combination, within MSC aggregates from three healthy porcine donors to induce sustained production of these transgenes. Three donor populations were investigated in this work due to the noted MSC donor-to-donor variability in differentiation capacity documented in the literature. Delivery of pBMP-2 within Donor 1 aggregates promoted chondrogenesis at week 2, followed by an enhanced osteogenic phenotype at week 4. Donor 2 and 3 aggregates did not promote robust glycosaminoglycan (GAG) production at week 2, but by week 4, Donor 2 aggregates with pTGF-ß1/pBMP-2 and Donor 3 aggregates with both unloaded MCM and pBMP-2 enhanced osteogenesis compared to controls. These results demonstrate the ability to promote osteogenesis in stem cell aggregates through controlled, non-viral gene delivery within the cell masses. These findings also indicate the need to screen donor MSC regenerative potential in response to gene transfer prior to clinical application. Taken together, this work demonstrates a promising gene therapy approach to control stem cell fate in biomimetic 3D condensations for treatment of bone defects.


Assuntos
Engenharia Tecidual/métodos , Animais , Proteína Morfogenética Óssea 2/administração & dosagem , Proteína Morfogenética Óssea 2/farmacologia , Osso e Ossos/citologia , Células Cultivadas , Condrogênese/efeitos dos fármacos , Durapatita/química , Técnicas de Transferência de Genes , Glicosaminoglicanos , Humanos , Células-Tronco Mesenquimais/citologia , Suínos , Fator de Crescimento Transformador beta1/administração & dosagem , Fator de Crescimento Transformador beta1/farmacologia
19.
Mar Pollut Bull ; 129(2): 469-473, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29079302

RESUMO

Understanding which factors influence the invasion of alien seaweed has become a central concern in ecology. Increasing evidence suggests that the feeding preferences of native herbivores influence the success of alien seaweeds in the new community. We investigated food selection of a generalist native grazer Paracentrotus lividus, in the presence of two alien seaweeds (Caulerpa cylindracea and Caulerpa taxifolia var. distichophylla) and two native seaweeds (Dictyopteris membranacea and Cystoseira compressa). Sea urchins were fed with six experimental food items: C. cylindracea, C. taxifolia var. distichophylla, a mixture of C. cylindracea and C. taxifolia var. distichophylla, D. membranacea, C. compressa and a mixture of D. membranacea and C. compressa. P. lividus ingested all the combinations of food offered, though it preferentially consumed the alien mixture, C. cylindracea and D. membranacea. The alien C. taxifolia var. distichophylla was consumed significantly less than the other food items and, interestingly, it was ingested in a greater amount when mixed with C. cylindracea than when on its own. This finding suggests that C. taxifolia var. distichophylla may become vulnerable to sea urchin grazing when it grows intermingled with C. cylindracea, which does not gain immediate protection from the presence of the very low palatable congeneric seaweed. The present study highlights the potential role of native grazers to indirectly affect the interspecific competition between the two alien seaweeds in the Mediterranean Sea.


Assuntos
Caulerpa , Preferências Alimentares/fisiologia , Herbivoria/fisiologia , Espécies Introduzidas , Paracentrotus/fisiologia , Alga Marinha , Animais , Caulerpa/crescimento & desenvolvimento , Mar Mediterrâneo , Alga Marinha/crescimento & desenvolvimento
20.
Biol Lett ; 13(11)2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29093176

RESUMO

Predation occurs when an organism completely or partially consumes its prey. Partial consumption is typical of herbivores but is also common in some marine microbenthic carnivores that feed on colonial organisms. Associations between nudibranch molluscs and colonial hydroids have long been assumed to be simple predator-prey relationships. Here we show that while the aeolid nudibranch Cratena peregrina does prey directly on the hydranths of Eudendrium racemosum, it is stimulated to feed when hydranths have captured and are handling prey, thus ingesting recently captured plankton along with the hydroid polyp such that plankton form at least half of the nudibranch diet. The nudibranch is thus largely planktivorous, facilitated by use of the hydroid for prey capture. At the scale of the colony this combines predation with kleptoparasitism, a type of competition that involves the theft of already-procured items to form a feeding mode that does not fit into existing classifications, which we term kleptopredation. This strategy of subsidized predation helps explain how obligate-feeding nudibranchs obtain sufficient energy for reproduction from an ephemeral food source.


Assuntos
Gastrópodes/fisiologia , Hidrozoários/fisiologia , Plâncton , Animais , Comportamento de Escolha , Dieta , Comportamento Alimentar , Comportamento Predatório
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