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In mechanotransduction studies, flow-induced shear stress (FSS) is often applied to two-dimensional (2D) cultured cells with a parallel-plate flow chamber (PPFC) due to its simple FSS estimation. However, cells behave differently under FSS inside a 3D scaffold (e.g. 10 mPa FSS was shown to induce osteogenesis of human mesenchymal stem cells (hMSC) in 3D but over 900 mPa was needed for 2D culture). Here, a simple in vitro biomimetic perfusion system using borosilicate glass capillary tubes has been developed to study the cellular behaviour under low-level FSS that mimics 3D culture. It has been shown that, compared to cells in the PPFC, hMSC in the capillary tubes had upregulated Runx-2 expression and osteogenic cytoskeleton actin network under 10 mPa FSS for 24 h. Also, an image analysis method based on Haralick texture measurement has been used to identify osteogenic actin network. The biomimetic perfusion system can be a valuable tool to study mechanotransduction in 3D for more clinical relevant tissue-engineering applications.
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Osteochondral tissue engineering aims to regenerate functional tissue-mimicking physiological properties of injured cartilage and its subchondral bone. Given the distinct structural and biochemical difference between bone and cartilage, bilayered scaffolds, and bioreactors are commonly employed. We present an osteochondral culture system which cocultured ATDC5 and MC3T3-E1 cells on an additive manufactured bilayered scaffold in a dual-chamber perfusion bioreactor. Also, finite element models (FEM) based on the microcomputed tomography image of the manufactured scaffold as well as on the computer-aided design (CAD) were constructed; the microenvironment inside the two FEM was studied and compared. In vitro results showed that the coculture system supported osteochondral tissue growth in terms of cell viability, proliferation, distribution, and attachment. In silico results showed that the CAD and the actual manufactured scaffold had significant differences in the flow velocity, differentiation media mixing in the bioreactor and fluid-induced shear stress experienced by the cells. This system was shown to have the desired microenvironment for osteochondral tissue engineering and it can potentially be used as an inexpensive tool for testing newly developed pharmaceutical products for osteochondral defects.
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Osso e Ossos , Cartilagem , Técnicas de Cultura de Células , Microambiente Celular , Simulação por Computador , Alicerces Teciduais/química , Microtomografia por Raio-X , Animais , Osso e Ossos/química , Osso e Ossos/diagnóstico por imagem , Osso e Ossos/metabolismo , Cartilagem/citologia , Cartilagem/diagnóstico por imagem , Cartilagem/metabolismo , Diferenciação Celular , Linhagem Celular , Proliferação de Células , Técnicas de Cocultura , CamundongosRESUMO
The management of flexor tendon injury has seen many iterations over the years, but more substantial innovations in practice have been sadly lacking. The aim of this study was to investigate the current practice of flexor tendon injury management, and variation in practice from the previous reports, most troublesome complications, and whether there was a clinical interest in potential innovative tendon repair technologies. An online survey was distributed via the British Society for Surgery of the Hand (BSSH) and a total of 132 responses were collected anonymously. Results showed that although most surgeons followed the current medical recommendation based on the literature, a significant number of surgeons still employed more conventional treatments in clinic, such as general anesthesia, ineffective tendon retrieval techniques, and passive rehabilitation. Complications including adhesion formation and re-rupture remained persistent. The interest in new approaches such as use of minimally invasive instruments, biodegradable materials and additive manufactured devices was not strong, however the surgeons were potentially open to more effective and economic solutions.
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The successful translation of organ-on-a-chip devices requires the development of an automated workflow for device fabrication, which is challenged by the need for precise deposition of multiple classes of materials in micro-meter scaled configurations. Many current heart-on-a-chip devices are produced manually, requiring the expertise and dexterity of skilled operators. Here, we devised an automated and scalable fabrication method to engineer a Biowire II multiwell platform to generate human iPSC-derived cardiac tissues. This high-throughput heart-on-a-chip platform incorporated fluorescent nanocomposite microwires as force sensors, produced from quantum dots and thermoplastic elastomer, and 3D printed on top of a polystyrene tissue culture base patterned by hot embossing. An array of built-in carbon electrodes was embedded in a single step into the base, flanking the microwells on both sides. The facile and rapid 3D printing approach efficiently and seamlessly scaled up the Biowire II system from an 8-well chip to a 24-well and a 96-well format, resulting in an increase of platform fabrication efficiency by 17,5000-69,000% per well. The device's compatibility with long-term electrical stimulation in each well facilitated the targeted generation of mature human iPSC-derived cardiac tissues, evident through a positive force-frequency relationship, post-rest potentiation, and well-aligned sarcomeric apparatus. This system's ease of use and its capacity to gauge drug responses in matured cardiac tissue make it a powerful and reliable platform for rapid preclinical drug screening and development.
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The lack of organic electron donors limits the potential utility of aerobic denitrification in treatment of oligotrophic source water. Here, reduced manganese (Mn) was used as an inorganic electron donor to improve the denitrification of oligotrophic source water under the high dissolved oxygen condition (7-9 mg L-1). Over 30 days, the total nitrogen removed by the treatment with reduced Mn was 76.21 ± 2.11% (maximum), substantially higher than that of the control treatment, which was 41.48 ± 2.33%. Furthermore, the addition of Mn resulted in the directional evolution of the microbial community. Water samples with Mn added showed a higher abundance of Limnohabitans, the dominant denitrifying genus, reaching 51.02%, 36.79%, and 20.19% (with 30, 50, and 70 g Mn, respectively), versus only 5.54% in the control. In biofilm, Mn promoted Hydrogenophaga and Brevundimonas growth while Pseudarthrobacter growth was promoted by 30 and 50 g Mn, but inhibited by 70 g Mn. This study demonstrates an improved performance in aerobic denitrification of water sources through the use of inorganic electron donors.
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Microbiota , Água , Manganês , Desnitrificação , Bactérias Aeróbias , Nitrogênio , NitratosRESUMO
Metalimnetic oxygen minima has been reported in many lakes and reservoirs, but the double metalimnetic oxygen minima (DMOM) is so far poorly understood. In this work, we first reported DMOM in the Sanhekou Reservoir, and investigated its formation reason and influence on the bacterial community composition (BCC). The results showed that the two anaerobic layers were formed in DMOM, located at 10 m and 45 m approximately. The rapid water storage process and thermal stratification resulted in the double metalimnions. Algal accumulation, decomposition and oxygen consumption in these regions during the sedimentation process eventually leaded to the formation of DMOM. Water temperature and DO gradients made outstanding contributions to the spatiotemporal environmental heterogeneity and significantly affected the BCC. Depending on the distribution of dissolved oxygen (DO), the storage process could be divided into three periods: DMOM, single MOM period and mixed period. Exiguobacterium and Ralstonia were dominated in DMOM due to the soil discharge and plant decomposition. Besides, BCC presented the largest vertical difference in DMOM and existed the interlayer-similar phenomenon (BCC in the two anaerobic layers were more similar). This study explained the formation of DMOM and its influence on BCC, which was helpful to understand the response of BCC to the storage process and unique DO structure in a moderate eutrophication reservoir.
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Água Potável , Oxigênio/análise , Qualidade da Água , Bactérias , Temperatura , Eutrofização , ChinaRESUMO
Electrical stimulation of cells allows exogenous electric signals as stimuli to manipulate cell growth, preferential orientation and bone remodelling. In this study, commercially pure titanium discs were utilised in combination with a custom-built bioreactor to investigate the cellular responses of human mesenchymal stem cells via in-vitro functional assays. Finite element analysis revealed the homogeneous delivery of electric field in the bioreactor chamber with no detection of current density fluctuation in the proposed model. The custom-built bioreactor with capacitive stimulation delivery system features long-term stimulation with homogeneous electric field, biocompatible, sterilisable, scalable design and cost-effective in the manufacturing process. Using a continuous stimulation regime of 100 and 200 mV/mm on cp Ti discs, viability tests revealed up to an approximately 5-fold increase of cell proliferation rate as compared to non-stimulated controls. The human mesenchymal stem cells showed more elongated and differentiated morphology under this regime, with evidence of nuclear elongation and cytoskeletal orientation perpendicular to the direction of electric field. The continuous stimulation did not cause pH fluctuations and hydrogen peroxide production caused by Faradic reactions, signifying the suitability for long-term toxic free stimulation as opposed to the commonly used direct stimulation regime. An approximate of 4-fold increase in alkaline phosphatase production and approximately 9-fold increase of calcium deposition were observed on 200 mV/mm exposed samples relative to non-stimulated controls. It is worth noting that early stem cell differentiation and matrix production were observed under the said electric field even without the presence of chemical inductive growth factors. STATEMENT OF SIGNIFICANCE: This manuscript presents a study on combining pure titanium (primarily preferred as medical implant materials) and electrical stimulation in a purpose-built bioreactor with capacitive stimulation delivery system. A continuous capacitive stimulation regime on titanium disc has resulted in enhanced stem cell orientation, nuclei elongation, proliferation and differentiation as compared to non-stimulated controls. We believe that this manuscript creates a paradigm for future studies on the evolution of healthcare treatments in the area of targeted therapy on implantable and wearable medical devices through tailored innovative electrical stimulation approach, thereby influencing therapeutic conductive and electroactive biomaterials research prospects and development.
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Osteogênese , Titânio , Diferenciação Celular , Estimulação Elétrica/métodos , Humanos , Células-Tronco , Titânio/farmacologiaRESUMO
Electroactive materials based on conductive polymers are promising options for tissue engineering and regenerative medicine applications. In the present work, the conducting copolymers of poly (3,4-ethylenedioxythiophene) and poly (d, l-lactic acid) (PEDOT-co-PDLLA) with PEDOT:PDLLA molar ratios of 1:50, 1:25, and 1:5 were synthesized and compared to the insulating macromonomer of EDOT-PDLLA as an experimental control. Bone marrow-derived human mesenchymal stem cells (hMSC-BM) were cultured on the copolymers and the macromonomer thin films inside a bioreactor that induced a capacitive electrical stimulation (CES) with an electric field of 100 mV/mm for 2 h per day for 21 days. Under CES, the copolymers exhibited good cell viability and promoted the differentiation from hMSC-BM to osteogenic lineages, revealed by higher mineralization mainly when the contents of conducting segments of PEDOT (i.e., copolymer with 1:25 and 1:5 PEDOT:PDLLA ratios) were increased. The results indicate that the intrinsic electrical conductivity of the substrates is an important key point for the effectiveness of the electric field generated by the CES, intending to promote the differentiation effect for bone cells.
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Células-Tronco Mesenquimais , Osteogênese , Condutividade Elétrica , Estimulação Elétrica/métodos , Humanos , Polímeros/químicaRESUMO
Electrical stimulation (ES) has potential to be an effective tool for bone injury treatment in clinics. However, the therapeutic mechanism associated with ES is still being discussed. This study aims to investigate the initial mechanism of action by characterising the physical and chemical changes in the extracellular environment during ES and correlate them with the responses of mesenchymal stem/stromal cells (MSCs). Computational modelling was used to estimate the electrical potentials relative to the cathode and the current density across the cell monolayer. We showed expression of phosphorylated ERK1/2, c-FOS, c-JUN, and SPP1 mRNAs, as well as the increased metabolic activities of MSCs at different time points. Moreover, the average of 2.5 µM of H2O2 and 34 µg/L of dissolved Pt were measured from the electrically stimulated media (ES media), which also corresponded with the increases in SPP1 mRNA expression and cell metabolic activities. The addition of sodium pyruvate to the ES media as an antioxidant did not alter the SPP1 mRNA expression, but eliminated an increase in cell metabolic activities induced by ES media treatment. These findings suggest that H2O2 was influencing cell metabolic activity, whereas SPP1 mRNA expression was regulated by other faradic by-products. This study reveals how different electrical stimulation regime alters cellular regenerative responses and the roles of faradic by-products, that might be used as a physical tool to guide and control cell behaviour.