RESUMO
Focusing on the defects in the lighting color of LED lamps and the chip heat exists in the traditional LED package which caused phosphor performance degradation, color temperature drift and the uneven light, the remote phosphor package which is emerging in recent years is used in this paper. With yellow-green YGG phosphor and nitrogen red phosphors mixing with silica gel, the remote phosphor is made and then encapsulated as the LED lamps. A lot of experiments were made to determine the best ratio of yellow green phosphor, red phosphor and silica gel, LED lamps with different color temperature was prepared. The lamps were also tested and analyzed with some parameters such as e color coordinates, luminous efficiency, color rendering index (CRI), R9, color quality scale (CQS), color temperature, and the gamut area index (GAI), which provide a more objective and comprehensive evaluation to the high quality LED lighting. Experimental results show that the optimum ratio of red and yellow-green phosphor is 1â¶7.6, and total phosphor with silica gel is 1â¶5ï¼at this time the white LED lighting color temperature is 4 113 K, the color coordinate (x, y) is (0.375 4, 0.373 1), luminous efficiency is 52.33 lm·w-1, color gamut is 0.981, color rendering index is up to 96, R9 is 97, color quality scale Qa is up to 93, and the gamut area index is 79. Compared with the traditional packagingï¼the surface temperature ofthe remote phosphor encapsulated fluorescent plate is much lower than that of adhesive dispensing encapsulation, which can effectively avoid the harmful effect caused by high temperature on the LED.
Assuntos
Iluminação , Semicondutores , Cor , TemperaturaRESUMO
Endothelial cells (ECs) play a crucial role in regulating various physiological and pathological processes. The behavior of ECs is modulated by physical (e.g., substrate stiffness) and biochemical cues (e.g., growth factors). However, the synergistic influence of these cues on EC behavior has rarely been investigated. In this study, we constructed poly(l-lysine)/hyaluronan (PLL/HA) multilayer films with different stiffness and exposed ECs to these substrates with and without hepatocyte growth factor (HGF)-supplemented culture medium. We demonstrated that EC adhesion, migration, and proliferation were positively correlated with substrate stiffness and that these behaviors were further promoted by HGF. Interestingly, ECs on the lower stiffness substrates showed stronger responses to HGF in terms of migration and proliferation, suggesting that HGF can profoundly influence stiffness-dependent EC behavior correlated with EC growth. After the formation of an EC monolayer, EC behaviors correlated with endothelial function were evaluated by characterizing monolayer integrity, nitric oxide production, and gene expression of endothelial nitric oxide synthase. For the first time, we demonstrated that endothelial function displayed a negative correlation with substrate stiffness. Although HGF improved endothelial function, HGF was not able to change the stiffness-dependent manner of endothelial functions. Taken together, this study provides insights into the synergetic influence of physical and biochemical cues on EC behavior and offers great potential in the development of optimized biomaterials for EC-based regenerative medicine.
Assuntos
Endotélio Vascular/efeitos dos fármacos , Fator de Crescimento de Hepatócito/farmacologia , Ácido Hialurônico/química , Polilisina/química , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , Dureza , Humanos , Propriedades de SuperfícieRESUMO
The development of gene therapy puts forward the requirements for efficient delivery of genetic information into diverse cells. However, in some cases of transfection, especially those for transfecting some primary cells and for delivering large size plasmid DNA (pDNA), the existing conventional transfection methods show poor efficiency. How to further improve transfection efficiency in these hard-to-achieve issues remains a crucial challenge. Here, we report a photothermal-assisted surface-mediated gene delivery based on a polydopamine-polyethylenimine (PDA-PEI) surface. The PDA-PEI surface was prepared through PEI-accelerated dopamine polymerization, which showed efficiency in the immobilization of PEI/pDNA polyplexes and remarkable photothermal properties. Upon IR irradiation, we observed improved transfection efficiencies of two important hard-to-achieve transfection issues, namely the transfection of primary endothelial cells, which are kinds of typical hard-to-transfect cells, and the transfection of cells with large-size pDNA. We demonstrate that the increases of transfection efficiency were due to the hyperthermia-induced pDNA release, the local cell membrane disturbance, and the polyplex internalization. This work highlights the importance of local immobilization and release of pDNA to gene deliveries, showing great potential applications in medical devices in the field of gene therapy.
Assuntos
Células Endoteliais/química , Indóis/química , Plasmídeos/genética , Polietilenoimina/química , Polímeros/química , Terapia Genética , Células HEK293 , Temperatura Alta , Células Endoteliais da Veia Umbilical Humana , Humanos , Raios Infravermelhos , Tamanho da Partícula , Plasmídeos/efeitos da radiação , Propriedades de Superfície , TransfecçãoRESUMO
Achievement of an endothelial cell (EC) monolayer (re-endothelialization) on the vascular implant surface with competent and functioning features is critical for long-term safety after implantation. Oriented EC monolayer is beneficial to improve endothelial function such as enhanced athero-resistant property. However, the information about antithrombotic property of oriented EC monolayer is limited. Here, we used the microgrooved polydimethylsiloxane substrates to guide EC orientation and obtain oriented EC monolayer. The effects of anisotropic topography on EC behaviors and antithrombotic function of the EC monolayer were then evaluated. Our data demonstrated that ECs responded to grooves in a size-dependent way as shown in oriented cell cytoskeleton and nuclei, enhanced directed migration, and overall velocity. Furthermore, compared to the EC monolayer on the flat surface, the oriented EC monolayer formed on the grooved substrates exhibited improved antithrombotic capability as indicated by higher expression of functional related genes, production of prostacyclin and tissue plasminogen activator, and prolonged activated coagulation time. The improvement of antithrombotic function was especially notable on the smaller-size groove. These findings reveal the responses of ECs to varisized topography and antithrombotic function of the oriented EC monolayer, providing insights into optimal design of vascular implants.
RESUMO
Endothelialization has proved to be critical for maintaining long-term success of implantable vascular devices. The formation of monolayer of endothelial cells (ECs) on the implant surfaces is one of the most important factors for the endothelialization. However, endothelial function of regenerated EC monolayer, which plays a much more important role in preventing the complications of post-implantation, has not received enough attention. Here, a vascular endothelial growth factor (VEGF)-incorporated poly(l-lysine)/hyaluronan (PLL/HA) polyelectrolyte multilayer film was fabricated. Through varying the crosslinking degree, stiffness of the film was manipulated, offering either soft or stiff film. We demonstrated that ECs were able to adhere and proliferate on both soft and stiff films, subsequently forming an integrated EC monolayer. Furthermore, endothelial functions were evaluated by characterizing EC monolayer integrity, expression of genes correlated with the endothelial functions, and nitric oxide production. It demonstrated that EC monolayer on the soft film displayed higher endothelial function compared to that on the stiff film. Our study highlights the influence of substrate stiffness on endothelial function, which offers a new criterion for surface design of vascular implants.
Assuntos
Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Ácido Hialurônico/farmacologia , Polieletrólitos/farmacologia , Polilisina/farmacologia , Fator A de Crescimento do Endotélio Vascular/farmacologia , Antígenos CD/genética , Antígenos CD/metabolismo , Biglicano/genética , Biglicano/metabolismo , Caderinas/genética , Caderinas/metabolismo , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Colágeno Tipo IV/genética , Colágeno Tipo IV/metabolismo , Módulo de Elasticidade , Fibronectinas/genética , Fibronectinas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Dureza , Células Endoteliais da Veia Umbilical Humana/citologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Ácido Hialurônico/química , Membranas Artificiais , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase Tipo III/genética , Óxido Nítrico Sintase Tipo III/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/genética , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Polieletrólitos/química , Polilisina/química , Propriedades de Superfície , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Extracellular matrix and cells are inherent in coordinating and adapting to each other during all physiological and pathological processes. Synthetic materials, however, show rarely reciprocal and spatiotemporal responses to cells, and lacking self-adapting properties as well. Here, a mechanical adaptability based on the matrix metalloproteinase (MMPs) sensitive polyelectrolyte film is reported. Poly-lysine (PLL) and methacrylated hyaluronic acid (HA-MA) nanolayers are employed to build the thin film through the layer-by-layer assembly, and it is further crosslinked using MMP sensitive peptides, which endows the films with changeable mechanical properties in response to MMPs. It is demonstrated that stiffness of the (PLL/HA-MA) films increases with the crosslinking, and then decreases in response to a treatment of enzyme. Consequently, the crosslinked (PLL/HA-MA) films reveal effective growth of endothelial cells (ECs), leading to fast formation of EC monolayer. Importantly, significantly improved endothelial function of the EC monolayer, which is characterized by integrity, biomolecules release, expression of function related gene, and antithrombotic properties, is achieved along with the decrosslinking of the film because of EC-secreted MMPs. These results suggest that mechanical adaptability of substrate in Young's modulus plays a significant role in endothelial progression, which shows great application potential in tissue engineering, regenerative medicine, and organ-on-a-chip.
Assuntos
Colagenases/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Ácido Hialurônico/química , Membranas Artificiais , Polilisina/química , Células Endoteliais da Veia Umbilical Humana/citologia , HumanosRESUMO
Substrate-mediated delivery of functional plasmid DNA (pDNA) has been proven to be a promising strategy to promote competitiveness of endothelial cells (ECs) over smooth muscle cells (SMCs), which is beneficial to inducing fast endothelialization of implanted vascular devices. Thus, it is of great importance to develop universal approaches with simplicity and easiness to immobilize DNA complex nanoparticles on substrates. In this study, the bioinspired polydopamine (PDA) coating was employed in immobilization of DNA complex nanoparticles, which were composed of protamine (PrS) and plasmid DNA encoding with hepatocyte growth factor (HGF-pDNA) gene. We demonstrated that the DNA complex nanoparticles can be successfully immobilized onto the PDA surface. Consequently, the HGF expression of both ECs and SMCs were significantly improved when they cultured on the DNA complex nanoparticles-immobilized substrates. Furthermore, EC proliferation was specifically promoted due to bioactivity of HGF, leading to an enhancement of EC competitiveness over SMCs. Our findings demonstrated the substrate-mediated functional gene nanoparticle delivery through PDA coating as a simple and efficient approach. It may hold great potential in the field of interventional cardiovascular implants.
Assuntos
DNA/química , Técnicas de Transferência de Genes , Fator de Crescimento de Hepatócito/farmacologia , Células Endoteliais da Veia Umbilical Humana/citologia , Indóis/química , Miócitos de Músculo Liso/citologia , Nanopartículas/química , Polímeros/química , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , DNA/metabolismo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Miócitos de Músculo Liso/efeitos dos fármacos , Plasmídeos , Protaminas/metabolismo , Propriedades de SuperfícieRESUMO
Endothelialization on the vascular implants is of great importance for prevention of undesired postimplantation symptoms. However, endothelial dysfunction of regenerated endothelial cell (EC) monolayer has been frequently observed, leading to severe complications, such as neointimal hyperplasia, late thrombosis, and neoatherosclerosis. It has significantly impeded long-term success of the therapy. So far, very little attention has been paid on endothelial function of EC monolayer. Bioinspired by the microenvironment of the endothelium in a blood vessel, this study described a soft polyelectrolyte multilayer film (PEM) through layer-by-layer assembly of poly(l-lysine) (PLL) and hyaluronan (HA). The (PLL/HA) PEM was chemically cross-linked and further incorporated with vascular endothelial growth factor. It demonstrated that this approach could promote EC adhesion and proliferation, further inducing formation of EC monolayer. Further, improved endothelial function of the EC monolayer was achieved as shown with the tighter integrity, higher production of nitric oxide, and expression level of endothelial function related genes, compared to EC monolayers on traditional substrates with high stiffness (e.g., glass, tissue culture polystyrene, and stainless steel). Our findings highlighted the influence of substrate stiffness on endothelial function of EC monolayer, giving a new strategy in the surface design of vascular implants.
Assuntos
Células Endoteliais/metabolismo , Técnicas de Cultura de Tecidos/métodos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Células Cultivadas , Ácido Hialurônico/química , Polieletrólitos/química , Fator A de Crescimento do Endotélio Vascular/químicaRESUMO
The stiffness of the substrates has been found to have a strong effect on cell behaviors, especially on cell adhesion, which is the first cellular event when cells contact materials. Much effort has been made to develop the materials with controlled stiffness for regulating cell adhesion. However, most available strategies for controlling the stiffness of material surfaces are generally limited to be static, which means that the stiffness is fixed during cell adhesion. Herein, we developed polyelectrolyte multilayer films (PEMs), and their stiffness can be dynamically modulated by mild stimuli. The PEMs were made by alternative deposition of poly-l-lysine (PLL) and thiol group modified hyaluronan (HA-SH) using the layer-by-layer assembly technique. The (PLL/HA-SH) multilayers can be cross-linked via oxidation of thiol groups. After crosslinking, the stiffness was increased and the adhesion of fibroblast cells was promoted. The stiffness of the multilayer films can be down-regulated dynamically by adding glutathione (GSH) in the medium, leading to in situ reduction of cell adhesion. Our study provides a promising strategy for the development of material surfaces with dynamically changeable stiffness, which is of great potential in the field of cell-based biomaterials.
RESUMO
Inspired by the adhesion strategy of marine mussels, self-polymerization of dopamine under alkaline condition has been proven to be a simple and effective method for surface modification of biomaterials. However, this method still has many drawbacks, such as the use of alkaline aqueous medium, low poly(dopamine) deposition rate, and inefficient utilization of dopamine, which greatly hinder its practical application. In the present study, we demonstrate that electropolymerization of dopamine is a facile and versatile approach to surface tailoring of metallic cardiovascular stents, such as small and complex-shaped coronary stent. Electropolymerization of dopamine leads to the formation of a continuous and smooth electropolymerized poly(dopamine) (ePDA) coating on the substrate surface. This electrochemical method exhibits a higher deposition rate and is more efficient in dopamine utilization compared with the typical self-polymerization method. The ePDA coating facilitates the immobilization of biomolecules onto substrates to engineer biomimetic microenvironments. In vitro and in vivo experiments demonstrate that ePDA coating functionalized with vascular endothelial growth factor can greatly enhance the desired cellular responses of endothelial cells and prevent the neointima formation after stent implantation. The proposed methodology may find applications in the area of metallic surface engineering, especially for the cardiovascular stents and potentially all biomedical devices with electroconductive surface as well.
Assuntos
Prótese Vascular , Dopamina/química , Técnicas Eletroquímicas/métodos , Polimerização , Stents , Actinas/metabolismo , Animais , Biomarcadores/metabolismo , Movimento Celular/efeitos dos fármacos , Materiais Revestidos Biocompatíveis/farmacologia , Eletrodos , Artéria Femoral/efeitos dos fármacos , Artéria Femoral/patologia , Imunofluorescência , Células Endoteliais da Veia Umbilical Humana/citologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Proteínas Imobilizadas/farmacologia , Masculino , Microscopia Eletrônica de Varredura , Polimerização/efeitos dos fármacos , Coelhos , Soluções , Coloração e Rotulagem , Propriedades de SuperfícieRESUMO
Mussel-inspired poly(dopamine) (PDA) coating is proven to be a simple, versatile, and effective strategy to promote cell adhesion onto various substrates. In this study, the initial adhesive behavior of human umbilical vein endothelial cells (HUVECs) is evaluated on a PDA coating under serum-free conditions. It is found that HUVECs can attach directly to and spread with well-organized cytoskeleton and fibrillar adhesions on the PDA surface, whereas cells adhere poorly to and barely spread on the control polycaprolactone surface. Endogenous fibronectin and α5 ß1 integrin are found to be involved in the cell adhesion process. These findings will lead to a better understanding of interactions between cells and PDA coating, paving the way for the further development of PDA.