RESUMEN
The growing world population and increasing life expectancy are driving the need to improve the quality of blood transfusion, organ transplantation, and preservation. Here, to improve the ability of red blood cells (RBCs) for normothermic machine perfusion, a biocompatible blood silicification approach termed "shielding-augmenting RBC-in-nanoscale amorphous silica (SARNAS)" has been developed. The key to RBC surface engineering and structure augmentation is the precise control of the hydrolysis form of silicic acid to realize stabilization of RBC within conformal nanoscale silica-based exoskeletons. The formed silicified RBCs (Si-RBCs) maintain membrane/structural integrity, normal cellular functions (e.g., metabolism, oxygen-carrying capability), and enhance resistance to external stressors as well as tunable mechanical properties, resulting in nearly 100% RBC cryoprotection. In vivo experiments confirm their excellent biocompatibility. By shielding RBC surface antigens, the Si-RBCs provide universal blood compatibility, the ability for allogeneic mechanical perfusion, and more importantly, the possibility for cross-species transfusion. Being simple, reliable, and easily scalable, the SARNAS strategy holds great promise to revolutionize the use of engineered blood for future clinical applications.
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Materiales Biocompatibles , Eritrocitos , Dióxido de Silicio , Eritrocitos/metabolismo , Dióxido de Silicio/química , Materiales Biocompatibles/química , Animales , Humanos , Perfusión/métodos , Conservación de la Sangre/métodos , Transfusión Sanguínea/métodos , RatonesRESUMEN
OBJECTIVE: To prepare porous core-shell composite particles (PCPs) in order to improve the flowability and compactibility of powder materials for direct compaction (DC), as well as the dissolution of tablets. SIGNIFICANCE: The results obtained are meaningful to boosting the development and further research of PCPs on DC. Methods: In this study, hydroxypropyl methylcellulose (HPMC E3) and polyvinylpyrrolidone (PVP K30) were selected as shell materials, the Xiao Er Xi Shi formulation powder (XEXS) was used as the core materials, ammonium bicarbonate (NH4HCO3), and sodium bicarbonate (NaHCO3) were employed as pore-forming agent. Using co-spray drying method to prepare composite particles (CPs). Then, the physical properties and comparison between different CPs were characterized comprehensively. Finally, the different CPs were directly compacted as tablets to explore the effect on the dissolution behavior of DC tablets, respectively. RESULTS: (i) The XEXS PCPs were prepared successfully by co-spray drying, and the yield of PCPs is almost 80%; (ii) The TS values of PCP-X-P-Na, PCP-X-P-NH4, PCP-X-H-Na and PCP-X-P-Na were 5.70, 7.56, 3.98, and 6.88 times higher than that of raw material (X); (iii) The disintegration time of PCPs tablets decreased 10-25% when compared with CPs tablets; (iv) The values of Carr's index (CI), Hausner ratio (HR), Caking strength (CS), and Cohesion index (CoI) of PCP-X-H-NH4 were 19.16%, 19.29%, 40.14%, and 6.39% lower than that of X, respectively. CONCLUSIONS: The PCPs prepared by co-spray drying did improve the flowability and compactibility of powder, as well as the dissolution of tablets.
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Povidona , Polvos , Porosidad , Composición de Medicamentos/métodos , Comprimidos , SolubilidadRESUMEN
This study aims to improve the solubility and bioavailability of daidzein by preparing the ß-cyclodextrin-daidzein/PEG_(20000)/Carbomer_(940) nanocrystals. Specifically, the nanocrystals were prepared with daidzein as a model drug, PEG_(20000), Carbomer_(940), and NaOH as a plasticizer, a gelling agent, and a crosslinking agent, respectively. A two-step method was employed to prepare the ß-cyclodextrin-daidzein/PEG_(20000)/Carbomer_(940) nanocystals. First, the insoluble drug daidzein was embedded in ß-cyclodextrin to form inclusion complexes, which were then encapsulated in the PEG_(20000)/Carbomer_(940) nanocrystals. The optimal mass fraction of NaOH was determined as 0.8% by the drug release rate, redispersability, SEM morphology, encapsulation rate, and drug loading. The inclusion status of daidzein nanocrystals was determined by Fourier transform infrared spectroscopy(FTIR), thermogravimetric analysis(TGA), and X-ray diffraction(XRD) analysis to verify the feasibility of the preparation. The prepared nanocrystals showed the average Zeta potential of(-30.77±0.15)mV and(-37.47±0.64)mV and the particle sizes of(333.60±3.81)nm and(544.60±7.66)nm before and after daidzein loading, respectively. The irregular distribution of nanocrystals before and after daidzein loading was observed under SEM. The redispersability experiment showed high dispersion efficiency of the nanocrystals. The in vitro dissolution rate of nanocrystals in intestinal fluid was significantly faster than that of daidzein, and followed the first-order drug release kinetic model. XRD, FTIR, and TGA were employed to determine the polycrystalline properties, drug loading, and thermal stability of the nanocrystals before and after drug loading. The nanocrystals loaded with daidzein demonstrated obvious antibacterial effect. The nanocrystals had more significant inhibitory effects on Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa than daidzein because of the improved solubility of daidzein. The prepared nanocrystals can significantly increase the dissolution rate and oral bioavailability of the insoluble drug daidzein.
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Resinas Acrílicas , Nanopartículas , Hidróxido de Sodio , Escherichia coliRESUMEN
Since the advent of additive manufacturing, known commonly as 3D printing, this technology has revolutionized the biofabrication landscape and driven numerous pivotal advancements in tissue engineering and regenerative medicine. Many 3D printing methods were developed in short course after Charles Hull first introduced the power of stereolithography to the world. However, materials development was not met with the same enthusiasm and remained the bottleneck in the field for some time. Only in the past decade has there been deliberate development to expand the materials toolbox for 3D printing applications to meet the true potential of 3D printing technologies. Herein, we review the development of biomaterials suited for light-based 3D printing modalities with an emphasis on bioprinting applications. We discuss the chemical mechanisms that govern photopolymerization and highlight the application of natural, synthetic, and composite biomaterials as 3D printed hydrogels. Because the quality of a 3D printed construct is highly dependent on both the material properties and processing technique, we included a final section on the theoretical and practical aspects behind light-based 3D printing as well as ways to employ that knowledge to troubleshoot and standardize the optimization of printing parameters.
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Materiales Biocompatibles/química , Luz , Impresión Tridimensional , Ingeniería de Tejidos , Materiales Biocompatibles/síntesis química , Investigación Biomédica , Humanos , Procesos Fotoquímicos , PolimerizacionRESUMEN
Towards repairing bone defects, calcium sulfate and calcium phosphate cement have been recognized as promising bone grafts. However, the current bone cements are generally lack of proper porosity for cell migration and new tissue formation. On the other hand, porous scaffold cannot be delivered by injection, which limits its use its clinical use. Herein, we develop a novel tricalcium phosphate/calcium sulfate granule to overcome the limitations of injectable cements and traditional scaffolds. The biocompatible granule underwent in situ self-setting to form scaffold with porous structure after injection. It contributes to calcium deposition and upregulation of osteogenic genes of mesenchymal stem cells in a time-dependent manner. Within three months, cavitary bone defects of distal rabbit femurs implanted the granules exhibited better bone formation than those with those implanted with autologous bone.
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Cementos para Huesos/química , Regeneración Ósea/efectos de los fármacos , Fosfatos de Calcio/farmacología , Sulfato de Calcio/farmacología , Osteogénesis/efectos de los fármacos , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Animales , Humanos , Masculino , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/ultraestructura , Microscopía Electrónica de Rastreo , Osteoblastos/efectos de los fármacos , Osteoblastos/ultraestructura , Porosidad , Conejos , Difracción de Rayos XRESUMEN
BACKGROUND: An endovascular covered-stent has unique advantages in treating complex intracranial aneurysms; however, in-stent stenosis and late thrombosis have become the main factors affecting the efficacy of covered-stent treatment. Smooth-muscle-cell phenotypic modulation plays an important role in late in-stent stenosis and thrombosis. Here, we determined the efficacy of using covered stents loaded with drugs to inhibit smooth-muscle-cell phenotypic modulation and potentially lower the incidence of long-term complications. METHODS: Nanofiber-covered stents were prepared using coaxial electrospinning, with the core solution prepared with 15% heparin and 20 µM rosuvastatin solution (400: 100 µL), and the shell solution prepared with 120 mg/mL hexafluoroisopropanol. We established a rabbit carotid-artery aneurysm model, which was treated with covered stents. Angiography and histology were performed to evaluate the therapeutic efficacy and incidence rate of in-stent stenosis and thrombosis. Phenotype, function, and inflammatory factors of smooth-muscle cells were studied to explore the mechanism of rosuvastatin action in smooth-muscle cells. RESULT: Heparin-rosuvastatin-loaded nanofiber scaffold mats inhibited the proliferation of synthetic smooth-muscle cells, and the nanofiber-covered stent effectively treated aneurysms in the absence of notable in-stent stenosis. Additionally, in vitro experiments showed that rosuvastatin inhibited the smooth-muscle-cell phenotypic modulation of platelet-derived growth factor-BB induction and decreased synthetic smooth-muscle-cell viability, as well as secretion of inflammatory cytokines. CONCLUSION: Rosuvastatin inhibited the abnormal proliferation of synthetic smooth-muscle cells, and heparin-rosuvastatin-loaded covered stents reduced the incidence of stenosis and late thrombosis, thereby improving the healing rates of stents used for aneurysm treatment.
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Supervivencia Celular/efectos de los fármacos , Constricción Patológica/tratamiento farmacológico , Heparina/farmacología , Músculos/efectos de los fármacos , Nanofibras/química , Poliésteres/química , Rosuvastatina Cálcica/farmacología , Trombosis/tratamiento farmacológico , Animales , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Citocinas/metabolismo , Aneurisma Intracraneal/terapia , Masculino , Ratones , Poliésteres/farmacología , Conejos , Stents , Trombosis/patologíaRESUMEN
Stimulated Raman scattering (SRS) microscopy in combination with innovative tagging strategies offers great potential as a universal high-throughput biomedical imaging tool. Here, we report rationally tailored small molecular monomers containing triple-bond units with large Raman scattering cross-sections, which can be polymerized at the nanoscale for enhancement of SRS contrast with smaller but brighter optical nanotags with artificial fingerprint output. From this, a class of triple-bond rich polymer nanoparticles (NPs) was engineered by regulating the relative dosages of three chemically different triple-bond monomers in co-polymerization. The bonding strategy allowed for 15 spectrally distinguishable triple-bond combinations. These accurately structured nano molecular aggregates, rather than long-chain macromolecules, could establish a universal method for generating small-sized biological SRS imaging tags with high sensitivity for high-throughput multi-color biomedical imaging.
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Nanopartículas/química , Imagen Óptica , Polímeros/química , Humanos , Células MCF-7 , Estructura Molecular , Espectrometría RamanRESUMEN
BACKGROUND: HA modified by bisphosphonate (BP) (HA-BP) was synthesized by chemical reaction and possessed promising properties such as self-healing, injection ability, and strong adhesion. The main aim of this study was to confirm its role in promoting osteogenic differentiation in vitro and bone regeneration in vivo. METHODS: The cell biocompatibility of this material was determined using the CCK-8 assay. Alkaline phosphatase (ALP), osteocalcin (OT), vascular endothelial growth factor (VEGF), and collagen I were assessed by quantitative real-time polymerase chain reaction (Q-PCR) in the treated group. The number and density of calcium nodules and ALP were evaluated by Alizarin Red staining and ALP staining. We have successfully developed an animal model simulating osteonecrosis of the femoral head (ONFH). Utilizing this animal model, the impact of HA-BP/CaP on bone formation was assessed. The amount of bone regeneration at 1 and 2 months after HA-BP/CaP injection was estimated by micro-computed tomography (micro-CT) analysis and H&E, collagen I, and periostin staining. RESULTS: The number of cells gradually increased in the experimental group over time and was close to that of the blank control group. ALP, collagen I, and VEGF expression was significantly higher in the experimental group than in the blank group (VEGF, ALP, both **p < 0.01; collagen I, ***p<0.001). In addition, the number and density of calcium nodules and ALP was clearly greater in the material group than in the control group. The quantification analysis showed that the mineral contents of regenerated bone at 1 and 2 months after HA-BP/CaP injection were significantly greater than those in the control group, according to micro-CT evaluation (**p<0.01). The amount of organic components in the HA-BP/CaP group was greater than that in the control group after decalcification and H&E staining. In addition, collagen I and periostin staining further confirmed the results of H&E staining. CONCLUSION: This material can boost proliferation and osteogenic differentiation of MC3T3-E1 cells in vitro. It can intensely accelerate bone regeneration in vivo, which is a promising strategy for tissue engineering.
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Regeneración Ósea/efectos de los fármacos , Necrosis de la Cabeza Femoral/tratamiento farmacológico , Ácido Hialurónico/análogos & derivados , Células 3T3 , Animales , Materiales Biocompatibles/administración & dosificación , Fosfatos de Calcio/administración & dosificación , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Colecistoquinina/metabolismo , Difosfonatos/administración & dosificación , Modelos Animales de Enfermedad , Femenino , Necrosis de la Cabeza Femoral/patología , Necrosis de la Cabeza Femoral/fisiopatología , Humanos , Ácido Hialurónico/administración & dosificación , Hidrogeles , Ensayo de Materiales , Ratones , Osteogénesis/efectos de los fármacos , Fragmentos de Péptidos/metabolismo , Conejos , Ingeniería de Tejidos , Microtomografía por Rayos XRESUMEN
In order to increase the stability and solubility of essential oil in Jieyu Anshen Formula, this study was to prepare the essential oil into liposomes. In this experiment, the method for the determination of encapsulation efficiency of liposomes was established by ultraviolet spectrophotometer and dextran gel column. The encapsulation efficiency and particle size of liposomes were used as evaluation indexes for single factor investigation and Box-Behnken design-response surface method was used to optimize the design. Then the optimal formulation of volatile oil liposome was characterized using methyleugenol, elemin, ß-asarone and α-asarone as index components. Finally, the in vitro transdermal properties of liposomes were studied by modified Franz diffusion cell. The results showed that the concentration of lecithin, the mass ratio of lecithin to volatile oil, and the stirring speed were the three most significant factors affecting the liposome preparation. The optimum formulation of volatile oil liposome was as follows: the concentration of lecithin was 7 g·L~(-1); mass ratio of lecithin to volatile oil was 5â¶1; and the stirring speed was 330 r·min~(-1). Under such conditions, the prepared liposomes had blue emulsion light, good fluidity, half translucent, with particle size of(102.6±0.35) nm, Zeta potential of(-17.8±0.306) mV, permeability of(1.67±1.01)%, and stable property if liposome was stored at 4 â. 24 h after percutaneous administration, the cumulative osmotic capacity per unit time was(30.485 2±1.238 9),(34.794 8±0.928 3),(26.677 1±1.171 7),(3.066 2±0.175 3) µg·cm~(-2)respectively for methyleugenol, elemin, ß-asarone and α-asarone. In vitro transdermal behaviors of methyleugenol, elemin, ß-asarone and α-asarone in liposomes were all consistent with Higuchi equation. The prepared volatile oil liposomes met the relevant quality requirements, providing a reference for further research on preparation of multi-component Chinese medicine essential oil liposomes.
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Medicamentos Herbarios Chinos/análisis , Liposomas , Aceites Volátiles/análisis , Administración Cutánea , Tamaño de la Partícula , SolubilidadRESUMEN
BACKGROUND: Vertebrates use different pigmentation strategies to adapt to various environments. A large amount of research has been done on disclosing the mechanisms of pigmentation strategies in vertebrates either under light, or, living in constant darkness. However, less attention has been paid to non-obligate, darkness dwellers. Red-spotted toothed toads Oreolalax rhodostigmatus (Megophryidae; Anura) from the karst mountainous region of southwestern China are non-obligate cave dwellers. Most tadpoles of the species possess transparent skin as they inhabit the dark karst caves. But remarkably, the transparent tadpoles can darken just within 15 h once exposed to light. Obviously, it is very significant to reveal molecular mechanisms of the unexpected rapid-darkening phenomenon. RESULTS: We compared the transcriptomes of O. rhodostigmatus tadpoles with different durations of light exposure to investigate the cellular processes and potential regulation signals for their light-induced rapid darkening. Genes involved in melanogenesis (i.e. TYR, TYRP1 and DCT) and melanocyte proliferation, as well as their transcriptional factor (MITF), showed light-induced transcription, suggesting a dominating role of morphological color change (MCC) in this process. Transcription of genes related to growth factor, MAPK and PI3K-Akt pathways increased with time of light exposure, suggesting that light could induce significant growth signal, which might facilitate the rapid skin darkening. Most importantly, an in-frame deletion of four residues was identified in O. rhodostigmatus melanocortin-1 receptor (MC1R), a critical receptor in MCC. This deletion results in a more negatively charged ligand pocket with three stereo-tandem aspartate residues. Such structural changes likely decrease the constitutive activity of MC1R, but increase its ligands-dependent activity, thus coordinating pigment regression and rapid melanogenesis in the dark and light, respectively. CONCLUSION: Our study suggested that rapid MCC was responsible for the light-induced rapid darkening of O. rhodostigmatus tadpoles. Genetic mutations of MC1R in them could explain how these non-obligate cave dwellers coordinate pigment regression and robust melanogenesis in darkness and light, respectively. To our knowledge, this is the first study that reports the association between pigmentation phenotype adaptation and MC1R mutations in amphibians and/or in non-obligate cave dwellers.
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Anuros/genética , Anuros/metabolismo , Cuevas , Perfilación de la Expresión Génica , Luz , Pigmentación/genética , Pigmentación/efectos de la radiación , Animales , Cinética , ARN Mensajero/genéticaRESUMEN
Nitroxyl (HNO) is a derivative of nitric oxide (NO) that plays an essential role in various biological and pharmacological events. Until now, the in situ trapping and specific detection of HNO in living samples is still challenging. In this project, we fabricated a novel BODIPY-based micellar nanoprobe for monitoring nitroxyl in vitro and in vivo in ratiometric mode in aqueous solution. The probe (P-BODIPY-N) contains an asymmetrical BODIPY dye for fluorescent signaling and a diphenylphosphinobenzoyl as the trigger moiety; then we encapsulated P-BODIPY-N into the hydrophobic interior of an amphiphilic copolymer (mPEG-DSPE) and prepared a novel BODIPY-based micellar nanoprobe: NP-BODIPY-N. As far as we know, this probe is the first reported ratiometric fluorescent nanoprobe for HNO, which exhibits ultrasensitivity, high selectivity, and good biocompatibility. Above all, this nanoprobe shows favorable cellular uptaken and was successfully used to detect intracellular HNO released by Angeli's salt in living cells and zebrafish larvae. These results indicate that our newly designed nanoprobe will provide a promising tool for the studies of HNO in living system.
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Compuestos de Boro/química , Colorantes Fluorescentes/química , Óxidos de Nitrógeno/análisis , Imagen Óptica/métodos , Animales , Células Hep G2 , Humanos , Micelas , Fosfatidiletanolaminas/química , Fosfinas/química , Polietilenglicoles/química , Pez CebraRESUMEN
Cartilage tissue is prone to degradation and has little capacity for self-healing due to its avascularity. Tissue engineering, which provides artificial scaffolds to repair injured tissues, is a novel and promising strategy for cartilage repair. 3D bioprinting offers even greater potential for repairing degenerative tissue by simultaneously integrating living cells, biomaterials, and biological cues to provide a customized scaffold. With regard to cell selection, mesenchymal stem cells (MSCs) hold great capacity for differentiating into a variety of cell types, including chondrocytes, and could therefore be utilized as a cartilage cell source in 3D bioprinting. In the present study, we utilize a tabletop stereolithography-based 3D bioprinter for a novel cell-laden cartilage tissue construct fabrication. Printable resin is composed of 10% gelatin methacrylate (GelMA) base, various concentrations of polyethylene glycol diacrylate (PEGDA), biocompatible photoinitiator, and transforming growth factor beta 1 (TGF-ß1) embedded nanospheres fabricated via a core-shell electrospraying technique. We find that the addition of PEGDA into GelMA hydrogel greatly improves the printing resolution. Compressive testing shows that modulus of the bioprinted scaffolds proportionally increases with the concentrations of PEGDA, while swelling ratio decreases with the increase of PEGDA concentration. Confocal microscopy images illustrate that the cells and nanospheres are evenly distributed throughout the entire bioprinted construct. Cells grown on 5%/10% (PEGDA/GelMA) hydrogel present the highest cell viability and proliferation rate. The TGF-ß1 embedded in nanospheres can keep a sustained release up to 21 d and improve chondrogenic differentiation of encapsulated MSCs. The cell-laden bioprinted cartilage constructs with TGF-ß1-containing nanospheres is a promising strategy for cartilage regeneration.
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Bioimpresión , Cartílago/fisiología , Células Madre Mesenquimatosas/citología , Nanosferas/química , Impresión Tridimensional , Ingeniería de Tejidos/métodos , Cartílago/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Proliferación Celular/efectos de los fármacos , Condrogénesis/efectos de los fármacos , Condrogénesis/genética , Preparaciones de Acción Retardada , Liberación de Fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacología , Tinta , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Nanosferas/ultraestructura , Estrés Mecánico , Andamios del Tejido/química , Factor de Crecimiento Transformador beta1/farmacologíaRESUMEN
Osteonecrosis of the femoral head (ONFH) is a major cause of morbidity, and total hip arthroplasty is both traumatic and expensive. Here, we created a gelatine scaffold embedded in uniquely shaped, 3D-printed porous titanium parts, which could attract and promote the proliferation of osteoblasts as well as bone regeneration, as the extracellular matrix (ECM) does in vivo. Interestingly, after hybridisation with platelets, the scaffold exhibited a low yet considerable rate of stable, safe and long-term growth factor release. Additionally, a novel ONFH model was constructed and verified. Scaffolds implanted in this model were found to accelerate bone repair. In conclusion, our scaffold successfully simulates the ECM and considerably accelerates bone regeneration, in which platelets play an indispensable role. We believe that platelets should be emphasised as carriers that may be employed to transport drugs, cytokines and other small molecules to target locations in vivo. In addition, this novel scaffold is a useful material for treating ONFH. An overview of the novel scaffold mimicking the extracellular environment in bone repair. a and b: A gelatine scaffold was cross-linked and freeze-dried within 3D-printed porous titanium. c: Platelets were coated onto the gelatine microscaffold after freeze-drying platelet-rich plasma. d: The microscaffold supported the migration of cells into the titanium pores and their subsequent growth, while the platelets slowly released cell factors, exerting bioactivity.
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Cabeza Femoral/irrigación sanguínea , Neovascularización Fisiológica , Osteogénesis , Impresión Tridimensional , Andamios del Tejido/química , Titanio/química , Animales , Materiales Biocompatibles , Plaquetas/metabolismo , Regeneración Ósea , Huesos/irrigación sanguínea , Adhesión Celular , Proliferación Celular , Materiales Biocompatibles Revestidos/metabolismo , Citocinas/metabolismo , Matriz Extracelular/metabolismo , Cabeza Femoral/patología , Gelatina/química , Humanos , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Ensayo de Materiales , Osteoblastos/metabolismo , Porosidad , Diseño de Prótesis , Conejos , Cicatrización de HeridasRESUMEN
Lonicera japonica Thunb. flower is a well-known medicinal plant that has been widely used for the treatment of human disease. To explore the molecular mechanisms underlying the biological activities of L. japonica immature flower buds, a gel-free/label-free proteomic technique was used in combination with combinatorial peptide ligand libraries (CPLL) and polyethylene glycol (PEG) fractionation for the enrichment of low-abundance proteins and removal of high-abundance proteins, respectively. A total of 177, 614, and 529 proteins were identified in crude protein extraction, CPLL fractions, and PEG fractions, respectively. Among the identified proteins, 283 and 239 proteins were specifically identified by the CPLL and PEG methods, respectively. In particular, proteins related to the oxidative pentose phosphate pathway, signaling, hormone metabolism, and transport were highly enriched by CPLL and PEG fractionation compared to crude protein extraction. A total of 28 secondary metabolism-related proteins and 25 metabolites were identified in L. japonica immature flower buds. To determine the specificity of the identified proteins and metabolites for L. japonica immature flower buds, Cerasus flower buds were used, which resulted in the abundance of hydroxymethylbutenyl 4-diphosphate synthase in L. japonica immature flower buds being 10-fold higher than that in Cerasus flower buds. These results suggest that proteins related to secondary metabolism might be responsible for the biological activities of L. japonica immature flower buds.
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Flores/metabolismo , Lonicera/metabolismo , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Precipitación Química , Ligandos , Malus/metabolismo , Redes y Vías Metabólicas , Biblioteca de Péptidos , Proteínas de Plantas/aislamiento & purificación , Polietilenglicoles/química , Proteoma/aislamiento & purificación , Proteómica , Espectrometría de Masas en TándemRESUMEN
H2S produced in small amounts by mammalian cells has been identified in mediating biological signaling functions. However, the in situ trapping of endogenous H2S generation is still handicapped by a lack of straightforward methods with high selectivity and fast response. Here, we encapsulate a semi-cyanine-BODIPY hybrid dye (BODInD-Cl) and its complementary energy donor (BODIPY1) into the hydrophobic interior of an amphiphilic copolymer (mPEG-DSPE), especially for building up a ratiometric fluorescent H2S nanoprobe with extraordinarily fast response. A remarkable red-shift in the absorption band with a gap of 200 nm in the H2S response can efficiently switch off the Förster resonance energy transfer (FRET) from BODIPY1 to BODInD-Cl, subsequently recovering the donor fluorescence. Impressively, both the interior hydrophobicity of supramolecular micelles and electron-withdrawing nature of indolium unit in BODInD-Cl can sharply increase aromatic nucleophilic substitution with H2S. The ratiometric strategy based on the unique self-assembled micellar aggregate NanoBODIPY achieves an extremely fast response, enabling in situ imaging of endogenous H2S production and mapping its physiological and pathological consequences. Moreover, the amphiphilic copolymer renders the micellar assembly biocompatible and soluble in aqueous solution. The established FRET-switchable macromolecular envelope around BODInD-Cl and BODIPY1 enables cellular uptake, and makes a breakthrough in the trapping of endogenous H2S generation within raw264.7 macrophages upon stimulation with fluvastatin. This study manifests that cystathione γ-lyase (CSE) upregulation contributes to endogenous H2S generation in fluvastatin-stimulated macrophages, along with a correlation between CSE/H2S and activating Akt signaling pathway.
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Ácidos Grasos Monoinsaturados/química , Transferencia Resonante de Energía de Fluorescencia/métodos , Sulfuro de Hidrógeno/química , Indoles/química , Nanopartículas/química , Animales , Compuestos de Boro/química , Cistationina gamma-Liasa/química , Colorantes Fluorescentes/química , Fluvastatina , Macrófagos/metabolismo , Ratones , Micelas , Microscopía Confocal , Microscopía Fluorescente/métodos , Polímeros/química , Células RAW 264.7 , Regulación hacia ArribaRESUMEN
A circular hydrophobic-hydrophilic-Ti(4+) immobilized phosphate polymer is patterned on a silicon wafer. Such a wafer is used as a novel sample support to allow fast selective enrichment, wash-free self-desalting and mass spectroscopy (MS) analysis of phosphopeptides, thanks to the high Ti(4+) loading amount, pure phosphate polymer-Ti(4+) interface, and strong hydrophobic-hydrophilic attraction pattern. The detection sensitivity was enhanced 300 folds compared with what was obtained using the common MALDI plate. Remarkable selectivity for phosphopeptides can be achieved at a molar ratio as low as 1 : 500 of phosphopeptides (casein digest)/nonphosphopeptides (BSA). High-quality mass spectra can be obtained even in the presence of NaCl (1 M), NH4HCO3 (100 mM), or urea (1 M). These microspots were also used to selectively capture phosphopeptides from milk and human serum, which further demonstrated that they were capable of identifying low-abundance phosphopeptides from real complex samples. They provide a low detection limit (3 fmol µL(-1)), small sample size, and excellent enrichment and desalting efficiency. Such a method significantly simplifies the analytical procedures, reduces possible sample loss, and is relatively low cost. Therefore, this on-plate patterned technique is very promising in the high-throughput phosphoproteomic research, especially for the detection of tiny amounts of samples.
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Leche/química , Fosfatos/química , Fosfopéptidos/análisis , Fosfopéptidos/sangre , Silicio/química , Titanio/química , Animales , Humanos , Límite de Detección , Fosfopéptidos/aislamiento & purificación , Polímeros/química , Ácidos Polimetacrílicos/química , Sales (Química)/química , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodosRESUMEN
OBJECTIVE: To evaluate the toxicity of extracts from disposable chopsticks, toothpicks, and paper cups on L-929 cells. METHODS: We followed national standards to prepare the extracts from disposable chopsticks, toothpicks, and paper cups used for the cell culture media, and the morphology of L-929 cells was observed with an optical microscope. The loss rate for adherent cells was evaluated with the trypan blue exclusion method, and cell proliferation was determined using the WST-1 assay. RESULTS: Compared with the control group, the cells cultured in media containing the extracts showed signs of apoptosis and necrosis after culturing for 4 or 7 days, and the loss rate for adherent cells was significantly increased (P < 0.05). An obvious decrease in cell viability was also observed (P < 0.05). CONCLUSION: The extracts from disposable chopsticks, toothpicks, and paper cups can affect the growth and proliferation of L-929 cells and are potentially toxic to humans.
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Apoptosis/efectos de los fármacos , Seguridad de Productos para el Consumidor , Utensilios de Comida y Culinaria , Fibroblastos/efectos de los fármacos , Papel , Extractos Vegetales/toxicidad , Madera/química , Animales , Adhesión Celular/efectos de los fármacos , Línea Celular , Proliferación Celular/efectos de los fármacos , Forma de la Célula/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , China , Células Clonales , Dispositivos para el Autocuidado Bucal/efectos adversos , Equipos Desechables , Fibroblastos/citología , Fibroblastos/patología , Ratones , Necrosis , Pruebas de Toxicidad , Madera/efectos adversosRESUMEN
Neural tissue engineering offers a promising avenue for repairing neural injuries. Advancement in nanotechnology and neural scaffold manufacturing strategies has shed light on this field into a new era. In this study, a novel tissue engineered scaffold, which possesses highly aligned poly-ε-caprolactone microfibrous framework and adjustable bioactive factor embedded poly (d, l-lactide-co-glycolide) core-shell nanospheres, was fabricated by combining electrospinning and electrospraying techniques. The fabricated nanocomposite scaffold has cell favorable nanostructured feature and improved hydrophilic surface property. More importantly, by incorporating core-shell nanospheres into microfibrous scaffold, a sustained bioactive factor release was achieved. Results show rat pheochromocytoma (PC-12) cell proliferation was significantly promoted on the nanocomposite scaffold. In addition, confocal microscope images illustrated that the highly aligned scaffold increased length of neurites and directed neurites extension along the fibers in both PC-12 and astrocyte cell lines, which indicates that the scaffold is promising for guiding neural tissue growth and regeneration. From the clinical editor: In an attempt to direct neural cell growth, biomimetic neural scaffold was produced by electrospinning integrated with co-axial electrospraying techniques. In-vitro data provided a framework for future designs for neuronal regeneration.
Asunto(s)
Proliferación Celular , Nanocompuestos/química , Neuritas/metabolismo , Poliglactina 910/química , Regeneración , Andamios del Tejido/química , Animales , Células PC12 , RatasRESUMEN
Leukocytes can cross intact blood-brain barrier under healthy conditions and in many neurological diseases, including psychiatric diseases. In present study, a cyclic RGD (cRGD) peptide with high affinity for integrin receptors of leukocytes was used to modify liposomes. The cRGD-modified liposomes (cRGDL) showed high affinity for monocytes in vitro and in vivo and co-migrated across in vitro BBB model with THP-1. The trefoil factor 3 (TFF3), a macromolecular drug, was rapidly and persistently delivered to brain for at least 12 h when loaded into cRGDL while 2.8-fold increase in drug concentration in basolateral amygdala regions related to depression was observed. A systemic administration of cRGDL-TFF3 mimicked antidepressant-like effect of direct intra-basolateral amygdala administration of TFF3 solution in rats subjected to chronic mild stress. The effective dual-brain targeting delivery resulting from the combination and co-migration of cRGDL with leukocyte cross BBB may be a promising strategy for targeted brain delivery. FROM THE CLINICAL EDITOR: In an effort to treat depression, brain targeted delivery via monocyte-cRGD liposome complexes capable of crossing the intact BBB was performed in this study in a murine model. Similar approaches may be helpful in the treatment of other neuropsychiatric conditions.
Asunto(s)
Barrera Hematoencefálica/efectos de los fármacos , Depresión/tratamiento farmacológico , Sistemas de Liberación de Medicamentos , Péptidos Cíclicos/administración & dosificación , Péptidos/administración & dosificación , Animales , Encéfalo/efectos de los fármacos , Encéfalo/patología , Depresión/patología , Sinergismo Farmacológico , Humanos , Leucocitos/efectos de los fármacos , Liposomas/administración & dosificación , Masculino , Ratones , Ratas , Factor Trefoil-3RESUMEN
To improve a fast and high-quality isolation method for culturing the primary cardiomyocyte and fibroblast in vitro, the neonatal Wistar rats were decapitated accordingly and left ventricles were isolated under the sterile condition. The ventricles were chopped and digested in the enzyme solution containing 0.5 mg/mL type II collagenase. During this process, the digesting time, frequency and stirring speed, centrifuging frequency and speed were strictly controlled. The cardiomyocytes were separated from the cardiac fibroblast by using the Percoll density gradient centrifugation. The cell viability was tested by staining with 0.2% trypan blue. The purity of cardiomyocytes and fibroblasts were determined by immunoflourescent staining with anti-cTnI, anti-Vimentin and anti-α-SMA antibodies. The results indicated that with this protocol, the viability and purity of cardiomyocytes were 92% and 95%. The automobile pulse of the adhered cardiomyocyte was visible. For fibroblasts, the cell viability and purity were 96% and 94%. Our results demonstrate that this advanced isolation method is reproducible, and can simultaneously produce high-quality primary cardiomyocytes and fibroblasts for the future study.