用于蛋白质BSA识别的温度/pH双敏印迹聚合物

用N-异丙基丙烯酰胺(NIPAM)和壳聚糖为功能单体,牛血清白蛋白（BSA）为模板蛋白,在改性SiO₂表面制备温度/pH双敏蛋白质印迹聚合物。TEM、FTIR和TG等结果证明印迹层已成功接枝在载体表面。系统研究了聚合物的温度/pH双敏性、吸附容量、吸附动力学、特异性、竞争吸附性及重复性。结果表明,印迹聚合物（MIP）的溶胀率和吸附容量受温度和pH影响较大,高温碱性收缩,低温酸性溶胀。在pH 4.6和35℃下,对0.6mg/mL BSA吸附4h时获得较大的吸附容量（83.74mg/g）,印迹因子为2.02。同时MIP也有较好的特异性和竞争吸附性。重复5次后,吸附容量仍能维持在88%,说明重复性良好。这种新型的温度/pH双敏蛋白质分子印迹合成方法简单,在蛋白质的分离和识别方面有较好的应用前景。     

Studies on the interaction of CdTe QDs with bovine serum albumin

BACKGROUND: Quantum dots (QDs) have attracted much attention in biological and medical applications. In particular, the interaction of QDs with bovine serum albumin (BSA) is crucial, and has been systematically investigated by various spectroscopic techniques under the physiological conditions. RESULTS: The effects of ionic strength and pH on the interaction of CdTe QDs with BSA were studied by changing NaCl concentration and pH in mixed solution and making fluorescence spectroscopic measurements. The Stern‐Volmer quenching constant (K_a) of different ionic strength and pH were calculated, and information on the structural features of BSA were discussed by means of circular dichroism (CD) spectrum. CONCLUSION: Both fluorescence (FL) and circular dichroism (CD) results indicated that hydrophobic and electrostatic interactions play a major role in the binding reaction, and the nature of quenching is static, resulting in forming QDs‐BSA complexes. Copyright © 2012 Society of Chemical Industry     

Influence of sodium dodecyl sulfate on the characteristics of bovine serum albumin solutions and foams

Results from surface tension measurements on mixed solutions of the protein bovine serum alburnin (BSA) and an anionic surfactant (SDS: sodium dodecyl sulfate) suggested that at an air-liquid interface, adsorption was affected by the protein-surfactant interaction and by the relative concentration of each component in solution. Two plateaus corresponding to the critical aggregation concentration (CAC) and the critical micelle concentration (CMC) of SDS, respectively, were observed in the surface tension isotherms of SDS in the presence of BSA. The CAC and CMC depended on the concentration of BSA. Effects of SDS concentration on the conformational changes of BSA were investigated by Fourier transform-Raman spectroscopy. The results showed that the contents of α-helix decreased while the contents of random coil increased. The presence of the anionic surfactant SDS had a negative influence on the way that proteins adsorb at an air-liquid interface, leading to the change of behavior of protein-stabilized film.     

Interface/interphase engineering of polymers for adhesion enhancement: Part II. Theoretical and technological aspects of surface-engineered interphase-interface systems for adhesion enhancement

Part I of this paper reviewed the theoretical principles of the macromolecular design of polymer interface/interphase systems for obtaining maximum adhesion and fracture performance of adhesively bonded assemblies. In Part II a novel, relatively simple and industry-feasible technology for surface-grafting connector molecules is demonstrated and discussed in detail and supported by a range of experimental examples. It is shown, in agreement with contemporary theory, that the use of chemically attached graft chemicals of controlled spatial geometry and chemical functionality enables a significant increase in the strength and fracture energy of the interphase, to the point of cohesive fracture of the substrate, or that of an adjacent medium such as adhesive, elastomer, or other material. This occurs even after prolonged exposure of investigated systems to adverse environments such as hot water.     

Study of the adsorption process of bovine serum albumin on passivated surfaces of CoCrMo biomedical alloy

Adsorption of bovine serum albumin (BSA) on CoCrMo surface was studied by electrochemical techniques in order to determine the mechanism of protein adsorption at different surface conditions (which are reached by considering different passivation times) in phosphate buffered solution (PBS). At open circuit potential (OCP), adsorption kinetic was influenced by surface passivation when passivation time was lower than 1 h, whereas, at higher passivation times, no apparent modification in the kinetic mechanisms of adsorption was observed. On the other hand, at a passive applied potential, the BSA addition decreased the passivation kinetics of the process at all the passivation times.     

Simple fabrication of functionalized surface with polyethylene glycol microstructure and glycidyl methacrylate moiety for the selective immobilization of proteins and cells

This study reported simple surface modification for the immobilization of biomolecules such as proteins and cells onto desired area at micron-scale level. First, thin film composed of glycidyl methacrylate (GMA) was prepared by UV-photopolymerization. Then, the polyethylene glycol (PEG) microstructures which played a role in the prevention of nonspecific binding of biomolecules were fabricated by using micromolding in capillaries (MIMIC). Thus, we could easily obtain an orthogonal surface having biomolecular attraction and repulsion areas. In addition, we could control of the height of prepared PEG microstructures with spin coating or not. For the investigation of feasibility of biomolecule patterning onto the functionalized surface, FITC-BSA and HEK 293 were examined as representative biomolecule models. A functionalized surface with GMA promotes the strong adhesion of biomolecules, and PEG microstructures located on the background prevent nonspecific binding of biomolecules at micron-scale level. The orthogonal difference in surface functionality showed strong possibility of simple patterning of biomolecules. In addition, the proposed method could easily control the size, shape, and height of patterns. It will be useful platform technology for the construction of a biomolecule array.     

Interaction of tetradecyltrimethylammonium bromide with bovine serum albumin in different compositions: Effect of temperatures and electrolytes/urea

Herein, the interaction of a protein, bovine serum albumin (BSA), with tetradecyltrimethylammonium bromide (TTAB, a cationic surfactant), has been investigated using the conductivity measurement technique in pure water and some sodium salts (NaCl, Na₂SO₄, Na₂CO₃, and Na₃PO₄) solutions at temperature range of 295.15-320.15 K. Results reveal that, in the plot of specific conductivity versus the concentration of TTAB, only a single critical micelle concentration (cmc) was found for the TTAB + BSA mixed system in all solvents media studied. The addition of BSA in aqueous TTAB solution, the value of cmc undergoes a change from its pure form, which indicates the presence of strong interaction operating between the BSA and TTAB molecules. In aqueous system, the cmc values of the TTAB + BSA mixtures are obtained higher compared to the values found for single TTAB surfactant. However, the addition of salt decreases the cmc value of mixed TTAB + BSA system. The values of cmc of the BSA + TTAB mixed system at 310.15 K and 1.00 mmol·kg^-1 ionic strength of salt followed the order: cmc_Na2CO3 > cmc_Na3PO4 > cmc_NaCl > cmc_Na2SO4. The cmc values of TTAB + BSA mixture were found to be lowered in urea solution within the concentrations studied. The values of degree of dissociation (α) and fraction of counter ion binding (β) were found to be dependent on additives and temperature. The free energy of micellization (△G_m^o) is negative for all the systems, which manifests that the micellization phenomenon is energetically spontaneous. The enhancement of the negative value of △G_m^o in aqueous salt solutions reveals an increase of spontaneity of the TTAB + BSA micellization process. The values of △G_m^o also reveal that the spontaneity of micelle formation is enhanced at higher temperatures in all media studied. The values of free energy of transfer (△G_{m, t}^o) were also determined for numerous solvent media used in the present study and described with appropriate reasoning.     

泡沫相塔壁对泡沫分离牛血清蛋白分离性能的影响

In order to enhance foam drainage, the column with an inner sleeve in the foam phase was designed for studying effect of the column wall of foam phase on foam separation performances using bovine serum albumin(BSA) as the research system. The effects of the wall on the liquid holdup out of the top column, bubble size, enrichment and recovery percentage were investigated. The results indicated that the experimental column with the inner sleeve decreased the liquid holdup, accelerated the coarsening and coalescence of bubbles and increased enrichment of BSA compared the contrasted column without the inner sleeve. Under the conditions of the initial concentration 0.2 g?L-1 of BSA, air flow rate 400 ml?min-1, the experimental column achieved up to a 2.06 fold increase in enrichment compared to the contrasted column. The enrichment of BSA increased with the increase of inner sleeve length. Channel theoretical analysis showed that the ratio of exterior channels to interior channels increased with the increase of bubble diameter. So the experiment column obtained the better performances at the lower concentration and the lower air flow rate. The better performances obtained by experimental column showed that the drainage rate of plateau borders on wall was greater than that of plateau borders between bubbles. So the inner sleeve provided more plateau borders on wall and improved foam drainage.     

Preparation of pH and temperature dual‐sensitive molecularly imprinted polymers based on chitosan and N‐isopropylacrylamide for recognition of bovine serum albumin

pH and temperature dual‐sensitive protein imprinted microspheres with high absorption capacity have been successfully synthesized on the surface of SiO₂ using chitosan grafted N‐isopropylacrylamide (CS‐g‐NIPAM) as the pH and temperature sensitive monomer, with acrylamide as comonomer, N,N′‐methylenebisacrylamide as the crosslinking agent and bovine serum albumin (BSA) as the template protein. The pH and temperature dual‐sensitivity was also investigated. The results showed that the adsorption capacity and imprinting factor improved slowly with increasing incubation pH from 4.6 to 7.0, and then decreased sharply in alkaline conditions due to the reduction of non‐specific binding from electrostatic and hydrogen bonding interactions. Fourier transform infrared spectroscopy, thermogravimetric analysis and transmission electron microscopy were used to characterize the polymers. The as‐prepared SiO₂@BSA molecularly imprinted polymers were also found to have high adsorption capacity (119.88 mg g^?1) within 2 h, an excellent imprinting factor (α = 2.25), specific selectivity and good reusability. © 2019 Society of Chemical Industry     

Photocross-linked polymers based on plant-derived monomers for potential application in optical 3D printing

Photocross linking of the resins composed of plant-derived monomers, acrylated epoxidized soybean oil (AESO), myrcene (MYR) and vanillin dimethacrylate (VDM) or divinylbenzene (DVB, for comparison), was performed using 2,2-dimethoxy-2-phenylacetophenone as photoinitiator. Photocross-linking rate and properties of the crosslinked polymers depended on the resin compositions. The higher amount of MYR caused not only the better homogenization and lower viscosity of the resin but also the reduction of polymerization rate and the worse mechanical and thermal properties of the resulting polymers. The higher amount of aromatic component (VDM or DVB) improved mechanical and thermal properties of polymers. Moreover, the use of VDM instead of DVB in the system led to the higher photocross-linking rate and higher yield of insoluble fraction. The resin composed of only plant-derived monomers AESO/MYR/VDM, molar ratio 1:1:3, showed characteristics comparable to those of commercial petroleum-derived photoresins and was selected as a potential renewable photoresin for application in optical 3D printing. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 137, 48708.     

Measurements and predictive modeling of water diffusion coefficients in bovine serum albumin/polymer blends for biosensors

The knowledge of mass transport parameters in polymer/protein blend films is of substantial interest for the preparation of glucose biosensor test strips where drying and rehydration need to be controlled. In this study, the diffusion of water in bovine serum albumin (BSA) and poly(vinylpyrrolidone) (PVP) films and blends of BSA with PVP and polyvinylalcohol is investigated and the applicability of predictive models for the diffusion coefficient in mixtures is explored. Water concentration profiles are measured with Raman spectroscopy during drying experiments under controlled conditions. Concentration‐dependent diffusion coefficients of water in the films are then determined by fitting simulated drying curves to measured data. Comparison of the resulting diffusion coefficients in the blends with a predictive logarithmic model shows good agreement. The application of such predictive models could greatly facilitate the development of polymer/protein blends in the future. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 45368.     

基于牛血清白蛋白偶联微球的蛋白-药物结合常数快速测定方法研究

以氨基化多孔微粒为固相介质,牛血清白蛋白(BSA)为模型蛋白,通过戊二醛交联反应,制备牛血清白蛋白偶联微球,与药物结合,通过离心沉降微球,高效液相色谱法测定上清液中药物浓度的变化,并代入Rosenthal模型,研发出一种新型方法用于药物蛋白结合常数(Ka)的快速测定。利用该方法测定了瑞替加滨与BSA的结合常数,并与传统荧光光谱法对比,两种方法结果一致。键合蛋白微球经PBS缓冲液充分洗脱后,可重复利用。     

A micro- and nano-structured drug carrier based on biocompatible,hybrid polymeric nanoparticles for potential application in dry powder inhalation therapy

We herein describe the development of a micro-scale powder containing hybrid polymeric nanoparticles, for pulmonary application. The nanoparticles, composed of chitosan, carboxy methyl-β-cyclodextrin and a gelling counterion, were prepared, by ionotropic gelation, and characterized using differential scanning calorimetry and infrared spectroscopy besides other techniques. Nanoparticles transformation into powders was achieved, by co-spray drying with thermo-protectant, resulting in microspheres with adequate aerodynamic properties. Besides morphological and aerodynamic analysis, the developed powder was subjected to in-depth profiling of surface composition, using X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectroscopy for investigating nanoparticles spatial distribution. After fluorescent labelling of both nanoparticles and microsphere matrix, confocal microscopic examination was further performed, for structure analysis. Overall, the applied analytical techniques evidence the homogeneous and almost complete encapsulation of the nanoparticles within microspheres' matrix, emphasizing the efficiency of the microencapsulation method and the potential of the hybrid nanoparticles, as a solid formulation, for lung delivery.     

Chemical synthesis of Co and Mn co-doped NiO nanocrystalline materials as high-performance electrode materials for potential application in supercapacitors

Co and Mn co-doped with NiO nanostructued materials, such as, Ni_0.95Co_0.01Mn_0.04O_1−δ, Ni_0.95Co_0.04Mn_0.01O_1−δ and Ni_0.95Co_0.025Mn_0.025O_1−δ were synthesized by chemical synthesis route and studied for potential application as electrode materials for supercapacitors. The phase structure of the materials was characterized by X-ray diffraction (XRD) and the crystallographic parameters were found out and reported. FTIR (Fourier Transform Infrared) spectroscopy revealed the presence of M–O bond in the compounds. The particle size of the materials was found to be in the range of 291.5–336.5 nm. The morphological phenomenon of the materials was studied by scanning electron microscopy (SEM) and the particles were found to be in spherical shape with average grain size of 14–28 nm. EDAX analysis confirmed the presence of appropriate levels of elements in the samples. The in-depth morphological characteristics were also studied by HR-TEM (High Resolution Tunneling Electron Microscopy). Cyclic voltammetry, chronopotentiometry and electrochemical impedance measurements were applied in an aqueous electrolyte (6 mol L⁻¹ KOH) to investigate the electrochemical performance of the Co and Mn co-doped NiO nanostructured electrode materials. The results indicate that the doping level of Co and Mn in NiO had a significant role in revealing the capacitive behaviors of the materials. Among the three electrode materials studied, Ni_0.95Co_0.025Mn_0.025O_1−δ electrode material shows a maximum specific capacitance of 673.33 F g⁻¹ at a current density of 0.5 A g⁻¹. The electrochemical characteristics of blank graphite sheet were studied and compared with the performance of Co/Mn co-doped NiO based electrode materials. Also, Ni_0.95Co_0.025Mn_0.025O_1−δ has resulted in a degradation level of 4.76% only after 1000 continuous cycles, which shows its excellent electrochemical performance, indicating a kind of potential candidate for supercapacitors.     

The role of titania on the microstructure,biocorrosion and mechanical properties of Mg/HA-based nanocomposites for potential application in bone repair

Bone defects are very challenging in orthopedic practice. The ideal bone grafts should provide mechanical support and enhance the bone healing. Biodegradable magnesium (Mg)–based alloys demonstrate good biocompatibility and osteoconductive properties, which are promising biomaterials for bone substitutes. However, the high rate of their biodegradation in human body environment is still challenging. For this scope, synthesis Mg-based composites with bioceramic additives such as HA and titania (TiO₂) is a routine to solve this problem. The aim of this study was to evaluate the effect of addition TiO₂ nanopowders on the corrosion behavior and mechanical properties of Mg/HA-based nanocomposites fabricated using a milling-pressing-sintering technique for medical applications. The microstructure of Mg/HA/TiO₂ nanocomposites, in vitro degradation and biological properties including in vitro cytocompatibility were investigated. The corrosion resistance of Mg/HA-based nanocomposites was significantly improved by addition 15 wt% of TiO₂ and decrease HA amount to 5 wt% this was inferred from the lower corrosion current; 4.8 µA/cm² versus 285.3 µA/cm² for the Mg/27.5 wt%HA, the higher corrosion potential; −1255.7 versus −1487.3 mV_SCE, the larger polarization resistance; 11.86 versus 0.25 kΩ cm² and the significantly lower corrosion rate; 0.1 versus 4.28 mm/yr. Compressive failure strain significantly increased from 1.7% in Mg/27.5HA to 8.1% in Mg/5HA/15TiO₂ (wt%). The Mg/5HA/15TiO₂ (wt%) nanocomposite possessed high corrosion resistance, cytocompatibility and mechanical properties and can be considered as a promising material for implant applications.     

A comprehensive review on the preparation and application of calcium hydroxyapatite: A special focus on atomic doping methods for bone tissue engineering

Hydroxyapatite (HAP) has been considered to be one of the most preferred scaffold materials among many in the last decade for the bone tissue engineering. Be it prosthetic implants, scaffolds or artificial bone cement, hydroxyapatite has received highest attraction among all due to its chemical and physical properties similar to that of human bone. Although it can be used in the bone tissue engineering as the original composition; for enhancing its different properties relevant to in vivo applications, the calcium in HAP may also be replaced by other atomic dopants depending on usage. Here, we review various HAP coating agents and methods, their merits and demerits. We also review various HAP doping materials, including both cationic as well as anionic materials. We discuss the effects and usage of substitution of hydroxyapatite and their subsequent usage in both bone tissue engineering and maxillofacial surgeries. We consider various research articles published in recent times to accomplish detailed discussion on the subject.     

An in situ study of the deposition of a calcium phosphate mineralized layer on a silicon-substituted hydroxyapatite sensor modulated by bovine serum albumin using QCM-D technology

The deposition of a calcium phosphate (Ca-P) mineralized layer on Si-HA surface (the blank group, Au and the control group, HA) with and without BSA preadsorption was observed in real time using a quartz crystal microbalance with dissipation (QCM-D) technique. The effect of silicon doping on the adsorption of BSA onto a HA sensor was investigated. Approximately 1.5-fold more BSA adsorbed onto HA than Si-HA, indicating the inhibition of BSA adsorption onto the HA sensor by silicon doping. The decreasing ΔD_sat/Δf_sat value in the plots of ΔD–Δf versus adsorption time was predominantly attributed to BSA adsorption, and the decreasing ΔD value with adsorption time during adsorption equilibrium could be explained by the structural rearrangement of BSA on different substrates. Specifically, BSA preadsorption could be a trigger for the formation of nucleation sites for the mineralized layer, promoting nucleation but inhibiting growth. The growth rate was greater on bare substrates than on BSA adsorption substrates. The growth rate of the mineralized layer on the adsorbed BSA layer depended on the substrates in the following manner: Au>Si-HA>HA. The morphology of the mineralized layer depended on the surfaces, and a greater number and a more uniform distribution of smaller nanoparticles were observed on surfaces with preadsorbed BSA. Therefore, BSA could modulate the growth of the mineralized layer at the interface, which is advantageous for fabricating advanced biomaterials.     

Composite scaffolds for cartilage tissue engineering based on natural polymers of bacterial origin,thermoplastic poly(3‐hydroxybutyrate) and micro‐fibrillated bacterial cellulose

Cartilage tissue engineering is an emerging therapeutic strategy that aims to regenerate damaged cartilage caused by disease, trauma, ageing or developmental disorder. Since cartilage lacks regenerative capabilities, it is essential to develop approaches that deliver the appropriate cells, biomaterials and signalling factors to the defect site. Materials and fabrication technologies are therefore critically important for cartilage tissue engineering in designing temporary, artificial extracellular matrices (scaffolds), which support 3D cartilage formation. Hence, this work aimed to investigate the use of poly(3‐hydroxybutyrate)/microfibrillated bacterial cellulose (P(3HB)/MFC) composites as 3D‐scaffolds for potential application in cartilage tissue engineering. The compression moulding/particulate leaching technique employed in the study resulted in good dispersion and a strong adhesion between the MFC and the P(3HB) matrix. Furthermore, the composite scaffold produced displayed better mechanical properties than the neat P(3HB) scaffold. On addition of 10, 20, 30 and 40 wt% MFC to the P(3HB) matrix, the compressive modulus was found to have increased by 35%, 37%, 64% and 124%, while the compression yield strength increased by 95%, 97%, 98% and 102% respectively with respect to neat P(3HB). Both cell attachment and proliferation were found to be optimal on the polymer‐based 3D composite scaffolds produced, indicating a non‐toxic and highly compatible surface for the adhesion and proliferation of mouse chondrogenic ATDC5 cells. The large pores sizes (60 ‐ 83 µm) in the 3D scaffold allowed infiltration and migration of ATDC5 cells deep into the porous network of the scaffold material. Overall this work confirmed the potential of P(3HB)/MFC composites as novel materials in cartilage tissue engineering. © 2016 Society of Chemical Industry