Plasma Protein Adsorption Research Articles

In vitro and in silico protein corona formation evaluation of curcumin and capsaicin loaded-solid lipid nanoparticles

Nanotechnology has been an important tool for the production of nanoparticles with controlled release of drugs for therapeutic applications. Here, we produced solid lipid nanoparticles (SLN) loaded with curcumin and capsaicin (NCC) following the overarching goals of green chemistry. Currently, besides evaluating the composition, and size of these, it is necessary to understand the interactions between nanoparticles and the biomolecules present in the biological medium. For this, assays were conducted in order to evaluate the potential formation of the protein ‘corona’, and to better understand the results obtained in vitro, we also performed an interaction study, in silico, between the NCC components and the main serum protein, albumin. In the first hour of contact between the NCC and the culture medium showed fluctuation in the diameter of the NCC. However, after 24 and 48 h of the incubation period, all NCC concentrations showed an increase in size, which can be attributed to plasma protein adsorption. Since, hard corona takes a few seconds, while the soft corona can be formed in minutes up to a few hours. On the other hand, best docking binding-poses of interaction for the formed docking complexes evaluated suggest interactions following the docking affinity like free energy FEB (Tween 80-bovine serum albumin) ≈ FEB (Span 80-bovine serum albumin) showing a pharmacodynamic pattern based in non-covalent hydrophobic interactions with the bovine serum albumin binding-site. Our in silico results clarify and reinforce our in vitro findings of corona formation, which represents the real interaction with cell membranes in vivo.

In situ apolipoprotein E-enriched corona guides dihydroartemisinin-decorating nanoparticles towards LDLr-mediated tumor-homing chemotherapy

The therapeutic efficiency of active targeting nanoparticulate drug delivery systems (nano-DDS) is highly compromised by the plasma proteins adsorption on nanoparticles (NPs) surface, which significantly hinders cell membrane receptors to recognize the designed ligands, and provokes the off-target toxicity and rapid clearance of NPs in vivo. Herein, we report a novel dihydroartemisinin (DHA)-decorating nano-DDS that in situ specifically recruits endogenous apolipoprotein E (apoE) on the NPs surface. The apoE-anchored corona is able to prolong PLGA-PEG2000-DHA (PPD) NPs circulation capability in blood, facilitate NPs accumulating in tumor cells by the passive enhanced permeability and retention (EPR) effect and low-density lipoprotein receptor (LDLr)-mediated target transport, and ultimately improve the in vivo antitumor activity. Our findings demonstrate that the strategy of in situ regulated apoE-enriched corona ensures NPs an efficient LDLr-mediated tumor-homing chemotherapy.

Plasma protein adsorption on TiO2 nanoparticles: Impact of surface adsorption on temperature-dependent structural changes

Protein adsorption on metal oxide nanoparticle surfaces is crucial to the behavior of oxide nanoparticles in biological systems. In this study, attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy was used for the first time to probe differences in the temperature-dependent conformational changes of two plasma proteins, bovine serum albumin (BSA) and fibrinogen (Fib), in solution and adsorbed on titanium dioxide nanoparticle (ca. 20 nm in diameter) surfaces. ATR-FTIR spectra and autocorrelation moving window two-dimensional correlation (MW2D) spectroscopy were used to identify the denaturation temperature range of these two proteins in aqueous solution and adsorbed. Generalized two-dimensional correlation spectroscopy (2DCOS) was done to investigate the protein unfolding pathway. The results show that the thermostability of BSA changed when adsorbed on the surface whereas there was no difference for solution phase Fib and adsorbed on TiO2. The most notable fact was that solution BSA underwent significant structural changes upon adsorption whereas adsorbed Fib did not. Upon heating adsorbed BSA, no further changes occurred while BSA in solution thermally denatured. In contrast, solution and adsorbed Fib had identical denaturation temperatures and similar sequential secondary structural changes with increasing temperature. These studies show quantitatively, for the first time, that there are differences in the temperature-dependent structural changes of proteins adsorbed on nanoparticles surfaces and these changes depend on the initial interaction with the nanoparticle surface. Overall, the results of this study provide new insights into understanding the effects of temperature on the structure of adsorbed protein on nanoparticle surfaces and the range of different behaviors that can occur.

TiO2 Coating and UV Photofunctionalization Enhance Blood Coagulation on Zirconia Surfaces.

This in vitro study was designed to evaluate the effect of sol-gel derived TiO2 coating on blood coagulation, blood protein adsorption, and platelet response on zirconia surfaces. Square-shaped zirconia (n=96) (10x10x2 mm) was cut, ground, sintered, and finally cleansed ultrasonically in each of acetone and ethanol for 5 minutes. Three experimental groups (n=32) were fabricated: (a) zirconia coated with sol-gel derived TiO2, (b) zirconia coated with sol-gel derived TiO2 and treated with ultraviolet (UV) irradiation for 1 hour, and (c) non-coated zirconia as control. The coatings were prepared from tetraisopropyl orthotitanate solution by dip-coating. The thrombogenicity of the specimens was evaluated using a whole blood kinetic clotting time method where the extent of blood clotting was evaluated at 10, 20, 30, 40, 50, and 60 minutes (n=4/time point, total n=24/group). Scanning electron microscope images were taken to observe platelet morphologies after 1-hour incubation with platelet-rich plasma (PRP) (n=5/group). Surface characteristics were visualized using atomic force microscopy (n=1/group). Adsorption of plasma proteins and fibronectin on each surface was studied by gel electrophoresis (n=2/group). Significant differences were observed in blood coagulation between the test groups at 20-, 30-, 40-, and 50-minute time points (p<0.005). UV treated TiO2 coated specimens showed fastest blood coagulation followed by TiO2 coated and non-coated specimens. Furthermore, platelets appeared at a higher activation state on coated specimens. Gel electrophoresis revealed no difference in protein adsorption among the experimental groups. In summary, TiO2 coatings promoted blood coagulation, and it was further enhanced by UV treatment, which has the potential to hasten the wound healing process in vivo.

One step bulk modification of poly(L-lactic acid) composites with functional additives to improve mechanical and biological properties for cardiovascular implant applications

Poly(L-lactic acid) (PLLA) has been widely used as a promising biomaterial in biomedical applications due to its biodegradability and high mechanical strength. However, because of the inherent brittleness, low impact resistance, and weak thermal stability of PLLA, the modification process is usually required to utilize it for biomedical devices. Furthermore, acidic byproducts resulting from the hydrolysis of PLLA after implantation reduce the pH of the surrounding environment and cause inflammatory responses in the implanted area, leading to the failure of their clinical applications. To this end, here, we demonstrate a novel modification process for the PLLA composite with various functional additives, such as cis-aconitic anhydride (AA), triacetin (TA), isosorbide derivative (ISB), and/or Pluronic® F127 (F). The modified PLLA composite with TA and F (PLLA/TF) showed significantly improved elongation at break and Young’s modulus and retained tensile strength. Moreover, incorporating magnesium hydroxide (MH) nanoparticles (PLLA/TFMH) significantly reduced acid-induced inflammation responses caused by the acidic degradation products of PLLA. Reduced plasma protein adsorption was observed in the PLLA/TFMH. These results suggest that the one step bulk modification of biodegradable PLLA using TA, F, and MH will have great potential in cardiovascular implant applications.

Clopidogrel eluting electrospun polyurethane/polyethylene glycol thromboresistant, hemocompatible nanofibrous scaffolds.

Biomaterials used as blood-contacting material must be hemocompatible and exhibit lower thrombotic potential while maintaining hemostasis and angiogenesis. With the aim of developing thromboresistant, hemocompatible nanofibrous scaffolds, polyurethane/polyethylene glycol scaffolds incorporated with 1, 5, and 10 wt% Clopidogrel were fabricated and evaluated for their physiochemical properties, biocompatibility, hemocompatibility, and antithrombotic potential. The results of physicochemical characterization revealed the fabrication of nanometer-sized scaffolds with smooth surfaces. The incorporation of both polyethylene glycol and Clopidogrel to polyurethane enhanced the hydrophilicity and water uptake potential of polyurethane/polyethylene glycol/Clopidogrel scaffolds. The dynamic mechanical analysis revealed the enhancement in mechanical strength of the polyurethane/polyethylene glycol scaffolds on incorporation of Clopidogrel. The polyurethane/polyethylene glycol/Clopidogrel scaffolds showed a tri-phasic drug release pattern. The results of hemocompatibility assessment demonstrated the excellent blood compatibility of the polyurethane/polyethylene glycol/Clopidogrel scaffolds, with the developed scaffolds exhibiting lower hemolysis, increased albumin and plasma protein adsorption while reduction in fibrinogen adsorption. Further, the platelet adhesion was highly suppressed and significant increase in coagulation period was observed for Clopidogrel incorporated scaffolds. The results of cell adhesion and cell viability substantiate the biocompatibility of the developed nanofibrous scaffolds with the HUVEC cell viability on polyurethane/polyethylene glycol, polyurethane/polyethylene glycol/Clopidogrel-1, 5, and 10% at day 7 found to be 12.35, 13.36, 14.85, and 4.18% higher as compared to polyurethane scaffolds, and the NIH/3T3 cell viability found to be 35.27, 70.82, 36.60, and 7.95% higher as compared to polyurethane scaffolds, respectively. Altogether the results of the study advocate the incorporation of Clopidogrel to the polyurethane/polyethylene glycol blend in order to fabricate scaffolds with appropriate antithrombotic property, hemocompatibility, and cell proliferation capacity and thus, might be successfully used as antithrombotic material for biomedical application.

Plasma protein adsorption on Fe3O4-PEG nanoparticles activates the complement system and induces an inflammatory response.

BackgroundUnderstanding of iron oxide nanoparticles (IONP) interaction with the body milieu is crucial to guarantee their efficiency and biocompatibility in nanomedicine. Polymer coating to IONP, with polyethyleneglycol (PEG) and polyvinylpyrrolidone (PVP), is an accepted strategy to prevent toxicity and excessive protein binding.AimThe aim of this study was to investigate the feature of IONP adsorption of complement proteins, their activation and consequent inflammatory response as a strategy to further elucidate their biocompatibility.MethodsThree types of IONP with different surface characteristics were used: bare (IONP-bare), coated with PVP (IONP-PVP) and PEG-coated (IONP-PEG). IONPs were incubated with human plasma and adsorbed proteins were identified. BALB/c mice were intravenously exposed to IONP to evaluate complement activation and proinflammatory response.ResultsProtein corona fingerprinting showed that PEG surface around IONP promoted a selective adsorption of complement recognition molecules which would be responsible for the complement system activation. Furthermore, IONP-PEG activated in vitro, the complement system and induced a substantial increment of C3a and C4a anaphylatoxins while IONP-bare and IONP-PVP did not. In vivo IONP-PEG induced an increment in complement activation markers (C5a and C5b-9), and proinflammatory cytokines (IL-1β, IL-6, TNF-α).ConclusionThe engineering of nanoparticles must incorporate the association between complement proteins and nanomedicines, which will regulate the immunostimulatory effects through a selective adsorption of plasma proteins and will enable a safer application of IONP in human therapy.

Hemostatic wound dressings: Predicting their effects by in vitro tests.

Application of controlled in vitro techniques can be used as a screening tool for the development of new hemostatic agents allowing quantitative assessment of overall hemostatic potential. Several tests were selected to evaluate the efficacy of cotton gauze, collagen, and oxidized regenerated cellulose for enhancing blood clotting, coagulation, and platelet activation. Visual inspection of dressings after blood contact proved the formation of blood clots. Scanning electron microscopy demonstrated the adsorption of blood cells and plasma proteins. Significantly enhanced blood clot formation was observed for collagen together with β-thromboglobulin increase and platelet count reduction. Oxidized regenerated cellulose demonstrated slower clotting rates not yielding any thrombin generation; yet, led to significantly increased thrombin-anti-thrombin-III complex levels compared to the other dressings. As hemostyptica ought to function without triggering any adverse events, induction of hemolysis, instigation of inflammatory reactions, and initiation of the innate complement system were also tested. Here, cotton gauze provoked high PMN elastase and elevated SC5b-9 concentrations. A range of tests for desired and undesired effects of materials need to be combined to gain some degree of predictability of the in vivo situation. Collagen-based dressings demonstrated the highest hemostyptic properties with lowest adverse reactions whereas gauze did not induce high coagulation activation but rather activated leukocytes and complement.

Biological Response of and Blood Plasma Protein Adsorption on Silver-Doped Hydroxyapatite.

Hydroxyapatite (HAP) is an extensively used orthopedic biomaterial because of its high biocompatibility and osteoconductivity. Implant-related infection is a major cause of orthopedic device failure. Previous research showed that silver-doped hydroxyapatite nanoparticles (Ag-HAP NPs) have prominent antimicrobial activity, but their biocompatibility and plasma protein response remained unexplored. Here we investigated the effects of synthesis conditions on Ag-HAP NP antimicrobial (E. coli and S. epidermidis) activity, biocompatibility, and the adsorption of two blood plasma proteins, human serum albumin (HSA) and fibrinogen (Fib). It was found that synthesis pH affected the Ag content of Ag-HAP NPs and subsequent Ag+ release from the NPs in solution. This, in turn, affected antimicrobial efficiency and cytotoxicity to murine preosteoblast cells (MC3T3-E1). More HSA than Fib was adsorbed on a molar basis. The conformation of HSA changed drastically from predominantly α-helix and minor β-sheet content in solution to greater β-sheet than α-helix content when adsorbed. Correspondingly, the melting temperature Tm of HSA changed significantly from 76 °C in solution to ∼65-66 °C when adsorbed. Fib exhibited a modest decrease in α-helix content while theβ-sheet content increased modestly upon adsorption and its Tm remained unchanged at ∼60 °C. These differences in behavior of HSA and Fib are ascribed to the much smaller size of HSA, which allows a greater molecular packing density on the surface, which induces greater conformational changes. The protein adsorption behavior on Ag-HAP was similar to that on pure HAP. Thus, we show that Ag-HAP NPs have antimicrobial activity without deleterious effects on biocompatibility and blood plasma protein adsorption.

Phycocyanin functionalized single-walled carbon nanohorns hybrid for near-infrared light-mediated cancer phototheranostics

Application of carbon nanomaterials to near-infrared light-mediated phototherapy has attracted great interest. However, their practical use in biological systems is severely limited by their poor water dispersibility, inevitable adsorption of plasma proteins, and their single therapeutic function. Herein, we prepared a phycocyanin (PC)-functionalized single-walled carbon nanohorns (PC@SWNHs) hybrid, where PC is used to preform a protein corona around the SWNHs through non-covalent interface interactions. In the PC@SWNHs, the preformed PC corona effectively improves the water dispersibility of the SWNHs and protects them from plasma protein adsorption. In addition, the excellent photodynamic activity of PC overcomes the inability of SWNHs to generate reactive oxygen species. The SWNHs not only give the hybrid a photothermal therapy (PTT) function, but they also enhance the photostability of PC. These characteristics of the PC@SWNHs are confirmed by in vitro and in vivo experiments, where synergistic photodynamic therapy (PDT)/PTT growth inhibition of tumors is achieved. The PC@SWNHs also act as a photoacoustic/thermal imaging agent to visualize the in vivo treatment process. Hence, this work not only paves the way to use the complementarity between SWNHs and proteins for multifunctional cancer theranostics, but it also allows for a better understanding of bio–nano interface interactions between them.

Impact of Branching on the Solution Behavior and Serum Stability of Starlike Block Copolymers.

The size control of nanomedicines for tumor diagnosis and therapy is of high importance, since it enables or disables deep and sufficient tumor penetration. Amphiphilic star-shaped block copolypept(o)ides offer substantial promise to precisely adjust the hydrophobic core and the hydrophilic corona, independent of each other, and therefore simultaneously control the size dimension in the interesting size range from 10 to 30 nm. To gain access to core-shell structures of such sizes, 3-arm and 6-arm PeptoStars, based on poly(γ- tert-butyloxycarbonyl-l-glutamate)- b-polysarcosine (pGlu(O tBu)- b-pSar), were prepared via controlled living ring-opening polymerization (ROP) of the corresponding N-carboxyanhydrides. Moreover, size exclusion chromatography (SEC) proves the presence of well-defined star shaped polymers with molecular weights from 38 to 88 kg/mol with low polymer dispersities of 1.16 to 1.23. By varying the α-helical peptide core and maintain a constant polysarcosine corona, hydrodynamic size analyses revealed the importance of using a sufficiently large and dense hydrophilic shielding corona to prevent aggregation of the hydrophobic core and obtain uniform-sized spherical-shaped particles with hydrodynamic diameters below 24 nm. Fluorescence correlation spectroscopy (FCS) additionally demonstrates the absence of protein adsorption in human plasma for 6-arm polypept(o)ide stars and thus confirms polysarcosine as stealthlike material.

Magnetic nano capture agent with enhanced anion internal layer diffusion performance for removal of arsenic from human blood

Magnetic nano capture agent (MNCA)-based magnetic separation can remove heavy metals from blood efficiently and safely. It has been considered as the next-generation therapy for heavy metal intoxication. In this work, we report the first use of MNCA to remove arsenic (As) with high efficiency from blood. MNCA with a flexible, non-covalent and biocompatible surface layer has been synthesized by packaging cetyltrimethylammonium bromide (CTAB) and bovine serum albumin (BSA) together with oleic acid coated magnetic iron oxide nanoparticles (MNP@OA) via Van der Waals force. This surface layer enhances arsenic anion diffusion from the surface to the binding sites, leading to increased arsenic removal performance of MNCA. The adsorption capacities of BSA and CTAB enfolded MNP@OA (MNP@CTAB-BSA) for As (III) and As (V) are 131.00 mg g−1 and 119.58 mg g−1, respectively. 89.02% of As (III) and 81.78% of As (V) are removed from blood using MNP@CTAB-BSA, and the residual concentrations of As (III) and As (V) in blood decrease from 400.00 μg L−1 to 43.93 μg L−1 and 72.88 μg L−1, respectively. The as-synthesized MNP@CTAB-BSA shows excellent biocompatibility. Under operation condition, only 0.80% of hemolysis and 1.64% of plasma protein adsorption are induced by MNP@CTAB-BSA, and the cell viability of peripheral blood mononuclear cells (PBMCs) is more than 95.53%. This study paves a fast, safe and effective way for arsenic intoxication therapy.

Adsorption of Plasma Proteins on Single-Walled Carbon Nanotubes Reduced Cytotoxicity and Modulated Neutrophil Activation.

Proteins in the bloodstream bind to carbon nanotubes (CNTs) through noncovalent interactions to form a protein corona, thereby effectively influencing the biological properties and blood biocompatibility of the CNTs. Here, we investigated the binding of common plasma proteins (i.e., human immunoglobulin G (IgG), human serum albumin (HSA), and fibrinogen (FG)) to carboxylated single-walled CNTs (SWCNTs), and evaluated the effects of these different protein coronas on cytotoxicity to endothelial cells and immune response to neutrophils in the bloodstream. Measurements of adsorption parameters revealed tight binding of proteins to SWCNTs, and the SWCNTs adsorption capacities followed the order FG > HSA > IgG. In addition, the basic residues (Arg, Lys, His) were found to play an important role in the formation of protein-SWCNTs corona complexes and determine their adsorption capacity. Consistent with the higher protein adsorption capacity, FG more significantly reduced the cytotoxicity of CNTs to human umbilical vein endothelial cells than the other two proteins. However, only treatment of SWCNTs with IgG resulted in the enhancement of CNT-induced myeloperoxidase (MPO) release (i.e., neutrophil activation) in neutrophils, while MPO-dependent degradation of CNTs induced less cytotoxicity than initial nanomaterials. Consistent with these effects of protein coronas, the presence of serum attenuated the cytotoxicity of CNTs and CNTs could induce neutrophil activation in human blood plasma. Our study demonstrates the ability of adsorbed plasma proteins to influence cytotoxicity and neutrophil response caused by CNTs in the bloodstream.

Surface fluorination of polylactide as a path to improve platelet associated hemocompatibility

Surface-induced thrombosis is still a significant clinical concern for many types of blood-contacting medical devices. In particular, protein adsorption and platelet adhesion are important events due to their ability to trigger the coagulation cascade and initiate thrombosis. Poly(lactic acid) (PLA) has been the predominant polymer used for making bioresorbable stents. Despite long-term advantages, these stents are associated with higher rates of early thrombosis compared with permanent metallic stents. To address this issue, we modified the surface of PLA with a perfluoro compound facilitated by surface activation using radio frequency (RF) plasma. Fluoropolymers have been extensively used in blood contacting materials, such as blood vessel replacements due to their reduced thrombogenicity and reduced platelet reactivity. The compositions of plasma-treated surfaces were determined by electron spectroscopy for chemical analysis (ESCA). Also, contact angle measurements, cell cytotoxicity and the degradation profile of the treated polymers are presented. Finally, relevant blood compatibility parameters, including plasma protein adsorption, platelet adhesion and morphology, were evaluated. We hypothesized that tight binding of adsorbed albumin by fluoropolymers enhances its potential for blood-contacting applications. Statement of SignificanceAlthough bioresorbable stents made from poly(lactic acid) (PLA) may have long-term clinical advantages, they have shown higher rates of early thrombosis as compared with permanent metallic stents. To improve the thromboresistance of PLA, we developed a novel method for surface fluorination of this polymer with a perfluoro compound. Fluoropolymers (e.g., expanded polytetrafluoroethylene) have long been used in blood-contacting applications due to their satisfactory clinical performance. This is the first report of PLA surface fluorination which might be applied to the fabrication of a new generation of fluorinated PLA stents with improved platelet interaction, tunable degradability and drug release capabilities. Also, we describe a general strategy for improving the platelet interactions with biomaterials based on albumin retention.

Surface Nanostructuring of Parylene-C Coatings for Blood Contacting Implants.

This paper investigates the effects on the blood compatibility of surface nanostructuring of Parylene-C coating. The proposed technique, based on the consecutive use of O2 and SF6 plasma, alters the surface roughness and enhances the intrinsic hydrophobicity of Parylene-C. The degree of hydrophobicity of the prepared surface can be precisely controlled by opportunely adjusting the plasma exposure times. Static contact angle measurements, performed on treated Parylene-C, showed a maximum contact angle of 158°. The nanostructured Parylene-C retained its hydrophobicity up to 45 days, when stored in a dry environment. Storing the samples in a body-mimicking solution caused the contact angle to progressively decrease. However, at the end of the measurement, the plasma treated surfaces still exhibited a higher hydrophobicity than the untreated counterparts. The proposed treatment improved the performance of the polymer as a water diffusion barrier in a body simulating environment. Modifying the nanotopography of the polymer influences the adsorption of different blood plasma proteins. The adsorption of albumin—a platelet adhesion inhibitor—and of fibrinogen—a platelet adhesion promoter—was studied by fluorescence microscopy. The adsorption capacity increased monotonically with increasing hydrophobicity for both studied proteins. The effect on albumin adsorption was considerably higher than on fibrinogen. Study of the proteins simultaneous adsorption showed that the albumin to fibrinogen adsorbed ratio increases with substrate hydrophobicity, suggesting lower thrombogenicity of the nanostructured surfaces. Animal experiments proved that the treated surfaces did not trigger any blood clot or thrombus formation when directly exposed to the arterial blood flow. The findings above, together with the exceptional mechanical and insulation properties of Parylene-C, support its use for packaging implants chronically exposed to the blood flow.

Revealing the immune perturbation of black phosphorus nanomaterials to macrophages by understanding the protein corona

The increasing number of biological applications for black phosphorus (BP) nanomaterials has precipitated considerable concern about their interactions with physiological systems. Here we demonstrate the adsorption of plasma protein onto BP nanomaterials and the subsequent immune perturbation effect on macrophages. Using liquid chromatography tandem mass spectrometry, 75.8% of the proteins bound to BP quantum dots were immune relevant proteins, while that percentage for BP nanosheet–corona complexes is 69.9%. In particular, the protein corona dramatically reshapes BP nanomaterial–corona complexes, influenced cellular uptake, activated the NF-κB pathway and even increased cytokine secretion by 2–4-fold. BP nanomaterials induce immunotoxicity and immune perturbation in macrophages in the presence of a plasma corona. These findings offer important insights into the development of safe and effective BP nanomaterial-based therapies.

Influence of dynamic flow conditions on adsorbed plasma protein corona and surface-induced thrombus generation on antifouling brushes

The information regarding the nature of protein corona (and its changes) and cell binding on biomaterial surface under dynamic conditions is critical to dissect the mechanism of surface-induced thrombosis. In this manuscript, we investigated the nature of protein corona and blood cell binding in heparinized recalcified human plasma, platelet rich plasma and whole blood on three highly hydrophilic antifouling polymer brushes, (poly(N, N-dimethylacrylamide) (PDMA), poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) and poly[N-(2-hydroxypropyl) methacrylamide] (PHPMA) using an in vitro blood loop model at comparable arterial and venous flow, and static conditions. A fluid dynamics model was used initially to better understand the resulting flow patterns in a vertical channel containing the substrates to arrive at the placement of the substrates within the blood loop. The protein binding on the brush modified substrates was determined using ellipsometry, fluorescence microscopy and the nature of the protein corona was investigated using mass spectrometry based proteomics. The flow elevated fouling on brush coated surface from blood. The extent of plasma protein adsorption and platelet adhesion onto PDMA brush was lower than other surfaces in both static and flow conditions. The profiles of adsorbed protein corona showed strong dependence on the test conditions (static vs. flow), and the chemistry of the polymer brushes. Specially, the PDMA brush under flow conditions was more enriched with coagulation proteins, complement proteins, vitronectin and fibronectin but was less enriched with serum albumin. Apolipoprotein B-100 and complement proteins were the most abundant proteins seen on PMPC and PHPMA surfaces under both flow and static conditions, respectively. Unlike PDMA brush, the flow conditions did not affect the composition of protein corona on PMPC and PHPMA brushes. The nature of the protein corona formed in flow conditions influenced the platelet and red blood cell binding. The dependence of shear stress on platelet adhesion from platelet rich plasma and whole blood highlights the contribution of red blood cells in enhancing platelet adhesion on the surface under high shear condition.

Influence of ligands property and particle size of gold nanoparticles on the protein adsorption and corresponding targeting ability

Nanoparticulated vesicles were widely used for carriers of drugs and imaging probes. To improve the targeting delivery efficiency of these vesicles, ligands were often functionalized onto their surfaces. However, the interaction between vesicles and plasma proteins may cover the ligands and hinder the targeting delivery. It is important to address the potential influence of ligands modification on plasma protein adsorption and the following targeting delivery. In this study, two common used ligands were chosen as the model: transferrin and RGD peptide. Gold nanoparticles were utilized as model particles. Sodium dodecyl sulfate polyacrylamide gel electrophoresis data demonstrated that higher PEG modification and smaller particle size could reduce the plasma protein adsorption, while ligand modification could increase. The cellular uptake results showed that the targeting ability of smaller ligand RGD peptide would be more easily influenced by the proteins corona.

A substrate-independent ultrathin hydrogel film as an antifouling and antibacterial layer for a microfiltration membrane anchored via a layer-by-layer thiol-ene click reaction.

Herein, a substrate-independent ultrathin hydrogel film was constructed on a microfiltration membrane through layer-by-layer (LbL) thiol-ene click chemistry to improve the antifouling and antibacterial properties. In our strategy, ene-functionalized dopamine was synthesized and coated onto a model substrate (polyethersulfone membrane) to introduce double bonds as anchoring sites for the hydrogel film; thiol-functionalized poly[oligo(ethylene glycol)mercaptosuccinate] (POEGMS) and ene-functionalized P(SBMA-co-AA) were synthesized as hydrogel precursors. The membrane was alternately immersed in the precursor solutions to form the ultrathin hydrogel film. Finally, Ag nanoparticles (AgNPs) were loaded into the hydrogel layer by adsorption and reduction procedures. By coating the hydrogel films, the loaded AgNPs could kill almost all the contacting bacteria and the bacteria in the surroundings, and the enhanced hydrophilicity of the modified membrane could effectively prevent the attachment of the bacteria. The membrane flux showed no significant decrease, the rejection ratio of BSA increased from 51% to 89%, and the FRR increased from 36% to 90%. Moreover, the improvement of the hemocompatibility was confirmed by the decline in the plasma protein adsorption, prolonged clotting times, low hemolysis ratio, and prevention of platelet adhesion. Compared with that of other techniques for attaching hydrogel films, the main advantage of the current technique is that the hydrogel film thickness could be well controlled within the nanometer range; thus, it could significantly improve the antifouling and antibacterial properties of the membrane, but without compromising its permeability. Another advantage is that it is versatile for various substrates such as PVDF, PAN, and CA. This study opens up a facile and versatile route for anchoring ultrathin hydrogel film onto polymeric membranes to achieve excellent antifouling, antibacterial and hemocompatible properties.

Evaluating the biocompatibility of marine-derived chitosan–collagen polymeric blends for biomedical applications

The capability of biomaterials such as chitosan and collagen to support cell growth and proliferation makes them promising candidates in biomedical applications. Chitosan and collagen from marine world have already been proved to be better alternatives to those from terrestrial world. In the current study, chitosan and collagen were isolated from shrimp shell and fish skin, respectively. The polymers were characterized by ultraviolet-visible spectra analysis, Fourier transform infrared-attenuated total reflectance analysis, CHN analysis, and sodium dodedcyl sulfate polyacrylamide gel electrophoresis analysis. Interpenetrating blends of these polymers were synthesized in the form of films in two different ratios. Glutaraldehyde was used as an additional cross-linker to provide more stability to the blends. The polymeric blends were also characterized by Fourier transform infrared-attenuated total reflectance, scanning electron microscopy analysis, and swelling studies. The biocompatibility evaluation included hemocompatibility and cytocompatibility studies. Fourier transform infrared-attenuated total reflectance analysis of films confirmed the presence of characteristic functional groups and molecular interactions of the two polymers in the two blends. Homogenous blending of the two biopolymers in both film compositions was confirmed by the smooth surface images in scanning electron microscopy analysis. The swelling study revealed that both the films can effectively transfer water across it, hence nutrients and waste materials. During hemocompatibility evaluations, no red blood cell aggregation was observed and both the films adsorbed plasma proteins, predominantly albumin, when they made contact with blood. Although one of the films showed slightly higher hemolysis, the value was within the acceptable range. More than 90% viability obtained in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay shows the non-toxic nature of the two films. No sign of morphological changes to L929 cells was seen when they were in direct contact with both films. Live/dead assay using acridine orange/ethidium bromide cocktail showed that the films have not induced apoptosis to the L929 cells, which further asserts their biocompatible nature.