Poly Carboxybetaine Research Articles

New Nano-Crystalline Hydroxyapatite-Polycarboxy/Sulfo Betaine Hybrid Materials: Synthesis and Characterization.

Hybrid materials based on calcium phosphates and synthetic polymers can potentially be used for caries protection due to their similarity to hard tissues in terms of composition, structure and a number of properties. This study is focused on the biomimetic synthesis of hybrid materials consisting of hydroxiapatite and the zwitterionic polymers polysulfobetaine (PSB) and polycarboxybetaine (PCB) using controlled media conditions with a constant pH of 8.0-8.2 and Ca/P = 1.67. The results show that pH control is a dominant factor in the crystal phase formation, so nano-crystalline hydroxyapatite with a Ca/P ratio of 1.63-1.71 was observed as the mineral phase in all the materials prepared. The final polymer content measured for the synthesized hybrid materials was 48-52%. The polymer type affects the final microstructure, and the mineral particle size is thinner and smaller in the synthesis performed using PCB than using PSB. The final intermolecular interaction of the nano-crystallized hydroxyapatite was demonstrated to be stronger with PCB than with PSB as shown by our IR and Raman spectroscopy analyses. The higher remineralization potential of the PCB-containing synthesized material was demonstrated by in vitro testing using artificial saliva.

Polycarboxy/Sulfo Betaine-Calcium Phosphate Hybrid Materials with a Remineralization Potential.

Biomacromolecules control mineral formation during the biomineralization process, but the effects of the organic components' functionality on the type of mineral phase is still unclear. The biomimetic precipitation of calcium phosphates in a physiological medium containing either polycarboxybetaine (PCB) or polysulfobetaine (PSB) was investigated in this study. Amorphous calcium phosphate (ACP) or a mixture of octacalcium phosphate (OCP) and dicalcium phosphate dihydrate (DCPD) in different ratios were identified depending on the sequence of initial solution mixing and on the type of the negative functional group of the polymer used. The more acidic character of the sulfo group in PSB than the carboxy one in PCB determines the dominance of the acidic solid phases, namely, an acidic amorphous phase or DCPD. In the presence of PCB, the formation of ACP with acicular particles arranged in bundles with the same orientation was observed. A preliminary study on the remineralization potential of the hybrid material with the participation of PSB and a mixture of OCP and DCPD did not show an increase in enamel density, contrary to the materials based on PCB and ACP. Moreover, the latter showed the creation of a newly formed crystal layer similar to that of the underlying enamel. This defines PCB/ACP as a promising material for enamel remineralization.

Evaluation Of Differential Response Of Herbal Priming For Augmenting Drought Tolerant Potentiality In Few Commercially Important Indian Rice Varieties (Oryza Sativa L, Var Indica)

Abiotic stress (drought)-a prime threat to the farming community, causing intolerance and less crop productivity in arid and semi-arid regions around the globe.Seed priming is an important hydration technique whereseeds are soaked at a controlled manner and at different condition that helps in increasing crop yield and greater emergence of root and shoot under drought stress and also allow pre- germination of seeds.This priming technique imparts greater seed germination performance, ensuring uniformity,greater tolerance against abiotic stress. Priming method should be cost effective,simple,easy, affordable to the user and must be ecofriendly. To develop crop plant with enhanced tolerance of drought stress, various conventional and genetic approach has been performed, which require great power, cost, skill, and time. The use of natural primer (herbal extract) rather than synthetic one is very much reliable as it is environment friendly and are known to accumulate in plants under stress condition and can act as potent antioxidant that helps in scavenging Reactive Oxygen Species (ROS) produced during abiotic stress and later one causes physiological changes by reducing the fertility of soil and is costly.An experiment was conducted in Randomized Block Design to analyse the efficiency of flower extracts of Marigold (Tagetes patula L), Family (Asteraceae) , Rose ( Rosa rubiginosa), family (Rosaceae), Hibiscus (Hibiscus- roa sinensis) , Family(Malvaceae)—using as priming agent to different rice seed varieties viz Satabdi (IET-4786), Jaya (IET-723), Swarna (MTU-7029) and Cottondora Sannalu (MTU- 1010) against.In the present experiment artificially, drought was mimicked upon the varieties using Poly carboxy Betaine(PCB) and the artificially drought induced seed used for priming with the herbal extract. Data were recorded in terms of seed germination, seedling development, free radicle scavenging potentiality and assessment of alpha amylase as germination indicator.From this study it was revealed that herbal priming shows differential response in different rice variety in all morphological attributes. Satabdi (IET-4786) showed most promising response when compared with check variety , while CottondoraSannalu (MTU-1010) showed least response.These comparative resultsindicated thatSatabdi (IET-4786)seeds primed with petal extract of T. petula may exhibit drought tolerant potentiality in augmenting drought stress and therefore can be a convincing measure for boosting plant’s intrinsic drought tolerant potentiality at the time of early development.

A low-immunogenic genetically-fusible zwitterionic polypeptide

Protein, nucleic acid, and small-molecule drugs frequently require chemical modification with polymers such as polyethylene glycol (PEG), to increase their in vivo circulation time and reduce their immunogenicity. Previously, we developed low-immunogenic zwitterionic polycarboxybetaine (PCB) to replace PEGylation. Herein, we delineate the design principles of zwitterionic polypeptide to mimic PCB. These studies establish an EKP polypeptide composed of glutamic acid (E), lysine (K), and proline (P) as a low-immunogenic, unstructured, zwitterionic peptide. Then, we demonstrate EKP peptide as a genetically encodable fusion protein platform. The unique zwitterionic design ensures high hydration, thereby increasing hydrodynamic size and improving stability. EKP fusion proteins achieve prolonged circulation, low immunogenicity, and maintained circulation time over multiple injections, thus avoiding accelerating blood clearance (ABC). Its performance substantiates EKP as a valuable platform technology for drugs and biologics.

Combination of polycarboxybetaine coating and factor XII inhibitor reduces clot formation while preserving normal tissue coagulation during extracorporeal life support

Blood contact with high surface area medical devices, such as dialysis and extracorporeal life support (ECLS), induces rapid surface coagulation. Systemic anticoagulation, such as heparin, is thus necessary to slow clot formation, but some patients suffer from bleeding complications. Both problems might be reduced by 1) replacing heparin anticoagulation with artificial surface inhibition of the protein adsorption that initiates coagulation and 2) selective inhibition of the intrinsic branch of the coagulation cascade. This approach was evaluated by comparing clot formation and bleeding times during short-term ECLS using zwitterionic polycarboxybetaine (PCB) surface coatings combined with either a potent, selective, bicyclic peptide inhibitor of activated Factor XII (FXII900) or standard heparin anticoagulation. Rabbits underwent venovenous ECLS with small sham oxygenators for 60 min using three means of anticoagulation (n = 4 ea): (1) PCB coating + FXII900 infusion, (2) PCB coating + heparin infusion with an activated clotting time of 220–300s, and (3) heparin infusion alone. Sham oxygenator blood clot weights in the PCB + FXII900 and PCB + heparin groups were 4% and 25% of that in the heparin group (p < 10−6 and p < 10−5), respectively. At the same time, the bleeding time remained normal in the PCB + FXII900 group (2.4 ± 0.2 min) but increased to 4.8 ± 0.5 and 5.1 ± 0.7 min in the PCB + heparin and heparin alone groups (p < 10−4 and 0.01). Sham oxygenator blood flow resistance was significantly lower in the PCB + FXII900 and PCB + heparin groups than in the heparin only group (p < 10−6 and 10−5). These results were confirmed by gross and scanning electron microscopy (SEM) images and fibrinopeptide A (FPA) concentrations. Thus, the combined use of PCB coating and FXII900 markedly reduced sham oxygenator coagulation and tissue bleeding times versus the clinical standard of heparin anticoagulation and is a promising anticoagulation method for clinical ECLS.

Charge-switchable zwitterionic polycarboxybetaine particle as an intestinal permeation enhancer for efficient oral insulin delivery.

Insulin, a peptide hormone, is one of the most common and effective antidiabetic drugs. Although oral administration is considered to be the most convenient and safe choice for patients, the oral bioavailability of insulin is very low due to the poor oral absorption into blood circulation. Intestinal epithelium is a major barrier for the oral absorption of insulin. Therefore, it is vital to develop intestinal permeation enhancer to increase the antidiabetic efficacy of insulin after oral administration.Methods: Charge-switchable zwitterionic polycarboxybetaine (PCB) was used to load insulin to form PCB/insulin (PCB/INS) particles through the electrostatic interaction between positively charged PCB in pH 5.0 and negatively charged insulin in 0.01 M NaOH. The opening effect of PCB/INS particles on intestinal epithelium was evaluated by detecting the changes of claudin-4 (CLDN4) protein and transepithelial electrical resistance (TEER) after incubation or removal. The mechanism was further elucidated based on the results of Western blot and fluorescence images. The PCB/INS particles were then used for type 1 diabetes mellitus therapy after oral administration.Results: PCB could load insulin with the loading efficiency above 86% at weight ratio of 8:1. PCB/INS particles achieved sustained release of insulin at pH 7.4 due to their charge-switchable ability. Surprisingly, PCB/INS particles induced the open of the tight junctions of intestinal epithelium in endocytosis-mediated lysosomal degradation pathway, which resulted in increased intestinal permeability of insulin. Additionally, the opening effect of PCB/INS particles was reversible, and the decreased expression of CLDN4 protein and TEER values were gradually recovered after particles removal. In streptozotocin-induced type 1 diabetic rats, oral administration of PCB/INS particles with diameter sub-200 nm, especially in capsules, significantly enhanced the bioavailability of insulin and achieved longer duration of hypoglycemic effect than the subcutaneously injected insulin. Importantly, there was no endotoxin and pathological change during treatment, indicating that PCB/INS particles were safe enough for in vivo application.Conclusion: These findings indicate that this system can provide a platform for oral insulin and other protein drugs delivery.

Engineering polyzwitterion with acylsulfonamide-based betaine structure for protonated switch of surface chemistry at tumoral pH and reductive responsive drug release of polymeric micelles

The surface chemistry of stimulus-responsive nanoparticles plays an important role in mediating nano-bio interactions in cancer nanomedicine by targeting the unique features of the tumor microenvironment, such as low extracellular pH. To develop therapeutic nanoparticles with high sensitivity and instant response to slight pH differences in the systemic circulation, we produced a novel polyzwitterion with acylsulfonamide-based betaine structure by one-step modification of polycarboxybetaine (PCB) with benzene sulfonamide. The zwitterionic micellar shells show high antifouling in the blood circulation and acutely convert into positive charge via acylsulfonamide protonation, thereby improving cell affinity at tumor sites. Moreover, a disulfide bond between the shell and poly-ε-caprolactone (PCL) core allows for reductive-responsive release of doxorubicin (DOX) after internalization of the polymeric micelles. Finally, in vitro and in vivo competition assays demonstrated that dual responsive drug-loaded micelles have better anticancer efficiency than free DOX or micelles without zwitterionic pH-responsive properties. Thus, we have developed a simple and valuable strategy to enhance pH sensitivity of micellar carriers for cancer treatment.

MOF-Based Antibiofouling Hemoadsorbent for Highly Efficient Removal of Protein-Bound Bilirubin.

A metal-organic framework (MOF)-based antibiofouling hemoadsorbent (PCB-MIL101) was developed through a facile encapsulation of MIL-101(Cr) in zwitterionic poly carboxybetaine (PCB) hydrogel. PCB-MIL101 possessed strong mechanical strength and superior hemocompatibility, ensuring its safety in hemoperfusion applications. In addition, it showed efficient and effective adsorption toward bilirubin (BR), and its maximum adsorption capacity was ∼583 mg g-1. Moreover, due to the protection of antibiofouling PCB hydrogel, PCB-MIL101 showed ability to resist protein adsorption, thus working effectively to remove BR molecules from their binding albumin in biological solutions. The finding in this study provides a novel insight into developing MOF-based hemoadsorbents for the improvement of hemoperfusion therapies.

Developing mechanically robust, triazole-zwitterionic hydrogels to mitigate foreign body response (FBR) for islet encapsulation

Zwitterionic hydrogels such as those based on polycarboxybetaine (PCB) or polysulfobetaine (PSB) have potential for various biomedical applications, due to their biocompatibility and low biofouling properties. However, the poor mechanical properties of zwitterionic hydrogels developed to date remain a challenge, severely limiting their practical uses. To improve the mechanical properties without compromising their zwitterionic feature or biocompatibility, we designed a new class of zwitterionic hydrogels by introducing triazole moieties into the hydrogel monomers that could form energy-dissipating π-π stacking. Compared to conventional zwitterionic hydrogels, the triazole-zwitterionic (TR-ZW) ones exhibited similarly excellent antifouling properties, but were much more mechanically robust with higher stretchability (250% tensile strain), better compression-resistance (89% compressive strain and 65% compression for at least 10 cycles without any crack) and better folding-resistance. In addition, upon subcutaneous implantation in mice, the TR-ZW hydrogels induced significantly lower foreign body responses (FBR) (i.e. less fibrosis and more blood vessel formation relative to a poly(2-hydroxyethyl methacrylate) hydrogel control). As an example of their potential applications, we showed the use of the TR-ZW hydrogels for islet encapsulation and transplantation and demonstrated diabetes correction up to ~1 month in mice in the convenient subcutaneous site.

A Multifunctional Pro‐Healing Zwitterionic Hydrogel for Simultaneous Optical Monitoring of pH and Glucose in Diabetic Wound Treatment

AbstractDiabetic ulcer is the most common kind of chronic wound worldwide. Though great efforts have been devoted, diabetic ulcer still remains as a challenge that requires constant monitoring and management. In this work, a multifunctional zwitterionic hydrogel is developed to simultaneously detect two fluctuant wound parameters, pH and glucose level, to monitor the diabetic wound status. A pH indicator dye (phenol red) and two glucose sensing enzymes, glucose oxidase (GOx) and horseradish peroxidase (HRP), are encapsulated in the anti‐biofouling and biocompatible zwitterionic poly‐carboxybetaine (PCB) hydrogel matrix. The visible images are collected by a smartphone and transformed into RGB signals to quantify the wound parameters. Results show that the activity and stability of both two enzymes are improved within PCB hydrogel, and the Kcat/Km value of PCB‐HRP is ≈5.5 fold of free HRP in artificial wound exudate. This novel wound dressing can successfully monitor the pH range of 4–8 and glucose level of 0.1–10 × 10−3 m. Meanwhile, it also provides a moist healing environment that can promote diabetic wound healing. This multifunctional wound dressing may open vistas in chronic wound management and guide the diabetes treatment in clinical applications.

Simultaneously Antimicrobial, Protein-repellent and Cell-compatible Polyzwitterion Networks: More Insight on Bioactivity and Physical Properties.

A poly(oxanorbornene)-based polyzwitterion with primary ammonium and carboxylate groups (PZI) has been reported previously as the first simultaneously antimicrobial and protein-repellent polyzwitterion. Here, additional physical and biological properties of three poly(oxanorbornene)-based polyzwitterions with different functional groups (PZI, the polycarboxybetaine PCB, and the polysulfobetaine PSB) are compared to understand the molecular origins of this unusual bioactivity. Additionally, the three polyzwitterions and the antimicrobial, polycationic SMAMP are exposed to proteins, bacteria suspensions, human plasma and serum. These interactions are investigated by surface plasmon resonance spectroscopy. In protein adhesion studies, neither fibrinogen nor lysozyme adhere irreversibly to PZI, yet reversible interaction with lysozyme is observed at pH 7 and 8. In the presence of bivalent cations, reversible fibrinogen adhesion on PZI and PSB is observed, but not on PCB. This might explain why mammalian cells grow on PZI and PSB, but not on PCB. PZI does not show human plasma adhesion, while PCB and PSB have 0.27 and 0.48 ng mm-2 adhered plasma, and SMAMP even 6.3 ng mm-2. Both PZI and SMAMP show strong serum adhesion, while no serum adhered to PCB, and only little to PSB. This could be related to the pH difference between serum and plasma, to which the pH-responsive primary ammonium groups are susceptible, while the permanently charged NR4 + groups are unaffected. Both PZI and PCB showed none or only little bacterial adhesion. PCB is also intrinsically antimicrobial against E. coli and S. aureus bacteria and thus is also simultaneously protein-repellent and antimicrobially active. Thus, while the carboxylate groups of PZI and PCB seems to be a prerequisite for the dual antimicrobial activity and protein-repellency, the pH-responsiveness of the primary ammonium group seems to make the PZI molecule vulnerable for protein adhesion in fluids that are slightly out of the physiological range.

Antifouling zwitterionic hydrogel coating improves hemocompatibility of activated carbon hemoadsorbent

Activated carbon has been widely used in hemoperfusion treatments. However, its performance has been significantly compromised by their poor hemocompatibility. In this work, we developed a novel antifouling adsorbent based on zwitterionic poly-carboxybetaine (PCB) hydrogel and powdered activated carbon (PAC) to improve hemocompatibility. We found this new adsorbent (PCB-PAC) was highly stable with negligible leakage of activated carbon debris. It could efficiently resist protein adsorption and avoid any hemolysis effect. The adsorption performance of PCB-PAC for methylene blue was not influenced in a single protein solution or even in 100% fetal bovine serum (FBS), in which pristine PAC lost 50% of its adsorption ability. The isotherms results showed that the adsorption process of PCB-PAC fitted the Langmuir isotherm well, indicating that the PAC particles were homogenously distributed in the PCB hydrogel matrix. Moreover, PCB-PAC could also adsorb bilirubin molecules bound to albumin in solution, while pristine PAC showed no discernible adsorption effect. Findings in this work hold great potential to significantly improve the performance and efficiency of current extracorporeal devices for removing toxins from blood directly.

Zwitterionic hydrogels promote skin wound healing.

Skin traumas are among the most common health problems in the world, and are routinely treated with wound dressings such as bandages and gauze. Traditional dressings are typically made of dry cotton, which tends to adhere to the wound, causing scab formation and bacterial infections. Ideally, a dressing should accelerate wound healing while avoiding any side effects or complications. Recent studies have found that wet dressings, especially hydrogels, can provide a moist environment for wounds that mimics the in vivo environment, thus facilitating debridement of necrotic tissue, enhancing tissue regeneration, and avoiding scab formation. Zwitterionic hydrogels exhibit strong hydration, are biomimetic in nature, and show excellent anti-fouling properties; their resulting high water content, excellent biocompatibility, and negligible interactions with proteins and cells make them ideal wet wound dressings. In this work, we demonstrated that zwitterionic hydrogels, especially poly-carboxybetaine (PCB) hydrogels, can efficiently promote skin wound healing. Skin wounds treated with zwitterionic hydrogels healed significantly better than those treated with PHEMA hydrogels and the commercial product DuoDerm. Moreover, these zwitterionic hydrogels can be easily coated on cotton gauze or bandage pads for easy handling and application. The findings in this work hold great promise for the development of next-generation wound dressings to improve healthcare.

Zwitterionic gel encapsulation promotes protein stability, enhances pharmacokinetics, and reduces immunogenicity

Advances in protein therapy are hindered by the poor stability, inadequate pharmacokinetic (PK) profiles, and immunogenicity of many therapeutic proteins. Polyethylene glycol conjugation (PEGylation) is the most successful strategy to date to overcome these shortcomings, and more than 10 PEGylated proteins have been brought to market. However, anti-PEG antibodies induced by treatment raise serious concerns about the future of PEGylated therapeutics. Here, we demonstrate a zwitterionic polymer network encapsulation technology that effectively enhances protein stability and PK while mitigating the immune response. Uricase modified with a comprehensive zwitterionic polycarboxybetaine (PCB) network exhibited exceptional stability and a greatly prolonged circulation half-life. More importantly, the PK behavior was unchanged, and neither anti-uricase nor anti-PCB antibodies were detected after three weekly injections in a rat model. This technology is applicable to a variety of proteins and unlocks the possibility of adopting highly immunogenic proteins for therapeutic or protective applications.

Structure-Function Relationships of a Tertiary Amine-Based Polycarboxybetaine.

Zwitterionic polycarboxybetaine (PCB) materials have attracted noticeable interest for biomedical applications, such as wound healing/tissue engineering, medical implants, and biosensors, due to their excellent antifouling properties and design flexibility. Antifouling materials with buffering capability are particularly useful for many biomedical applications. In this work, an integrated zwitterionic polymeric material, poly(2-((2-hydroxyethyl)(2-methacrylamidoethyl)ammonio)acetate) (PCBMAA-1T), was synthesized to carry desired properties (antifouling, switchability and buffering capability). A tertiary amine was used to replace quaternary ammonium as the cation to endow the materials with buffering capability under neutral pH. Through this study, a better understanding on the structure-property relationship of zwitterionic materials was obtained. The tertiary amine cation does not compromise antifouling properties of zwitterionic materials. The amount of adsorbed proteins on PCBMAA-1T polymer brushes is less than 0.8 ng/cm(2) for fibrinogen and 0.3 ng/cm(2) (detection limit of the surface plasmon resonance sensor) for both undiluted blood plasma and serum. It is found that the tertiary amine is favorable to obtain good lactone ring stability in switchable PCB materials. Titration study showed that PCBMAA-1T could resist pH changes under both acidic (pH 1-3) and neutral/basic (pH 7-9) conditions. To the best of our knowledge, such an all-in-one material has not been reported. We believe this material might be potentially used for a variety of applications, including tissue engineering, chronic wound healing and medical device coating.

Enhanced endosomal/lysosomal escape by distearoyl phosphoethanolamine-polycarboxybetaine lipid for systemic delivery of siRNA

Cationic liposome based siRNA delivery system has improved the efficiencies of siRNA. However, cationic liposomes are prone to be rapidly cleared by the reticuloendothelial system (RES). Although modification of cationic liposomes with polyethylene glycol (PEG) could prolong circulation lifetime, PEG significantly inhibits siRNA entrapment efficiency, cellular uptake and endosomal/lysosomal escape process, resulting in low gene silencing efficiency of siRNA. In this study, we report the synthesis of zwitterionic polycarboxybetaine (PCB) based distearoyl phosphoethanolamine-polycarboxybetaine (DSPE-PCB) lipid for cationic liposome modification. The DSPE-PCB20 cationic liposome/siRNA complexes (lipoplexes) show an excellent stability in serum medium. The siRNA encapsulation efficiency of DSPE-PCB20 lipoplexes could reach 92% at N/P ratio of 20/1, but only 73% for DSPE-PEG lipoplexes. The zeta potential of DSPE-PCB20 lipoplexes is 8.19±0.53mV at pH 7.4, and increases to 24.6±0.87mV when the pH value is decreased to 4.5, which promotes the endosomal/lysosomal escape of siRNA. The DSPE-PCB20 modification could enhance the silencing efficiency of siRNA by approximately 20% over the DSPE-PEG 2000 lipoplexes at the same N/P ratio in vitro. Furthermore, DSPE-PCB20 lipoplexes could efficiently mediate the down-regulation of Apolipoprotein B (ApoB) mRNA in the liver and consequently decrease the total cholesterol in the serum in vivo, suggesting therapeutic potentials for siRNA delivery in hypercholesterolemia-related diseases.

Tunable disintegration of layer-by-layer assembly multilayer films based on hydrolytical-polybetaine at wide-range time

A cationic hydrolytical-polycarboxybetaine (HPCB), poly(N-ethyl acetate-4-vinylpyridinium bromide) was synthesized by incorporating ester group into the side chain of polycarboxybetaine (PCB). The hydrolytic behaviors of HPCB samples in pH 7.4 phosphate buffer saline (PBS) were investigated by FT-IR and 1H NMR. The layer-by-layer (LbL) assembly of HPCB/poly (sodium 4-styrenesulfonate) PSS and the disintegration of HPCB/PSS multilayer films were monitored by UV–vis absorption spectroscopy, quartz crystal microbalance (QCM) and atomic force microscopy (AFM). The disintegrated behavior of multilayer films was studied in detail by changing the cationic degree of HPCB and the pH of the immersion solution (PBS) in the disintegration process. The disintegration time of HPCB/PSS multilayer films could be controlled widely from 2min to 30days in PBS.

Detection Mechanism of Metallized Carbon Epoxy Oxidase Enzyme Based Sensors

AbstractThe coenzyme FAD has been identified to play an important role in the detection mechanism of oxidase enzyme based biosensors. Incorporating FAD into the carbon composite improved sensitivity to H2O2 consequently increasing sensitivity to the respective analyte. The amount of active enzyme also increased thus enhancing the overall performance of the sensors. Polycarboxybetaine (PCB) has been used as a biocompatible membrane coating. The PCB coated sensors gave reproducible calibrations in protein solutions, which has been shown to be a valid protocol for testing biocompatibility. The importance of reporting selectivity in a manner which indicates the “fitness of purpose” of biosensors has been discussed.