ABSTRACT
In recent years, there has been a growing interest in multifunctional theranostic agents capable of delivering therapeutic payloads while facilitating simultaneous diagnostic imaging of diseased sites. This approach offers a comprehensive strategy particularly valuable in dynamically evolving diseases like cancer, where combining therapy and diagnostics provides crucial insights for treatment planning. Nanoscale platforms, specifically nanogels, have emerged as promising candidates due to their stability, tunability, and multifunctionality as carriers. As a well-studied subgroup of soft polymeric nanoparticles, nanogels exhibit inherent advantages due to their size and chemical compositions, allowing for passive and active targeting of diseased tissues. Moreover, nanogels loaded with therapeutic and diagnostic agents can be designed to respond to specific stimuli at the disease site, enhancing their efficacy and specificity. This capability enables fine-tuning of theranostic platforms, garnering significant clinical interest as they can be tailored for personalized treatments. The ability to monitor tumor progression in response to treatment facilitates the adaptation of therapies according to individual patient responses, highlighting the importance of designing theranostic platforms to guide clinicians in making informed treatment decisions. Consequently, the integration of therapy and diagnostics using theranostic platforms continues to advance, offering intelligent solutions to address the challenges of complex diseases such as cancer. In this context, nanogels capable of delivering therapeutic payloads and simultaneously armed with diagnostic modalities have emerged as an attractive theranostic platform. This review focuses on advances made toward the fabrication and utilization of theranostic nanogels by highlighting examples from recent literature where their performances through a combination of therapeutic agents and imaging methods have been evaluated.
Subject(s)
Nanogels , Neoplasms , Theranostic Nanomedicine , Humans , Nanogels/chemistry , Neoplasms/diagnostic imaging , Neoplasms/drug therapy , Animals , Diagnostic Imaging/methods , Drug Carriers/chemistry , Drug Delivery Systems , Multifunctional Nanoparticles/chemistryABSTRACT
Macroporous cryogels are attractive scaffolds for biomedical applications, such as biomolecular immobilization, diagnostic sensing, and tissue engineering. In this study, thiol-reactive redox-responsive cryogels with a porous structure are prepared using photopolymerization of a pyridyl disulfide poly(ethylene glycol) methacrylate (PDS-PEG-MA) monomer. Reactive cryogels are produced using PDS-PEG-MA and hydrophilic poly(ethylene glycol) methyl ether methacrylate (PEGMEMA) monomers, along with a PEG-based cross-linker and photoinitiator. Functionalization of cryogels using a fluorescent dye via the disulfide-thiol exchange reactions is demonstrated, followed by release under reducing conditions. For ligand-mediated protein immobilization, first, thiol-containing biotin or mannose is conjugated onto the cryogels. Subsequently, fluorescent dye-labeled proteins streptavidin and concanavalin A (ConA) are immobilized via ligand-mediated conjugation. Furthermore, we demonstrate that the mannose-decorated cryogel could capture ConA selectively from a mixture of lectins. The efficiency of protein immobilization could be easily tuned by changing the ratio of the thiol-sensitive moiety in the scaffold. Finally, an integrin-binding cell adhesive peptide is attached to cryogels to achieve successful attachment, and the on-demand detachment of integrin-receptor-rich fibroblast cells is demonstrated. Redox-responsive cryogels can serve as potential scaffolds for a variety of biomedical applications because of their facile synthesis and modification.
Subject(s)
Cryogels , Oxidation-Reduction , Polyethylene Glycols , Cryogels/chemistry , Polyethylene Glycols/chemistry , Animals , Concanavalin A/chemistry , Concanavalin A/metabolism , Methacrylates/chemistry , Mice , Mannose/chemistry , Immobilized Proteins/chemistry , Immobilized Proteins/metabolism , Sulfhydryl Compounds/chemistry , Streptavidin/chemistry , Streptavidin/metabolism , Proteins/chemistry , Proteins/metabolism , Biotin/chemistry , Biotin/metabolism , Biotin/analogs & derivatives , PorosityABSTRACT
Increasing interest in the utilization of hydrogels in various areas of biomedical sciences ranging from biosensing and drug delivery to tissue engineering has necessitated the synthesis of these materials using efficient and benign chemical transformations. In this regard, the advent of "click" chemistry revolutionized the design of hydrogels and a range of efficient reactions was utilized to obtain hydrogels with increased control over their physicochemical properties. The ability to apply the "click" chemistry paradigm to both synthetic and natural polymers as hydrogel precursors further expanded the utility of this chemistry in network formation. In particular, the ability to integrate clickable handles at predetermined locations in polymeric components enables the formation of well-defined networks. Although, in the early years of "click" chemistry, the copper-catalyzed azide-alkyne cycloaddition was widely employed, recent years have focused on the use of metal-free "click" transformations, since residual metal impurities may interfere with or compromise the biological function of such materials. Furthermore, many of the non-metal-catalyzed "click" transformations enable the fabrication of injectable hydrogels, as well as the fabrication of microstructured gels using spatial and temporal control. This review article summarizes the recent advances in the fabrication of hydrogels using various metal-free "click" reactions and highlights the applications of thus obtained materials. One could envision that the use of these versatile metal-free "click" reactions would continue to revolutionize the design of functional hydrogels geared to address unmet needs in biomedical sciences.
Subject(s)
Hydrogels , Polymers , Hydrogels/chemistry , Polymers/chemistry , Click Chemistry , Metals , Drug Delivery SystemsABSTRACT
Polymeric microgels, fabricated via microfluidic techniques, have garnered significant interest as versatile drug delivery carriers. Despite the advances, the loading and release of hydrophobic drugs such as curcumin from polymeric microgels is not trivial. Herein, we report that effective drug loading can be achieved by the design of porous particles and the use of supramolecular cyclodextrin-based curcumin complexes. The fabrication of porous microgels through the judicious choice of chemical precursors under flow conditions was established. The evaluation of the curcumin loading dependence on the porosity of the microgels was performed. Microgels with higher porosity exhibited better curcumin loading compared to those with lower porosity. Curcumin-loaded microgels released the drug, which, upon internalization by U87 MG human glioma cancer cells, induced cytotoxicity. The findings reported here provide valuable insights for the development of tailored drug delivery systems using a microfluidics-based platform and outline a strategy for the effective delivery of hydrophobic therapeutic agents such as curcumin through supramolecular complexation.
ABSTRACT
Polymeric surface coatings capable of effectively integrating desired functional molecules and ligands are attractive for fabricating bio-interfaces necessary for various applications. Herein, we report the design of a polymeric platform amenable to such modifications in a modular fashion through host-guest chemistry. Copolymers containing adamantane (Ada) moieties, diethylene glycol (DEG) units, and silyloxy groups to provide functionalization handles, anti-biofouling character, and surface attachment, respectively, were synthesized. These copolymers were employed to modify silicon/glass surfaces to enable their functionalization using beta-cyclodextrin (ßCD) containing functional molecules and bioactive ligands. Moreover, surface functionalization could be spatially controlled using a well-established technique like microcontact printing. Efficient and robust functionalization of polymer-coated surfaces was demonstrated by immobilizing a ßCD-conjugated fluorescent rhodamine dye through the specific noncovalent binding between Ada and ßCD units. Furthermore, biotin, mannose, and cell adhesive peptide-modified ßCD were immobilized onto the Ada-containing polymer-coated surfaces to direct noncovalent conjugation of streptavidin, concanavalin A (ConA), and fibroblast cells, respectively. It was demonstrated that the mannose-functionalized coating could selectively bind to the target lectin ConA, and the interface could be regenerated and reused several times. Moreover, the polymeric coating was adaptable for cell attachment and proliferation upon noncovalent modification with cell-adhesive peptides. One can envision that the facile synthesis of the Ada-based copolymers, mild conditions for coating surfaces, and their effective transformation to various functional interfaces in a modular fashion offers an attractive approach to engineering functional interfaces for several biomedical applications.
Subject(s)
Mannose , Polymers , Ligands , Polymers/chemistryABSTRACT
Recent years have witnessed increased attention to the use of droplet-based microfluidics as a tool for the fabrication of microparticles due to this method's ability to exploit fluid mechanics to create materials with a narrow range of sizes. In addition, this approach offers a controllable way to configure the composition of the resulting micro/nanomaterials. To date, molecularly imprinted polymers (MIPs) in particle form have been prepared using various polymerization methods for several applications in biology and chemistry. However, the traditional approach, that is, the production of microparticles through grinding and sieving, generally leads to poor control over particle size and distribution. Droplet-based microfluidics offers an attractive alternative for the fabrication of molecularly imprinted microparticles. This mini-review aims to present recent examples highlighting the application of droplet-based microfluidics to fabricate molecularly imprinted polymeric particles for applications in the chemical and biomedical sciences.
ABSTRACT
Bioorthogonal activation of pro-dyes and prodrugs using transition-metal catalysts (TMCs) provides a promising strategy for imaging and therapeutic applications. TMCs can be loaded into polymeric nanoparticles through hydrophobic encapsulation to generate polymeric nanocatalysts with enhanced solubility and stability. However, biomedical use of these nanostructures faces challenges due to unwanted tissue accumulation of nonbiodegradable nanomaterials and cytotoxicity of heavy-metal catalysts. We report here the creation of fully biodegradable nanocatalysts based on an engineered FDA-approved polymer and the naturally existing catalyst hemin. Stable nanocatalysts were generated through kinetic stabilization using flash nanoprecipitation. The therapeutic potential of these nanocatalysts was demonstrated through effective treatment of bacterial biofilms through the bioorthogonal activation of a pro-antibiotic.
Subject(s)
Nanoparticles , Nanostructures , Transition Elements , Polymers/chemistry , Nanoparticles/chemistry , Transition Elements/chemistry , Anti-Bacterial Agents/pharmacologyABSTRACT
In recent years, the bottom-up approach has emerged as a powerful tool in the fabrication of functional nanomaterials through the self-assembly of nanoscale building blocks. The cues embedded at the molecular level provide a handle to control and direct the assembly of nano-objects to construct higher-order structures. Molecular recognition among the building blocks can assist their precise positioning in a predetermined manner to yield nano- and microstructures that may be difficult to obtain otherwise. A well-orchestrated combination of top-down fabrication and directed self-assembly-based bottom-up approach enables the realization of functional nanomaterial-based devices. Among the various available molecular recognition-based "host-guest" combinations, cyclodextrin-mediated interactions possess an attractive attribute that the interaction is driven in aqueous environments, such as in biological systems. Over the past decade, cyclodextrin-based specific host-guest interactions have been exploited to design and construct structural and functional nanomaterials based on cyclodextrin-coated metal nanoparticles. The focus of this review is to highlight recent advances in the self-assembly of cyclodextrin-coated metal nanoparticles driven by the specific host-guest interaction.
Subject(s)
Cyclodextrins , Nanoparticles , Nanostructures , Cyclodextrins/chemistry , Nanostructures/chemistry , Nanoparticles/chemistry , WaterABSTRACT
Electrospun nanofibers are a 3D scaffold of choice for many drug delivery devices due to their high surface area, significant capacity for drug payload, ease of in situ placement, and scalable manufacture. Herein, we report the synthesis of polymeric, pH-responsive nanofiber buttresses via electrospinning. The homopolymer is comprised of an acrylic backbone with acid-sensitive, hydrolyzable, trimethoxybenzaldehyde-protected side chains that lead to buttress transformation from a hydrophobic to a hydrophilic state under physiologically relevant pH conditions (e.g., extracellular tumor environment with pH = 6.5). Hydrolysis of the side chains leads to an increase in fiber diameter from approximately 350 to 900 nm and the release of the encapsulated drug cargo. In vitro drug release profiles demonstrate that significantly more drug is released at pH 5.5 compared to pH 7.4, thereby limiting the release to the target site, with docetaxel releasing over 20 days and doxorubicin over 7 days. Drug burst release, defined as >50% within 24 hours, does not occur at either pH or with either drug. Drug-loaded buttresses preserve drug activity and are cytotoxic to multiple human cancer lines, including breast and lung. Important to their potential application in surgical applications, the tensile strength of the buttresses is 6.3 kPa and, though weaker than commercially available buttresses, they provide sufficient flexibility and mechanical integrity to serve as buttressing materials via the application with a conventional surgical cutting stapler.
Subject(s)
Nanofibers , Neoplasms , Humans , Nanofibers/chemistry , Drug Delivery Systems , Polymers/chemistry , Doxorubicin/pharmacology , Hydrogen-Ion Concentration , Drug LiberationABSTRACT
Molecularly imprinted polymers (MIPs) continue to gain increasing attention as functional materials due to their unique characteristics such as higher stability, simple preparation, robustness, better binding capacity, and low cost. In particular, MIP-coated inorganic nanoparticles have emerged as a promising platform for various biomedical applications ranging from drug delivery to bioimaging. The integration of MIPs with inorganic nanomaterials such as silica (SiO2), iron oxide (Fe3O4), gold (Au), silver (Ag), and quantum dots (QDs) combines several attributes from both components to yield highly multifunctional materials. These materials with a multicomponent hierarchical structure composed of an inorganic core and an imprinted polymer shell exhibit enhanced properties and new functionalities. This review aims to provide a general overview of key recent advances in the fabrication of MIPs-coated inorganic nanoparticles and highlight their biomedical applications, including drug delivery, biosensor, bioimaging, and bioseparation.
ABSTRACT
Facile and effective functionalization of the interface of polymer-coated surfaces allows one to dictate the interaction of the underlying material with the chemical and biological analytes in its environment. Herein, we outline a modular approach that would enable installing a variety of "clickable" handles onto the surface of polymer brushes, enabling facile conjugation of various ligands to obtain functional interfaces. To this end, hydrophilic anti-biofouling poly(ethylene glycol)-based polymer brushes are fabricated on glass-like silicon oxide surfaces using reversible addition-fragmentation chain transfer (RAFT) polymerization. The dithioester group at the chain-end of the polymer brushes enabled the installation of azide, maleimide, and terminal alkene functional groups, using a post-polymerization radical exchange reaction with appropriately functionalized azo-containing molecules. Thus, modified polymer brushes underwent facile conjugation of alkyne or thiol-containing dyes and ligands using alkyne-azide cycloaddition, Michael addition, and radical thiol-ene conjugation, respectively. Moreover, we demonstrate that the radical exchange approach also enables the installation of multivalent motifs using dendritic azo-containing molecules. Terminal alkene groups containing dendrons amenable to functionalization with thiol-containing molecules using the radical thiol-ene reaction were installed at the interface and subsequently functionalized with mannose ligands to enable sensing of the Concanavalin A lectin.
Subject(s)
Dendrimers , Polymers , Alkenes , Alkynes/chemistry , Azides/chemistry , Coloring Agents , Concanavalin A , Ligands , Maleimides , Mannose , Polyethylene Glycols/chemistry , Polymers/chemistry , Silicon Dioxide , Sulfhydryl Compounds/chemistryABSTRACT
Fast-forming yet easily dissolvable hydrogels (HGs) have potential applications in wound healing, burn incidences, and delivery of therapeutic agents. Herein, a combination of a thiol-maleimide conjugation and thiol-disulfide exchange reaction is employed to fabricate fast-forming HGs which rapidly dissolve upon exposure to dithiothreitol (DTT), a nontoxic thiol-containing hydrophilic molecule. In particular, maleimide disulfide-terminated telechelic linear poly(ethylene glycol) (PEG) polymer and PEG-based tetrathiol macromonomers are employed as gel precursors, which upon mixing yield HGs within a minute. The selectivity of the thiol-maleimide conjugation in the presence of a disulfide linkage was established through 1H NMR spectroscopy and Ellman's test. Rapid degradation of HGs in the presence of thiol-containing solution was evident from the reduction in storage modulus. HGs encapsulated with fluorescent dye-labeled dextran polymers and bovine serum albumin were fabricated, and their cargo release was investigated under passive and active conditions upon exposure to DTT. One can envision that the rapid gelation and fast on-demand dissolution under relatively benign conditions would make these polymeric materials attractive for a range of biomedical applications.
Subject(s)
Hydrogels , Sulfhydryl Compounds , Disulfides/chemistry , Dithiothreitol , Hydrogels/chemistry , Maleimides/chemistry , Oxidation-Reduction , Polyethylene Glycols/chemistry , Polymers/chemistry , Sulfhydryl Compounds/chemistryABSTRACT
In recent years, stimuli-responsive degradation has emerged as a desirable design criterion for functional hydrogels to tune the release of encapsulated payload as well as ensure degradation of the gel upon completion of its function. Herein, redox-responsive hydrogels with a well-defined network structure were obtained using a highly efficient thiol-disulfide exchange reaction. In particular, gelation occurred upon combining thiol-terminated tetra-arm polyethylene glycol (PEG) polymers with linear telechelic PEG-based polymers containing pyridyl disulfide units at their chain ends. Rapid gelation proceeds with good conversions (>85%) to yield macroporous hydrogels possessing high water uptake. Furthermore, due to the presence of the disulfide linkages, the thus-obtained hydrogels can self-heal. The obtained hydrogels undergo complete degradation when exposed to environments rich in thiol-containing agents such as dithiothreitol (DTT) and L-glutathione (GSH). Also, the release profile of encapsulated protein, namely, bovine serum albumin, can be tuned by varying the molecular weight of the polymeric precursors. Additionally, it was demonstrated that complete dissolution of the hydrogel to rapidly release the encapsulated protein occurs upon treating these hydrogels with DTT. Cytotoxicity evaluation of the hydrogels and their degradation products indicated the benign nature of these hydrogels. Additionally, the cytocompatible nature of these materials was also evident from a live/dead cell viability assay. One can envision that the facile fabrication and their ability to degrade on-demand and release their payload will make these benign polymeric scaffolds attractive for various biomedical applications.
Subject(s)
Hydrogels , Polyethylene Glycols , Disulfides/chemistry , Dithiothreitol , Hydrogels/chemistry , Oxidation-Reduction , Polyethylene Glycols/chemistry , Sulfhydryl Compounds/chemistryABSTRACT
Electrospun fiber mats loaded with therapeutics have gained considerable attention as a versatile tool in the biomedical field. While these bandages are largely based on fast-dissolving polymers to release the cargo, stimuli-responsive fiber mats have the advantages of providing a timely and spatially controlled drug delivery platform, which can be refilled and reused several times. These benefits make electrospun fiber patches original platforms for painless and convenient on-demand hormone release. Because of the high need of more convenient and non-invasive methods for delivering insulin, a hormone that is currently used to treat hundred million people with diabetes worldwide, we have investigated the tremendous potential of reduced graphene oxide modified poly(acrylic acid) based fiber mats as an original platform for buccal and corneal insulin delivery on-demand. The PAA@rGO hydrogel-like fibers rendered water-insoluble by incorporating ß-cyclodextrin, followed by thermal cross-linking, which showed adequate tensile strength along with high adsorption capacity of insulin at pH 7 and good recyclability. The fiber mats maintained good fibrous morphology and high loading efficiency even after five loading-release cycles. The mucoadhesive nature of the fibers allowed their application for insulin delivery via the eye cornea and the buccal mouth lining, as evidenced in ex vivo studies. Insulin loaded PAA@rGO hydrogel-like fibers showed an insulin flux via buccal lining of pigs of 16.6 ± 2.9 µg cm-2 h-1 and 24.3 ± 3.1 µg cm-2 h-1 for porcine cornea. Testing on healthy adult volunteers confirmed the excellent, mucoadhesive nature of the bandage, with three out of six volunteers feeling completely comfortable (note 8.3) while wearing the patches in the buccal cavity.
Subject(s)
Insulin , Mouth Mucosa , Administration, Buccal , Animals , Cornea , Humans , Hydrogels , Insulin, Regular, Human , SwineABSTRACT
Biofilm infections caused by multidrug-resistant (MDR) bacteria are an urgent global health threat. Incorporation of natural essential oils into biodegradable oil-in-water cross-linked polymeric nanoemulsions (X-NEs) provides effective eradication of MDR bacterial biofilms. The X-NE platform combines the degradability of functionalized poly(lactic acid) polymers with the antimicrobial activity of carvacrol (from oregano oil). These X-NEs exhibited effective penetration and killing of biofilms formed by pathogenic bacteria. Biofilm-fibroblast coculture models demonstrate that X-NEs selectively eliminate bacteria without harming mammalian cells, making them promising candidates for antibiofilm therapeutics.
Subject(s)
Anti-Bacterial Agents/pharmacology , Biofilms/drug effects , Cymenes/pharmacology , Drug Carriers/chemistry , Emulsions/chemistry , Polyesters/chemistry , Animals , Drug Carriers/toxicity , Drug Resistance, Multiple, Bacterial/drug effects , Emulsions/toxicity , Gram-Negative Bacteria/drug effects , Gram-Negative Bacteria/physiology , Gram-Positive Bacteria/drug effects , Gram-Positive Bacteria/physiology , Mice , Microbial Sensitivity Tests , NIH 3T3 Cells , Polyesters/toxicityABSTRACT
There has been significant interest in the use of peptides as antimicrobial agents, and peptide containing hydrogels have been proposed as biological scaffolds for various applications. Limited stability and rapid clearance of small molecular weight peptides pose challenges to their widespread implementation. As a common approach, antibacterial peptides are physically loaded into hydrogel scaffolds, which leads to continuous release through the passive mode with spatial control but provides limited control over drug dosage. Although utilization of peptide covalent linkage onto hydrogels addresses partially this problem, the peptide release is commonly too slow. To alleviate these challenges, in this work, maleimide-modified antimicrobial peptides are covalently conjugated onto furan-based cryogel (CG) scaffolds via the Diels-Alder cycloaddition at room temperature. The furan group offers a handle for specific loading of the peptides, thus minimizing passive and burst drug release. The porous nature of the CG matrix provides rapid loading and release of therapeutic peptides, apart from high water uptake. Interfacing the peptide adduct containing a CG matrix with a reduced graphene oxide-modified Kapton substrate allows "on-demand" photothermal heating upon near-infrared (NIR) irradiation. A fabricated photothermal device enables tunable and efficient peptide release through NIR exposure to kill bacteria. Apart from spatial confinement offered by this CG-based bandage, the selective ablation of planktonic Staphylococcus aureus is demonstrated. It can be envisioned that this modular "on-demand" peptide-releasing device can be also employed for other topical applications by appropriate choice of therapeutic peptides.
Subject(s)
Anti-Bacterial Agents/pharmacology , Antimicrobial Cationic Peptides/pharmacology , Cryogels/chemistry , Staphylococcal Infections/drug therapy , Anti-Bacterial Agents/chemistry , Antimicrobial Cationic Peptides/chemistry , Cryogels/chemical synthesis , Cryogels/radiation effects , Cycloaddition Reaction , Drug Liberation , Escherichia coli/drug effects , Furans/chemical synthesis , Furans/chemistry , Furans/radiation effects , HeLa Cells , Heating , Humans , Infrared Rays , Methacrylates/chemical synthesis , Methacrylates/chemistry , Methacrylates/radiation effects , Microbial Sensitivity Tests , Polyethylene Glycols/chemical synthesis , Polyethylene Glycols/chemistry , Polyethylene Glycols/radiation effects , Staphylococcus aureus/drug effectsABSTRACT
A myriad of topical therapies and dressings are available to the clinicians for wound healing skin, but only a very few have shown their effectiveness in promoting wound repair due to challenges in controlling drug release. To address this issue, in this work, a near infrared (NIR)-light activable cryogel based on butyl methacrylate (BuMA) and poly(ethylene glycol) methyl ether methacrylate (PEGMEMA) incorporated with reduced graphene oxide (rGO) was fabricated. The obtained cryogel provides the required hydrophilicity beneficial for wound treatment. The excellent photo-thermal properties of rGO allow for heating the cryogel, which results in subsequent swelling of the cryogel (CG) followed by release of the encapsulated drug load, cefepime in our case. Without photothermal activation, no release of payload was observed. The potential of this bandage for wound healing was examined using an ex vivo human skin model infected with Staphylococcus aureus (S. aureus). Apart from the efficacy of the cryogel based wound healing system, our results also suggest that the ex vivo wound model evaluated here provides a rapid and valuable tool to study superficial skin infections in humans and test the efficacy of antimicrobial agents.
Subject(s)
Cryogels , Wound Infection , Anti-Bacterial Agents , Humans , Skin , Staphylococcus aureusABSTRACT
Macroporous cryogels that are amenable to facile functionalization are attractive platforms for biomolecular immobilization, a vital step for fabrication of scaffolds necessary for areas like tissue engineering and diagnostic sensing. In this work, thiol-reactive porous cryogels are obtained via photopolymerization of a furan-protected maleimide-containing poly(ethylene glycol) (PEG)-based methacrylate (PEGFuMaMA) monomer. A series of cryogels are prepared using varying amounts of the masked hydrophilic PEGFuMaMA monomer, along with poly(ethylene glycol) methyl ether methacrylate and poly(ethylene glycol) dimethacrylate, a hydrophilic monomer and cross-linker, respectively, in the presence of a photoinitiator. Subsequent activation to the thiol-reactive form of the furan-protected maleimide groups is performed through the retro Diels-Alder reaction. As a demonstration of direct protein immobilization, bovine serum albumin is immobilized onto the cryogels. Furthermore, ligand-directed immobilization of proteins is achieved by first attaching mannose- or biotin-thiol onto the maleimide-containing platforms, followed by ligand-directed immobilization of concanavalin A or streptavidin, respectively. Additionally, we demonstrate that the extent of immobilized proteins can be controlled by varying the amount of thiol-reactive maleimide groups present in the cryogel matrix. Compared to traditional hydrogels, cryogels demonstrate enhanced protein immobilization/detection. Additionally, it is concluded that utilization of a longer linker, distancing the thiol-reactive maleimide group from the gel scaffold, considerably increases protein immobilization. It can be envisioned that the facile fabrication, conjugation, and control over the extent of functionalization of these cryogels will make these materials desirable scaffolds for numerous biomedical applications.
Subject(s)
Cryogels/chemistry , Immobilized Proteins/chemistry , Methacrylates/chemistry , Polyethylene Glycols/chemistry , Serum Albumin, Bovine/chemistry , Smart Materials/chemistry , Sulfhydryl Compounds/chemistry , Animals , Cattle , Click Chemistry , Cryogels/chemical synthesis , Cycloaddition Reaction , Maleimides/chemical synthesis , Maleimides/chemistry , Methacrylates/chemical synthesis , Polyethylene Glycols/chemical synthesis , Porosity , Smart Materials/chemical synthesisABSTRACT
Multifunctionalizable hydrogel coatings on titanium interfaces are useful in a wide range of biomedical applications utilizing titanium-based materials. In this study, furan-protected maleimide groups containing multi-clickable biocompatible hydrogel layers are fabricated on a titanium surface. Upon thermal treatment, the masked maleimide groups within the hydrogel are converted to thiol-reactive maleimide groups. The thiol-reactive maleimide group allows facile functionalization of these hydrogels through the thiol-maleimide nucleophilic addition and Diels-Alder cycloaddition reactions, under mild conditions. Additionally, the strained alkene unit in the furan-protected maleimide moiety undergoes radical thiol-ene reaction, as well as the inverse-electron-demand Diels-Alder reaction with tetrazine containing molecules. Taking advantage of photo-initiated thiol-ene 'click' reactions, we demonstrate spatially controlled immobilization of the fluorescent dye thiol-containing boron dipyrromethene (BODIPY-SH). Lastly, we establish that the extent of functionalization on hydrogels can be controlled by attachment of biotin-benzyl-tetrazine, followed by immobilization of TRITC-labelled ExtrAvidin. Being versatile and practical, we believe that the described multifunctional and transformable 'clickable' hydrogels on titanium-based substrates described here can find applications in areas involving modification of the interface with bioactive entities.
ABSTRACT
Incorporation of a therapeutic antibody into nanosized drug delivery systems can improve their target specificity. This work reports an antibody-conjugated targeted delivery system composed of polymer-dendron conjugates. Trastuzumab is chosen as the targeting moiety, since it is clinically used against tumor cells expressing HER2 receptors. A micellar delivery system was generated using amphiphilic polymer-dendron conjugates containing a fourth-generation polyester dendron as the hydrophobic block and a linear poly(ethylene glycol) (PEG) chain as the hydrophilic block. After preparation of docetaxel loaded (ca. 10% wt) micelles, trastuzumab was conjugated onto the micellar shell using an amidation reaction. Micelles remained stable after conjugation of the antibody, with a slight increase in size from 179 nm to 185 nm upon functionalization. Docetaxel release was determined to be responsive to acidic pH, and over the course of 30 h, 54% drug release was measured in acidic media, whereas it was around 30% under neutral conditions. Cytotoxicity experiments on MCF-7 and SK-OV-3 cell lines displayed improved toxicity levels for targeted micelles in comparison with the non-targeted counterparts, whereas pulse-chase experiments indicated effectiveness of micellar formulations and the presence of targeting groups. Cellular internalization experiments using fluorescence microscopy and flow cytometry further demonstrated the enhanced cellular uptake of antibody conjugated targeted micelles.