RESUMO
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.
Assuntos
Ciclodextrinas , Nanopartículas , Nanoestruturas , Ciclodextrinas/química , Nanoestruturas/química , Nanopartículas/química , ÁguaRESUMO
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.
Assuntos
Criogéis/química , Proteínas Imobilizadas/química , Metacrilatos/química , Polietilenoglicóis/química , Soroalbumina Bovina/química , Materiais Inteligentes/química , Compostos de Sulfidrila/química , Animais , Bovinos , Química Click , Criogéis/síntese química , Reação de Cicloadição , Maleimidas/síntese química , Maleimidas/química , Metacrilatos/síntese química , Polietilenoglicóis/síntese química , Porosidade , Materiais Inteligentes/síntese químicaRESUMO
Nanogels that are amenable to facile multi-functionalization with imaging, therapeutic, and targeting agents are attractive theranostic platforms for addressing challenges in conventional diagnostics and therapy. In this work, reactive copolymers containing poly(ethylene glycol), maleimide, and pendant hydroxyl groups as side chains are used to construct nanogels by employing their thermoresponsive self-assembly in aqueous media. Subsequent cross-linking of these nanosized aggregates with dithiols using thiol-maleimide chemistry yields nanogels containing maleimide, thiol, and hydroxyl groups. The hydroxyl groups are readily activated to N-hydroxysuccinimide based carbonates that undergo conjugation with amine-containing molecules through carbamate linkage under mild conditions. As a demonstration of multi-functionalization, the maleimide, thiol, and activated carbonate groups were functionalized with a thiol-containing cancer cell targeting peptide, a maleimide-containing fluorescent indocyanine Cy5 dye, and an anticancer drug doxorubicin, respectively. It was observed that enhanced drug release from nanogels occurs under acidic conditions. While the parent nanogel vehicles did not possess any toxicity, drug conjugated constructs with and without targeting group were cytotoxic against MDA-MB-231 breast cancer cells. The cyclic peptide containing targeted nanogel exhibited slightly higher cytotoxicity than its counterpart devoid of any targeting group. Furthermore, higher level of drug internalization into MDA-MB-231 cells was observed for the targeting group containing construct. It can be envisioned that facile fabrication and multi-functionalization of these reactive nanogels simultaneously through nonreversible and reversible linkages offers a modular platform that can be configured as a theranostic agent for addressing challenges in conventional therapy of various diseases.
Assuntos
Antineoplásicos/administração & dosagem , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/tratamento farmacológico , Doxorrubicina/administração & dosagem , Corantes Fluorescentes/administração & dosagem , Géis/química , Nanoestruturas/química , Antineoplásicos/química , Compostos de Boro/administração & dosagem , Compostos de Boro/química , Carbocianinas/administração & dosagem , Carbocianinas/química , Linhagem Celular Tumoral , Doxorrubicina/química , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos , Feminino , Corantes Fluorescentes/química , Humanos , Imagem Óptica/métodos , Polietilenoglicóis/química , Ácidos Polimetacrílicos/química , Nanomedicina Teranóstica/métodosRESUMO
Photosensitizing nanogels were obtained through a surfactant-free single-step protocol by using a porphyrin-based cross-linker for stabilizing self-assembled nanosized aggregates of thermoresponsive copolymers. Nanogels with varying amounts of porphyrin retained the singlet oxygen generation ability of the porphyrin core and were also capable of inducing temperature increase upon irradiation at 635 nm. Photoinduced killing efficiency was tested against three cell lines: human breast adenocarcinoma (MDA-MB-231 and MCF7) and pancreatic adenocarcinoma (AsPC-1) cells, and a predominant photodynamic mechanism at 450 nm and a mixed photodynamic and photothermal effect at 635 nm was observed. This innovative access to photosensitizing nanogels is a proof of concept, and opens new perspectives toward the preparation of optimized nanophotosensitizers.
Assuntos
Géis/química , Nanoestruturas/química , Porfirinas/química , Tensoativos/química , Linhagem Celular Tumoral , Reagentes de Ligações Cruzadas/química , Humanos , Hipertermia Induzida/métodos , Fotoquimioterapia/métodosRESUMO
Multifunctionalizable nanogels are fabricated using thermally driven self-assembly and cross-linking of reactive thermoresponsive copolymers. Nanogels thus fabricated can be easily conjugated with various appropriately functionalized small molecules and/or ligands to tailor them for various applications in delivery and imaging. In this study, a poly(ethylene glycol)-methacrylate-based maleimide-bearing copolymer was cross-linked with a dithiol-based cross-linker to synthesize nanogels. Because of lower critical solution temperature (LCST) around 55 °C in aqueous media, these copolymers assemble into nanosized aggregates when heated to this temperature, and they are cross-linked using the thiol-maleimide conjugation. Nanogels thus fabricated contain both thiol and maleimide groups in the same cross-linked nanogels. Postgelation functionalization of the residual maleimide and thiol groups is demonstrated through conjugation of a thiol-bearing hydrophobic dye (BODIPY-SH) and N-(fluoresceinyl) maleimide, respectively. In addition, to demonstrate the utility of multifunctionality of these nanogels, a thiol-bearing cyclic-peptide-based targeting group, cRGDfC, and N-(fluoresceinyl)-maleimide-based fluorescent tag was conjugated to nanogels in aqueous media. Upon treatment with breast cancer cell lines, MDA-MB-231, it was deduced from cellular internalization studies using fluorescence microscopy and flow cytometry that the peptide carrying constructs were preferentially internalized. Overall, a facile synthesis of multifunctionalizable nanogels that can be tailored using effective conjugation chemistry under mild conditions can serve as promising candidates for various applications.
Assuntos
Neoplasias da Mama/diagnóstico por imagem , Corantes Fluorescentes/química , Maleimidas/química , Polietilenoglicóis/química , Polietilenoimina/química , Polímeros/química , Compostos de Sulfidrila/química , Neoplasias da Mama/patologia , Feminino , Humanos , Interações Hidrofóbicas e Hidrofílicas , Microscopia de Fluorescência , Imagem Molecular , Nanogéis , Polimerização , Temperatura , Células Tumorais CultivadasRESUMO
Advances in medical science have led to diverse new therapeutic modalities, as well as enhanced understanding of the progression of various disease states. These findings facilitate the design and development of more customized and exquisite drug delivery systems that aim to improve therapeutic indices of drugs to treat a variety of conditions. Synthetic polymer-based drug carriers have often been the focus of such research. However, these structures suffer from challenges with heterogeneity of the starting material, limited chemical features, complex functionalization methods, and in some cases a lack of biocompatibility. Consequently, protein-based polymers have garnered much attention in recent years due to their monodisperse features, ease of production and functionalization, and biocompatibility. Genetic engineering techniques enable the advancement of protein-based drug delivery systems with finely tuned physicochemical properties, and thus an expanded level of customization unavailable with synthetic polymers. Of these genetically engineered proteins, elastin-like proteins (ELP), silk-like proteins (SLP), and silk-elastin-like proteins (SELP) provide a unique set of alternatives for designing drug delivery systems due to their inherent chemical and physical properties and ease of engineering afforded by recombinant DNA technologies. In this review we examine the advantages of genetically engineered drug delivery systems with emphasis on ELP and SLP constructions. Methods for fabrication and relevant biomedical applications will also be discussed.
Assuntos
Sistemas de Liberação de Medicamentos/métodos , Elastina/química , Engenharia de Proteínas/métodos , Seda/química , Materiais Biocompatíveis/química , Técnicas de Transferência de Genes , Humanos , Hidrogéis/química , Nanopartículas , Tamanho da Partícula , Proteínas Recombinantes/químicaRESUMO
The sustained release of levonorgestrel, a contraceptive, from silk-based microneedle patches was demonstrated for transdermal delivery. Modifications in the formulation of the silk protein and drug loading enabled the tuning of drug loading and release rates from the microneedle patches over time. Sustained drug release reached up to 100 days when the drug was loaded directly inside the microneedles, while release continued for more than a year when the drug was loaded inside microparticles prior to casting inside the microneedle patches. When coupled with the shelf-stable, refrigeration-less features of the silk protein matrix utilized in the microneedle fabrication, these findings suggest that long-acting contraception patches are feasible. This advance could provide practical options for women to have access to new options for protection against unwanted pregnancy.
RESUMO
Sustained, local delivery of the antibiotic ciprofloxacin under different formats from porous silk protein-based memory foam systems was studied. Similarly, protease XIV was incorporated during processing to provide control of the degradation kinetics of the silk materials. In vitro antibiotic release studies combined with degradation assessments were utilized to assess the mechanisms and kinetics of release from the silk materials. The sequestered protease XIV affected the degradation profiles of the silk foams yet did not impact the release kinetics of the ciprofloxacin, which was controlled by solubility and diffusion of the drug. The ability to tune the release of ciprofloxacin between 1 and 200 days, combined with the option to modulate the degradation rate up to 80% in 2 weeks via incorporation of a protease, suggests utility for drug release devices. Further, we anticipate that this approach could also be extended to other medical implant needs and other drugs.
RESUMO
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.
Assuntos
Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Criogéis/química , Infecções Estafilocócicas/tratamento farmacológico , Antibacterianos/química , Peptídeos Catiônicos Antimicrobianos/química , Criogéis/síntese química , Criogéis/efeitos da radiação , Reação de Cicloadição , Liberação Controlada de Fármacos , Escherichia coli/efeitos dos fármacos , Furanos/síntese química , Furanos/química , Furanos/efeitos da radiação , Células HeLa , Calefação , Humanos , Raios Infravermelhos , Metacrilatos/síntese química , Metacrilatos/química , Metacrilatos/efeitos da radiação , Testes de Sensibilidade Microbiana , Polietilenoglicóis/síntese química , Polietilenoglicóis/química , Polietilenoglicóis/efeitos da radiação , Staphylococcus aureus/efeitos dos fármacosRESUMO
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.
Assuntos
Criogéis , Infecção dos Ferimentos , Antibacterianos , Humanos , Pele , Staphylococcus aureusRESUMO
INTRODUCTION: Silk is a promising biomaterial for controlled delivery of therapeutics and has a unique protein chemistry that can be tuned to form different carrier formats. The protein has been studied for sustained release depot systems for the targeted or localized delivery of drugs. AREAS COVERED: An overview of natural silk proteins for controlled delivery of therapeutics is provided, with a focus on the features of silk proteins that allow them to be useful tools for controlled delivery. Recent applications of natural silk proteins as controlled delivery systems are also summarized. EXPERT OPINION: The versatility of silk proteins makes them desirable biomaterials for a broad range of applications for controlled delivery of both small and large molecules. Further, the degradation profile leading to peptides and amino acids provides compatibility with pH-sensitive therapeutics. While silk sericin and spider silks are under study, silk fibroin extracted from silkworms (e.g. Bombyx mori) dominates pharmaceutical studies with silk. Silk fibroin can be formed into drug delivery tools for systemic or local injections, topical and transdermal applications, and implantation; depending on the target disease and therapeutic molecule. In vitro to in vivo correlations and scale-up needs are the next steps towards clinical applications.