RESUMO
The nonlinear elasticity of many tissue-specific extracellular matrices is difficult to recapitulate without the use of fibrous architectures, which couple strain-stiffening with stress relaxation. Herein, bottlebrush polymers are synthesized and crosslinked to form poly(ethylene glycol)-based hydrogels and used to study how strain-stiffening behavior affects human mesenchymal stromal cells (hMSCs). By tailoring the bottlebrush polymer length, the critical stress associated with the onset of network stiffening is systematically varied, and a unique protrusion-rich hMSC morphology emerges only at critical stresses within a biologically accessible stress regime. Local cell-matrix interactions are quantified using 3D traction force microscopy and small molecule inhibitors are used to identify cellular machinery that plays a critical role in hMSC mechanosensing of the engineered, strain-stiffening microenvironment. Collectively, this study demonstrates how covalently crosslinked bottlebrush polymer hydrogels can recapitulate strain-stiffening biomechanical cues at biologically relevant stresses and be used to probe how nonlinear elastic matrix properties regulate cellular processes.
Assuntos
Actomiosina , Elasticidade , Hidrogéis , Células-Tronco Mesenquimais , Polietilenoglicóis , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/efeitos dos fármacos , Hidrogéis/química , Hidrogéis/farmacologia , Humanos , Actomiosina/metabolismo , Polietilenoglicóis/química , Polímeros/química , Polímeros/farmacologia , Matriz Extracelular/metabolismo , Matriz Extracelular/químicaRESUMO
Nanodroplet formation is important to achieve supersaturation of active pharmaceutical ingredients (APIs) in an amorphous solid dispersion. The aim of the current study was to explore how polymer composition, architecture, molar mass, and surfactant concentration affect polymer-drug nanodroplet morphology with the breast cancer API, GDC-0810. The impact of nanodroplet size and morphology on dissolution efficacy and drug loading capacity was explored using polarized light microscopy, dynamic light scattering, and cryogenic transmission electron microscopy. Poly(N-isopropylacrylamide-stat-N,N-dimethylacrylamide) (PND) was synthesized as two linear derivatives and two bottlebrush derivatives with carboxylated or PEGylated end-groups. Hydroxypropyl methylcellulose acetate succinate grade MF (HPMCAS-MF) and poly(vinylpyrrolidone-co-vinyl acetate) (PVPVA) were included as commercial polymer controls. We report the first copolymerization synthesis of a PVPVA bottlebrush copolymer, which was the highest performing excipient in this study, maintaining 688 µg/mL GDC-0810 concentration at 60 wt % drug loading. This is likely due to strong polymer-drug noncovalent interactions and the compaction of GDC-0810 along the PVPVA bottlebrush backbone. Overall, it was observed that the most effective formulations had a hydrodynamic radius less than 25 nm with tightly compacted nanodroplet morphologies.
Assuntos
Cinamatos , Indazóis , Polímeros , Povidona/análogos & derivados , SolubilidadeRESUMO
A cationic unimolecular bottlebrush polymer with chemically modified end-groups was synthesized to understand the impact of hydrophilicity on colloidal stability, nucleic acid delivery performance, and toxicity. The bottlebrush polymer template was synthesized using grafting-through techniques and was therefore composed of a polynorbornene backbone with poly(2-(dimethylamino)ethyl methacrylate) side chains with dodecyl trithiocarbonate end-groups. Postpolymerization modification was performed to fully remove the end-groups or install hydroxy and methoxy poly(ethylene glycol) functional groups on the bottlebrush exterior. The bottlebrush family was preformulated with biological payloads of pDNA and CRISPR-Cas9 RNP in both water and PBS to understand binding, aggregation kinetics, cytotoxicity, and delivery efficacy. Increasing end-group hydrophilicity and preformulation of bottleplexes in PBS increased colloidal stability and cellular viability; however, this did not always result in increased transfection efficiency. The bottlebrush family exemplifies how formulation conditions, polymer loading, and end-group functionality of bottlebrushes can be tuned to balance expression with cytotoxicity ratios and result in enhanced overall performance.
Assuntos
Metacrilatos , Polímeros , Interações Hidrofóbicas e Hidrofílicas , Cátions , Transfecção , Polímeros/química , Metacrilatos/químicaRESUMO
Bottlebrush polymers have great potential as vehicles to noncovalently sequester, stabilize, and deliver hydrophobic small molecule actives. To this end, we synthesized a poly(N-isopropylacrylamide-stat-N,N-dimethylacrylamide) bottlebrush copolymer using ring-opening metathesis polymerization and developed a facile method to control the thermoresponsive properties using postpolymerization modification. Six increasingly hydrophilic end-groups were installed, yielding cloud point temperature control over a range of 22-42 °C. Solubility enhancement of the antiseizure medication, phenytoin, increased significantly with the hydrophilicity of the end-group moiety. Notably, carboxylated bottlebrush copolymers solubilized formulations with higher drug loadings than linear copolymers because they exist as unimolecular nanoparticles with a synthetically defined density of polymer chains that are more stable in solution. This work provides the first investigation of bottlebrush polymers for hydrophobic noncovalent sequestration and solubilization of pharmaceuticals.
Assuntos
Excipientes , Polímeros , Excipientes/química , Interações Hidrofóbicas e Hidrofílicas , Polimerização , Polímeros/química , SolubilidadeRESUMO
Thin-film formation and transport properties of two copper-paddlewheel metal-organic framework (MOF) -based systems (MOF-14 and MOF-399) are investigated for their potential integration into electrochemical device architectures. Thin-film analogs of these two systems are fabricated by the sequential, alternating, solution-phase deposition of the inorganic and organic ligand precursors that result in conformal films via van der Merwe-like growth. Atomic force microscopy reveals smooth film morphologies with surface roughnesses determined by the underlying substrates and linear film growth of 1.4 and 2.2 nm per layer for the MOF-14 and MOF-399 systems, respectively. Electrochemical impedance spectroscopy is used to evaluate the electronic transport properties of the thin films, finding that the MOF-14 analog films demonstrate low electronic conductivity, while MOF-399 analog films are electronically insulating. The intrinsic porosities of these ultrathin MOF analog films are confirmed by cyclic voltammetry redox probe characterization using ferrocene. Larger peak currents are observed for MOF-399 analog films compared to MOF-14 analog films, which is consistent with the larger pores of MOF-399. The layer-by-layer deposition of these systems provides a promising route to incorporate MOFs as thin films with nanoscale thickness control and low surface roughness for electrochemical devices.
RESUMO
Metal-organic frameworks (MOFs) are extremely porous, crystalline materials with high surface area for potential use in gas storage, sequestration, and separations. Toward incorporation into structures for these applications, this study compares three variations of surface-bound and free-standing HKUST-1 MOF structures: surface-anchored MOF (surMOF) thin film, drop-cast film, and bulk powder. Herein, effects of HKUST-1 ammonia interaction and framework activation, which is removal of guest molecules via heat, are investigated. Impact on morphology and crystal structure as a function of surface confinement and size variance are examined. Scanning probe microscopy, scanning electron microscopy, powder X-ray diffraction, Fourier-transform infrared spectroscopy, and energy dispersive X-ray spectroscopy monitor changes in morphology and crystal structure, track ammonia uptake, and examine elemental composition. After fabrication, ammonia uptake is observed for all MOF variations, but reveals dramatic morphological and crystal structure changes. However, activation of the framework was found to stabilize morphology. For activated surMOF films, findings demonstrate consistent morphology throughout uptake, removal, and recycling of ammonia over multiple exposures. To understand morphological effects, additional ammonia exposure experiments with controlled post-synthetic solvent adsorbates were conducted utilizing a HKUST-1 standard powder. These findings are foundational for determining the capabilities and limitation of MOF films and powders.
RESUMO
Thin films can integrate the versatility and great potential found in the emerging field of metal-organic frameworks directly into device architectures. For fabrication of smart interfaces containing surface-anchored metal-organic frameworks, it is important to understand how the foundational layers form to create the interface between the underlying substrate and porous framework. Herein, the formation and morphology of the first ten cycles of film deposition are investigated for the well-studied HKUST-1 system. Effects of processing variables, such as deposition temperature and substrate quality, are studied. Sequences of scanning probe microscopy images collected after cycles of alternating solution-phase deposition reveal the formation of a discontinuous surface with nucleating and growing crystallites consistent with a Volmer-Weber growth mechanism. Quantitative image analysis determines surface roughness and surface coverage as a function of deposition cycles, producing insight regarding growth and structure of foundational film layers. For carboxylic acid terminated self-assembled monolayers on gold, preferred crystal orientation is influenced by deposition temperature with crystal growth along [100] observed at 25 °C and [111] favored at 50 °C. This difference in crystal orientation results in reduced surface roughness and increased surface coverage at 50 °C. To properly fabricate and fully determine the potential of this material for industrial applications, fundamental understanding of film formation is crucial.