Customized hybrid and NIR-light triggered thermoresponsive drug delivery microparticles synthetized by photopolymerization in a one-step flow focusing continuous microreactor.

Photopolymerization is a selective technique that takes advantage of light-sensitive molecules to initiate and propagate monomeric structures to render covalently bonded macromolecular materials structures known as polymers. Herein, we present a novel one-step microfluidic synthesis of customized hybrid-thermoresponsive Poly(N-isopropylacrylamide) (PNIPAm) based microparticles (MPs) containing plasmonic hollow gold nanoparticles (HGNPs) and bupivacaine (BVP) used as a model drug. Those hybrid microparticles were prepared using a flow-focusing microreactor coupled to a UV LED device built with a simple outer PTFE tubing and an inner flexible capillary. Different tubing characteristics and flow rate ratios were altered in order to control the size of the resulting microparticles. In addition, components such as monomer, crosslinker and photoinitiator concentrations, as well as LED intensity and irradiation time were tuned to obtain different MPs and their characteristics and polymerization rates were compared by Gel Permeation Chromatography (GPC). Thermoresponsive properties were analyzed and the presence of HGNPs was confirmed in light-activated triggered drug release applications. Bupivacaine loading and release studies were evaluated with the resulting hollow and solid microparticles (which were obtained depending on the polymerization rate used) and their temperature responsiveness was assessed using a NIR laser when HGNPs were present in the constructs. Finally, cytotoxicity studies, cell-cycle arrest and apoptotic induction were carried out to certify their suitability for further biomedical applications to be used as triggerable drug depots.

Solvation dynamics of N-substituted acrylamide polymers and the importance for phase transition behavior.

Functional groups present in thermo-responsive polymers are known to play an important role in aqueous solutions by manifesting their coil-to-globule conformational transition in a specific temperature range. Understanding the role of these functional groups and their interactions with water is of great interest as it may allow us to control both the nature and temperature of this coil-to-globule transition. In this work, polyacrylamide (PAAm), poly(N-isopropylacrylamide) (PNIPAm), and poly(N-isopropylmethacrylamide) (PNIPMAm) solvated in water are studied with the goal of discovering the structure of the solvent and its interaction with these polymers in determining the polymer conformations. Specifically, all-atom molecular dynamics (MD) simulations were performed on polymer chains with 30 monomer units (30-mers) at 295 K, 310 K and 320 K, which is below and above the lower critical solution temperature (LCST) of PNIPAm (LCST = 305 K) and PNIPMAm (LCST = 315 K), respectively. The MD simulation trajectories suggest that changes in the functional groups in the backbone and side-chains alter the water solvation shell around the polymer. This results in a change in the residence time probability and hydrogen bond characteristics of water at simulated temperatures. Specifically, water molecules reside for longer times near PAAm (no LCST) and PNIPMAm (LCST = 315 K) chains as compared to PNIPAm. This might be one of the possible causes for the higher LCST of PNIPMAm as compared to that of PNIPAm. These results can guide experimentalists and theoreticians to design new polymer structures with tailor-made LCST transitions while controlling the water solvation shell around the functional group.

Cleavable and thermo-responsive hybrid nanoparticles for on-demand drug delivery.

By combining the photothermal ability of copper sulphide nanoparticles (NPs) upon excitation with Near Infrared (NIR) Light and the thermo-responsive properties of the homemade oligo (ethylene glycol) methyl ether methacrylate copolymer we have obtained fragmentable nanocomposites able to release a carried drug on-demand after NIR-light triggering. A complete physico-chemical characterization of the resulting nanoparticles has been carried out and their degradation assessed at different temperatures. Herein, we have also evaluated the drug loading capacity of those nanoparticles and the temperature dependence in their drug release kinetics using bupivacaine hydrochloride as a model drug. For those hybrid nanoparticles, subcytotoxic doses on four different cell lines and their potential interference in cell metabolism, induction of apoptosis, and cell cycle have been evaluated by Alamar Blue fluorometry and flow cytometry.

Near infrared dye-labelled polymeric micro- and nanomaterials: in vivo imaging and evaluation of their local persistence.

The use of micro- and nanomaterials as carriers of therapeutic molecules can enhance the efficiency of treatments while avoiding side effects thanks to the development of controlled drug delivery systems. The binding of a dye to a drug or to a drug carrier has opened up a wide range of possibilities for an effective in vivo optical tracing of drug biodistribution by using non-invasive real-time technologies prior to their potential use as therapeutic vectors. Here, we describe the fluorescent tagging of polymeric micro- and nanomaterials based on poly(lactic-co-glycolic) acid and on the thermoresponsive poly(N-isopropylacrylamide) with the fluorescent probe IR-820 which was chemically modified for its covalent coupling to the materials. The chemical modification of the dye and the polymers yielded micro- and nanoparticulated labelled materials to be potentially used as drug depots of different therapeutic molecules. In vitro biological studies revealed their reduced cytotoxicity. A spatiotemporal in vivo micro- and nanoparticle tracking allowed the evaluation of the biodistribution of materials showing their local persistence and high biocompatibility after pathological studies. These results underline the suitability of these materials for the local, sustained, not harmful and/or on-demand drug delivery and the remarkable importance of evaluating the biodistribution of materials and tissue persistence for their use as local drug depots.

Microfluidic Synthesis and Biological Evaluation of Photothermal Biodegradable Copper Sulfide Nanoparticles.

The continuous synthesis of biodegradable photothermal copper sulfide nanoparticles has been carried out with the aid of a microfluidic platform. A comparative physicochemical characterization of the resulting products from the microreactor and from a conventional batch reactor has been performed. The microreactor is able to operate in a continuous manner and with a 4-fold reduction in the synthesis times compared to that of the conventional batch reactor producing nanoparticles with the same physicochemical requirements. Biodegradation subproducts obtained under simulated physiological conditions have been identified, and a complete cytotoxicological analysis on different cell lines was performed. The photothermal effect of those nanomaterials has been demonstrated in vitro as well as their ability to generate reactive oxygen species.

Continuous synthesis of drug-loaded nanoparticles using microchannel emulsification and numerical modeling: effect of passive mixing.

By using interdigital microfluidic reactors, monodisperse poly(d,l lactic-co-glycolic acid) nanoparticles (NPs) can be produced in a continuous manner and at a large scale (~10 g/h). An optimized synthesis protocol was obtained by selecting the appropriated passive mixer and fluid flow conditions to produce monodisperse NPs. A reduced NP polydispersity was obtained when using the microfluidic platform compared with the one obtained with NPs produced in a conventional discontinuous batch reactor. Cyclosporin, an immunosuppressant drug, was used as a model to validate the efficiency of the microfluidic platform to produce drug-loaded monodisperse poly(d,l lactic-co-glycolic acid) NPs. The influence of the mixer geometries and temperatures were analyzed, and the experimental results were corroborated by using computational fluid dynamic three-dimensional simulations. Flow patterns, mixing times, and mixing efficiencies were calculated, and the model supported with experimental results. The progress of mixing in the interdigital mixer was quantified by using the volume fractions of the organic and aqueous phases used during the emulsification-evaporation process. The developed model and methods were applied to determine the required time for achieving a complete mixing in each microreactor at different fluid flow conditions, temperatures, and mixing rates.