Point-of-care antimicrobial coating protects orthopaedic implants from bacterial challenge.

Implant related infections are the most common cause of joint arthroplasty failure, requiring revision surgeries and a new implant, resulting in a cost of $8.6 billion annually. To address this problem, we created a class of coating technology that is applied in the operating room, in a procedure that takes less than 10 min, and can incorporate any desired antibiotic. Our coating technology uses an in situ coupling reaction of branched poly(ethylene glycol) and poly(allyl mercaptan) (PEG-PAM) polymers to generate an amphiphilic polymeric coating. We show in vivo efficacy in preventing implant infection in both post-arthroplasty infection and post-spinal surgery infection mouse models. Our technology displays efficacy with or without systemic antibiotics, the standard of care. Our coating technology is applied in a clinically relevant time frame, does not require modification of implant manufacturing process, and does not change the implant shelf life.

Click by Click Microporous Annealed Particle (MAP) Scaffolds.

Macroporous scaffolds are being increasingly used in regenerative medicine and tissue repair. While the recently developed microporous annealed particle (MAP) scaffolds have overcome issues with injectability and in situ hydrogel formation, limitations with respect to tunability to be able to manipulate hydrogel strength and rigidity for broad applications still exist. To address these key issues, here hydrogel microparticles (HMPs) of hyaluronic acid (HA) are synthesized using the thiol-norbornene click reaction and then HMPs are subsequently annealed into a porous scaffold using the tetrazine-norbornene click reaction. This assembly method allows for straightforward tuning of bulk scaffold rigidity by varying the tetrazine to norbornene ratio, with increasing tetrazine resulting in increasing scaffold storage modulus, Young's modulus, and maximum stress. These changes are independent of void fraction. Further incorporation of human dermal fibroblasts throughout the porous scaffold reveals the biocompatibility of this annealing strategy as well as differences in proliferation and cell-occupied volume. Finally, injection of porous HA-Tet MAP scaffolds into an ischemic stroke model shows this chemistry is biocompatible in vivo with reduced levels of inflammation and astrogliosis as previously demonstrated for other crosslinking chemistries.

The Use of a Novel Antimicrobial Implant Coating In Vivo to Prevent Spinal Implant Infection.

STUDY DESIGN: A controlled, interventional animal study. OBJECTIVE: Spinal implant infection (SII) is a devastating complication. The objective of this study was to evaluate the efficacy of a novel implant coating that has both a passive antibiotic elution and an active-release mechanism triggered in the presence of bacteria, using an in vivo mouse model of SII. SUMMARY OF BACKGROUND DATA: Current methods to minimize the frequency of SII include local antibiotic therapy (vancomycin powder), betadine irrigation, silver nanoparticles, and passive release from antibiotic-loaded poly(methyl methacrylate) cement beads, all of which have notable weaknesses. A novel implant coating has been developed to address some of these limitations but has not been tested in the environment of a SII. METHODS: A biodegradable coating using branched poly(ethylene glycol)-poly(propylene sulfide) (PEG-PPS) polymer was designed to deliver antibiotics. The in vivo performance of this coating was tested in the delivery of either vancomycin or tigecycline in a previously established mouse model of SII. Noninvasive bioluminescence imaging was used to quantify the bacterial burden, and implant sonication was used to determine bacterial colony-forming units (CFUs) from the implant and surrounding bone and soft tissue. RESULTS: The PEG-PPS-vancomycin coating significantly lowered the infection burden from postoperative day 3 onwards (P < 0.05), whereas PEG-PPS-tigecycline only decreased the infection on postoperative day 5 to 10 (P < 0.05). CFUs were lower on PEG-PPS-vancomycin pins than PEG-PPS-tigecycline and PEG-PPS pins alone on both the implants (2.4 × 10, 8.5 × 10, and 1.0 × 10 CFUs, respectively) and surrounding bone and soft tissue (1.3 × 10, 4.8 × 10, and 5.4 × 10 CFUs, respectively) (P < 0.05). CONCLUSION: The biodegradable PEG-PPS coating demonstrates promise in decreasing bacterial burden and preventing SII. The vancomycin coating outperformed the tigecycline coating in this model compared to prior work in arthroplasty models, highlighting the uniqueness of the paraspinal infection microenvironment. LEVEL OF EVIDENCE: N/A.

Microporous annealed particle hydrogel stiffness, void space size, and adhesion properties impact cell proliferation, cell spreading, and gene transfer.

Designing scaffolds for polyplex-mediated therapeutic gene delivery has a number of applications in regenerative medicine, such as for tissue repair after wounding or disease. Microporous annealed particle (MAP) hydrogels are an emerging class of porous biomaterials, formed by annealing microgel particles to one another in situ to form a porous bulk scaffold. MAP gels have previously been shown to support and enhance proliferative and regenerative behaviors both in vitro and in vivo. Therefore, coupling gene delivery with MAP hydrogels presents a promising approach for therapy development. To optimize MAP hydrogels for gene delivery, we studied the effects of particle size and stiffness as well as adhesion potential on cell surface area and proliferation and then correlated this information with the ability of cells to become transfected while seeded in these scaffolds. We find that the void space size as well as the presentation of integrin ligands influence transfection efficiency. This work demonstrates the importance of considering MAP material properties for guiding cell spreading, proliferation, and gene transfer. STATEMENT OF SIGNIFICANCE: Microporous annealed particle (MAP) hydrogels are an emerging class of porous biomaterials, formed by annealing spherical microgels together in situ, creating a porous scaffold from voids between the packed beads. Here we investigated the effects of MAP physical and adhesion properties on cell spreading, proliferation, and gene transfer in fibroblasts. Particle size and void space influenced spreading and proliferation, with larger particles improving transfection. MAP stiffness was also important, with stiffer scaffolds increasing proliferation, spreading, and transfection, contrasting studies in nonporous hydrogels that showed an inverse response. Last, RGD ligand concentration and presentation modulated spreading similar to non-MAP hydrogels. These findings reveal relationships between MAP properties and cell processes, suggesting how MAP can be tuned to improve future design approaches.

Accelerated wound healing by injectable star poly(ethylene glycol)-b-poly(propylene sulfide) scaffolds loaded with poorly water-soluble drugs.

Injectable hydrogel matrices take the shape of a wound cavity and serve as scaffold for tissue repair and regeneration. Yet these materials are generally hydrophilic, limiting the incorporation of poorly water soluble, hydrophobic drugs. Here we show this shortcoming is circumvented through a star-shaped amphiphilic block copolymer comprising poly(ethylene glycol) and poly (propylene sulfide). This star-shaped amphiphilic polymer self-assembles in an aqueous medium into a physically stable hydrogel and effectively dissolves hydrophobic molecules delivering them at therapeutic doses. The self assembled hydrogel is a robust three-dimensional scaffold in vivo effectively promoting cellular infiltration, reducing inflammation, and wound clsoure. When combined with a hydrophobic BRAF inhibitor that promotes paradoxical mitogen-activated protein kinase (MAPK) activation in keratinocytes and wound closure, our self assembled scaffold supported dermal wound closure at a reduced drug dosage compared to administering the drug in dimethyl sulfoxide (DMSO) without a polymeric matrix. This family of star-shaped amphiphilic polymers delivers poorly water soluble active agents at a fraction of generally required dosage for efficacy and supports three-dimensional cell growth at tissue wounds, showing great promise for novel uses of hydrophobic drugs in tissue repair applications.

o-Nitrobenzyl-Based Photobase Generators: Efficient Photoinitiators for Visible-Light Induced Thiol-Michael Addition Photopolymerization.

In this contribution, three o-nitrobenzyl-based photobase systems were synthesized and evaluated for visible light initiated thiol-Michael addition polymerizations. With a modified structure, the (3,4-methylenedioxy-6-nitrophenyl)-propyloxycarbonyl (MNPPOC) protected base performance exceeds that of the nonsubstituted 2-(2-nitrophenyl)-propyloxycarbonyl (NPPOC) protected base and an ITX sensitized photobase system, with respect to both long-wavelength light sensitivity and photolytic efficiency. In material synthesis, MNPPOC-TMG is capable of initiating photo thiol-Michael polymerization efficiently and orthogonally with only limited visible light exposure and generating a highly homogeneous cross-linked polymer network. This approach enables the thiol-Michael "click" reaction to be conducted with a low-energy, visible light irradiation and, thus, expands its applications in biocompatible and UV sensitive materials.

Productive Exchange of Thiols and Thioesters to Form Dynamic Polythioester-Based Polymers.

Polymers that possess dynamic covalent bonds activated at ambient conditions are ideal platforms for smart, responsive materials. Herein, a class of dynamic covalent polymerizations is developed based on the thiol-thioester exchange, that is, transthioesterification, reaction. Shifts in the equilibrium extent of the exchange reactions are deliberately utilized to drive the formation of oligomers and polymers. In particular, a series of AB and A2-B2 monomers, including amino acid derivatives, were polymerized rapidly by catalytic amounts of mild bases in various organic solvents under ambient conditions. Thioester backbone oligopeptides, including cysteine and glycine, were obtained with an average repeating unit of four and a PDI of 1.8. Further, structurally dynamic hydrogels were obtained by such reactions between four-armed poly(ethylene glycol)-10000 based monomers in neutral water. These hydrogels showed dynamic, frequency-dependent flow behavior between sol and gel states.

A Novel Synthetic UV-Curable Fluorinated Siloxane Resin for Low Surface Energy Coating.

Low surface energy materials have attracted much attention due to their properties and various applications. In this work, we synthesized and characterized a series of ultraviolet (UV)-curable fluorinated siloxane polymers with various fluorinated acrylates-hexafluorobutyl acrylate, dodecafluoroheptyl acrylate, and trifluorooctyl methacrylate-grafted onto a hydrogen-containing poly(dimethylsiloxane) backbone. The structures of the fluorinated siloxane polymers were measured and confirmed by proton nuclear magnetic resonance and Fourier transform infrared spectroscopy. Then the polymers were used as surface modifiers of UV-curable commercial polyurethane (DR-U356) at different concentrations (1, 2, 3, 4, 5, and 10 wt %). Among three formulations of these fluorinated siloxane polymers modified with DR-U356, hydrophobic states (91°, 92°, and 98°) were obtained at low concentrations (1 wt %). The DR-U356 resin is only in the hydrophilic state at 59.41°. The fluorine and siloxane element contents were investigated by X-ray photoelectron spectroscopy and the results indicated that the fluorinated and siloxane elements were liable to migrate to the surface of resins. The results of the friction recovering assays showed that the recorded contact angles of the series of fluorinated siloxane resins were higher than the original values after the friction-annealing progressing.

Implementation of two distinct wavelengths to induce multistage polymerization in shape memory materials and nanoimprint lithography.

Here, a process is introduced for forming dual stage thiol-Michael/acrylate hybrid networks photocured by two different wavelengths, demonstrating its use in nanoimprint lithography (NIL) and shape memory materials. Initiated with a visible light sensitive photobase and a UV-sensitive radical initiator, thiol-Michael-acrylate hybrid polymerizations were programmed to proceed sequentially and orthogonally, with base-catalyzed thiol-Michael photopolymerization as the first stage and radical mediated acrylate photopolymerization as the second stage. By regulating the photopolymerization formulations, i.e. thiol-to-acrylate ratios, initiator loadings and irradiation conditions, a series of materials with highly tunable mechanical performance was achieved, with ultimate Tg values ranging from 23 to 70 °C. With a photopatternable first stage and a readily reconfigurable second stage, its implementation in nanoimprint lithography (NIL) enabled surface features on the scale of 10 nm to be formed on a photopatterned substrate. Additionally, the dual stage polymer results in a relatively homogenous polymer network with a narrow glass transition temperature (Tg), which enables rapid response in applications as shape memory materials, with shape-fixity values above 95% and shaperecovery values above 99%. With its unique photocuring process and programmable mechanical properties, the two color light controlled photopolymerization can be exploited as a useful tool in a wide range of materials science applications.

Wavelength-Selective Sequential Polymer Network Formation Controlled with a Two-Color Responsive Initiation System.

We report a wavelength-selective polymerization process controlled by visible/UV light, whereby a base is generated for anion-mediated thiol-Michael polymerization reaction upon exposure at one wavelength (400-500 nm), while radicals are subsequently generated for a second stage radical polymerization at a second, independent wavelength (365 nm). Dual wavelength, light controlled sequential polymerization not only provides a relatively soft intermediate polymer that facilitates optimum processing and modification under visible light exposure but also enables a highly cross-linked, rigid final material after the UV-induced second stage radical polymerization. A photobase generator, NPPOC-TMG, and a photo-radical initiator, Irgacure 2959, were selected as the appropriate initiator pair for sequential thiol-Michael polymerization and acrylate homopolymerization. FT-IR and rheological tests were utilized to monitor the dual cure photo-polymerization process, and mechanical performance of the polymer was characterized at each distinct stage by dynamic mechanical analysis (DMA). By demonstrating complete light control in another sequential polymerization system (thiol-Michael and thiol-ene hybrid polymerization), this initiator pair exhibits great potential to regulate many other coupled anion and radical hybrid polymerizations in both a sequential and controllable manner.

Single-Dose, Preoperative Vitamin-D Supplementation Decreases Infection in a Mouse Model of Periprosthetic Joint Infection.

BACKGROUND: Despite recent advances, infection remains the most common etiology of arthroplasty failure. Recent work suggests that 25-hydroxyvitamin D (25D) deficiency correlates with the frequency of periprosthetic joint infection (PJI). We endeavored to examine whether 25D3 deficiency leads to increased bacterial burden in vivo in an established mouse model of PJI and, if so, whether this effect can be reversed by preoperative 25D3 supplementation. METHODS: Mice (lys-EGFP) possessing fluorescent neutrophils were fed a vitamin D3-sufficient (n = 20) or deficient (n = 40) diet for 6 weeks. A group of 25D3-deficient mice (n = 20) were "rescued" with 1 intraperitoneal dose of 25D3 at 3 days before surgery. A stainless steel implant was inserted into the knee joint and the joint space was inoculated with bioluminescent Staphylococcus aureus (1 × 10 colony forming units [CFUs]). In vivo imaging was used to monitor bacterial burden and neutrophil infiltration. Blood was drawn to confirm 25D3 levels 3 days before surgery and on postoperative days (PODs) 0 and 14. Mice were killed at POD 21, and CFUs were quantified after culture. Myeloperoxidase (MPO) and ß-N-acetylglucosaminidase (NAG) were assayed to look at neutrophil infiltration and activated tissue macrophage recruitment, respectively. RESULTS: Serum values confirmed 25D3 deficiency and repletion of the 25D3-rescued group. Bacterial bioluminescence and neutrophil fluorescence were significantly greater (p < 0.05) in the 25D3-deficient group. CFU counts from the joint tissue and implant were also significantly greater in this group (p < 0.05). Rescue treatment significantly decreased bacterial burden and neutrophil infiltration (p < 0.05). Compared with the 25D3-sufficient and 25D3-rescued groups, MPO activity was higher (p < 0.02) and NAG activity was lower (p < 0.03) in the 25D3-deficient group. CONCLUSIONS: This study demonstrated in vivo in a mouse model of PJI that (1) 25D3 deficiency results in increased bacterial burden and neutrophil infiltration, and (2) this effect can be reversed with preoperative repletion of 25D3. CLINICAL RELEVANCE: Considering that >65% of patients undergoing arthroplasty have insufficient or low levels of total 25D and that 25D levels can be replenished with ease using a U.S. Food and Drug Administration (FDA)-approved, oral 25D3 product, 25D deficiency may be an important modifiable risk factor in humans undergoing joint replacement.

Nucleobase-Containing Polymers: Structure, Synthesis, and Applications.

Nucleobase interactions play a fundamental role in biological functions, including transcription and translation. Natural nucleic acids like DNA are also widely implemented in material realm such as DNA guided self-assembly of nanomaterials. Inspired by that, polymer chemists have contributed phenomenal endeavors to mimic both the structures and functions of natural nucleic acids in synthetic polymers. Similar sequence-dependent responses were observed and employed in the self-assembly of these nucleobase-containing polymers. Here, the structures, synthetic approaches, and applications of nucleobase-containing polymers are highlighted and a brief look is taken at the future development of these polymers.

Visible-Light-Initiated Thiol-Michael Addition Polymerizations with Coumarin-Based Photobase Generators: Another Photoclick Reaction Strategy.

An efficient visible-light-sensitive photobase generator for thiol-Michael addition reactions was synthesized and evaluated. This highly reactive catalyst was designed by protecting a strong base (tetramethyl guanidine, TMG) with a visible-light-responsive group which was a coumarin derivative. The coumarin-coupled TMG was shown to exhibit extraordinary catalytic activity toward initiation of the thiol-Michael reaction, including thiol-Michael addition-based polymerization, upon visible-light irradiation, leading to a stoichiometric reaction of both thiol and vinyl functional groups. Owing to its features, this visible-light photobase generator enables homogeneous network formation in thiol-Michael polymerizations and also has the potential to be exploited in other visible-light-induced, base-catalyzed thiol-click processes such as thiol-isocynate and thiol-epoxy network-forming reactions.

Clickable Nucleic Acids: Sequence-Controlled Periodic Copolymer/Oligomer Synthesis by Orthogonal Thiol-X Reactions.

Synthetic polymer approaches generally lack the ability to control the primary sequence, with sequence control referred to as the holy grail. Two click chemistry reactions were now combined to form nucleobase-containing sequence-controlled polymers in simple polymerization reactions. Two distinct approaches are used to form these click nucleic acid (CNA) polymers. These approaches employ thiol-ene and thiol-Michael reactions to form homopolymers of a single nucleobase (e.g., poly(A)n ) or homopolymers of specific repeating nucleobase sequences (e.g., poly(ATC)n). Furthermore, the incorporation of monofunctional thiol-terminated polymers into the polymerization system enables the preparation of multiblock copolymers in a single reaction vessel; the length of the diblock copolymer can be tuned by the stoichiometric ratio and/or the monomer functionality. These polymers are also used for organogel formation where complementary CNA-based polymers form reversible crosslinks.

Experimental and theoretical photoluminescence studies in nucleic acid assembled gold-upconverting nanoparticle clusters.

Combinations of rare earth doped upconverting nanoparticles (UCNPs) and gold nanostructures are sought as nanoscale theranostics due to their ability to convert near infrared (NIR) photons into visible light and heat, respectively. However, because the large NIR absorption cross-section of the gold coupled with their thermo-optical properties can significantly hamper the photoluminescence of UCNPs, methods to optimize the ratio of gold nanostructures to UCNPs must be developed and studied. We demonstrate here nucleic acid assembly methods to conjugate spherical gold nanoparticles (AuNPs) and gold nanostars (AuNSs) to silica-coated UCNPs and probe the effect on photoluminescence. These studies showed that while UCNP fluorescence enhancement was observed from the AuNPs conjugated UCNPs, AuNSs tended to quench fluorescence. However, conjugating lower ratios of AuNSs to UCNPs led to reduced quenching. Simulation studies both confirmed the experimental results and demonstrated that the orientation and distance of the UCNP with respect to the core and arms of the gold nanostructures played a significant role in PL. In addition, the AuNS-UCNP assemblies were able to cause rapid gains in temperature of the surrounding medium enabling their potential use as a photoimaging-photodynamic-photothermal agent.

Spatial and Temporal Control of Thiol-Michael Addition via Photocaged Superbase in Photopatterning and Two-Stage Polymer Networks Formation.

Photochemical processes enable spatial and temporal control of reactions, which can be implemented as an accurate external control approach in both polymer synthesis and materials applications. "Click" reactions have also been employed as efficient tools in the same field. Herein, we combined photochemical processes and thiol-Michael "click" reactions to achieve a "photo-click" reaction that can be used in surface patterning and controlled polymer network formation, owing to the ease of spatial and temporal control through use of photolabile amines as appropriate catalysts.

A new photoclick reaction strategy: photo-induced catalysis of the thiol-Michael addition via a caged primary amine.

The utilization of 2-(2-nitrophenyl)propyloxycarbonyl (NPPOC) as a photolabile primary amine cage enables the thiol-Michael 'click' reaction to be photo-triggered. The photolabile amine exhibits efficient catalytic activity upon UV irradiation and is shown to initiate the photopolymerization of tetrathiol and diacrylate comonomers via Michael addition.

Nitrogen-Centered Nucleophile Catalyzed Thiol-Vinylsulfone Addition, Another Thiol-ene "Click" Reaction.

A new group of nitrogen-centered nucleophilic catalysts for the thiol-Michael addition "click" reactions is examined. These nucleophiles showed efficient catalytic activities as compared with traditional base catalysts, such as triethylamine, and are demonstrated to be a viable strategy for cross-linking polymerization reactions. Additionally, an experimental and computational mechanistic study was performed, suggesting a pathway for the nitrogen-centered catalyst to undergo the nucleophilic addition mechanism.