Research of Global Tilt and Functional Independence: Insights into Spinal Health of Older Women.

Spinal alignment intricately influences functional independence, particularly in older women with osteopenia experiencing mild neck and back pain. This study elucidates the interplay between spinal alignment, bone mineral density (BMD), and muscle strength in elderly women presenting with mild neck and back pain. Focusing on a cohort of 189 older women, we examined the associations among global tilt (GT), coronal and sagittal alignment, BMD, grip strength, and functional independence as gauged by the Barthel index. Our findings indicate significant associations between functional capacity and grip strength, bone density, GT, and pelvic tilt (PT). Elderly women with a Barthel Index above 80 demonstrated higher grip strength and better bone quality, reflected by less negative average T scores. These individuals also exhibited lower values of GT and PT, suggesting a better sagittal alignment compared to those with a Barthel index of 80 or below. The results highlight that deviations in GT and PT are significantly associated with decreased functional independence. These insights emphasize the importance of maintaining optimal spinal alignment and muscle strength to support functional independence in elderly women. This study underscores the potential for targeted interventions that improve postural stability and manage pain effectively in this vulnerable population.

Myopia progression after cessation of atropine in children: a systematic review and meta-analysis.

Purpose: To comprehensively assess rebound effects by comparing myopia progression during atropine treatment and after discontinuation. Methods: A systematic search of PubMed, EMBASE, Cochrane CENTRAL, and ClinicalTrials.gov was conducted up to 20 September 2023, using the keywords "myopia," "rebound," and "discontinue." Language restrictions were not applied, and reference lists were scrutinized for relevant studies. Our study selection criteria focused on randomized control trials and interventional studies involving children with myopia, specifically those treated with atropine or combination therapies for a minimum of 6 months, followed by a cessation period of at least 1 month. The analysis centered on reporting annual rates of myopia progression, considering changes in spherical equivalent (SE) or axial length (AL). Data extraction was performed by three independent reviewers, and heterogeneity was assessed using I2 statistics. A random-effects model was applied, and effect sizes were determined through weighted mean differences with 95% confidence intervals Our primary outcome was the evaluation of rebound effects on spherical equivalent or axial length. Subgroup analyses were conducted based on cessation and treatment durations, dosage levels, age, and baseline SE to provide a nuanced understanding of the data. Results: The analysis included 13 studies involving 2060 children. Rebound effects on SE were significantly higher at 6 months (WMD, 0.926 D/y; 95%CI, 0.288-1.563 D/y; p = .004) compared to 12 months (WMD, 0.268 D/y; 95%CI, 0.077-0.460 D/y; p = .006) after discontinuation of atropine. AL showed similar trends, with higher rebound effects at 6 months (WMD, 0.328 mm/y; 95%CI, 0.165-0.492 mm/y; p < .001) compared to 12 months (WMD, 0.121 mm/y; 95%CI, 0.02-0.217 mm/y; p = .014). Sensitivity analyses confirmed consistent results. Shorter treatment durations, younger age, and higher baseline SE levels were associated with more pronounced rebound effects. Transitioning or stepwise cessation still caused rebound effects but combining optical therapy with atropine seemed to prevent the rebound effects. Conclusion: Our meta-analysis highlights the temporal and dose-dependent rebound effects after discontinuing atropine. Individuals with shorter treatment durations, younger age, and higher baseline SE tend to experience more significant rebound effects. Further research on the rebound effect is warranted. Systematic Review Registration: [https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=463093], identifier [registration number].

Optimizing Dental Bond Strength: Insights from Comprehensive Literature Review and Future Implications for Clinical Practice.

This review examines the modifying factors affecting bond strength in various bonding scenarios, particularly their relevance to the longevity of dental restorations. Understanding these factors is crucial for improving clinical outcomes in dentistry. Data were gathered from the PubMed database, ResearchGate, and Google Scholar resources, covering studies from 1992 to 2022. The findings suggest that for dentin-resin bonds, minimizing smear layers and utilizing MMP inhibitors to prevent hybrid layer degradation are essential. In the case of resin-resin bonds, reversing blood contamination is possible, but preventing saliva contamination is more challenging, underscoring its critical importance during clinical procedures. Additionally, while pretreatment on ceramics has minimal impact on bond strength, the influence of specific colorings should be carefully considered in treatment planning. This comprehensive review highlights that although established practices recognize significant bond strength factors, ongoing research provides valuable insights to enhance the clinical experience for patients. Once confirmed through rigorous experimentation, these emerging findings should be swiftly integrated into dental practice to improve patient outcomes.

Conformal Conductive Features on Curvilinear Surfaces with Self-Assembled Silver Nanoplate Thin Films.

In this study, a water transfer method was developed to fabricate conducive thin-film patterns on 3D curvilinear surfaces. Crystalline silver nanoplates (AgNPLs) with a dimension of 700 nm and a thickness of 35 nm were suspended in ethanol with an anionic surfactant, sodium dodecyl sulfate, to improve the suspension stability. The prepared AgNPL suspension was then spread over the water surface via the Langmuir-Blodgett approach to generate a self-assembled thin film. By dipping an accepting object with a robotic arm, the floating AgNPL thin film with nanometer thickness can be effectively transferred to the object surfaces and exhibited a superior conductivity up to 15% of bulk silver without thermal sintering. Besides good conductivity, the AgNPL conductive thin films can also be transferred efficiently on any curvilinear (concave and convex) surface. Moreover, with the help of masks, conductive patterns can be produced on water surfaces and transferred to curvilinear surfaces for electronic applications. As a proof of concept, several examples were demonstrated to display the capability of this approach for radiofrequency identification and other printed circuit applications.

Systematic Review of Myopia Progression after Cessation of Optical Interventions for Myopia Control.

Despite high discontinuation rates for myopia optical interventions, limited attention has been given to the potential rebound effects post-discontinuation. This systematic review aims to assess the extent of the rebound effects following the cessation of common clinical optical myopia-control interventions in children. A comprehensive search of PubMed, Embase, Cochrane CENTRAL, and ClinicalTrials.gov was conducted from inception to October 2023. The rebound effects, defined as changes in the axial length or spherical equivalent during and after treatment cessation, were categorized into four levels. These studies encompassed 703 participants and spanned from 2019 to 2023, with durations of treatment and cessation ranging from 6 months to 3.5 years and from 2 weeks to 5 years, respectively. This review, encompassing 14 studies, revealed a predominant strong rebound effect in orthokeratology (8 studies), a weak rebound effect in multifocal soft contact lenses (4 studies), and a variable rebound effect in peripheral-plus spectacle lenses (2 studies). Notably, with the increasing cessation duration, the rebound effects diminished, potentially linked to the reversal of choroidal thickening and the disappearance of peripheral myopic defocus. In conclusion, a temporal trend of rebound effects exists in all three myopia optical interventions, possibly contributing to their myopia control mechanisms.

Enhancing Stability of High-Concentration ß-Tricalcium Phosphate Suspension for Biomedical Application.

We propose a novel process to efficiently prepare highly dispersed and stable Tricalcium Phosphate (ß-TCP) suspensions. TCP is coupled with a polymer to enhance its brittleness to be used as an artificial hard tissue. A high solid fraction of ß-TCP is mixed with the polymer in order to improve the mechanical strength of the prepared material. The high solid fractions led to fast particle aggregation due to Van der Waals forces, and sediments appeared quickly in the suspension. As a result, we used a dispersant, dispex AA4040 (A40), to boost the surface potential and steric hindrance of particles to make a stable suspension. However, the particle size of ß-TCP is too large to form a suspension, as the gravity effect is much more dominant than Brownian motion. Hence, ß-TCP was subjected to wet ball milling to break the aggregated particles, and particle size was reduced to ~300 nm. Further, to decrease sedimentation velocity, cellulose nanocrystals (CNCs) are added as a thickening agent to increase the overall viscosity of suspension. Besides the viscosity enhancement, CNCs were also wrapped with A40 micelles and increase the stability of the suspension. These CNC/A40 micelles further facilitated stable suspension of ß-TCP particles with an average hydration radius of 244.5 nm. Finally, ß-TCP bone cement was formulated with the suspension, and the related cytotoxicity was estimated to demonstrate its applicability for hard tissue applications.

Microwave-Assisted Synthesis for Silver Nanoplates with a High Aspect Ratio.

In this work, a simple and rapid synthesis method was developed to prepare silver nanoplates (AgNPLs) with a high aspect ratio. A microwave heating process with a high heating rate and uniform heating was used to promote the silver reduction reaction. Silver nitrate (AgNO3) was used as the precursor of AgNPLs, and N,N-dimethylformamide (DMF) played the role of a solvent and reducing agent. Poly(vinylpyrrolidone) (PVP) with a molecular weight of 29,000 and a PVP/AgNO3 ratio of 10 were used to control the shape of synthesized AgNPLs. By adjusting the optimal microwave heating parameters, temperature ramping rate, reaction time, and reaction temperature, triangular AgNPLs with high aspect ratios could be produced. The synthesized AgNPLs had an edge length up to 700 nm and a thickness of 35 nm with aspect ratios up to 20. The AgNPLs were also used to produce conductive patterns via pen writing with a conductivity of 2 × 106 S/m to demonstrate the feasibility of applying the synthesized nanomaterials for electronic applications.

A Leaf Disc Assay for Evaluating the Response of Tea (Camellia sinensis) to PEG-Induced Osmotic Stress and Protective Effects of Azoxystrobin against Drought.

Tea (Camellia sinensis), a globally cultivated beverage crop, is sensitive to drought, which can have an adverse effect on the yield and quality of tea. Azoxystrobin (AZ) is one kind of fungicide considered as an agent to relieve damage caused by stress. Initially, the response of tea plant to osmotic-gradient stress was evaluated using leaf disc assays with PEG-induced osmotic stress. The decline of the maximum quantum yield of PSII (Fv/Fm), actual photosynthetic efficiency of PS II (Y(II)), total chlorophylls, carotenoids, DPPH radical scavenging capacity, reducing power, total phenols, and the increase in MDA was observed in leaf discs treated with a gradient of PEG solutions (22.8, 33.2, 41.1% PEG, and blank). These results revealed that efficiency of photosystem II (PSII), photosynthetic pigments, and antioxidant ability in leaf discs were inhibited with an aggravated lipid peroxidation under PEG-induced osmotic stress, and indicated leaf disc assay with moderate PEG iso-osmotic condition would reflect a portion of tea plant response to drought stress. Therefore, the protective effect of AZ (0.125 and 1.25 g a.i. L-1) on tea plants suffering from drought was evaluated using leaf disc assays with 22.8% PEG iso-osmotic condition. Pretreatment of AZ (0.125 a.i. g L-1) reversed Fv/Fm, Y(II), DPPH radical scavenging capacity, and reducing power with reduced MDA in PEG-treated leaf discs, but photosynthetic pigments, total phenols, and ascorbate peroxidase activity were irresponsive to AZ. An Alleviated physiological damage in tea leaf with AZ applying was preliminarily revealed in this study. A Rapid screening of agents for tea plants against drought was developed to assist in the selection of protective agents.

An Ultraviolet Sensor and Indicator Module Based on p-i-n Photodiodes.

The monolithic integration of an ultraviolet (UV) sensor and warning lamp would reduce the cost, volume, and footprint, in comparison to a hybrid combination of discrete components. We constructed a module comprising a monolithic sensor indicator device based on basic p-i-n (PIN) photodiodes and a transimpedance amplifier. GaN-based light-emitting diodes (LEDs) with an indium-tin oxide (ITO) current-spreading layer and PIN photodiodes without ITO deposition on the light-receiving area, were simultaneously fabricated. The resultant incident photon-to-electron conversion efficiencies of the PIN photodiodes at UV wavelengths were significantly higher than those of the reverse-biased LEDs. The photocurrent signals of the PIN photodiode were then converted to voltage signals to drive an integrated visible LED, which functioned as an indicator. The more the ambient UV-light intensity exceeded a specified level, the brighter the glow of the LED. The responsivities of 0.20 and 0.16 A/W were obtained at 381 and 350 nm, respectively, under a bias voltage of 5 V. We also addressed the epitaxial structural details that can affect the collection efficiency of the photocurrent generated by UV light absorption. The crosstalk between the PIN photodiode and LEDs (of various center-to-center distances) was measured.

Monolithic integration of GaN-based phototransistors and light-emitting diodes.

Monolithic integration of GaN-based phototransistors and light-emitting diodes (LEDs) is reported. Starting with an LED epitaxial wafer, selective Si diffusion was performed to produce an n-p-i-n structure for the phototransistor. A traditional AlGaN bulk electron-blocking layer (EBL) can block electron injection from an emitter to a collector, thereby hindering the photocurrent amplification process. We used an LED wafer with a superlattice EBL; blocking can be removed under a bias of approximately 7 V and above. External quantum efficiencies of more than 100% and 600% at approximately 380 nm and 330 nm, respectively, were achieved at room temperature and a bias of 11 V, corresponding to responsivities of 0.31 and 1.6 A/W, respectively, significantly higher than commercially available ultraviolet (UV) detectors. Furthermore, we demonstrated an integrated operation of the device. UV light was detected using a phototransistor that sent signals to drive an integrated LED as an indicator.

Low-dose controlled release of mTOR inhibitors maintains T cell plasticity and promotes central memory T cells.

An important goal for improving vaccine and immunotherapy technologies is the ability to provide further control over the specific phenotypes of T cells arising from these agents. Along these lines, frequent administration of rapamycin (Rapa), a small molecule inhibitor of the mammalian target of rapamycin (mTOR), exhibits a striking ability to polarize T cells toward central memory phenotypes (TCM), or to suppress immune function, depending on the concentrations and other signals present during administration. TCM exhibit greater plasticity and proliferative capacity than effector memory T cells (TEFF) and, therefore, polarizing vaccine-induced T cells toward TCM is an intriguing strategy to enhance T cell expansion and function against pathogens or tumors. Here we combined biodegradable microparticles encapsulating Rapa (Rapa MPs) with vaccines composed of soluble peptide antigens and molecular adjuvants to test if this approach allows polarization of differentiating T cells toward TCM. We show Rapa MPs modulate DC function, enhancing secretion of inflammatory cytokines at very low doses, and suppressing function at high doses. While Rapa MP treatment reduced - but did not stop - T cell proliferation in both CD4+ and CD8+ transgenic T cell co-cultures, the expanding CD8+ T cells differentiated to higher frequencies of TCM at low doses of MP Rapa MPs. Lastly, we show in mice that local delivery of Rapa MPs to lymph nodes during vaccination either suppresses or enhances T cell function in response to melanoma antigens, depending on the dose of drug in the depots. In particular, at low Rapa MP doses, vaccines increased antigen-specific TCM, resulting in enhanced T cell expansion measured during subsequent booster injections over at least 100days.

In Vivo Expansion of Melanoma-Specific T Cells Using Microneedle Arrays Coated with Immune-Polyelectrolyte Multilayers.

Microneedles (MNs) are micron-scale polymeric or metallic structures that offer distinct advantages for vaccines by efficiently targeting skin-resident immune cells, eliminating injection-associated pain, and improving patient compliance. These advantages, along with recent studies showing therapeutic benefits achieved using traditional intradermal injections in human cancer patients, suggest MN delivery might enhance cancer vaccines and immunotherapies. We recently developed a new class of polyelectrolyte multilayers based on the self-assembly of model peptide antigens and molecular toll-like receptor agonists (TLRa) into ultrathin, conformal coatings. Here, we reasoned that these immune polyelectrolyte multilayers (iPEMs) might be a useful platform for assembling cancer vaccine components on MN arrays for intradermal delivery from these substrates. Using conserved human melanoma antigens and a potent TLRa vaccine adjuvant, CpG, we show that iPEMs can be assembled on MNs in an automated fashion. These films, prepared with up to 128 layers, are approximately 200 nm thick but provide cancer vaccine cargo loading >225 µg/cm2. In cell culture, iPEM cargo released from MNs is internalized by primary dendritic cells, promotes activation of these cells, and expands T cells during coculture. In mice, application of iPEM-coated MNs results in the codelivery of tumor antigen and CpG through the skin, expanding tumor-specific T cells during initial MN applications and resulting in larger memory recall responses during a subsequent booster MN application. This study support MNs coated with PEMs built from tumor vaccine components as a well-defined, modular system for generating tumor-specific immune responses, enabling new approaches that can be explored in combination with checkpoint blockade or other combination cancer therapies.

Sustained delivery of recombinant human bone morphogenetic protein-2 from perlecan domain I - functionalized electrospun poly (ε-caprolactone) scaffolds for bone regeneration.

BACKGROUND: Biomaterial scaffolds that deliver growth factors such as recombinant human bone morphogenetic proteins-2 (rhBMP-2) have improved clinical bone tissue engineering by enhancing bone tissue regeneration. This approach could be further improved if the controlled delivery of bioactive rhBMP-2 were sustained throughout the duration of osteogenesis from fibrous scaffolds that provide control over dose and bioactivity of rhBMP-2. In nature, heparan sulfate attached to core proteoglycans serves as the co-receptor that delivers growth factors to support tissue morphogenesis. METHODS: To mimic this behavior, we conjugated heparan sulfate decorated recombinant domain I of perlecan/HSPG2 onto an electrospun poly(ε-caprolactone) (PCL) scaffold, hypothesizing that the heparan sulfate chains will enhance rhBMP-2 loading onto the scaffold and preserve delivered rhBMP-2 bioactivity. RESULTS: In this study, we demonstrated that covalently conjugated perlecan domain I increased loading capacity of rhBMP-2 onto PCL scaffolds when compared to control unconjugated scaffolds. Additionally, rhBMP-2 released from the modified scaffolds enhanced alkaline phosphatase activity in W20-17 mouse bone marrow stromal cells, indicating the preservation of rhBMP-2 bioactivity indicative of osteogenesis. CONCLUSIONS: We conclude that this platform provides a sophisticated and efficient approach to deliver bioactive rhBMP-2 for bone tissue regeneration applications.

Design of Polyelectrolyte Multilayers to Promote Immunological Tolerance.

Recent studies demonstrate that excess signaling through inflammatory pathways (e.g., toll-like receptors, TLRs) contributes to the pathogenesis of human autoimmune diseases, including lupus, diabetes, and multiple sclerosis (MS). We hypothesized that codelivery of a regulatory ligand of TLR9, GpG oligonucleotide, along with myelin-the "self" molecule attacked in MS-might restrain the pro-inflammatory signaling typically present during myelin presentation, redirecting T cell differentiation away from inflammatory populations and toward tolerogenic phenotypes such as regulatory T cells. Here we show that myelin peptide and GpG can be used as modular building blocks for co-assembly into immune polyelectrolyte multilayers (iPEMs). These nanostructured capsules mimic attractive features of biomaterials, including tunable cargo loading and codelivery, but eliminate all carriers and synthetic polymers, components that often exhibit intrinsic inflammatory properties that could exacerbate autoimmune disease. In vitro, iPEMs assembled from myelin and GpG oligonucleotide, but not myelin and a control oligonucleotide, restrain TLR9 signaling, reduce dendritic cell activation, and polarize myelin-specific T cells toward tolerogenic phenotype and function. In mice, iPEMs blunt myelin-triggered inflammatory responses, expand regulatory T cells, and eliminate disease in a common model of MS. Finally, in samples from human MS patients, iPEMs bias myelin-triggered immune cell function toward tolerance. This work represents a unique opportunity to use PEMs to regulate immune function and promote tolerance, supporting iPEMs as a carrier-free platform to alter TLR function to reduce inflammation and combat autoimmunity.

Reprogramming the Local Lymph Node Microenvironment Promotes Tolerance that Is Systemic and Antigen Specific.

Many experimental therapies for autoimmune diseases, such as multiple sclerosis (MS), aim to bias T cells toward tolerogenic phenotypes without broad suppression. However, the link between local signal integration in lymph nodes (LNs) and the specificity of systemic tolerance is not well understood. We used intra-LN injection of polymer particles to study tolerance as a function of signals in the LN microenvironment. In a mouse MS model, intra-LN introduction of encapsulated myelin self-antigen and a regulatory signal (rapamycin) permanently reversed paralysis after one treatment during peak disease. Therapeutic effects were myelin specific, required antigen encapsulation, and were less potent without rapamycin. This efficacy was accompanied by local LN reorganization, reduced inflammation, systemic expansion of regulatory T cells, and reduced T cell infiltration to the CNS. Our findings suggest that local control over signaling in distinct LNs can promote cell types and functions that drive tolerance that is systemic but antigen specific.

Assembly and Immunological Processing of Polyelectrolyte Multilayers Composed of Antigens and Adjuvants.

While biomaterials provide a platform to control the delivery of vaccines, the recently discovered intrinsic inflammatory characteristics of many polymeric carriers can also complicate rational design because the carrier itself can alter the response to other vaccine components. To address this challenge, we recently developed immune-polyelectrolyte multilayer (iPEMs) capsules electrostatically assembled entirely from peptide antigen and molecular adjuvants. Here, we use iPEMs built from SIINFEKL model antigen and polyIC, a stimulatory toll-like receptor agonist, to investigate the impact of pH on iPEM assembly, the processing and interactions of each iPEM component with primary immune cells, and the role of these interactions during antigen-specific T cell responses in coculture and mice. We discovered that iPEM assembly is pH dependent with respect to both the antigen and adjuvant component. Controlling the pH also allows tuning of the relative loading of SIINFEKL and polyIC in iPEM capsules. During in vitro studies with primary dendritic cells (DCs), iPEM capsules ensure that greater than 95% of cells containing at least one signal (i.e., antigen, adjuvant) also contained the other signal. This codelivery leads to DC maturation and SIINFEKL presentation via the MHC-I antigen presentation pathway, resulting in antigen-specific T cell proliferation and pro-inflammatory cytokine secretion. In mice, iPEM capsules potently expand antigen-specific T cells compared with equivalent admixed formulations. Of note, these enhancements become more pronounced with successive booster injections, suggesting that iPEMs functionally improve memory recall response. Together our results reveal some of the features that can be tuned to modulate the properties of iPEM capsules, and how these modular vaccine structures can be used to enhance interactions with immune cells in vitro and in mice.

Modular Vaccine Design Using Carrier-Free Capsules Assembled from Polyionic Immune Signals.

New vaccine adjuvants that direct immune cells toward specific fates could support more potent and selective options for diseases spanning infection to cancer. However, the empirical nature of vaccines and the complexity of many formulations has hindered design of well-defined and easily characterized vaccines. We hypothesized that nanostructured capsules assembled entirely from polyionic immune signals might support a platform for simple, modular vaccines. These immune-polyelectrolyte (iPEM) capsules offer a high signal density, selectively expand T cells in mice, and drive functional responses during tumor challenge. iPEMs incorporating clinically relevant antigens could improve vaccine definition and support more programmable control over immunity.

Polyelectrolyte Multilayers Assembled Entirely from Immune Signals on Gold Nanoparticle Templates Promote Antigen-Specific T Cell Response.

Materials that allow modular, defined assembly of immune signals could support a new generation of rationally designed vaccines that promote tunable immune responses. Toward this goal, we have developed the first polyelectrolyte multilayer (PEM) coatings built entirely from immune signals. These immune-PEMs (iPEMs) are self-assembled on gold nanoparticle templates through stepwise electrostatic interactions between peptide antigen and polyanionic toll-like receptor (TLR) agonists that serve as molecular adjuvants. iPEMs do not require solvents or mixing, offer direct control over the composition and loading of vaccine components, and can be coated on substrates at any scale. These films also do not require other structural components, eliminating the potentially confounding effects caused by the inherent immune-stimulatory characteristics of many synthetic polymers. iPEM loading on gold nanoparticle substrates is tunable, and cryoTEM reveals iPEM shells coated on gold cores. These nanoparticles are efficiently internalized by primary dendritic cells (DCs), resulting in activation, selective triggering of TLR signaling, and presentation of the antigens used to assemble iPEMs. In coculture, iPEMs drive antigen-specific T cell proliferation and effector cytokines but not cytokines associated with more generalized inflammation. Compared to mice treated with soluble antigen and adjuvant, iPEM immunization promotes high levels of antigen-specific CD8(+) T cells in peripheral blood after 1 week. These enhancements result from increased DC activation and antigen presentation in draining lymph nodes. iPEM-immunized mice also exhibit a potent recall response after boosting, supporting the potential of iPEMs for designing well-defined vaccine coatings that provide high cargo density and eliminate synthetic film components.

Controlled delivery of a metabolic modulator promotes regulatory T cells and restrains autoimmunity.

Autoimmune disorders occur when the immune system abnormally recognizes and attacks self-molecules. Dendritic cells (DCs) play a powerful role in initiating adaptive immune response, and are therefore a recent target for autoimmune therapies. N-Phenyl-7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxamide (PHCCC), a small molecule glutamate receptor enhancer, alters how DCs metabolize glutamate, skewing cytokine secretion to bias T cell function. These effects provide protection in mouse models of multiple sclerosis (MS) by polarizing T cells away from inflammatory TH17 cells and toward regulatory T cells (TREG) when mice receive daily systemic injections of PHCCC. However, frequent, continued treatment is required to generate and maintain therapeutic benefits. Thus, the use of PHCCC is limited by poor solubility, the need for frequent dosing, and cell toxicity. We hypothesized that controlled release of PHCCC from degradable nanoparticles (NPs) might address these challenges by altering DC function to maintain efficacy with reduced treatment frequency and toxicity. This idea could serve as a new strategy for harnessing biomaterials to polarize immune function through controlled delivery of metabolic modulators. PHCCC was readily encapsulated in nanoparticles, with controlled release of 89% of drug into media over three days. Culture of primary DCs or DC and T cell co-cultures with PHCCC NPs reduced DC activation and secretion of pro-inflammatory cytokines, while shifting T cells away from TH17 and toward TREG phenotypes. Importantly, PHCCC delivered to cells in NPs was 36-fold less toxic compared with soluble PHCCC. Treatment of mice with PHCCC NPs every three days delayed disease onset and decreased disease severity compared with mice treated with soluble drug at the same dose and frequency. These results highlight the potential to promote tolerance through controlled delivery of metabolic modulators that alter DC signaling to polarize T cells, and suggest future gains that could be realized by engineering materials that provide longer term release.

The effect of glutathione as chain transfer agent in PNIPAAm-based thermo-responsive hydrogels for controlled release of proteins.

PURPOSE: To control degradation and protein release using thermo-responsive hydrogels for localized delivery of anti-angiogenic proteins. METHODS: Thermo-responsive hydrogels derived from N-isopropylacrylamide (NIPAAm) and crosslinked with poly(ethylene glycol)-co-(L-lactic acid) diacrylate (Acry-PLLA-b-PEG-b-PLLA-Acry) were synthesized via free radical polymerization in the presence of glutathione, a chain transfer agent (CTA) added to modulate their degradation and release properties. Immunoglobulin G (IgG) and the recombinant proteins Avastin® and Lucentis® were encapsulated in these hydrogels and their release was studied. RESULTS: The encapsulation efficiency of IgG was high (75-87%) and decreased with CTA concentration. The transition temperature of these hydrogels was below physiological temperature, which is important for minimally invasive therapies involving these materials. The toxicity from unreacted monomers and free radical initiators was eliminated with a minimum of three buffer extractions. Addition of CTA accelerated degradation and resulted in complete protein release. Glutathione caused the degradation products to become solubilized even at 37°C. Hydrogels prepared without glutathione did not disintegrate nor released protein completely after 3 weeks at 37°C. PEGylation of IgG postponed the burst release effect. Avastin® and Lucentis® released from degraded hydrogels retained their biological activity. CONCLUSIONS: These systems offer a promising platform for the localized delivery of proteins.