Integrated Multi-Omics Analysis to Reveal the Molecular Mechanisms of Inflorescence Elongation in Medicago sativa.

The morphological architecture of inflorescence influences seed production. The regulatory mechanisms underlying alfalfa (Medicago sativa) inflorescence elongation remain unclear. Therefore, in this study, we conducted a comparative analysis of the transcriptome, proteome, and metabolome of two extreme materials at three developmental stages to explore the mechanisms underlying inflorescence elongation in alfalfa. We observed the developmental processes of long and short inflorescences and found that the elongation capacity of alfalfa with long inflorescence was stronger than that of alfalfa with short inflorescences. Furthermore, integrative analysis of the transcriptome and proteome indicated that the phenylpropanoid biosynthesis pathway was closely correlated with the structural formation of the inflorescence. Additionally, we identified key genes and proteins associated with lignin biosynthesis based on the differential expressed genes and proteins (DEGs and DEPs) involved in phenylpropanoid biosynthesis. Moreover, targeted hormone metabolome analysis revealed that IAA, GA, and CK play an important role in the peduncle elongation of alfalfa inflorescences. Based on omics analysis, we detected key genes and proteins related to plant hormone biosynthesis and signal transduction. From the WGCNA and WPCNA results, we furthermore screened 28 candidate genes and six key proteins that were correlated with lignin biosynthesis, plant hormone biosynthesis, and signaling pathways. In addition, 19 crucial transcription factors were discovered using correlation analysis that might play a role in regulating candidate genes. This study provides insight into the molecular mechanism of inflorescence elongation in alfalfa and establishes a theoretical foundation for improving alfalfa seed production.

Evaluation of dentinal tubule occlusion and pulp tissue response after using 980-nm diode laser for dentin hypersensitivity treatment.

OBJECTIVES: To evaluate the effectiveness of the 980-nm diode laser for dentinal tubule occlusion, measure the intrapulpal temperature, and investigate the dental pulp response. MATERIALS AND METHODS: The dentinal samples were randomly divided into G1-G7 groups: control; 980-nm laser irradiation (0.5 W, 10 s; 0.5 W, 10 s × 2; 0.8 W, 10 s; 0.8 W, 10 s × 2; 1.0 W, 10 s; 1.0 W, 10 s × 2). The dentin discs were applied for laser irradiation and analyzed by scanning electron microscopy (SEM). The intrapulpal temperature was measured on the 1.0-mm and 2.0-mm thickness samples, and then divided into G2-G7 groups according to laser irradiation. Moreover, forty Sprague Dawley rats were randomly divided into the laser-irradiated group (euthanized at 1, 7, and 14 days after irradiation) and the control group (non-irradiated). qRT-PCR, histomorphology, and immunohistochemistry analysis were employed to evaluate the response of dental pulp. RESULTS: SEM indicated the occluding ratio of dentinal tubules in the G5 (0.8 W, 10 s × 2) and G7 (1.0 W, 10 s × 2) were significantly higher than the other groups (p < 0.05). The maximum intrapulpal temperature rises in the G5 were lower than the standard line (5.5 ℃). qRT-PCR showed that the mRNA expression level of TNF-α and HSP-70 upregulated significantly at 1 day (p < 0.05). Histomorphology and immunohistochemistry analysis showed that, compared with the control group, the inflammatory reaction was slightly higher at the 1 and 7 days (p < 0.05) and decreased to the normal levels at 14 days (p > 0.05). CONCLUSIONS: A 980-nm laser at a power of 0.8 W with 10 s × 2 defines the best treatment for dentin hypersensitivity in terms of compromise between the efficacy of the treatment and the safety of the pulp. CLINICAL RELEVANCE: The 980-nm laser is an effective option for treating dentin sensitivity. However, we need to ensure the safety of the pulp during laser irradiation.

3D-printed swab with cover for precision diagnosis.

The collection capacity of common nasopharyngeal swabs and irregularities of medical personnel limit the accuracy of PCR testing. This study describes a newly designed 3D-printed swab that is combined with a 3D-printed cover to prevent the extraction of undesired nasal secretions. This swab improved the accuracy of PCR test results. The results of a series of experiments showed that, because of the mucus extraction effect, 3D-printed swabs can replace ordinary cotton swabs. The crisis of the worldwide medical supply shortage can be ameliorated to a certain extent by applying 3D printing technology.

Effects of active and passive smoking on salivary cytokines levels in rats: A pilot study.

Cigarette smoking is an established risk factor for some oral diseases. As an essential fluid in the oral cavity, saliva is crucial to maintain oral health. Relative to active smoking, there are very few studies assessing the effect of passive smoking on salivary cytokines levels. In the present study, we established the rat models by the means of the intraoral cigarette smoking or whole body cigarette smoke exposure to simulate human active or passive smoking, respectively. The effects of active or passive smoking on salivary cytokines levels were assessed by using ProcartaPlex multiplex immunoassays. The results of the current study indicated that both active and passive smoking diminished the body weights of rats and increased the levels of some blood counts. Intriguingly, active smoking enhanced the salivary levels of IL-6 and IL-12 p70 and passive smoking elevated the salivary IL-6 level. Moreover, active smoking appeared to have a more prominent activation effect on the salivary IL-6 level. It was noted that active or passive smoking had no significant effect on the salivary IFN-γ level. Active or passive smoking could have potential effects on the salivary levels of some pro-inflammatory cytokines.

Nitrilotriacetic Acid-Functionalized Glucose-Responsive Complex Micelles for the Efficient Encapsulation and Self-Regulated Release of Insulin.

Insulin plays a significant role in diabetes treatment. Although a huge number of insulin-loaded, glucose-responsive nanocarriers have been developed in past decades, most of them showed a lower loading capacity and efficiency due to the weak interaction between insulin and nanocarriers. In this work, a novel insulin-encapsulated glucose-responsive polymeric complex micelle (CM) is devised, showing (i) enhanced insulin-loading efficiency owing to the zinc ions' chelation by nitrilotriacetic acid (NTA) groups of NTA-functioned glycopolymer and the histidine imidazole of insulin, (ii) the glucose-triggered pulse release of insulin, and (iii) long stability under physiological conditions. This CM was fabricated by the self-assembly of block copolymer PEG- b-P(Asp- co-AspPBA) and glycopolymer P(Asp- co-AspGA- co-AspNTA), resulting in complex micelles with a PEG shell and a cross-linked core composed of phenylboronic acid (PBA)/glucose complexations. Notably, the modified nitrilotriacetic acid (NTA) groups of CM could specifically bind insulin via chelated zinc ions, thus enhancing the loading efficacy of insulin compared to that of nonmodified CM. The dynamic PBA/glucose complexation core of CM dissociates under the trigger of high glucose concentration (>2 g/L) while being quite stable in low glucose concentrations (<2 g/L), as demonstrated by the pulse release of insulin in vitro. Finally, in a murine model of type 1 diabetes, NTA-modified complex micelles loading an insulin (NTA-CM-INS) group exhibited a long hypoglycemic effect which is superior to that of free insulin in the PBS (PBS-INS) group and insulin-loaded complex micelles without an NTA modification (CM-INS) group. This long-term effect benefited from Zn(II) chelation by NTA-modified complex micelles and could avoid hypoglycemia caused by the burst release of insulin. Taken together, this constitutes a highly effective way to encapsulate insulin and release insulin via an on-demand manner for blood glucose control in diabetes.

Maintenance of amyloid ß peptide homeostasis by artificial chaperones based on mixed-shell polymeric micelles.

The disruption of Aß homeostasis, which results in the accumulation of neurotoxic amyloids, is the fundamental cause of Alzheimer's disease (AD). Molecular chaperones play a critical role in controlling undesired protein misfolding and maintaining intricate proteostasis in vivo. Inspired by a natural molecular chaperone, an artificial chaperone consisting of mixed-shell polymeric micelles (MSPMs) has been devised with tunable surface properties, serving as a suppressor of AD. Taking advantage of biocompatibility, selectivity toward aberrant proteins, and long blood circulation, these MSPM-based chaperones can maintain Aß homeostasis by a combination of inhibiting Aß fibrillation and facilitating Aß aggregate clearance and simultaneously reducing Aß-mediated neurotoxicity. The balance of hydrophilic/hydrophobic moieties on the surface of MSPMs is important for their enhanced therapeutic effect.

Temperature-responsive mixed-shell polymeric micelles for the refolding of thermally denatured proteins.

We have fabricated a mixed-shell polymeric micelle (MSPM) that closely mimics the natural molecular chaperone GroEL-GroES complex in terms of structure and functionality. This MSPM, which possesses a shared PLA core and a homogeneously mixed PEG and PNIAPM shell, is constructed through the co-assembly of block copolymers poly(lactide-b-poly(ethylene oxide) (PLA-b-PEG) and poly(lactide)-b-poly(N-isopropylacryamide) (PLA-b-PNIPAM). Above the lower critical solution temperature (LCST) of PNIPAM, the MSPM evolves into a core-shell-corona micelle (CSCM), as a functional state with hydrophobic PNIPAM domains on its surface. Light scattering (LS), TEM, and fluorescence and circular dichroism (CD) spectroscopy were performed to investigate the working mechanism of the chaperone-like behavior of this system. Unfolded protein intermediates are captured by the hydrophobic PNIPAM domains of the CSCM, which prevent harmful protein aggregation. During cooling, PNIPAM reverts into its hydrophilic state, thereby inducing the release of the bound unfolded proteins. The refolding process of the released proteins is spontaneously accomplished by the presence of PEG in the mixed shell. Carbonic anhydrase B (CAB) was chosen as a model to investigate the refolding efficiency of the released proteins. In the presence of MSPM, almost 93 % CAB activity was recovered during cooling after complete denaturation at 70 °C. Further results reveal that this MSPM also works with a wide spectrum of proteins with more-complicated structures, including some multimeric proteins. Given the convenience and generality in preventing the thermal aggregation of proteins, this MSPM-based chaperone might be useful for preventing the toxic aggregation of misfolded proteins in some diseases.

Influence of calcium-based desensitizing toothpastes on the bonding performance of universal adhesive.

The study evaluated the effect of calcium-based desensitizing toothpastes on the dentinal tubule occlusion and its influence on the dentin bond strength of universal adhesive. Mid-coronal dentin samples were prepared for hypersensitivity model and treated by the following calcium-based desensitizing toothpastes: no treatment (Control), Clinpro (fTCP), Pro-Relief (Pro-Argin), and Repair & Protect (Novamin). Single Bond Universal adhesive was applied in self-etch or etch-and-rinse mode. The dentinal tubule occlusion and adhesion interface were evaluated under scanning electron microscope (SEM). A double-fluorescence technique was used to examine interfacial permeability under confocal laser scanning microscopy (CLSM). The micro-tensile bond strength (µTBS) was employed, followed by the fracture interface observation. SEM showed the toothpastes occluded dentinal tubules, and the occlusion exhibited stability against acid and abrasion. Hindered resin infiltration was observed in the adhesion interface after desensitization. CLSM showed more water permeation within or under the adhesion interface in etch-and-rinse mode than self-etch mode. Desensitization decreased the µTBS in self-etch mode. When using etch-and-rinse mode, the desensitized samples presented similar µTBS to the control group. No difference in µTBS was found between the two bonding modes, except for the control group. Calcium-based desensitizing toothpastes can effectively occlude the exposed dentinal tubules with acid-resistant and abrasion-resistant stability. The desensitization reduced the dentin bond strength of the universal adhesive system in self-etch mode but did not affect the bond strength of etch-and-rinse mode.

Bonding properties of mild universal adhesives to dentin pretreated with hydroxyapatite-based desensitizing agents.

OBJECTIVES: The purpose of the study was to evaluate the effect of hydroxyapatite (HA)-based desensiti-zing agents and determine their influence on the bonding performance of mild universal adhesives. METHODS: Mid-coronal dentin samples were sectioned from human third molars and prepared for a dentin-sensitive model. According to desensitizing applications, they were randomly divided into four groups for the following treatments: no desensitizing treatment (control), Biorepair toothpaste (HA-based desensitizing toothpaste) treatment, Dontodent toothpaste (HA-based desensitizing toothpaste) treatment, and HA paste treatment. Dentin tubular occlusion and occluded area ratios were evaluated by scanning electron microscopy (SEM). Furthermore, All-Bond Universal, Single Bond Universal, and Clearfil Universal Bond were applied to the desensitized dentin in self-etch mode. The wettability and surface free energy (SFE) of desensitized dentin were evaluated by contact angle measurements. Bonded specimens were sectioned into beams and tested for micro-tensile bond strength to analyze the effect of desensitizing treatment on the bond strength to dentin of universal adhesives. RESULTS: SEM revealed that the dentin tubule was occluded by HA-based desensitizing agents, and the area ratios for the occluded dentin tubules were in the following order: HA group>Biorepair group>Dontodent group (P<0.05). Contact angle analysis demonstrated that HA-based desensitizing agents had no statistically significant influence on the wettability of the universal adhesives (P>0.05). The SFE of dentin significantly increased after treatment by HA-based desensitizing agents (P<0.05). The micro-tensile bond strength test showed that HA-based desensitizing toothpastes always decreased the µTBS values (P<0.05), whereas the HA paste group presented similar bond strength to the control group (P>0.05), irrespective of universal adhesive types. CONCLUSIONS: HA-based desensitizing agents can occlude the exposed dentinal tubules on sensitive dentin. When mild and ultra-mild universal adhesives were used for subsequent resin restoration, the bond strength was reduced by HA-based desensitizing toothpastes, whereas the pure HA paste had no adverse effect on bond strength.

Stapled Liposomes Enhance Cross-Priming of Radio-Immunotherapy.

The release of tumor-associated antigens (TAAs) and their cross-presentation in dendritic cells (DCs) are crucial for radio-immunotherapy. However, the irradiation resistance of tumor cells usually results in limited TAA generation and release. Importantly, TAAs internalized by DCs are easily degraded in lysosomes, resulting in unsatisfactory extent of TAA cross-presentation. Herein, an antigen-capturing stapled liposome (ACSL) with a robust structure and bioactive surface is developed. The ACSLs capture and transport TAAs from lysosomes to the cytoplasm in DCs, thereby enhancing TAA cross-presentation. l-arginine encapsulated in ACSLs induces robust T cell-dependent antitumor response and immune memory in 4T1 tumor-bearing mice after local irradiation, resulting in significant tumor suppression and an abscopal effect. Replacing l-arginine with radiosensitizers, photosensitizers, and photothermal agents may make ACSL a universal platform for the rapid development of various combinations of anticancer therapies.

Bonding Performance of Universal Adhesives Applied to Nano-Hydroxyapatite Desensitized Dentin Using Etch-and-Rinse or Self-Etch Mode.

The study assessed the bonding performance of three universal adhesives on desensitized dentin with etch-and-rinse mode or self-etch mode after nano-hydroxyapatite (nHAp)-based desensitizers application. Simulated sensitive dentin specimens were prepared and separated into four groups: no treatment as the negative control, groups desensitized by Biorepair toothpaste, Dontodent toothpaste, or nHAp paste. Three universal adhesives of All-Bond Universal, Single Bond Universal, and Clearfil Universal Bond with etch-and-rinse or self-etch mode were bonded to the desensitized dentin specimens separately, followed by resin composite build-ups. Micro-tensile bond strength was measured using a micro-tensile tester. The wettability of desensitized dentin was evaluated by the contact angle of the adhesives. Resin infiltration was observed by confocal laser scanning microscopy. Dentin tubular occlusion and nanoleakage were observed by scanning electron microscope. The results showed that the etch-and-rinse or self-etch mode of each adhesive showed similar bond strength when bonding to nHAp-based desensitized dentin. The dentin surface was partially covered by desensitizers after desensitization. Compared with the self-etch mode, stronger demineralization and more reopened dentin tubules were observed in the etch-and-rinse mode after acid etching; longer resin tags and more nanoleakage in the resin-dentin interface were observed when using the etch-and-rinse mode. When bonding to nHAp-based desensitized dentin with universal adhesives, no significant difference in bond strength was found between self-etch mode or etch-and-rinse mode; while the latter produced more nanoleakage in the resin-dentin interfaces.

NIR-activated self-sensitized polymeric micelles for enhanced cancer chemo-photothermal therapy.

NIR-activated therapies based on light-responsive drug delivery systems are emerging as a remote-controlled method for cancer precise therapy. In this work, fluorescent dye indocyanine green (ICG)-conjugated and bioactive compound gambogic acid (GA)-loaded polymeric micelles (GA@PEG-TK-ICG PMs) were smoothly fabricated via the self-assembly of the reactive oxygen species (ROS)-responsive thioketal (TK)-linked amphiphilic polymer poly(ethyleneglycol)-thioketal-(indocyanine green) (PEG-TK-ICG). The resultant micelles demonstrated increased resistance to photobleaching, enhanced photothermal conversion efficiency, NIR-controlled drug release behavior, preferable biocompatibility, and excellent tumor accumulation performance. Moreover, upon an 808 nm laser irradiation, the micellar photoactive chromophore ICG converted the absorbed optical energy to both hyperthermia for photothermal therapy (PTT) and ROS as the feedback trigger to the micelles for the tumor-specific release of GA, which could serve as not only a chemotherapeutic drug to directly kill tumor cells but also a heat shock protein 90 (HSP90) inhibitor to realize the photothermal sensitization. As a result, an extremely high tumor inhibition rate (97.9%) of mouse 4 T1 breast cancer models was achieved with negligible side effects after the chemo-photothermal synergistic therapy. This NIR-activated nanosystem with photothermal self-sensitization function may provide a feasible option for the effective treatment of aggressive breast cancers.

ICG-Conjugated and 125 I-Labeled Polymeric Micelles with High Biosafety for Multimodality Imaging-Guided Photothermal Therapy of Tumors.

Noninvasive multimodality imaging-guided precision photothermal therapy (PTT) is proven to be an effective strategy for tumor theranostics by integrating diagnostics and therapeutics in one nanoplatform. In this study, indocyanine green (ICG)-conjugated and radionuclide iodine-125 (125 I)-labeled polymeric micelles (PEG-PTyr(125 I)-ICG PMs) are strategically prepared by the self-assembly of the ICG-decorated amphiphilic diblock polymer poly(ethylene glycol)-poly(l-tyrosine-125 I)-(indocyanine green) (PEG-PTyr(125 I)-ICG). The as-prepared polymeric micelles exhibit favorable biocompatibility, excellent size/photo/radiolabel stability, a high-photothermal conversion efficiency, a passive tumor-targeting ability, and a fluorescence (FL)/photoacoustic (PA)/single photon emission computed tomography (SPECT) imaging property. After tail intravenous injection, the polymeric micelles can efficiently accumulate at the tumor site and present comprehensive FL/PA/SPECT images with a high sensitivity, excellent spatial resolution, and unlimited tissue penetration under near-infrared (NIR) irradiation. Upon 808 nm laser irradiation, the subsequent precision PTT of tumors can be achieved with minimal cumulative side effects. Thus, this capable multifunctional nanoplatform with simple components and preparation procedures for FL/PA/SPECT multimodality imaging-guided PTT can be a potential candidate for clinical tumor theranostics.

Self-targeting, zwitterionic micellar dispersants enhance antibiotic killing of infectious biofilms-An intravital imaging study in mice.

Extracellular polymeric substances (EPS) hold infectious biofilms together and limit antimicrobial penetration and clinical infection control. Here, we present zwitterionic micelles as a previously unexplored, synthetic self-targeting dispersant. First, a pH-responsive poly(ε-caprolactone)-block-poly(quaternary-amino-ester) was synthesized and self-assembled with poly(ethylene glycol)-block-poly(ε-caprolactone) to form zwitterionic, mixed-shell polymeric micelles (ZW-MSPMs). In the acidic environment of staphylococcal biofilms, ZW-MSPMs became positively charged because of conversion of the zwitterionic poly(quaternary-amino-ester) to a cationic lactone ring. This allowed ZW-MSPMs to self-target, penetrate, and accumulate in staphylococcal biofilms in vitro. In vivo biofilm targeting by ZW-MSPMs was confirmed for staphylococcal biofilms grown underneath an implanted abdominal imaging window through direct imaging in living mice. ZW-MSPMs interacted strongly with important EPS components such as eDNA and protein to disperse biofilm and enhance ciprofloxacin efficacy toward remaining biofilm, both in vitro and in vivo. Zwitterionic micellar dispersants may aid infection control and enhance efficacy of existing antibiotics against remaining biofilm.

Characterization and sequencing of lithium cationized ß-O-4 lignin oligomers using higher-energy collisional dissociation mass spectrometry.

The prospect of developing new bio-based products from lignin partially depends on the development of effective analytical techniques to characterize the end products from lignin degradation experiments. To date, the most utilized mass spectrometric technique for characterizing lignin oligomers has been negative ion mass spectrometry. Positive ion mass spectrometry remains a relatively unexplored approach for lignin sequencing. Here we report on the sequencing of lignin oligomers using lithium cationization positive ion electrospray followed by higher-energy collisional dissociation (HCD) tandem mass spectrometry on a high-resolution accurate-mass Orbitrap mass spectrometer. Thirteen synthetic lignin model oligomers containing ß-O-4 and 4-O-α linkages were analyzed and sequenced based on their HCD tandem mass spectrometry fragmentation patterns. Proposed sequence-specific fragmentation pathways are provided. The method presented in this study provides a potential application of lithium cationization tandem mass spectrometry for the analysis of lignin degradation products.

Glucose-responsive complex micelles for self-regulated delivery of insulin with effective protection of insulin and enhanced hypoglycemic activity in vivo.

Large amounts of insulin-loaded glucose-responsive micelles based on poly(amino acid)s have been developed for diabetes treatment over last decades, but most of them could not effectively protect insulin from enzymatic degradation in vivo because the micellar core was biodegradable and lacked protective structure for insulin, which would lower the efficacy of insulin to a large extent. In this study, we fabricated a new type of insulin-loaded glucose-responsive complex micelles (CMs), which were self-assembled by a phenylboronic acid (PBA)-modified block copolymer PEG-b-P(Asp-co-AspPBA) and a glucosamine (GA)/nitrilotriacetic acid (NTA)-functionalized block copolymer PNIPAM-b-P(Asp-co-AspGA-co-AspNTA), for self-regulated delivery of insulin with effective protection of insulin and enhanced hypoglycemic activity in vivo. The CMs possessed mixed shell of PEG/PNIPAM and cross-linked core of PBA/GA complex, which could be disintegrated under the condition of high glucose concentration (5 g/L) while maintaining stable at low glucose concentration (1 g/L). The NTA groups of CMs greatly improved the loading content of insulin by specifically bind insulin via the chelated zinc ions. More importantly, PNIPAM chains in the mixed shell would collapse under 37 °C and form hydrophobic domains around the micellar core, which could significantly protect the micellar core as well as the encapsulated insulin from attacking by external proteases. In a murine model of type 1 diabetes, the CMs with insulin chelated by NTA showed a long hypoglycemic effect, which is superior to insulin-loaded simple micelles without PNIPAM and insulin in PBS buffer (pH 7.4). Therefore, this kind of CMs could be a potential candidate for insulin delivery in diabetes therapy.

Glucose and H2O2 dual-sensitive nanogels for enhanced glucose-responsive insulin delivery.

Diabetes is a chronic metabolic disorder disease characterized by high blood glucose levels and has become one of the most serious threats to human health. In recent decades, a number of insulin delivery systems, including bulk gels, nanogels, and polymeric micelles, have been developed for the treatment of diabetes. Herein, a kind of glucose and H2O2 dual-responsive polymeric nanogel was designed for enhanced glucose-responsive insulin delivery. The polymeric nanogels composed of poly(ethylene glycol) and poly(cyclic phenylboronic ester) (glucose and H2O2 dual-sensitive groups) were synthesized by a one-pot thiol-ene click chemistry approach. The nanogels displayed glucose-responsive release of insulin and the release rate could be promoted by the incorporation of glucose oxidase (GOx), which generated H2O2 at high glucose levels and H2O2 further oxidizes and hydrolyzes the phenylboronic ester group. The nanogels have characteristics of long blood circulation time, a fast response to glucose, and excellent biocompatibility. Moreover, subcutaneous delivery of insulin to diabetic mice with the insulin/GOx-loaded nanogels presented an effective hypoglycemic effect compared to that of injection of insulin or insulin-loaded nanogels. This kind of nanogel would be a promising candidate for the delivery of insulin in the future.

Nanocarriers with conjugated antimicrobials to eradicate pathogenic biofilms evaluated in murine in vivo and human ex vivo infection models.

Conventional antimicrobials are becoming increasingly ineffective for treating bacterial infection due to the emergence of multi-drug resistant (MDR) pathogens. In addition, the biofilm-mode-of-growth of infecting bacteria impedes antimicrobial penetration in biofilms. Here, we report on poly(ethylene)glycol-poly(ß-amino esters) (PEG-PAE) micelles with conjugated antimicrobials, that can uniquely penetrate biofilms, target themselves to bacterial cell surfaces once inside the low-pH environment of a biofilm and release conjugated antimicrobials through degradation of their ester-linkage with PAE by bacterial lipases. In vitro, PEG-PAE micelles with conjugated Triclosan (PEG-PAE-Triclosan) yielded no inadvertent leakage of their antimicrobial cargo and better killing of MDR Staphylococcus aureus, Escherichia coli and oral streptococcal biofilms than Triclosan in solution. In mice, PEG-PAE-Triclosan micelles with conjugated Triclosan yielded better eradication efficacy towards a MDR S. aureus-infection compared with Triclosan in solution and Triclosan-loaded micelles at equal Triclosan-equivalent concentrations. Ex vivo exposure of multi-species oral biofilms collected from orthodontic patients to PEG-PAE-Triclosan micelles, demonstrated effective bacterial killing at 30-40 fold lower Triclosan-equivalent concentrations than achieved by Triclosan in solution. Importantly, Streptococcus mutans, the main causative organism of dental caries, was preferentially killed by PEG-PAE-Triclosan micelles. Thus PEG-PAE-Triclosan micelles present a promising addendum to the decreasing armamentarium available to combat infection in diverse sites of the body. STATEMENT OF SIGNIFICANCE: pH-adaptive polymeric micelles with conjugated antimicrobials can efficiently eradicate infectious biofilms from diverse body sites in mice and men. An antimicrobial was conjugated through an ester-linkage to a poly(ethylene glycol) (PEG)/poly(ß-amino ester) block copolymer to create micellar nanocarriers. Stable micelle structures were formed by the hydrophobic poly(ß-amino ester) inner core and a hydrophilic PEG outer shell. Thus formed PEG-PAE-Triclosan micelles do not lose their antimicrobial cargo underway to an infection site through the blood circulation, but penetrate and accumulate in biofilms to release antimicrobials once inside a biofilm through degradation of its ester-linkage by bacterial lipases, to kill biofilm-embedded bacteria at lower antimicrobial concentrations than when applied in solution. PEG-PAE-Triclosan micelles effectively eradicate biofilms of multi-drug-resistant pathogens and oral bacteria, most notably highly cariogenic Streptococcus mutans, in mice and men respectively, and possess excellent clinical translation possibilities.

Silver-Decorated Polymeric Micelles Combined with Curcumin for Enhanced Antibacterial Activity.

Because of the mounting prevalence of complicated infections induced by multidrug-resistant bacteria, it is imperative to develop innovative and efficient antibacterial agents. In this work, we design a novel polymeric micelle for simultaneous decorating of silver nanoparticles and encapsulating of curcumin as a combination strategy to improve the antibacterial efficiency. In the constructed combination system, silver nanoparticles were decorated in the micellar shell because of the in situ reduction of silver ions, which were absorbed by the poly(aspartic acid) (PAsp) chains in the shell. Meanwhile, natural curcumin was encapsulated into the poly(ε-caprolactone) (PCL) core of the micelle through hydrophobic interaction. This strategy could prevent aggregation of silver nanoparticles and improve the water solubility of curcumin at the same time, which showed enhanced antibacterial activity toward Gram-negative P.aeruginosa and Gram-positive S.aureus compared with sliver-decorated micelle and curcumin-loaded micelle alone, due to the cooperative antibacterial effects of the silver nanoparticles and curcumin. Furthermore, the achieved combinational micelles had good biocompatibility and low hemolytic activity. Thus, our study provides a new pathway in the rational design of combination strategy for efficiently preventing the ubiquitous bacterial infections.

A biocompatible cobaltporphyrin-based complex micelle constructed via supramolecular assembly for oxygen transfer.

Herein, a complex micelle as an oxygen nano-carrier is constructed through the hierarchical assembly of the diblock copolymer poly(ethylene glycol)-block-poly(l-lysine) (PEG-b-PLys), tetrakis(4-sulfonatophenyl)porphinato cobalt(ii) (Co(ii)TPPS), a heptapeptide (Cys-His-His-His-His-His-His) and heptakis(2,3,6-tri-O-methyl)-ß-cyclodextrin (TM-ß-CD). Co(ii)TPPS was encapsulated into the cavities of TM-ß-CDs driven by the host-guest interaction so that the irreversible formation of a µ-oxo-dimer of Co(ii)TPPS can be effectively prevented. The imidazole groups of the heptapeptide were selected as good axial ligands coordinating to the centric cobalt of Co(ii)TPPS, which subtly constituted the five-coordinated precursor serving as an active functional centre for oxygen binding. The sixth position of Co(ii)TPPS can bind oxygen. Furthermore, the host-guest inclusion (TM-ß-CD/Co(ii)TPPS) was loaded into the hydrophobic core of the complex micelle and tightly fixed with PLys chains. The hydrophilic PEG blocks stretched in the aqueous solution constitute the shells which stabilize the structure of the complex micelle as well as impart the complex micelle sufficient blood circulation time. Moreover, the complex micelle exhibited excellent biocompatibility and cellular uptake. Therefore, the rationally designed amphiphilic structure can work as promising artificial O2 carriers in vivo. Potentially, the complex micelle can be expected to change the anaerobic microenvironment and find applications in the repair of the cells damaged by cellular hypoxia.