Affinity-Driven Aryl Diazonium Labeling of Peptide Receptors on Living Cells.

Peptide-receptor interactions play critical roles in a wide variety of physiological processes. Methods to link bioactive peptides covalently to unmodified receptors on the surfaces of living cells are valuable for studying receptor signaling, dynamics, and trafficking and for identifying novel peptide-receptor interactions. Here, we utilize peptide analogues bearing deactivated aryl diazonium groups for the affinity-driven labeling of unmodified receptors. We demonstrate that aryl diazonium-bearing peptide analogues can covalently label receptors on the surface of living cells using both the neurotensin and the glucagon-like peptide 1 receptor systems. Receptor labeling occurs in the complex environment of the cell surface in a sequence-specific manner. We further demonstrate the utility of this covalent labeling approach for the visualization of peptide receptors by confocal fluorescence microscopy and for the enrichment and identification of labeled receptors by mass spectrometry-based proteomics. Aryl diazonium-based affinity-driven receptor labeling is attractive due to the high abundance of tyrosine and histidine residues susceptible to azo coupling in the peptide binding sites of receptors, the ease of incorporation of aryl diazonium groups into peptides, and the relatively small size of the aryl diazonium group. This approach should prove to be a powerful and relatively general method to study peptide-receptor interactions in cellular contexts.

Polyphenol analysis in human milk by a rapid, cost-effective, and accurate micromethod: translational development.

OBJECTIVE: To develop a rapid method for analysing polyphenols, which are potentially active antioxidants against neonatal oxidative stress, from small human milk (HM) volumes. METHODS: Acid and alkaline extractions were compared using two dyes: Folin-Ciocalteu and Fast Blue BB. Linearity, sensitivity, recovery percentage, polyphenol content, precision, and stability were assessed in 14 HM samples and compared using the Kruskal-Wallis H test (p<0.05). The best technique was applied to 284 HM samples to determine their polyphenolic content and its association with maternal diet by multifactorial linear regression. RESULTS: Acidic extraction successfully recovered the gallic acid reference standard, whereas alkaline extraction overestimated it. Calibration curves for all methods were linear (R2>0.96) up to 500 mg/L. All bicarbonate-based Folin-Ciocalteu methods assayed were stable and repeatable, whereas Fast Blue BB-based variants were not. HM polyphenols (mean=94.68 mg/L) positively correlated to the dietary intake of hydroxycinnamic acids, the most consumed polyphenolic family in this population. CONCLUSIONS: A bicarbonate-based Folin-Ciocalteu micromethod allowed the accurate determination of polyphenols in HM, which might be useful for translational research settings and HM banks.

pH change accompanying long-distance electrical signal controls systemic jasmonate biosynthesis.

Local damaging stimuli cause a rapid increase in the content of the defense phytohormone jasmonic acid (JA) and its biologically active derivative jasmonoyl-L-isoleucine (JA-Ile) in undamaged distal tissues. The increase in JA and JA-Ile levels was coincident with a rapid decrease in the levels of the precursor 12-oxo-phytodienoic acid (OPDA). The propagation of a stimulus-induced long-distance electrical signal, variation potential (VP), which is accompanied by intracellular changes in pH and Ca2+ levels, preceded systemic changes in jasmonate content. The decrease in pH during VP, mediated by transient inactivation of the plasma membrane H+-ATPase, induced the conversion of OPDA to JA, probably by regulating the availability of the OPDA substrate to JA biosynthetic enzymes. The regulation of systemic synthesis of JA and JA-Ile by the Ca2+ wave accompanying VP most likely occurs by the same mechanism of pH-induced conversion of OPDA to JA due to Ca2+-mediated decrease in pH as a result of H+-ATPase inactivation. Thus, the transient increase in intracellular Ca2+ levels and the transient decrease in intracellular pH are most likely the key mechanisms of VP-mediated regulation of jasmonate production in systemic tissues upon local stimulation.

Colorimetric Detection of HER2-Overexpressing-Cancer-Derived Exosomes in Mouse Urine Using Magnetic-Polydiacetylene Nanoparticles.

Breast cancer (BC) is a major global health problem, with ≈20-25% of patients overexpressing human epidermal growth factor receptor 2 (HER2), an aggressive marker, yet access to early detection and treatment varies across countries. A low-cost, equipment-free, and easy-to-use polydiacetylene (PDA)-based colorimetric sensor is developed for HER2-overexpressing cancer detection, designed for use in low- and middle-income countries (LMICs). PDA nanoparticles are first prepared through thin-film hydration. Subsequently, hydrophilic magnetic nanoparticles and HER2 antibodies are sequentially conjugated to them. The synthesized HER2-MPDA can be concentrated and separated by a magnetic field while inheriting the optical characteristics of PDA. The specific binding of HER2 antibody in HER2-MPDA to HER2 receptor in HER2-overexpressing exosomes causes a blue-to-red color change by altering the molecular structure of the PDA backbone. This colorimetric sensor can simultaneously separate and detect HER2-overexpressing exosomes. HER2-MPDA can detect HER2-overexpressing exosomes in the culture medium of HER2-overexpressing BC cells and in mouse urine samples from a HER2-overexpressing BC mouse model. It can selectively isolate and detect only HER2-overexpressing exosomes through magnetic separation, and its detection limit is found to be 8.5 × 108 particles mL-1. This colorimetric sensor can be used for point-of-care diagnosis of HER2-overexpressing BC in LMICs.

Toxicity Analysis of Mesoporous Polydopamine on Intestinal Tissue and Microflora.

As a promising therapy, photothermal therapy (PTT) converts near-infrared (NIR) light into heat through efficient photothermal agents (PTAs), causing a rapid increase in local temperature. Considering the importance of PTAs in the clinical application of PTT, the safety of PTAs should be carefully evaluated before their widespread use. As a promising PTA, mesoporous polydopamine (MPDA) was studied for its clinical applications for tumor photothermal therapy and drug delivery. Given the important role that intestinal microflora plays in health, the impacts of MPDA on the intestine and on intestinal microflora were systematically evaluated in this study. Through biological and animal experiments, it was found that MPDA exhibited excellent biocompatibility, in vitro and in vivo. Moreover, 16S rRNA analysis demonstrated that there was no obvious difference in the composition and classification of intestinal microflora between different drug delivery groups and the control group. The results provided new evidence that MPDA was safe to use in large doses via different drug delivery means, and this lays the foundation for further clinical applications.

Dual-responsive and NIR-driven free radical nanoamplifier with glutathione depletion for enhanced tumor-specific photothermal/thermodynamic/chemodynamic synergistic Therapy.

The efficacy of free radical-based therapeutic strategies is severely hindered by nonspecific accumulation, premature release and glutathione (GSH) scavenging effects. Herein, a tumor microenvironment-responsive MPDA/AIPH@Cu-TA@HA (abbreviated as MACTH) nanoplatform was constructed by coating Cu2+ and tannic acid (TA) on the surface of azo initiator (AIPH)-loaded mesoporous polydopamine (MPDA) nanoparticles and further modifying them with hyaluronic acid (HA) to achieve tumor-specific photothermal/thermodynamic/chemodynamic synergistic therapy (PTT/TDT/CDT). Once accumulated and internalized into cancer cells through CD44 receptor-mediated active targeting and endocytosis, the HA shell of MACTH would be preliminarily degraded by hyaluronidase (HAase) to expose the Cu-TA metal-phenolic networks, which would further dissociate in response to an acidic lysosomal environment, leading to HAase/pH dual-responsive release of Cu2+ and AIPH. On the one hand, the released Cu2+ could deplete the overexpressed GSH via redox reactions and produce Cu+, which in turn catalyzes endogenous H2O2 into highly cytotoxic hydroxyl radicals (˙OH) for CDT. On the other hand, the local hyperthermia generated by MACTH under 808 nm laser irradiation could not only augment CDT efficacy through accelerating the Cu+-mediated Fenton-like reaction, but also trigger the decomposition of AIPH to produce biotoxic alkyl radicals (˙R) for TDT. The consumption of GSH and accumulation of oxygen-independent free radicals (˙OH/˙R) synergistically amplified intracellular oxidative stress, resulting in substantial apoptotic cell death and significant tumor growth inhibition. Collectively, this study provides a promising paradigm for customizing stimuli-responsive free radical-based nanoplatforms to achieve accurate and efficacious cancer treatment.

Mesoporous Polydopamine-Based Nanovehicles as a Versatile Drug Loading Platform to Enable Tumor-Sufficient Synergistic Therapy.

The combination of photothermal therapy and chemotherapy are developing as a promising clinical strategy but it urgently needs the high exploration of intelligent multifunctional drug delivery nanovectors. In this paper, we used a versatile method to construct mesoporous polydopamine nanovehicles (MPDA) with the dendritic mesopores loaded with a clinical chemotherapeutic drug, Doxorubicin (MPDA@DOX). The monodisperse nanoagents are spherical with a size of ∼160 nm and pore size of approximately 10 nm. MPDA could efficiently delivery DOX with π-π stacking interaction and acts as the potent photothermal agents. Importantly, MPDA@DOX are preferentially internalized by cancerous cells, then bursting drug release and local hyperthermia generation were observed in conditions representative of the cytoplasm in tumor cells that highly synergistic cell killing effect were found under 808 nm laser irradiation. The fluorescent imaging results of human breast tumor bearing murine model evidenced that MPDA delivery platform have excellent tumor precise targeting effect and in vivo tumor ablation experiment further revealed that MPDA@DOX showed markedly eradicated tumor growth capability under laser exposure. Therefore, this work provided a fascinating strategy based on biocompatible MPDA based drug delivery system for malignant tumors eradication via synergistic therapy.

Nickel-Metformin Ternary Complexes: Geometrical, Thermal, DNA Binding, and Molecular Docking Studies.

Novel three nickel(II) complexes of type [Ni(metf)(o-phen)2]Cl2 (1), [Ni(metf)(opda)2]Cl2 (2), [Ni(metf)(2-2'bipy)2]Cl2 (3), (Metf = metformin, o-phen = ortho-phenanthroline, opda = ortho-phenylenediamine, 2-2' bipy = 2-2' bipyridyl) were synthesized and characterized by various analytical and spectral techniques. Based on these studies, octahedral geometry is assigned to these complexes. The DNA binding properties of these complexes were investigated by absorption, emission, and viscosity studies. From the spectral data, it was concluded that the complexes bind to DNA through groove mode of binding. The intrinsic binding constants (Kb) from absorption spectroscopy were 1.60 × 104, 3.57 × 104, and 5.70 × 104 M-1 for 1, 2, and 3, respectively, and Stern-Volmer quenching constants (Ksv) from emission spectroscopy were 0.11, 0.87, and 0.24, respectively. Thermal degradation pattern of the compounds was studied and Coats-Redfern method is used to determine kinetic parameters for complexes 1, 2, and 3 from thermal studies. The software Discovery Studio 2.1 was used to assess the binding affinity and interaction pattern of complexes with the B-DNA receptor protein and complex 1 has the highest dock score.

Accurate determination of total biophenols in unfractionated extra-virgin olive oil with the fast blue BB assay.

The phenolic compounds of extra-virgin olive oil (EVOO) are key contributors of nutritional and sensory quality as well as chemical stability. The reference method for their determination is the HPLC-UV, which is cost-/time-expensive. In this work, total phenolic compounds were evaluated in unfractionated EVOO adapting the Fast Blue BB (FBBB) assay, which involves the spectrophotometric (absorbance at 420 nm) determination of azo derivatives resulting from the coupling of phenolic compounds with FBBB diazonium salt in alkali pH. When tested on 26 EVOO samples, the FBBB assay and HPLC-determinations were strikingly correlated (R2 = 0.9653), differently from FBBB and Folin-Ciocalteu assays, which showed poor correlation. The assay is simple, repeatable, robust, rapid and cheap, and results might be evaluated on a printed colorimetric scale. This protocol of the FBBB assay could be routinely used to categorize EVOO according to the health claim allowed by EFSA concerning the content of phenolic compounds.

A Clinical PET Imaging Tracer ([18F]DASA-23) to Monitor Pyruvate Kinase M2-Induced Glycolytic Reprogramming in Glioblastoma.

PURPOSE: Pyruvate kinase M2 (PKM2) catalyzes the final step in glycolysis, a key process of cancer metabolism. PKM2 is preferentially expressed by glioblastoma (GBM) cells with minimal expression in healthy brain. We describe the development, validation, and translation of a novel PET tracer to study PKM2 in GBM. We evaluated 1-((2-fluoro-6-[18F]fluorophenyl)sulfonyl)-4-((4-methoxyphenyl)sulfonyl)piperazine ([18F]DASA-23) in cell culture, mouse models of GBM, healthy human volunteers, and patients with GBM. EXPERIMENTAL DESIGN: [18F]DASA-23 was synthesized with a molar activity of 100.47 ± 29.58 GBq/µmol and radiochemical purity >95%. We performed initial testing of [18F]DASA-23 in GBM cell culture and human GBM xenografts implanted orthotopically into mice. Next, we produced [18F]DASA-23 under FDA oversight, and evaluated it in healthy volunteers and a pilot cohort of patients with glioma. RESULTS: In mouse imaging studies, [18F]DASA-23 clearly delineated the U87 GBM from surrounding healthy brain tissue and had a tumor-to-brain ratio of 3.6 ± 0.5. In human volunteers, [18F]DASA-23 crossed the intact blood-brain barrier and was rapidly cleared. In patients with GBM, [18F]DASA-23 successfully outlined tumors visible on contrast-enhanced MRI. The uptake of [18F]DASA-23 was markedly elevated in GBMs compared with normal brain, and it identified a metabolic nonresponder within 1 week of treatment initiation. CONCLUSIONS: We developed and translated [18F]DASA-23 as a new tracer that demonstrated the visualization of aberrantly expressed PKM2 for the first time in human subjects. These results warrant further clinical evaluation of [18F]DASA-23 to assess its utility for imaging therapy-induced normalization of aberrant cancer metabolism.

Radical-Mediated Activation of Esters with a Copper/Selectfluor System: Synthesis of Bulky Amides and Peptides.

Herein, we describe a new approach for the activation of esters via a radical-mediated process enabled by a copper/Selectfluor system. A variety of para-methoxybenzyl esters derived from bulky carboxylic acids and amino acids can be easily converted into the corresponding acyl fluorides, directly used in the one-pot synthesis of amides and peptides. As a proof of concept, this method was applied to the iterative formation of sterically hindered amide bonds.

Development of a label-free electrochemical aptasensor based on diazonium electrodeposition: Application to cadmium detection in water.

In this study we have developed a new aptasensor for cadmium (Cd2+) detection in water. Gold electrode surface has been chemically modified by electrochemical reduction of diazonium salt (CMA) with carboxylic acid outward from the surface. This was used for amino-modified cadmium aptamer immobilization through carbodiimide reaction. Chemical surface modification was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). This latter was also used for Cd2+ detection. The aptasensor has exhibited a good linear relationship between the logarithm of the Cd2+ concentration and the impedance changes in the range from 10-3 to 10-9 M with a correlation R2 of 0.9954. A high sensitivity was obtained with a low limit of detection (LOD) of 2.75*10-10 M. Moreover, the developed aptasensor showed a high selectivity towards Cd2+ when compared to other interferences such as Hg2+, Pb2+ and Zn2+. The developed aptasensor presents a simple and sensitive approach for Cd2+detection in aqueous solutions with application for trace Cd2+ detection in spring water samples.

Tumor treating fields (TTFields) impairs aberrant glycolysis in glioblastoma as evaluated by [18F]DASA-23, a non-invasive probe of pyruvate kinase M2 (PKM2) expression.

Despite the anti-proliferative and survival benefits from tumor treating fields (TTFields) in human glioblastoma (hGBM), little is known about the effects of this form of alternating electric fields therapy on the aberrant glycolysis of hGBM. [18F]FDG is the most common radiotracer in cancer metabolic imaging, but its utility in hGBM is impaired due to high glucose uptake in normal brain tissue. With TTFields, radiochemistry, Western blot, and immunofluorescence microscopy, we identified pyruvate kinase M2 (PKM2) as a biomarker of hGBM response to therapeutic TTFields. We used [18F]DASA-23, a novel radiotracer that measures PKM2 expression and which has been shown to be safe in humans, to detect a shift away from hGBM aberrant glycolysis in response to TTFields. Compared to unexposed hGBM, [18F]DASA-23 uptake was reduced in hGBM exposed to TTFields (53%, P< 0.05) or temozolomide chemotherapy (33%, P > 0.05) for 3 d. A 6-d TTFields exposure resulted in a 31% reduction (P = 0.043) in 60-min uptake of [18F]DASA-23. [18F]DASA-23 was retained after a 10 but not 30-min wash-out period. Compared to [18F]FDG, [18F]DASA-23 demonstrated a 4- to 9-fold greater uptake, implying an improved tumor-to-background ratio. Furthermore, compared to no-TTFields exposure, a 6-d TTFields exposure caused a 35% reduction in [18F]DASA-23 30-min uptake compared to only an 8% reduction in [18F]FDG 30-min uptake. Quantitative Western blot analysis and qualitative immunofluorescence for PKM2 confirmed the TTFields-induced reduction in PKM2 expression. This is the first study to demonstrate that TTFields impairs hGBM aberrant glycolytic metabolism through reduced PKM2 expression, which can be non-invasively detected by the [18F]DASA-23 radiotracer.

Catalytic rhodium (Rh)-based (mesoporous polydopamine) MPDA nanoparticles with enhanced phototherapeutic efficiency for overcoming tumor hypoxia.

Catalytic nanomedicine with high oxygen-generation efficiency may find applications in alleviating tumor hypoxia and improving the efficiency of photodynamic therapy (PDT). In this study, a catalytic nanosystem (Ce6-Rh@MPDA) was developed using mesoporous polydopamine nanoparticles (MPDA) to encapsulate catalase-like rhodium nanoparticles (Rh NPs) and photosensitizer chlorine6 (Ce6) for photoacoustic/fluorescence dual imaging-guided tumor therapy. The Rh NPs can catalyze the production of O2 from tumor-enriched H2O2in situ, in which the mesoporous structure of MPDA plays an important role via improving the catalytic efficiency of Rh NPs. Moreover, the hyperthermia generated by both MPDA and Rh NPs under laser irradiation accelerates the O2 generation to promote the PDT. The Ce6-Rh@MPDA nanoparticles described herein represent a multifunctional metal-based catalytic nanomedicine which not only alleviates tumor hypoxia but also enables a synergistic antitumor treatment using PTT and PDT.

Characterization of the biosorption of fast black azo dye K salt by the bacterium Rhodopseudomonas palustris 51ATA strain

BACKGROUND: Removal of dyes from wastewater by microorganisms through adsorption, degradation, or accumulation has been investigated. Biological methods used for dye treatment are generally always effective and environmentally friendly. In this study, biosorption of the Fast Black K salt azo dye by the bacterium Rhodopseudomonas palustris 51ATA was studied spectrophotometrically, at various pH (2­10), temperatures (25°C, 35°C, and 45°C) and dye concentrations (25­400 mg L-1). RESULTS: The bacterial strain showed extremely good dye-removing potential at various dye concentrations. IR studies at different temperatures showed that the dye was adsorbed on the bacterial surface at lower temperatures. Characteristics of the adsorption process were investigated by Scatchard analysis at 25°C and 35°C. Scatchard analysis of the equilibrium binding data for the dye on this bacterium gave rise to linear plots, indicating that the Langmuir model could be applied. The regression coefficients obtained for the dye from the Freundlich and Langmuir models were significant and divergence from the Scatchard plot was observed. CONCLUSION: The adsorption behavior of the dye on this bacterium was expressed by the Langmuir, Freundlich, and Temkin isotherms. The adsorption data with respect to various temperatures provided an excellent fit to the Freundlich isotherm. However, when the Langmuir and Temkin isotherm models were applied to these data, a good fit was only obtained for the dye at lower temperatures, thus indicating that the biosorption ability of R. palustris 51ATA is dependent on temperature, pH, and dye concentration.

Exploring the Photodynamic Properties of Two Antiproliferative Benzodiazopyrrole Derivatives.

The identification of molecules whose biological activity can be properly modulated by light is a promising therapeutic approach aimed to improve drug selectivity and efficacy on the molecular target and to limit the side effects compared to traditional drugs. Recently, two photo-switchable diastereomeric benzodiazopyrrole derivatives 1RR and 1RS have been reported as microtubules targeting agents (MTAs) on human colorectal carcinoma p53 null cell line (HCT 116 p53-/-). Their IC50 was enhanced upon Light Emitting Diode (LED) irradiation at 435 nm and was related to their cis form. Here we have investigated the photo-responsive behavior of the acid derivatives of 1RR and 1RS, namely, d1RR and d1RS, in phosphate buffer solutions at different pH. The comparison of the UV spectra, acquired before and after LED irradiation, indicated that the trans→cis conversion of d1RR and d1RS is affected by the degree of ionization. The apparent rate constants were calculated from the kinetic data by means of fast UV spectroscopy and the conformers of the putative ionic species present in solution (pH range: 5.7-8.0) were modelled. Taken together, our experimental and theoretical results suggest that the photo-conversions of trans d1RR/d1RS into the corresponding cis forms and the thermal decay of cis d1RR/d1RS are dependent on the presence of diazonium form of d1RR/d1RS. Finally, a photo-reaction was detected only for d1RR after prolonged LED irradiation in acidic medium, and the resulting product was characterized by means of Liquid Chromatography coupled to High resolution Mass Spectrometry (LC-HRMS) and Nuclear Magnetic Resonance (NMR) spectroscopy.

Human biodistribution and radiation dosimetry of [18F]DASA-23, a PET probe targeting pyruvate kinase M2.

PURPOSE: To assess the safety, biodistribution, and radiation dosimetry of the novel positron emission tomography (PET) radiopharmaceutical 1-((2-fluoro-6-[[18F]]fluorophenyl)sulfonyl)-4-((4-methoxyphenyl)sulfonyl)piperazine ([18F]DASA-23) in healthy volunteers. METHODS: We recruited 5 healthy volunteers who provided a written informed consent. Volunteers were injected with 295.0 ± 8.2 MBq of [18F]DASA-23 intravenously. Immediately following injection, a dynamic scan of the brain was acquired for 15 min. This was followed by serial whole-body PET/MRI scans acquired up to 3 h post-injection. Blood samples were collected at regular intervals, and vital signs monitored pre- and post-radiotracer administration. Regions of interest were drawn around multiple organs, time-activity curves were calculated, and organ uptake and dosimetry were estimated with OLINDA/EXM (version 1.1) software. RESULTS: All subjects tolerated the PET/MRI examination, without adverse reactions to [18F]DASA-23. [18F]DASA-23 passively crossed the blood-brain barrier, followed by rapid clearance from the brain. High accumulation of [18F]DASA-23 was noted in organs such as the gallbladder, liver, small intestine, and urinary bladder, suggesting hepatobiliary and urinary clearance. The effective dose of [18F]DASA-23 was 23.5 ± 5.8 µSv/MBq. CONCLUSION: We successfully completed a pilot first-in-human study of [18F]DASA-23. Our results indicate that [18F]DASA-23 can be used safely in humans to evaluate pyruvate kinase M2 levels. Ongoing studies are evaluating the ability of [18F]DASA-23 to visualize intracranial malignancies, NCT03539731. TRIAL REGISTRATION: ClinicalTrials.gov , NCT03539731 (registered 28 May 2018).

Biodiesel synthesis from oil palm empty fruit bunch biochar derived heterogeneous solid catalyst using 4-benzenediazonium sulfonate.

In this research, biomass from oil palm empty fruit bunch was used as the carbon precursor and sulfonated by 4-benzenediazonium sulfonate (4-BDS) to produce solid acid catalyst. The as-synthesized catalysts were characterized and the performances were tested in esterification of palm fatty acid distillate (PFAD) for biodiesel production. Scanning Electron Microscopy (SEM) showed that clear porous and rough carbon surface was successfully developed after calcination which favored the attachment of sulfonic groups. Thermogravimetric Analysis (TGA) result showed that the catalyst was thermally stable up to 600 °C. Fourier Transform Infrared Spectroscopy (FTIR) proved that SO and SO3H sulfonic groups were successfully attached to the carbon catalyst. From the catalytic activity tests, the results showed that the catalyst which was calcined at 200 °C and sulfonated with 15:1 sulfanilic acid to AC ratio was the optimum catalyst as it provided the highest biodiesel yield. Further investigation showed that the reaction time of 7 h and 20 wt.% of catalyst loading were reported as optimum esterification conditions which provided the highest biodiesel yield at 98.1 %.

Remote eradication of biofilm on titanium implant via near-infrared light triggered photothermal/photodynamic therapy strategy.

Biofilm formation is a main challenge in treatment of bone-implant-associated infections, resulting in tolerance to immune system and antibiotics. However, smart non-surgical or non-invasive treatment methods of combating established biofilm on an implant have been less reported. Herein, a therapeutic system consisting of mesoporous polydopamine nanoparticles (MPDA) to combat biofilm is reported for the first time. We develop a synergistic photothermal/photodynamic therapy (PTT/PDT) strategy aiming for biofilms eradication on titanium (Ti) implant, which is integrated with MPDA loading with photosensitizer Indocyanine Green (ICG) by π-π stacking. Specifically, MPDA is functionalized with RGD peptide to endow the modified Ti sample (Ti-M/I/RGD) with good cytocompatibility. More importantly, Ti-M/I/RGD implant remarkably kills Staphylococcus aureus (S. aureus) biofilm with an efficiency of 95.4% in vivo upon near infrared (NIR). After biofilm eradication, implant still displays great performance regarding osteogenesis and osseointegration. Overall, this study provides a PTT/PDT strtategy for the development of antibacterial Ti implants for potential orthpediac application.

A Benzenediamine Analog FC-99 Drives M2 Macrophage Polarization and Alleviates Lipopolysaccharide- (LPS-) Induced Liver Injury.

Macrophages have variable functional phenotypes, high diversity, and plasticity and are involved in the pathogenesis of sepsis-induced liver injury. Alteration of macrophage polarization through activated (M1) macrophage to alternatively activated (M2) macrophage has emerged as a potential therapeutic strategy. This study was designed to explore the effect of a benzenediamine analog FC-99 on macrophage polarization in vitro and lipopolysaccharide- (LPS-) induced liver injury followed by the underlying mechanisms. For in vitro experiments, FC-99 inhibited M1-related macrophage factors and promoted M2-related markers induced by IL-4 in the mouse macrophage cell line RAW264.7. Moreover, FC-99-induced macrophages polarized to M2 phenotype which could be repressed by a PPAR-γ inhibitor but not STAT6 siRNA knockdown, indicating FC-99-induced M2 macrophage polarization through PPAR-γ rather than STAT6 signal. In LPS-induced septic mice, FC-99 pretreated mice displayed lower expression of M1 markers together with the increased M2 marker CD206 and improvement of liver injury. These findings illustrated that FC-99 could promote M2 macrophage polarization via PPAR-γ signaling and seemed to be a potential therapeutic candidate for inflammatory liver injury.