Suppressive stroma-immune prognostic signature impedes immunotherapy in ovarian cancer and can be reversed by PDGFRB inhibitors.

BACKGROUND: As the most lethal gynecologic cancer, ovarian cancer (OV) holds the potential of being immunotherapy-responsive. However, only modest therapeutic effects have been achieved by immunotherapies such as immune checkpoint blockade. This study aims to propose a generalized stroma-immune prognostic signature (SIPS) to identify OV patients who may benefit from immunotherapy. METHODS: The 2097 OV patients included in the study were significant with high-grade serous ovarian cancer in the III/IV stage. The 470 immune-related signatures were collected and analyzed by the Cox regression and Lasso algorithm to generalize a credible SIPS. Correlations between the SIPS signature and tumor microenvironment were further analyzed. The critical immunosuppressive role of stroma indicated by the SIPS was further validated by targeting the major suppressive stroma component (CAFs, Cancer-associated fibroblasts) in vitro and in vivo. With four machine-learning methods predicting tumor immune subtypes, the stroma-immune signature was upgraded to a 23-gene signature. RESULTS: The SIPS effectively discriminated the high-risk individuals in the training and validating cohorts, where the high SIPS succeeded in predicting worse survival in several immunotherapy cohorts. The SIPS signature was positively correlated with stroma components, especially CAFs and immunosuppressive cells in the tumor microenvironment, indicating the critical suppressive stroma-immune network. The combination of CAFs' marker PDGFRB inhibitors and frontline PARP inhibitors substantially inhibited tumor growth and promoted the survival of OV-bearing mice. The stroma-immune signature was upgraded to a 23-gene signature to improve clinical utility. Several drug types that suppress stroma-immune signatures, such as EGFR inhibitors, could be candidates for potential immunotherapeutic combinations in ovarian cancer. CONCLUSIONS: The stroma-immune signature could efficiently predict the immunotherapeutic sensitivity of OV patients. Immunotherapy and auxiliary drugs targeting stroma could enhance immunotherapeutic efficacy in ovarian cancer.

Comparison of microstructure, mechanical property, and degradation rate of Mg-1Li-1Ca and Mg-4Li-1Ca alloys.

Mg-1 wt.% Li-1 wt.% Ca (LX11) and Mg-4 wt.% Li-1 wt.% Ca (LX41) alloys share the same hexagonal closed-packed crystalline structure. However, the differences in microstructure, mechanical properties, and degradation rates between the two alloys are not well understood. Hereby, the above three aspects of LX11 and LX41 alloys were studied via optical microscopy, tensile tests, and electrochemical polarization and electrochemical impedance spectroscopy, together with hydrogen evolution. The concentration of the released Mg2+, Ca2+, and Li+ ions was analyzed using a flame atomic absorption spectrophotometer. Results demonstrated that the LX11 alloy was composed of finer α-Mg grains, fewer twins, and smaller volume fractions of the intermetallic phases Mg2Ca than the LX41 alloy. The increasing Li concentration generated a weak decrease in the yield strength of the Mg-Li-Ca alloys, a remarkable increase in elongation to failure, and a stable ultimate tensile strength. The LX11 alloy had better corrosion resistance than the LX41 alloy. The release rate of the cations (Mg2+, Ca2+, and Li+) varied significantly with time. The release rate of metallic ions in Hank's solution cannot reflect the true corrosion rate of Mg-Li-Ca alloys due to the formation of the precipitated corrosion products and their difference in solubility. The dealloying corrosion mechanism of the Mg2Ca phase in Mg-Li-Ca alloys was proposed.

Corrosion resistance and mechanisms of Nd(NO3)3 and polyvinyl alcohol organic-inorganic hybrid material incorporated MAO coatings on AZ31 Mg alloy.

This work reports the design and preparation of novel organic (polyvinyl alcohol, PVA)-inorganic (neodymium nitrate, Nd(NO3)3) hybrid coatings on micro-arc oxidation (MAO) coating for magnesium (Mg) alloy corrosion protection. X-ray diffractometer, X-ray photoelectron spectroscopy, fourier transform infrared spectroscopy, field emission scanning electron microscope, Energy Dispersive X-ray spectrometer and surface roughness were applied to characterize the chemical composition and surface morphology of the coatings. The corrosion resistance of the coatings was evaluated by electrochemical and salt spray tests. The results suggested that the formation of PVA-Nd3+ and PVA-Mg2+ complexes promoted the enrichment of Nd3+ on the surface, and thereby improved the sealing quality and compactness of the coating. Interestingly, when the coating was damaged, the Nd3+ ions were transformed to their carbonates and covered the active sites, and thus exhibiting self-healing function. Further, the corrosion resistance of PVA-Nd3+ modified MAO composite coating on AZ31 Mg alloy was improved.

In vitro corrosion resistance and dual antibacterial ability of curcumin loaded composite coatings on AZ31 alloy: Effect of amorphous calcium carbonate.

Rapid corrosion and bacterial infection are obstacles to put into use biodegradable magnesium (Mg) alloy as biomedical materials. In this research, an amorphous calcium carbonate (ACC)@curcumin (Cur) loaded poly-methyltrimethoxysilane (PMTMS) coating prepared by self-assembly method on micro-arc oxidation (MAO) coated Mg alloy has been proposed. Scanning electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy are adopted to analyze the morphology and composition of the obtained coatings. The corrosion behaviour of the coatings is estimated by hydrogen evolution and electrochemical tests. The spread plate method without or with 808 nm near-infrared irradiation is applied to evaluate the antimicrobial and photothermal antimicrobial ability of the coatings. Cytotoxicity of the samples is tested by 3-(4,5)-dimethylthiahiazo(-z-y1)-2,5-di- phenytetrazoliumromide (MTT) and live/dead assay culturing with MC3T3-E1 cells. Results show that the MAO/ACC@Cur-PMTMS coating exhibited favourable corrosion resistance, dual antibacterial ability, and good biocompatibility. Cur was employed as an antibacterial agent and photosensitizer for photothermal therapy. The core of ACC significantly improved the loading of Cur and the deposition of hydroxyapatite corrosion products during degradation, which greatly promoted the long-term corrosion resistance and antibacterial activity of Mg alloys as biomedical materials.

Corrosion self-warning and repair tracking of polymeric coatings based on stimulus responsive nanosensors.

Smart polymeric coatings with early corrosion self-warning and damage self-repairing characteristics have garnered tremendous interest due to their ability to sense corrosion reactions and repair coating defects. However, tracking the repair process and its underlying protection mechanism is highly challenging. Herein, we report the construction of a novel composite coating by incorporating multifunctional nanosensors (graphene oxide-zeolitic imidazole frameworks loaded with 1,10-phenanthroline) into a thermo-responsive polyurethane. Under damaging events, the localized acidity derived from metal corrosion stimulates the decomposition of the nanosensors to produce 1,10-phenanthroline and benzimidazole. The generated ferrous ions are rapidly sensed by the released 1,10-phenanthroline to produce a conspicuous red color, which warns of the corrosion occurrence. In profiting from the photothermal effect of graphene oxide, the composite coating exhibits efficient crack closure behavior under near-infrared light irradiation. Morphology observation indicates that a coating scratch (about 30 µm wide) almost closed with 20 s of irradiation. The photothermally activated crack closure combined with benzimidazole inhibition endows the prepared coating with superior self-repairing performance. Interestingly, the change in color intensity around the coating defect can assist in tracking the repair process. Therefore, this work provides a novel strategy to visualize microscopic behaviors during damage and repair processes.

In vitro degradation, photo-dynamic and thermal antibacterial activities of Cu-bearing chlorophyllin-induced Ca-P coating on magnesium alloy AZ31.

Surgical failures, caused by postoperative infections of bone implants, are commonly met, which cannot be treated precisely with intravenous antibiotics. Photothermal therapy (PTT) and photodynamic therapy (PDT) have attracted widespread attention due to their non-invasive antibacterial effects on tissues and no bacterial resistance, which may be an excellent approach to solve infections related to bone implants for biodegradable magnesium alloys. Herein, a sodium copper chlorophyllin (SCC) with a porphyrin ring induced Ca-P coating was prepared on AZ31 magnesium alloy via layer-by-layer (LbL) assembly. The morphology and composition of the samples were characterized through field emission scanning electron microscope (FE-SEM) with affiliated energy dispersive spectrometer (EDS), X-ray diffractometer (XRD), and Fourier infrared spectrometer (FTIR) and X-ray photoelectron spectrometer (XPS) as well. Potentiodynamic polarization, electrochemical impedance spectroscopy (EIS) and hydrogen evolution experiments were employed to evaluate the corrosion behavior of the samples. Atomic absorption spectrophotometer was used to measure Cu elemental content of different immersion periods. Cytocompatibility and antibacterial performance of the coatings were probed using in vitro cytotoxicity tests (MTT assay), live/dead cell staining and plate counting method. The results showed that the obtained (Ca-P/SCC)10 coating exhibited good corrosion resistance, antimicrobial activity (especially under 808 nm irradiation) and biocompatibility. The antibacterial rates for E. coli and S. aureus were 99.9% and 99.8%, respectively; and the photothermal conversion efficiency was as high as 42.1%. Triple antibacterial mechanisms including photodynamic, photothermal reactions and copper-ions release were proposed. This coating exhibited a promising application for biodegradable magnesium alloys.

In vitro corrosion resistance of a Ta2O5 nanofilm on MAO coated magnesium alloy AZ31 by atomic layer deposition.

Micro-arc oxidation (MAO) coating with outstanding adhesion strength to Mg alloys has attracted more and more attention. However, owing to the porous structure, aggressive ions easily invaded the MAO/substrate interface through the through pores, limiting long-term corrosion resistance. Therefore, a dense and biocompatible tantalum oxide (Ta2O5) nanofilm was deposited on MAO coated Mg alloy AZ31 through atomic layer deposition (ALD) technique to seal the micropores and regulate the degradation rate. Surface micrography, chemical compositions and crystallographic structure were characterized using FE-SEM, EDS, XPS and XRD. The corrosion resistance of all samples was evaluated through electrochemical and hydrogen evolution tests. Results revealed that the Ta2O5 film mainly existed in the form of amorphousness. Moreover, uniform deposition of Ta2O5 film and effective sealing of micropores and microcracks in MAO coating were achieved. The current density (i corr) of the composite coating decreased three orders of magnitude than that of the substrate and MAO coating, improving corrosion resistance. Besides, the formation and corrosion resistance mechanisms of the composite coating were proposed.

In vitro corrosion resistance of layer-by-layer assembled polyacrylic acid multilayers induced Ca-P coating on magnesium alloy AZ31.

Biodegradable magnesium (Mg)-based alloys have aroused great concern owing to their promising characteristics as temporary implants for orthopedic application. But their undesirably rapid corrosion rate under physiological conditions has limited the actual clinical application. This study reports the use of a novel biomimetic polyelectrolyte multilayer template, based on polyvinylpyrrolidone (PVP) and polyacrylic acid (PAA) via layer-by-layer (LbL) assembly, to improve the corrosion resistance of the alloy. Surface characterization techniques (field-emission scanning electron microscopy, Fourier transform infrared (FTIR) spectrophotometer and X-ray diffractometer) confirmed the formation of biomineralized Ca-P coating on AZ31 alloy. Both hydrogen evolution and electrochemical corrosion tests demonstrated that the corrosion protection of the polyelectrolyte-induced Ca-P coating on AZ31 alloy. The formation mechanism of biomineralized Ca-P coating was proposed.

In vitro degradation of pure magnesium-the synergetic influences of glucose and albumin.

The biocorrosion of magnesium in the external physiological environment is still difficult to accurately evaluate the degradation behavior in vivo, particularly, in the microenvironment of the patients with hyperglycemia or diabetes. Thus, we explored the synergistic effects of glucose and protein on the biodegradation of pure magnesium, so as to have a deeper understanding the mechanism of the degradation in vivo. The surface morphology and corrosion product composition of pure magnesium were investigated using SEM, EDS, FTIR, XRD and XPS. The effect of glucose and albumin on the degradation rate of pure magnesium was investigated via electrochemical and immersion tests. The adsorption of glucose and albumin on the sample surface was observed using fluorescence microscopy. The results showed that the presence of 2 g/L glucose changed the micromorphology of corrosion products on the magnesium surface by reacting with metal cations, thus inhibiting the corrosion of pure magnesium. Protein formed a barrier layer to protect the magnesium at early stage of immersion. The chelation reaction between protein and magnesium surface might accelerate the degradation at later stage. There may be a critical glucose (albumin) content. Biodegradation of pure magnesium was inhibited at low concentrations and promoted at high concentrations. The synergistic effect of glucose and protein restrained the adsorption of aggressive chloride ions to a certain extent, and thus inhibited the degradation of pure magnesium considerably. Moreover, XPS results indicated that glucose promoted the adsorption of protein on the sample surface.

Corrosion resistance and antibacterial activity of zinc-loaded montmorillonite coatings on biodegradable magnesium alloy AZ31.

A Zinc-loaded montmorillonite (Zn-MMT) coating was hydrothermally prepared using Zn2+ ion intercalated sodium montmorillonite (Na-MMT) upon magnesium (Mg) alloy AZ31 as bone repairing materials. Biodegradation rate of the Mg-based materials was studied via potentiodynamic polarization curves, electrochemical impedance spectroscopy (EIS) and hydrogen evolution tests. Results revealed that both Na-MMT and Zn-MMT coatings exhibited better corrosion resistance in Dulbecco's modified eagle medium (DMEM) + 10% calf serum (CS) than bare Mg alloy AZ31 counterparts. Hemolysis results demonstrated that hemocompatibility of the Na-MMT and Zn-MMT coatings were 5%, and lower than that of uncoated Mg alloy AZ31 pieces. In vitro MTT tests and live-dead stain of osteoblast cells (MC3T3-E1) indicated a significant improvement in cytocompatibility of both Na-MMT and Zn-MMT coatings. Antibacterial properties of two representative bacterial strains associated with device-related infection, i.e. Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus), were employed to explore the antibacterial behavior of the coatings. The measured inhibitory zone and bacterial growth rate confirmed that Zn-MMT coatings exhibited higher suppression toward both E. coli and S. aureus than that of Na-MMT coatings. The investigation on antibacterial mechanism through scanning electron microscopy (SEM) and lactate dehydrogenase (LDH) release assay manifested that Zn-MMT coating led to severe breakage of bacterial membrane of E. coli and S. aureus, which resulted in a release of cytoplasmic materials from the bacterial cells. In addition, the good inhibition of Zn-MMT coatings against E. coli and S. aureus might be attributed to the slow but sustainable release of Zn2+ ions (up to 144 h) from the coatings into the culture media. This study provides a novel coating strategy for manufacturing biodegradable Mg alloys with good corrosion resistance, biocompatibility and antibacterial activity for future orthopedic applications. STATEMENT OF SIGNIFICANCE: The significance of the current work is to develop a corrosion-resistant and antibacterial Zn-MMT coating on magnesium alloy AZ31 through a hydrothermal method. The Zn-MMT coating on magnesium alloy AZ31 shows better corrosion resistance, biocompatibility and excellent antibacterial ability than magnesium alloy AZ31. This study provides a novel coating on Mg alloys for future orthopedic applications.

In vitro corrosion of pure Mg in phosphate buffer solution-Influences of isoelectric point and molecular structure of amino acids.

Influences of proteins on degradation of magnesium alloys are of great significance but not well understood. In particular the roles of amino acids, the basic unit of proteins in regulating the progress of biodegradation of magnesium based materials remain unclear. This study aims to investigate the impacts of alanine, glutamic acid and lysine on degradation of pure magnesium in phosphate buffer solution through SEM, XPS, FTIR, potentiodynamic polarisation curves, electrochemical impedance spectroscopy and immersion tests. The changed contents of amino acids in solutions were detected by UV-vis spectrophotometer. Results demonstrate that the charges of the selected amino acids imposed significant contribution to suppressing the degradation of pure magnesium in phosphate buffer solution. The presence of amino acids led to the formation of phosphate-based corrosion products, increasing free corrosion potential, and reduction in corrosion current density and solution pH depending on their isoelectric points and molecular structures. A plausible corrosion mechanism organised by amino acids on pure magnesium was proposed.

Corrosion resistance of a novel SnO2-doped dicalcium phosphate coating on AZ31 magnesium alloy.

A SnO2-doped dicalcium phosphate coating was prepared on AZ31 alloy by means of hydrothermal deposition. The results showed that the coating possessed a globular morphology with a long lamellar crystalline structure and a thickness of approximately 40 µm. The surface of the coating became smooth with an increase additive amount of the SnO2 nanoparticles. The corrosion current density and hydrogen evolution rate of the coating prepared in presence of SnO2 were reduced compared to the coating without SnO2 and the bare AZ31 substrate, indicating an improvement in the corrosion resistance of the SnO2-doped coating.

Advances in functionalized polymer coatings on biodegradable magnesium alloys - A review.

Magnesium (Mg) and its alloys have become a research frontier in biodegradable materials owing to their superior biocompatibility and excellent biomechanical compatibility. However, their high degradation rate in the physiological environment should be well tackled prior to clinical applications. This review summarizes the latest progress in the development of polymeric coatings on biodegradable Mg alloys over the last decade, regarding preparation strategies for polylactic acid (PLA), poly (latic-co-glycolic) acid (PLGA), polycaprolactone (PCL), polydopamine (PDA), chitosan (CS), collagen (Col) and their composite, and their performance in terms of corrosion resistance and biocompatibility. Feasible perspectives and developing directions of next generation of polymeric coatings with respect to biomedical Mg alloys are briefly discussed. STATEMENT OF SIGNIFICANCE: Magnesium (Mg) and its alloys have become a research frontier in biodegradable materials owing to their superior biocompatibility and suitable biomechanical compatibility. However, the principal drawback of Mg-based implants is their poor corrosion resistance in physiological environments. Hence, it is vital to mitigate the degradation/corrosion behavior of Mg alloys for safe biomedical deployments. This review summarizes the latest progress in development of polymeric coatings on biomedical Mg alloys regarding preparation strategy, corrosion resistance and biocompatibility, including polylactic acid (PLA), poly (latic-co-glycolic) acid (PLGA), polycaprolactone (PCL), chitosan (CS), polydopamine (PDA), collagen (Col) and their composite. In addition, functionalized polymer coatings with Mg alloys exhibits a promising prospect owing to their ability of degradation along with biocompatibility, self-healing, drug-delivery and osteoinduction.

In Vitro Degradation of Pure Magnesium-The Effects of Glucose and/or Amino Acid.

The influences of glucose and amino acid (L-cysteine) on the degradation of pure magnesium have been investigated using SEM, XRD, Fourier transformed infrared (FTIR), X-ray photoelectron spectroscopy (XPS), polarization and electrochemical impedance spectroscopy and immersion tests. The results demonstrate that both amino acid and glucose inhibit the corrosion of pure magnesium in saline solution, whereas the presence of both amino acid and glucose accelerates the corrosion rate of pure magnesium. This may be due to the formation of -C=N- bonding (a functional group of Schiff bases) between amino acid and glucose, which restricts the formation of the protective Mg(OH)2 precipitates.

In Vitro Corrosion and Cytocompatibility of a Microarc Oxidation Coating and Poly(L-lactic acid) Composite Coating on Mg-1Li-1Ca Alloy for Orthopedic Implants.

Manipulating the degradation rate of biomedical magnesium alloys poses a challenge. The characteristics of a microarc oxidation (MAO), prepared in phytic acid, and poly(L-lactic acid) (PLLA) composite coating, fabricated on a novel Mg-1Li-1Ca alloy, were studied through field emission scanning electron microscopy (FE-SEM), electron probe X-ray microanalysis (EPMA), energy dispersive X-ray spectroscopy (EDS) and X-ray diffraction (XRD). The corrosion behaviors of the samples were evaluated via hydrogen evolution, potentiodynamic polarization and electrochemical impedance spectroscopy in Hanks' solution. The results indicated that the MAO/PLLA composite coatings significantly enhanced the corrosion resistance of the Mg-1Li-1Ca alloy. MTT and ALP assays using MC3T3 osteoblasts indicated that the MAO/PLLA coatings greatly improved the cytocompatibility, and the morphology of the cells cultured on different samples exhibited good adhesion. Hemolysis tests showed that the composite coatings endowed the Mg-1Li-1Ca alloys with a low hemolysis ratio. The increased solution pH resulting from the corrosion of magnesium could be tailored by the degradation of PLLA. The degradation mechanism of the composite coatings was discussed. The MAO/PLLA composite coating may be appropriate for applications on degradable Mg-based orthopedic implants.