Novel Mg-Incorporated Micro-Arc Oxidation Coatings for Orthopedic Implants Application.

In this study, Ti-6Al-4V alloy samples were processed by micro-arc oxidation (MAO) in phytic acid (H12Phy) electrolytes with the addition of different concentrations of EDTA-MgNa2 (Na2MgY) and potassium hydroxide (KOH). The surface characterization and cytocompatibility of MAO-treated samples were evaluated systematically. H12Phy is a necessary agent for MAO coating formation, and the addition of Na2MgY and KOH into the electrolytes increases the surface roughness, micropore size and Mg contents in the coatings. The MAO coatings are primarily composed of anatase, rutile, MgO and Mg3(PO4)2. Magnesium (Mg) ions in the electrolytes enter into MAO coatings by diffusion and electromigration. The MAO coatings containing 2.97 at% Mg show excellent cell viability, adhesion, proliferation, alkaline phosphatase activity, extracellular matrix (ECM) mineralization and collagen secretion, but the cytocompatibility of the MAO coatings containing 6.82 at% Mg was the worst due to the excessively high Mg content. Our results revealed that MAO coatings with proper Mg contents improve the cytocompatibility of the Ti-6Al-4V alloys and have large potential in orthopedic applications.

In vitro and in vivo antibacterial performance of Zr & O PIII magnesium alloys with high concentration of oxygen vacancies.

The effects of dual Zr and O plasma immersion ion implantation (Zr & O PIII) on antibacterial properties of ZK60 Mg alloys are systematically investigated. The results show that a hydrophobic, smooth, and ZrO2-containing graded film is formed. Electrochemical assessment shows that the corrosion rate of the plasma-treated Mg alloy decreases and the decreased degradation rate is attributed to the protection rendered by the surface oxide. In vitro and in vivo antibacterial tests reveal Zr & O PIII ZK60 presents higher antibacterial rate compared to Zr PIII ZK60 and untreated control. The hydrophobic and smooth surface suppresses bacterial adhesion. High concentration of oxygen vacancies in the surface films are determined by X-ray photoelectron spectroscopy (XPS), UV-vis diffuse reflectance spectra (UV-vis DRS) and electron paramagnetic resonance (EPR) and involved in the production of reactive oxygen species (ROS). The higher level of ROS expression inhibits biofilm formation by down-regulating the expression of icaADBC genes but up-regulating the expression of icaR gene. In addition, Zr & O PIII improves cell viability and initial cell adhesion confirming good cytocompatibility. Dual Zr & O PIII is a simple and practical means to expedite clinical acceptance of biodegradable magnesium alloys.

Determination of serum 25-hydroxyvitamin D status among population in southern China by a high accuracy LC-MS/MS method traced to reference measurement procedure.

OBJECTIVE: We aimed to describe the 25-hydroxyvitamin D (25(OH)D) status of southern Chinese individuals by a high-accuracy liquid chromatography tandem mass spectrometry (LC-MS/MS) method which can trace to reference measurement procedure. MATERIALS AND METHODS: From January 2018 to June 2019, a total of 4775 southern Chinese individuals were evaluated in our study. The serum levels of parathyroid hormone (PTH) were detected simultaneously in 162 cases. 25(OH)D was determined by LC-MS/MS, and PTH was detected using routine automated analysers. The distribution of the concentration, prevalence and seasonal variability of 25(OH)D in males and females of different age groups were studied. RESULTS: The mean 25(OH)D concentration in our study was 32.57 ng/mL (4.20-101.40 ng/mL). The global 25(OH)D concentration in males was higher than that in females of different age group. The prevalence of vitamin D deficiency (< 20 ng/mL) in females (16.65%) was higher than that in males (6.83%). The prevalence of vitamin D deficiency (< 20 ng/mL) was most common in winter (22.98% of all women and 15.49% of all men). 25(OH)D concentrations were higher in those from whom blood samples were collected in summer and autumn than in winter and spring. 25(OH)D2 was detected in 672 serum samples (14.07%). In addition, there was a negative correlation between the concentrations of 25(OH)D and serum PTH (r = - 0.149, P < 0.05). CONCLUSION: Our study demonstrated that the average serum 25(OH)D concentration in southern Chinese individuals was higher than that in other Chinese cohorts by a high-accuracy LC-MS/MS method. The global 25(OH)D concentration in males was higher than that in females of different ages, and the prevalence of vitamin D deficiency in females was higher than that in males. Seasonal change was an important aspect of 25(OH)D concentration in young and middle-aged people but became less relevant for that in older subjects. 25(OH)D2 detection was of minor practical significance in our study. In addition, we also found that there was a negative correlation between the serum levels of 25(OH)D and PTH in southern Chinese individuals.

Production and characterization of bacterial cellulose obtained by Gluconacetobacter xylinus utilizing the by-products from Baijiu production.

Bacterial cellulose (BC) has extensive application prospects in many fields in view of its unique characteristics. However, the large-scale applications of BC are severely limited because of relatively low BC productivity and high cost of culture medium. Herein, the distiller's grain enzymatic hydrolysate (DEH) and yellow water were successfully combined as an effective substitute (the best distiller's grains-yellow water medium, BDY medium) for traditional Hestrin-Schramm medium (HS medium) for BC production by Gluconacetobacter xylinus through the response surface methodology. The BC production in BDY medium was significantly enhanced to 7.42 g/l with BC conversion yield of 42.4% after 7 days static cultivation, which was 3.72-fold and 3.37-fold higher than that in HS medium, respectively. The structure and properties of BC membranes produced in HS and BDY medium were evaluated by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), field emission scanning electron microscopy (FE-SEM), thermogravimetric analysis (TGA) and hydrophilicity analysis. There was no significant difference between BC samples produced in the HS and BDY medium, indicating that BDY, as abundant and inexpensive substrates, can effectively replace HS medium to enhance BC production. The employment of distiller's grains and yellow water to BC production not only is conducive to achieve industrial production of BC, but also can effectively realize the recycling of waste from Baijiu distillery.

In vitro study on cytocompatibility and osteogenesis ability of Ti-Cu alloy.

Titanium implants easily suffer bacteria-related infections in clinic due to their inherent lack of self-protection ability. Therefore, a novel Ti-Cu alloy with good antibacterial activity has been developed as a new kind of implant material. This study focuses on a systematic evaluation of both cytocompatibility and osteogenesis activity of the Ti-Cu alloy in vitro. It was revealed that an addition of 5% Cu into pure Ti would not cause any negative effect on osteoblasts adhesion, proliferation and apoptosis cultured with Ti-Cu alloy. In addition, Ti-Cu alloy could significantly promote the osteogenic differentiation of MG 63 cells by upregulating the osteogenesis-related gene expressions including alkaline phosphatase (ALP), Collagen I (Colla I), osteopontin (OPN) and osteocalcin (OCN). These promising results suggest that the Ti-Cu alloy has great potential to be used as a multi-functional titanium implant for clinical applications.

In vitro and in vivo studies of anti-bacterial copper-bearing titanium alloy for dental application.

OBJECTIVE: A novel copper-bearing titanium alloy (Ti-Cu) was fabricated for dental application that is expected to efficiently restrain the growth of bacteria and discourage biofilm formation. The aim of this study was to investigate both the antibacterial activity and biofilm inhibition of Ti-Cu alloy in vitro, and the antibacterial effect of Ti-Cu implant in early stage of peri-implantitis in vivo. METHODS: Staphylococcus aureus and Escherichia coli were selected to evaluate the antibacterial activity of Ti-Cu alloy and Ti served as control. The antibacterial rate, attached bacteria and developed biofilms were studied from quantitative antibacterial test, biofilm observation and bacterial morphological examination. Electrochemical tests were used to investigate the corrosion property of Ti-Cu alloy. Furthermore, both Ti and Ti-Cu dental implants were manufactured and then implanted in the mandibular premolar sites of beagle dogs for 3 months with ligature-infected treatment. Implant-tissue samples were prepared for radiographic analysis, Micro-CT evaluation and histological examination. RESULTS: Ti-Cu alloy was found to efficiently kill the attached bacteria by ways of damaging cell membranes and cell walls and strongly inhibit the biofilm formation. However, Ti-Cu alloy had excellent corrosion resistance similar with Ti. Further, Ti-Cu dental implants showed superior capacities of inhibiting the bone resorption caused by bacterial infection and enhancing bone formation. SIGNIFICANCE: Ti-Cu alloy strongly inhibited biofilm formation in vitro and prevented bacterial infection associated with dental implant in vivo, making it great potential for application in dental implants with excellent antibacterial viability and positive effect against bone resorption induced by peri-implantitis.

Synthesis and antitumor, antityrosinase, and antioxidant activities of xanthone.

Ten substituted 1,3-dihydroxyxanthones were synthesized in one step. The yields ranged from 40 to 76%. Compounds 8-10 were first reported. Next, the compounds' in vitro anti-proliferative activities against nine human cancer cell lines, antityrosinase, and antioxidant activities were evaluated. Compounds 1, 4, 6-7, and 9-10 exhibited enhanced cytotoxicity against certain cancer cells. Compounds 2, 8, 9, and 10 inhibited tyrosinase activity to a certain extent. In addition, compound 4 exhibited the best antioxidant activity, which was consistent with theoretical calculations. These results demonstrated that compounds 1-2, 4, and 6-10 were promising leads for further investigation.

Investigation of Zinc and Phosphorus Elements Incorporated into Micro-Arc Oxidation Coatings Developed on Ti-6Al-4V Alloys.

In order to clarify the mechanism that zinc and phosphorus elements entering the micro-arc oxidation (MAO) coatings developed on Ti-6Al-4V alloys, anodic coatings containing different zinc and phosphorus were fabricated using an orthogonal experiment of four factors with three levels in an electrolyte containing EDTA-ZnNa2, KOH, and phytic acid. Surface morphology, element composition, chemical state and phase structure of MAO coatings were characterized by scanning electron microscope (SEM), energy dispersive X-ray spectrometer (EDS), X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD). The concentrations of zinc and phosphorus in the electrolyte were analyzed by an inductively coupled plasma optical emission spectrometry (ICP-OES). The results show that zinc and phosphorus elements in MAO coatings exist in the form of Zn3(PO4)2. Phytic acid is the most important factor on both zinc and phosphorus contents of MAO coatings. With the increase of phytic acid concentration or the decrease of KOH concentration, the contents of zinc and phosphorus in MAO coatings present a similarly increasing tendency. Our results indicate that phosphorus takes part in coating formation mainly by diffusion, while zinc enters into MAO coatings with phosphorus from phytic acid.

Designing Core-Shell Gold and Selenium Nanocomposites for Cancer Radiochemotherapy.

Radiotherapy is an important regime for treating malignant tumors. There is interest in the development of radiosensitizers to increase the local treatment efficacy under a relatively low and safe radiation dose. In this study, we designed Au@Se-R/A nanocomposites (Au@Se-R/A NCs) as nano-radiosensitizer to realize synergistic radiochemotherapy based on the radiotherapy sensitization property of Au nanorods (NRs) and antitumor activity of Se NPs. In vitro studies show that the combined treatment of A375 melanoma cells in culture with NCs and X-ray induces cell apoptosis through alteration in expression of p53 and DNA-damaging genes and triggers intracellular ROS overproduction, leading to greatly enhanced anticancer efficacy. Further studies using clinically used radiotherapy equipment demonstrate that the combined treatment of NCs and X-ray significantly inhibits the tumor growth in vivo and shows negligible acute toxicity to the major organs. Taken together, this study provides a strategy for clinical translation application of nanomedicne in cancer radiochemotherapy.

Nucleolin-targeted Extracellular Vesicles as a Versatile Platform for Biologics Delivery to Breast Cancer.

Small interfering RNAs (siRNA)/microRNAs (miRNA) have promising therapeutic potential, yet their clinical application has been hampered by the lack of appropriate delivery systems. Herein, we employed extracellular vesicles (EVs) as a targeted delivery system for small RNAs. EVs are cell-derived small vesicles that participate in cell-to-cell communication for protein and RNA delivery. We used the aptamer AS1411-modified EVs for targeted delivery of siRNA/microRNA to breast cancer tissues. Tumor targeting was facilitated via AS1411 binding to nucleolin, which is highly expressed on the surface membrane of breast cancer cells. This delivery vesicle targeted let-7 miRNA delivery to MDA-MB-231 cells in vitro as confirmed with fluorescent microscopic imaging and flow cytometry. Also, intravenously delivered AS1411-EVs loaded with miRNA let-7 labeled with the fluorescent marker, Cy5, selectively targeted tumor tissues in tumor-bearing mice and inhibited tumor growth. Importantly, the modified EVs were well tolerated and showed no evidence of nonspecific side effects or immune response. Thus, the RNAi nanoplatform is versatile and can deliver siRNA or miRNA to breast cancer cells both in vitro and in vivo. Our results suggest that the AS1411-EVs have a great potential as drug delivery vehicles to treat cancers.

SNHG1 lncRNA negatively regulates miR-199a-3p to enhance CDK7 expression and promote cell proliferation in prostate cancer.

Long noncoding RNAs (lncRNAs) have been reported to play vital roles in the development of human cancers, but our understandings of most lncRNAs in cancers are still limited. Recently, accumlating evidences have showed that many RNA transcripts could function as competing endogenous RNAs (ceRNAs) by competitively binding common microRNAs. In this study, we demonstrated that a lncRNA, Small Nucleolar RNA Host Gene 1 (SNHG1), as a ceRNA for miR-199a-3p, played a critical role in prostate cancer cell proliferation. We found that SNHG1 was aberrantly up-regulated in prostate carcinoma tissues; while, miR-199a-3p was abnormally down-regulated. The level of SNHG1 in prostate cancer was significantly negatively correlated with that of miR-199a-3p. Our data indicated that SNHG1 could interact with miR-199a-3p and inhibit the activity of miR-199a-3p in prostate cancer cells. In addition, miR-199a-3p could target the 3' UTR of CDK7 and suppress CDK7 expression. More importantly, SNHG1 increased CDK7 expression by competitively binding miR-199a-3p, and then promoted cell proliferation and cell cycle progression in prostate cancer. Taken together, these findings elucidated a novel mechanism of prostate cancer progression. Thus, SNHG1 might serve as a potential target for prostate cancer therapies.

Nucleolin-targeted selenium nanocomposites with enhanced theranostic efficacy to antagonize glioblastoma.

Glioblastoma is considered as the most lethal cancer, due to the inability of chemotherapeutic agents to reach the glioma core as well as the infiltration zone of the invasive glioma cells. Nanotechnology based delivery systems bring new hope to cancer targeted therapy and diagnosis owing to their enhancement of selective cellular uptake and cytotoxicity to cancer cells through various smart designs. We prepared a novel selenium-based composite nanosystem (QDs/Se@Ru(A)) surface functionalized with the AS1411 aptamer and loaded with quantum dots to realize selectivity against glioblastoma and enhance theranostic effects. This cancer targeted nanosystem significantly enhanced the cellular uptake in glioma cells through nucleolin mediated endocytosis, and increased selectivity between cancer and normal cells. The QDs/Se@Ru(A) nanosystem can also be used for spontaneous fluorescence of biological probes to explore their localization in cancer cells, because of the green fluorescent quantum dots loaded into the selenium nanoparticles. QDs/Se@Ru(A) promotes excess reactive oxygen species (ROS) production in glioma cells to induce DNA damage, thus activating diverse downstream signaling pathways, and inhibiting proliferation of U87 cells through the G2/M phase cycle. Thus, this study provides an effective strategy to design a theranostic agent to simultaneously realize cell imaging and therapy for glioblastoma treatment.

Functionalized Multiwalled Carbon Nanotubes as Carriers of Ruthenium Complexes to Antagonize Cancer Multidrug Resistance and Radioresistance.

Multidrug resistance and radioresistance are major obstacles for successful cancer therapy. Due to the unique characteristics of high surface area, improved cellular uptake, and the possibility to be easily bound with therapeutics, carbon nanotubes (CNTs) have attracted increasing attention as potential nanodrug delivery systems. In this study, a CNT-based radiosensitive nanodrug delivery system was rationally designed to antagonize the multidrug resistance in hepatocellular carcinoma. The nanosystem was loaded with a potent anticancer ruthenium polypyridyl complex (RuPOP) via π-π interaction and formation of a hydrogen bond. The functionalized nanosystem (RuPOP@MWCNTs) enhanced the cellular uptake of RuPOP in liver cancer cells, especially drug-resistant R-HepG2 cells, through endocytosis. Consistently, the selective cellular uptake endowed the nanosystem amplified anticancer efficacy against R-HepG2 cells but not in normal cells. Interestingly, RuPOP@MWCNTs significantly enhanced the anticancer efficacy of clinically used X-ray against R-HepG2 cells through induction of apoptosis and G0/G1 cell cycle arrest, with the involvement of ROS overproduction, which activated several downstream signaling pathways, including DNA damage-mediated p53 phosphorylation, activation of p38, and inactivation of AKT and ERK. Moreover, the nanosystem also effectively reduces the toxic side effects of loaded drugs and prolongs the blood circulation in vivo. Taken together, the results demonstrate the rational design of functionalized carbon nanotubes and their application as effective nanomedicine to overcome cancer multidrug resistance.

Rational design of cancer-targeted selenium nanoparticles to antagonize multidrug resistance in cancer cells.

Multidrug resistance is one of the greatest challenges in cancer therapy. Herein we described the synthesis of folate (FA)-conjugated selenium nanoparticles (SeNPs) as cancer-targeted nano-drug delivery system for ruthenium polypyridyl (RuPOP) exhibits strong fluorescence, which allows the direct imaging of the cellular trafficking of the nanosystem. This nanosystem could effectively antagonize against multidrug resistance in liver cancer. FA surface conjugation significantly enhanced the cellular uptake of SeNPs by FA receptor-mediated endocytosis through nystain-dependent lipid raft-mediated and clathrin-mediated pathways. The nanomaterials overcame the multidrug resistance in R-HepG2 cells through inhibition of ABC family proteins expression. Internalized nanoparticles triggered ROS overproduction and induced apoptosis by activating p53 and MAPKs pathways. Moreover, FA-SeNPs exhibited low in vivo acute toxicity, which verified the safety and application potential of FA-SeNPs as nanodrugs. This study provides an effective strategy for the design of cancer-targeted nanodrugs against multidrug resistant cancers. FROM THE CLINICAL EDITOR: In the combat against hepatocellular carcinoma, multidrug resistance remains one of the obstacles to be overcome. The authors designed and synthesized folate (FA)-conjugated selenium nanoparticles (SeNPs) with enhanced cancer-targeting capability. This system carried ruthenium polypyridyl (RuPOP), an efficient metal-based anti-cancer drug with strong fluorescence. It was shown that this combination was effective in antagonizing against multidrug resistance in vitro.

Construction of a cancer-targeted nanosystem as a payload of iron complexes to reverse cancer multidrug resistance.

Multidrug resistance has been identified as a major cause of failure of cancer treatment. Due to their relative non-toxicity, selenium nanoparticles (SeNPs) have been reported as excellent cancer therapeutic nanodrugs. In this study, we designed and prepared a novel nanosystem with borneol surface-functionalized and liver targeting to overcome the multidrug resistance. Borneol (Bor)-modified SeNPs can significantly improve the stability of SeNPs and their anticancer activity. Fe(PiP)3 (PiP = 2-phenylimidazo [4,5-f][1,10] phenanthroline) is a novel anticancer agent with low solubility and stability. In this study, we have constructed a functionalized SeNPs (GAL/Bor@SeNPs) by the surface decoration of galactosamine (GAL), which is a liver targeting ligand that significantly enhanced the cellular uptake of Fe(PiP)3-loaded nanosystem via dynamin-mediated lipid raft endocytosis and clathrin-mediated endocytosis in liver cancer cells overexpressing asialoglycoprotein receptor, thus achieving amplified anticancer efficacy. This multifunctional nanosystem exhibited excellent hemocompatibility and anticancer activity comparing with Fe(PiP)3 or SeNPs alone. Remarkably, GAL/Bor@SeNPs antagonized the multidrug resistance in R-HepG2 cells by inhibiting the expression of ABC family proteins, resulting in enhanced drug accumulation and retention. Internalized nanoparticles released free iron complexes into the cytoplasm, which triggered ROS down-regulation and induced apoptosis through activating AKT and MAPKs pathways. Moreover, this nanosystem effectively prolonged the circulation time of encapsulated drugs. Taken together, this study suggests that GAL and Bor functionalization could be an effective strategy to design cancer-targeted nanomaterials to antagonize multidrug resistance in cancers.

Gracilaria lemaneiformis polysaccharide as integrin-targeting surface decorator of selenium nanoparticles to achieve enhanced anticancer efficacy.

The poor permeability of glioma parenchyma represents a major limit for antiglioblastoma drug delivery. Gracilaria lemaneiformis polysaccharide (GLP), which has a high binding affinity to αvß3 integrin overexpressed in glioma cells, was employed in the present study to functionalize selenium nanoparticles (SeNPs) to achieve antiglioblastoma efficacy. GLP-SeNPs showed satisfactory size distribution, high stability, and selectivity between cancer and normal cells. In U87 glioma cell membrane, which has a high integrin expression level, GLP-SeNPs exhibited significantly higher cellular uptake than unmodified SeNPs. As expected, U87 cells exhibited a greater uptake of GLP-SeNPs than C6 cells with low integrin expression level. Furthermore, the internalization of GLP-SeNPs was inhibited by cyclo-(Arg-Gly-Asp-Phe-Lys) peptides, suggesting that cellular uptake into U87 cells and C6 cells occurred via αvß3 integrin-mediated endocytosis. For U87 cells, the cytotoxicity of SeNPs decorated by GLP was enhanced significantly because of the induction of various apoptosis signaling pathways. Internalized GLP-SeNPs triggered intracellular reactive oxygen species downregulation. Therefore, p53, MAPKs, and AKT pathways were activated to advance cell apoptosis. These findings suggest that surface decoration of nanomaterials with GLP could be an efficient strategy for design and preparation of glioblastoma targeting nanodrugs.