Pharmacokinetics and Biodegradation Performance of a Hydroxypropyl Chitosan Derivative

Hydroxypropyl chitosan(HP-chitosan) has been shown to have promising applications in a wide range of areas due to its biocompatibility, biodegradability and various biological activities, especially in the biomedical and pharmaceutical fields. However, it is not yet known about its pharmacokinetics and biodegradation performance, which are crucial for its clinical applications. In order to lay a foundation for its further applications and exploitations, here we carried out fluorescence intensity and GPC analyses to determine the pharmacokinetics mode of fluorescein isothiocyanate-labeled HP-chitosan(FITC-HP-chitosan) and its biodegradability. The results showed that after intraperitoneal administration at a dose of 10 mg per rat, FITC-HP-chitosan could be absorbed rapidly and distributed to liver, kidney and spleen through blood. It was indicated that FITC-HP-chitosan could be utilized effectively, and 88.47% of the FITC-HP-chitosan could be excreted by urine within 11 days with a molecular weight less than 10 k Da. Moreover, our data indicated that there was an obvious degradation process occurred in liver( 10 k Da at 24 h). In summary, HP-chitosan has excellent bioavailability and biodegradability, suggesting the potential applications of hydroxypropyl-modified chitosan as materials in drug delivery, tissue engineering and biomedical area.     

Establishing Gene Delivery Systems Based on Small-Sized Chitosan Nanoparticles

Chitosan is a natural cationic polysaccharide,which is often used for preparing biomedical materials because of its high biocompatibility.In this study,chitosan with a molecular weight of 160 k Da was chosen to prepare chitosan nanoparticles(CSNPs) as gene vectors by ionic cross-linking with tripolyphosphate(TPP).CSNPs were characterized in terms of particle size,zeta potential,and polydispersity index(PDI) using a Zetasizer,and morphology was evaluated by transmission electron microscopy(TEM).Furthermore,the cytotoxicity and biocompatibility of CSNPs were correspondingly examined by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay and histological examination.Agarose gel electrophoresis and UV spectrophotometric methods were performed to measure the loading capacity.The cell transfection efficiency of CSNPs loaded with plasmids or si RNA was analyzed by fluorescence microscopy or laser scanning confocal microscopy.The results showed that CSNPs were prepared successfully by the ionic gelation method,which had a smaller particle size(100 nm-200 nm),stable dispersibility,low cytotoxicity,good tissue-biocompatibility,and high gene-loading efficiency.These CSNPs could transfer the plasmids or si RNA to cells.However,CSNPs might have a much higher transfection efficiency for si RNAs than for plasmids,which implies that CSNPs might be a safer and more efficient vector for delivering si RNAs rather than plasmids.     

Researches on the Internal Molecular Weight Uniformity of Chitosan Biomaterials

Uniform molecular weight(Mw)chitosan(CS)is highly demanded in medical biomaterial industry.This present article described heterogeneous degradation of CS in aqueous HCl/ethanol solution,in which progress uniform Mw CS was successfully prepared.The Mw distribution of CS was measured by gel permeation chromatography(GPC)analysis.Moreover,the structure and properties of degraded CS were characterized by Fourier transform infrared spectroscopy(FT-IR),nuclear magnetic resonance spectroscopy(^1H NMR),X-ray diffraction(XRD)and thermogravimetric(TG)analysis.In addition,the biocompatibility of degraded CS was also assessed by hemolysis rate(HR)measurement.The Mw of CS dramatically decreased from 246 KDa to 76 k Da at the initial 30 min,and stabilized at 18 kDa after 24 h.GPC analysis results showed that the degraded CS molecular become homogenization.FT-IR and 1 H NMR analysis confirmed the basic structure of CS molecular backbone was not destroyed during this progress.Besides,the water solubility of CS was not significantly influenced by this reaction.Moreover,the XRD analysis revealed that crystallinity of degraded CS increased from 70.32% to 99.25%with time.The TG analysis showed improved thermal stability of degraded CS.HR measurement demonstrated the degraded CS possessed excellent biocompatibility.This simple and efficient heterogeneous degradation would open up a new route to produce uniform Mw CS.     

Pharmacokinetics and Biodegradation of Chitosan in Rats

Chitosan, an excellent biomedical material, has received a widespread in vivo application. In contrast, its metabolism and distribution once being implanted were less documented. In this study, the pharmacokinetics and biodegradation of fluorescein isothiocyanate(FITC) labeled and muscle implantation administrated chitosan in rats were investigated with fluorescence spectrophotometry, histological assay and gel chromatography. After implantation, chitosan was degraded gradually during its distribution to diverse organs. Among the tested organs, liver and kidney were found to be the first two highest in chitosan content, which was followed by heart, brain and spleen. Urinary excretion was believed to be the major pathway of chitosan elimination, yet 80% of chitosan administered to rats was not trackable in their urine. This indicated that the majority of chitosan was degraded in tissues. In average, the molecular weight of the degradation products of chitosan in diverse organs and urine was found to be 65 k Da. This further confirmed the in vivo degradation of chitosan. Our findings provided new evidences for the intensive and safe application of chitosan as a biomedical material.     

Construction and characterization of Gal-chitosan graft methoxy poly (ethylene glycol) (Gal-CS-mPEG) nanoparticles as efficient gene carrier

In the present study, galactosylated chitosan (Gal-CS) was conjugated with methoxy poly(ethylene glycol) (mPEG) as a hydrophilic group. The structure of Gal-CS-mPEG polymer was characterized and the nanoparticles (NPs) were prepared using ironic gelation method. The study was designed to investigate the characteristics and functions of Gal-CS-mPEG NPs. The morphology of Gal-CS-mPEG NPs was observed by SEM and it was a compact and spherical shape. The size of the NPs was approximately 200 nm in diameter under the ideal process parameters. The interaction between Gal-CS-mPEG NPs and pDNA, and the protection of pDNA against DNase I and serum degradation by Gal-CS-mPEG NPs were evaluated. Agarose gel electrophoresis results showed that Gal-CS-mPEG NPs had strong interaction with pDNA at the weight ratio of 12:1, 4:1 and 2:1 and could protect pDNA from DNase I and serum degradation. Gal-CS-mPEG NPs exhibited high loading efficiency and sustainable in vitro release. The blood compatibility studies demonstrated that Gal-CS-mPEG NPs had superior compatibility with erythrocytes in terms of aggregation degree and hemolysis level. Gal-CS-mPEG NPs showed no cytotoxicity on L929 cells, which is a normal mouse connective tissue fibroblast, but showed inhibitory effects on the proliferation of Bel-7402 cells, which is a liver cancer cell line. In conclusion, Gal-CS-mPEG NP is a bio-safe and efficient gene carrier with potential application in gene delivery.     

Effects of sulfate chitosan derivatives on nonalcoholic fatty liver disease

Sulfate chitosan derivatives have good solubility and therapeutic effect on the cell model of NAFLD. The aim of this study was to examine the therapeutic effect of sulfate chitosan derivatives on NAFLD. The male Wistar rats were orally fed high fat emulsion and received sulfate chitosan derivatives for 5 weeks to determine the pre-treatment effect of sulfate chitosan derivatives on NAFLD. To evaluate the therapeutic effect of sulfate chitosan derivatives on NAFLD, the rats were orally fed with high concentration emulsion for 5 weeks, followed by sulfate chitosan derivatives for 3 weeks. Histological analysis and biomedical assays showed that sulfate chitosan derivatives can dramatically prevent the development of hepatic steatosis in hepatocyte cells. In animal studies, pre-treatment and treatment with sulfate chitosan derivatives significantly protected against hepatic steatohepatitis induced by high fat diet according to histological analysis. Furthermore, increased TC, ALT, MDA, and LEP in NAFLD were significantly ameliorated by pre-treatment and treatment with sulfate chitosan derivatives. Furthermore, increased TG, AST, and TNF-α in NAFLD were significantly ameliorated by treatment with sulfate chitosan derivatives. Sulfate chitosan derivatives have good pre-treatment and therapeutic effect on NAFLD.     

新型壳聚糖季铵盐衍生物的合成与表征

采用甲磺酸作为溶剂兼作氨基保护试剂,对壳聚糖上的羟基选择性酯化,再游离出氨基并对其氯乙酰化,最后进行吡啶成盐,得到新型水溶性的壳聚糖季铵盐衍生物,产物的结构通过核磁共振氢谱和红外光谱进行了表征,从而为壳聚糖季铵盐衍生物的制备提供了一种新途径。     

壳聚糖希夫碱铜配合物的制备及其催化性能

采用微波辐射加热法使壳聚糖与水杨醛反应生成壳聚糖希夫碱,然后将其与铜盐配位得到壳聚糖希夫碱铜配合物,用UV、FTIR、TG表征产物的结构,并探讨该配合物对过氧化氢分解的催化性能。研究结果表明:壳聚糖希夫碱铜配合物对过氧化氢有良好的催化活性,常温(25℃)常压,pH=7.0的条件下放置12 h,催化H2O2的分解达99.8%,该配合物重复使用3次还保持较好的催化活性。     

Analgesis and wound healing effect of chitosan and carboxymethyl chitosan on scalded rats

Analgesis and wound healing effect of chitosan and carboxymethyl chitosan on scalded rats were investigated. A II degree scald model was established in rats, which was subsequently treated with chitosan and carboxymethyl chitosan solution, respectively. The concentration of bradykinin and 5-hydroxytryptophan was detected by assaying enzyme-linked immunosorbent. Healing condition was observed and pathological sections were made to determine the healing effect of chitosan and carboxymethyl chitosan. Results showed that the concentration of bradykinin and 5-hydroxytryptophan peaked at the third hour post-wound in all groups, while the concentration of hydroxyproline peaked at the seventh day post-wound in both chitosan and carboxymethyl chitosan group. The concentration of bradykinin and 5-hydroxytryptophan of carboxymethyl chitosan group was significantly lower than that of control (P < 0.05), while that of chitosan group was similar to that of control (P > 0.05). These findings indicated that carboxymethyl chitosan reduced the concentration of algogenic substances, resulting in analgesia. During the whole recovery process, the hydroxyproline concentration in chitosan and carboxymethyl chitosan group on day 3 and 7 was significantly higher than that of control (P < 0.01); however the significance of such a highness decreased on day 14 (P < 0.05). These findings indicated that chitosan and carboxymethyl chitosan accelerated tissue repair. Meanwhile, chitosan performed better in healing than carboxymethyl chitosan in both decrustation and healing time. In conclusion, carboxymethyl chitosan showed significant analgesis and wound-healing promotion effect, but chitosan only showed wound-healing promotion effect.     

Isolation,identification, and cytotoxicity of a new isobenzofuran derivative from marine Streptomyces sp. W007

A new isobenzofuran derivative( 1) was isolated from the marine Streptomyces sp. W007 and its structure was determined through extensive spectroscopic analyses, including 1D-NMR, 2D-NMR, and ESI-MS. The absolute configuration of compound 1 was determined by a combination of experimental analyses and comparison with reported data, including biogenetic reasoning, J-coupling analysis, NOESY, and 1 H- 1 HCOSY. Compound 1 exhibited no cytotoxicity against human cells of gastric cancer BGC-823, lung cancer A549, and breast cancer MCF7.     

壳聚糖对腐生葡萄球菌的作用机制

【目的】探究壳聚糖处理对腐生葡萄球菌(Staphylococcus saprophytic)的作用机制。【方法】通过光密度值结合菌液澄清度法综合确定壳聚糖对腐生葡萄球菌的最小抑菌浓度(Minimum Inhibitory Concentration,MIC),分析其在MIC与2MIC的壳聚糖作用下的微生物生长曲线、电导率值、碱性磷酸酶(Alkaline Phosphatase,AKP)含量、过氧化氢酶(Catalase,CAT)含量、苹果酸脱氢酶(Malatedehydrogenase,MDH)含量与生物膜生成量等指标变化,并结合扫描电镜(ScanningElectronMicroscopy,SEM)观察其作用后对菌体细胞形态变化影响,综合评价壳聚糖处理对腐生葡萄球菌的作用机制。【结果与结论】壳聚糖对腐生葡萄球菌的MIC为1.25 mg/mL;菌体经MIC和2MIC处理后,其正常生长受到抑制,细胞壁被破坏,细胞膜通透性改变,CAT与MDH含量降低,其作用效果与壳聚糖浓度正相关;SEM结果显示,与对照CK组对比,腐生葡萄球菌经MIC壳聚糖处理后,菌体形状不规则,表面皱褶;经2MIC壳聚糖处理后,菌体表面产生泡状物且黏附明显,表明壳聚糖使腐生葡萄球菌菌体细胞壁受损,菌体生长受到抑制,胞内保护酶与三羧酸循环中的关键酶活性降低,细胞代谢能力随之下降,细胞膜结构完整度受到破坏,最终导致菌体死亡。     

C_(17)-fengycin B,produced by deep-sea-derived Bacillus subtilis,possessing a strong antifungal activity against Fusarium solani

Root rot disease caused by Fusarium solani is the most devastating disease of the tomato and legume crops in China.The metabolites of Bacillus species can inhibit many fungal diseases.In this study,the metabolites of deep-sea-derived bacterium Bacillus subtilis 2 H11 can significantly inhibit the growth of F.solani.The metabolite C_(17)-fengycin B,one of the cyclic lipopeptides,was identified by the combination of silica column chromatography,high-performance liquid chromatography(HPLC),high-energy collision induced dissociation mass spectrometry(HCD-MS) and tandem mass spectrometry(HCD-MS/MS).The results of scanning electron microscopy(SEM) and transmission electron microscopy(TEM) showed that C_(17)-fengycin B could destroy the structure of the hyphae and spores of F.solani.The antifungal activities of C_(17)-fengycin B against F.solani were tested at concentrations ranging from 0.05 mg/mL to 0.20 mg/mL.The results indicated that C_(17)-fengycin B inhibited the growth of F.solani with antifungal index of 89.80%at 0.20 mg/mL,and the antifungal activity of C_(17)-fengycin B was further verified by the pot experiment.In addition,the cytotoxicity experiment showed that C_(17)-fengycin B had good biocompatibility and was a potential candidate for the development of biocontrol pesticide in the future.     

CYTOTOXICITY AND GENOTOXICITY OF POLYETHYLENIMINE AND NICKEL CHLORIDE IN RED SEA BREAM（Pagrosomus major）FIN CELL LINE RSBF

A continuous marine fish cell line RSBF(i.e.Red Sea Bream Fin)was utilized to screen the cytotoxicity and genotoxicity of polyethylenimine(PEI)and nickel cholride(NiCl2)in this study on the deleterious effects of aquatic genotoxins on fish.At the 0.01 to 1μgml concentration tested,PEI had acute toxicity to the treated RSBF cells(IC50=1.12,0.92,0.88 and 0.64μg/ml PEI for time 0 h,24 h,48 h and 72 h after treatment,respectively)and markedly inhibited their proliferation in a dose-dependent manner.At the 0.001 to 5 μmol/L concentration tested,NiCl2 posed no acute toxicity but significantly stimulated their growth(107?214?of control).Random amplified polymorphic DNA(RAPD)technique was used to detect the genotoxic effects of PEI and NiCl2 by comparing the RAPD banding patterns of the control and treated cells.RAPD analysis indicated that at the concentrations tested,PEI was more genotoxic than NiCl2 to RSBF cells;that there was a slight dose-dependent response in the genotoxic effect of PEI bue not NiCl2;and that RAPD technique might provide a sensitive,non-specif-ic gentoxic endpoint.And the potent cytotoxicity and genotoxicity of PEI on fish cells showed that we should be cautious in utilizing it as gene vector in fish gene transfer and human gene therapy.     

SYNTHESIS AND PROPERTIES OF SULFHYDRYL CHITOSAN

A new adsorbent for heavy metal ions, sulfhydryl chitosan (S-chitosan), was produced by treatment of chitosan with sulhydryl acetic acid in the presence of sulfuric acid as a catalyst. Its structure was confrrmed by elemental analysis and FI‘-IR spectra analysis. The adsorption properties of sulthydryl chitosan for Cu(Ⅱ ), Cd(Ⅱ ), Pb(Ⅱ), Cr(Ⅲ) and Ni(Ⅱ) were investigated, and the effect of pH value on adsorption, adsorption kinetics, and selective adsorption was examined. It was shown that S-chitosan has good adsorption for Pb(Ⅱ), Cu(Ⅱ) and Cd(Ⅱ) like chitosan, is also insoluble in acid solution; has good adsorption kinetic properties for heavy metal ions; and can be used in acid solution. The adsorption capacities of S-chitosan can be affected by media acidity. The adsorbed Cu(Ⅱ) Cd(Ⅱ) and Pb(Ⅱ) could be eluted by diluted chlorhydric acid.     

CYTOTOXICITY AND GENOTOXICITY OF POLYETHYLENIMINE AND NICKEL CHLORIDE IN RED SEA BREAM (Pagrosomus major) FIN CELL LINE RSBF

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Preparation of cationic chitosan-polyacrylamide flocculant and its properties in wastewater treatment

Chitosan derived from crab shells,was used to prepare the graft polymer in aqueous solution with acrylamide (AM) and methacrylatoethyl trimethyl ammonium chloride (DMC) as raw materials and ceric ammonium nitrate (CAN) as initiator.The floc-culation ability of the resulting polymer (PCAD) was studied in waste water treatment experiments.Its properties were determined on the basis of the transmittance of waste water after flocculation.The effects of chitosan and DMC content on PCAD’s flocculation ability wer...     

Chitosan removes toxic heavy metal ions from cigarette mainstream smoke

This study investigated the removal of heavy metal ions from cigarette mainstream smoke using chitosan. Chitosan of various deacetylation degrees and molecular weights were manually added to cigarette filters in different dosages. The mainstream smoke particulate matter was collected by a Cambridge filter pad, digested by a microwave digestor, and then analyzed for contents of heavy metal ions, including As(III/V), Pb(II), Cd(II), Cr(III/VI) and Ni(II), by graphite furnace atomic absorption spectrometry (GFAAS). The results showed that chitosan had a removal effect on Pb(II), Cd(II), Cr(III/VI) and Ni(II). Of these, the percent removal of Ni(II) was elevated with an increasing dosage of chitosan. Chitosan of a high deace tylation degree exhibited good binding performance toward Cd(II), Cr(III/VI) and Ni(II), though with poor efficiency for Pb(II). Except As(III/V), all the tested metal ions showed similar tendencies in the growing contents with an increasing chitosan molecular weight. Nonetheless, the percent removal of Cr(III/VI) peaked with a chitosan molecular weight of 200 kDa, followed by a dramatic decrease with an increasing chitosan molecular weight. Generally, chitosan had different removal effects on four out of five tested metal ions, and the percent removal of Cd(II), Pb(II), Cr(III/VI) and Ni(II) was approximately 55%, 45%, 50%, and 16%, respectively. In a word, chitosan used in cigarette filter can remove toxic heavy metal ions in the mainstream smoke, improve cigarette safety, and reduce the harm to smokers.     

Optimization of four types of antimicrobial agents to increase the inhibitory ability of marine Arthrobacter oxydans KQ11 dextranase mouthwash

We adopted the response surface methodology using single factor and orthogonal experiments to optimize four types of antimicrobial agents that could inhibit biofilm formation by Streptococcus mutans, which is commonly found in the human oral cavity and causes tooth decay. The objective was to improve the function of marine Arthrobacter oxydans KQ11 dextranase mouthwash(designed and developed by our laboratory). The experiment was conducted in a three-level, four-variable central composite design to determine the best combination of Zn SO_4, lysozyme, citric acid and chitosan. The optimized antibacterial agents were 2.16 g/L Zn SO_4, 14 g/L lysozyme, 4.5 g/L citric acid and 5 g/L chitosan. The biofilm formation inhibition reached 84.49%. In addition, microscopic observation of the biofilm was performed using scanning electron microscopy and confocal laser scanning microscopy. The optimized formula was tested in marine dextranase Arthrobacter oxydans KQ11 mouthwash and enhanced the inhibition of S. mutans. This work may be promoted for the design and development of future marine dextranase oral care products.     

Preparation and characterization of magnetic resin made from chitosan and cerium

In this study,the water-based ferromagnetic fluid and magnetic resin made from chitosan and cerium complex(MRCCC) were successfully prepared by using the chemical co-precipitation technique and by the reversed-phase suspension cross-linking polymerization.MRCCC presented uniform and narrow particle size distribution as determined by the Laser Particles Sizer.The Inductively Coupled Plasma-Atomic Emission Spectrometry(ICP-AES),Fourier transform infrared spectroscopy(FT-IR),differential scanning calorimetry(DSC)and X-ray powder diffraction(XRD)study demonstrated that there were iron and cerium existing in MRCCC.The movement of MRCCC under magnetic field proved its magnetic property.The swelling kinetics in water or solutions with different pH indicated that MRCCC could be applied in solutions with pH greater than 1.0.The ferromagnetic fluid particles were stable in MRCCC soaked in solutions with pH >2.0.In view of these results,MRCCC can be used as material for separation,clarification,adsorption,sustained release and hydrolysis activity.     

磺酸基改性梳状有机硅聚醚破乳剂的制备及性能分析

在铂催化剂条件下,利用含氢聚硅氧烷(PHMS)、烯丙基磺酸盐(COPS-1)和烯丙基聚醚(Y-1)的硅氢化反应生成磺酸盐有机硅聚醚破乳剂(COPESO),采用红外光谱、GPC、粒度、界面张力和破乳性能测试等,对破乳剂进行结构表征,分析破乳性能的影响因素.结果表明:当反应温度为90℃、pH=3、COPS-1质量分数为5%时,对破乳剂引入磺酸基活性基可以得到性能优良多支链高相对分子质量的COPESO,其表面活性高,出水快,水色透明且界面齐,对乳液破乳效果高达94.8%.