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Biological Properties of Chitosan Films with Different Degree of Deacetylation

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Journal of Materials Science & Technology 2008年第5期24卷 700-708页

作者： Jing ChanG Wanshun LIU baoqin han Bing LIU Department of Marine Life Science Ocean University of China Qingdao 266003 China

Chitin and chitosan films were prepared by solution casting method. Chitosan specimens used in this study were deacetylated by 50.4%, 69.2%, 85.5% and 96.3%. Their water content, protein adhesion ability, cytocompatibility, cell adhesion ability, in vitro and vivo degradability and biocompatibility were evaluated. Results indicated that with the degree of deacetylation （DD） between 50% and 70%, the chitosan showed higher water content. The higher the DD, the stronger protein adhesion ability the chitosan had. All the films have good cytocompatibility and the films with higher DD have better cell adhesion ability. Chitin films degraded more rapidly than others, which disappeared in 2 to 4 weeks after they were implanted in subcutaneous tissue and musculature. Their inflammatory reaction became weaker as the films degraded. As the DD got higher, the films degraded slower. The films of DD 85.5% and DD 90.3% even didn＇t disappeared in 12 weeks after they were implanted. Their inflammatory reaction was mild at the beginning of degradation, and became severe in 4 to 8 weeks, then weaken at last. This basic result can be very helpful for tissue engineering.

关键词： Chitosan Degree of deacetylation Biodegradability Cytocompatibility Celladhesion ability Biocompatibility

Antioxidant activity of N-acetyl-glucosamine based thiazolidine derivative

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High Technology Letters 2007年第4期13卷 441-446页

作者：李春雷 Yang Yan han baoqin Liu Wanshun College of Marine Life Sciences Ocean University of China Qingdao 266003 P.R. China

N-acetyl-glucosamine, the monomer of chitin, was cyclo-condensed with L-cysteine to prepare thiazolidine derivative： 2-N-acetyl-glucosamine-thiazolidine-4（R）-carboxylic acid （GlcNAcCys）. The stability of GlcNAcCys was evaluated by high performance liquid chromatography （HPLC） measurement. The results showed that GlcNAcCys was more stable than other TCA derivatives, especially in alkaline condition. The direct in vitro antioxidative properties of GlcNAcCys were investigated by using UV radiation-induced lipid peroxidation （LPO） in mitochondria and nuclei and . OH-induced LPO in red blood cell （RBC） ghosts models. UV radiation caused dose-dependent LPO in both mitochondria and nuclei. This effect was catalyzed by addition of Fe^2 ＋ while prevented by co-incubation with GlcNAcCys. When nuclei and mitochondria was treated with 100μl, 300μl, 500μl of GlcNAcCys and co-incubated at 37℃ for 30min, LPO was decreased to 96%, 72%, 68% in nuclei and 95%, 72%, 68% in mitochondria when compared to the UV radiation group respectively. Hydroxyl radicals （. OH） generated by Fenton reaction induced LPO in RBC ghosts. Pretreatment of RBC ghosts with GlcNAcCys could induce antioxidant RBC ghosts and inhibit concentration-dependent malondialdehyde （MDA） formation in antioxidant RBC ghosts. Its inhibition percent was 14%, 35%, 36%, 42% at 10, 20, 30, 40mg/ml respectively. In a conclusion, the data suggest that GlcNAcCys has antioxidant ability and can significantly inhibit lipid peroxidation in biological samples tested in vitro.

关键词： thiazolidine-4(R)-carboxylic acid N-acetyl-glucosamine antioxidant lipid pemxidation

Activities of the Deriviatives of Chitin on the Osbeoblast Proliferation and the Effect on Bone Strength in Ovariectomized Rats

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Wuhan University Journal of Natural Sciences 2006年第3期11卷 715-719页

作者： WEI Changzheng LIU Wanshun han baoqin DUAN Yixiang College of Marine Life Sciences Ocean University of China Qingdao 266003 Shandong China Orthopaedics of Qingdao Municipal Hospilal Qingdao 266003 Shandong China

This study intends to examine the effects of different concentrations of four kinds of degradations of chitin： glucosamine （GLC）, N-acetyl-D-glucosamine （NAG）, chitooligosaccharide（COS）, CM-chitooligosaccharide （CM-COS）—on the proliferation of MC3T3-E1 cell line cultured in vitro. Results suggest that all of the glucoses mentioned above promoted the proliferation of osteoblast, and various concentrations have different effects： the proliferation was remarkable when the concentration of GLC, NAG, COS, CM-COS was 100 , 100, 500, 500 μg/mL ,respectively. Furthermore we choose the glucosamine as the material and study the effect on the bone strength in ovariectomized rats. The results showed that the middle does of glucosamine can significantly increase the hone strength of femur in ovariectomized rats.

关键词： glucosamine chitin ovariectomized rats bone strength osteoporosis osteohlast proliferation

Construction and Characterization of Gal-Chitosan Graft Methoxy Poly(Ethylene Glycol)(Gal-CS-mPEG) Nanoparticles as Efficient Gene Carrier

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Journal of Ocean University of China 2017年第5期16卷 873-881页

作者： JIN Jiting FU Wandong LIAO Miaofei han baoqin ChanG Jing YANG Yan College of Marine Life Sciences Ocean University of China Qingdao 266003 P. R. China Zhejiang Research Institute of Marine Development Zhoushan 316100 P. R. China

In the present study, galactosylated chitosan(Gal-CS) was conjugated with methoxy poly(ethylene glycol)(m PEG) as a hydrophilic group. The structure of Gal-CS-m PEG 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-m PEG NPs. The morphology of Gal-CS-m PEG 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-m PEG NPs and p DNA, and the protection of p DNA against DNase I and serum degradation by Gal-CS-m PEG NPs were evaluated. Agarose gel electrophoresis results showed that Gal-CS-m PEG NPs had strong interaction with p DNA at the weight ratio of 12:1, 4:1 and 2:1 and could protect p DNA from DNase I and serum degradation. Gal-CS-m PEG NPs exhibited high loading efficiency and sustainable in vitro release. The blood compatibility studies demonstrated that Gal-CS-m PEG NPs had superior compatibility with erythrocytes in terms of aggregation degree and hemolysis level. Gal-CS-m PEG 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-m PEG NP is a bio-safe and efficient gene carrier with potential application in gene delivery.

关键词： DNA chitosan methoxy DNase compatibility Ethylene spherical hydrophilic conjugated sustainable

Effects of Low-Molecular-Weight-Chitosan on the AdenineInduced Chronic Renal Failure Rats in vitro and in vivo

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Journal of Ocean University of China 2015年第1期14卷 97-104页

作者： ZHI Xuan han baoqin SUI Xianxian HU Rui LIU Wanshun College of Marine Life Science Ocean University of China

The effects of low-molecular-weight-chitosan （LMWC） on chronic renal failure （CRF） rats induced by adenine were investigated in vivo and in vitro. Chitosan were hydrolyzed using chitosanase at pH 6-7 and 37℃ for 24h to obtain LMWC. In vitro, the effect of LMWC on the proliferation of renal tubular epithelial cells （RTEC） showed that it had no cytotoxic effect and could promote cell growtfi. For the in vivo experiment, chronic renal failure rats induced by adenine were randomly divided into control group, Niaoduqing group, and high-, medium- and low-dose LMWC groups. For each group, we detected serum creatinine （SCR）, blood urea nitrogen （BUN）, and total superoxide dismutase （T-SOD）, glutathione oxidase （GSH-Px） activities of renal tissue, and obtained the ratio of kidney weight/body weight, pathological changes of kidney. The levels of serum SCR, BUN were higher in the adenine-induced rats than those in the control group, indicating that the rat chronic renal failure model worked successfully. The re- suits after treatment showed that LMWC could reduce the SCR and BUN levels and enhance the activities/levels of T-SOD and GSH-PX in kidney compared to control group. Histopathological examination revealed that adenine-induced renal alterations were restored by LMWC at three tested dosages, especially at the low dosage of 100mgkg-1 d-1.

关键词： LMWC RTECs CRF adenine SCR BUN T-SOD GSH-PX renal histopathology

Analgesis and Wound Healing Effect of Chitosan and Carboxymethyl Chitosan on Scalded Rats

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Journal of Ocean University of China 2014年第5期13卷 837-841页

作者： HUANG Shuya han baoqin SHAO Kai YU Miao LIU Wanshun College of Marine Life Sciences Ocean University of China

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). 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 han 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.

关键词： bradykinin 5-hydroxytryptophan hydroxyproline carboxymethyl chitosan

The Evaluation on Biological Properties of Carboxymethyl-chitosan and Carboxymethyl-chitin

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Journal of Ocean University of China 2008年第4期7卷 404-410页

作者： ChanG Jing LIU Wanshun han baoqin LIU Bing College of Marine Life Sciences Ocean University of China Qingdao 266003P R. China

Carboxymethyl-chitosan and carboxymethyl-chitin were prepared with the methods developed in our laboratory. The DS （degree of substitution） and DD （degree of deacetylation） of the carboxymethyl-chitosan were 103.14% and 97.18% respectively, while the DS of the carboxymethyl-chitin was 96.37%. Their effects on human fibroblasts, intradermal irritation test, in vitro and vivo degradability, and biocompatibility were evaluated. The results indicate that the polysaccharides at low concentrations can facilitate the growth of human fibroblasts and the carboxymethyl-chitosan at 100 μg mL^-1 is the most effective. The polysaccharides at higher concentrations, however, inhibit the growth of fibroblasts. The PII （Primary Irritation Index） values of CM-chitosan and CM-chitin are both 0.0, which shows that they have no irritation reaction. Both of the polysaccharides show good degradability and biocompatibility. Carboxymethyl-chitin degrades faster in vitro than carboxymethyl-chitosan. The latter, however, has no inflammatory reaction after being implanted in vivo for 7 d and shows better biocompatibility. This study may provide a scientific basis for the use of earboxymethyl-ehitosan and carboxymethyl-chitin as biomaterials.

关键词： carboxymethyl-chitosan (CM-chitosan) carboxymethyl-chitin (CM-chitin) biocompatibility biodegradability

Pharmacokinetics and Biodegradation Performance of a Hydroxypropyl Chitosan Derivative

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Journal of Ocean University of China 2015年第5期14卷 888-896页

作者： SHAO Kai han baoqin DONG Wen SONG Fulai LIU Weizhi LIU Wanshun College of Marine Life Science Ocean University of China

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.

关键词： hydroxypropyl chitosan fluorescein isothiocyanate pharmacokinetics biodegradation performance biomedicalmaterial

Synthesis and Characterization of a Hydroxyethyl Derivative of Chitosan and Evaluation of Its Biosafety

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Journal of Ocean University of China 2015年第4期14卷 703-709页

作者： SHAO Kai han baoqin GAO Jinning SONG Fulai YANG Yan LIU Wanshun College of Marine Life Science Ocean University of China

Hydroxyethyl chitosan （HE-chitosan） is a water-soluble derivative of chitosan with many apparent biological properties. For example, it is non-toxic and rapidly biodegradable. Moreover, HE-chitosan has advantages in water-solubility, moisture retention and gelling property due to its hydroxyethyl group. However, the biocompatibility and biodegradability of this multifimctional de- rivative have rarely been documented although they are critical for its application in biomedical and clinical treatments. The purpose of this work was to evaluate the biosafety of HE-chitosan, and draw important clues for its diverse applications. HE-chitosan was synthesized and characterized its chemical structure with FTIR. Its molecular weight （Mw） was determined by gel permeation chro- matography （GPC）, and its deacetylation degree （DD） was investigated through potentiometric analysis. The cytotoxicity of HE-chitosan on mouse fibroblast cell L929 was tested. The biocompatibility and biodegradability of HE-chitosan in rat and rabbit were evaluated. The FTIR results indicated that the hydroxyethyl groups were linked to C6 of chitosan. The GPC analysis confirmed that its Mw was about 90.01 kDa. It was also demonstrated that HE-chitosan had excellent biocompatibility and biodegradability in vivo and had no cytotoxicity on L929. These findings indicated that HE-chitosan can potentially be applied as a biomaterial in tissue engineering, drug delivery, and other biomedical fields.

关键词： hydroxyethyl chitosan biomaterial cytotoxicity biocompatibility biodegradability