羧甲基壳聚糖基生物医用材料降解代谢行为的研究进展

作为一种水溶性多糖高分子材料,羧甲基壳聚糖(carboxymethyl chitosan, CMCTs)具有优异的生物相容性和生物降解性以及保湿、止血、抗菌、可吸收等一系列优良的功能特性,因而被广泛应用于医工交叉领域。羧甲基壳聚糖进入体内后,在酶、氧、微生物、水等环境适宜时能够被降解,并经吸收、代谢、排泄。其体内降解速率主要取决于材料的尺寸、脱乙酰度、取代度、分子量等。了解羧甲基壳聚糖在动物体内的降解代谢行为,对羧甲基壳聚糖在转化过程中的质量控制和临床应用至关重要。然而就目前而言,羧甲基壳聚糖在生物体内降解代谢的影响因素及其体内吸收、分布、代谢、排泄规律缺乏系统性总结,这一现状从基础层面严重制约了其在生物医药领域的进一步发展。基于上述问题,本文对近年来羧甲基壳聚糖基生物医用材料的降解、代谢相关研究进行梳理和总结,重点阐述了羧甲基壳聚糖作为可降解材料的生物学特性、降解方式、代谢过程等,系统揭示羧甲基壳聚糖体内降解、代谢的规律,并对植入物尺寸、脱乙酰度、取代度、分子量、交联度及成分比例等影响羧甲基壳聚糖降解速率的主要因素进行归纳,以期为羧甲基壳聚糖基生物医用材料的研发和转化研究提供参考。     

壳聚糖超声可控降解及降解动力学研究

通过正交实验法考察了壳聚糖溶液浓度、反应温度、超声强度以及醋酸溶液浓度对超声降解反应的影响,确定了最佳反应条件,制备了一系列不同分子量的壳聚糖.研究了壳聚糖溶液浓度、反应温度以及壳聚糖原料分子参数与降解速率常数的关系.通过红外光谱、X-射线衍射和凝胶渗透色谱对降解产物进行了表征.结果表明,超声降解壳聚糖的最佳条件为10℃,壳聚糖溶液浓度2.5g/L.降解速率常数随壳聚糖溶液浓度和反应温度的降低而增大.高分子量和低脱乙酰度的壳聚糖原料有较高的降解速率和降解速率常数,壳聚糖原料的分子量对降解速率和降解速率常数的影响大于脱乙酰度对其的影响.超声波导致了壳聚糖分子量的降低和产物晶体结构的破坏,但没有改变产物的脱乙酰度和糖残基结构.     

聚合度3～6甲壳寡糖的制备与TOF-MS分析

本文用自制酸性蛋白酶粗酶液降解壳聚糖,研究了降解过程中温度、pH值、反应时间、酶用量及壳聚糖脱乙酰度(DD)对酶降解能力的影响。以85.5%脱乙酰度(DD)、分子量(Mw)为56.0×104的壳聚糖为底物,45℃、pH4.6降解数小时,降解产物由红外光谱、分子量测定(GPC)和基体辅助激光解吸电离飞行时间质谱(MALDI TOF MS)表征。主要降解产物的脱乙酰度有所降低;酶解8h,其水解产物用NaOH调至pH9无沉淀,9h分子量降为0.93×103,经MALDI TOF MS分析产物为聚合度3～6的甲壳寡糖,3～6糖中各糖含量分别为13.3%、27.3%、49.0%、10.1%。     

不同脱乙酰度对壳聚糖表面物理及吸附性能的影响

壳聚糖是甲壳素部分脱乙酰化的产物,脱乙酰度的大小对壳聚糖的性能影响很大。利用碱液法,通过对反应时间的控制,制备了不同脱乙酰度的壳聚糖并对其性能进行了研究。结果表明,随着脱乙酰度的增大,壳聚糖膜的吸水率增加,而对其表面接触角的变化则没有影响。同时,壳聚糖海绵对Ca2+和牛血清蛋白(BSA)的吸附能力也随着脱乙酰度的增大而增加。     

光纤折射率传感用于壳聚糖脱乙酰度测定

建立了一种基于光纤折射率传感技术的壳聚糖脱乙酰度测定方法. 利用光纤折射率传感器监测酸碱滴定过程中溶液折射率的变化, 根据折射率变化转折点之间碱的用量来计算壳聚糖的脱乙酰度. 该方法测得的3种不同含量实际样品的脱乙酰度与氢核磁共振波谱(¹H NMR)方法测定结果相符, 验证了方法的可靠性. 该方法具有用量少、 结构简单、 准确、 重复性好和转折点明显等优点, 可应用于工业生产中壳聚糖脱乙酰度的测定.     

锑电极-电位滴定法测定壳聚糖的脱乙酰度

<正>壳聚糖又名脱乙酰甲壳素,是甲壳素经浓碱脱乙酰基后得到的衍生物,具有良好的生物相容性和生物可降解性,广泛应用于医药、食品、化妆品、轻工、印染、环保和生物工程等领域~[1-3]。而壳聚糖的脱乙酰度(DD)则是指壳聚糖多糖分子氨基上脱去乙酰基的百分比,其高低直接影响壳聚糖的溶解度、絮凝能力、螯合金属离子能力和N-选择性酰化能力等物化特性,是鉴定壳聚糖产品质量的一个重要指标。因此,如何快速准确地测定壳聚糖的脱乙酰度     

海洋烟曲霉菌产脱乙酰酶发酵条件优化

几丁质酶是一类复合酶,包括脱乙酰酶、几丁质酶、壳聚糖酶、几丁寡糖酶、几丁二糖酶等,将脱乙酰酶应用于壳聚糖的制备中,不仅可以有效地从甲壳素(几丁质)分子中脱除乙酰基,而且不会把甲壳素的长链降解为小分子,可以制备出高脱乙酰度且性能独特的壳聚糖,同时不存在排放废碱液而对环境造成严重污染.     

片状壳聚糖的脱乙酰化反应

本文采用一种新方法制备 10 0 %脱乙酰度的壳聚糖。低脱乙酰度壳聚糖溶解在酸溶液中并通过溶剂蒸发制备得到片状壳聚糖。片状壳聚糖进行一步碱处理就可以得到高脱乙酰度壳聚糖。     

壳聚糖折光指数增量的研究

本工作制备了具有不同胺基含量的系列壳聚糖样品,测定和研究了它们的脱乙酰度(D.D.)和溶液拆光指数增量(dn/dc)。发现不同D.D.壳聚糖的dn/dc值不同,而且两者间符合二元无规共聚物的dn/dc值与其化学组成之间的一般线性关系,从而建立了一种可从dn/dc值分析测定壳聚糖的脱乙酰度的新方法。     

碱量法测定壳聚糖脱乙酰度的研究

碱量法(包括酸碱滴定法和电位滴定法)是目前测定壳聚糖脱乙酰度较为常用的方法,其原理是用过量的稀酸与一定量的待测壳聚糖反应,然后用标准NaOH溶液滴定过量的酸来测量壳聚糖中自由氨基的量,从而计算壳聚糖的脱乙酰度(DD值)。计算脱乙酰度的公式多用:DD=(-NH2)%/0.094×100%(公式一)。而另外一个碱量法中未用过的脱乙酰度计算公式为:DD=[203n/(G 42n)]×100%(公式二),本文用计算公式一和公式二分别计算在酸碱滴定法和双突跃电位滴定法中壳聚糖的脱乙酰度,结果表明:在酸碱滴定法中,用公式一的计算的结果准确度更高,而在双突跃电位滴定法中用公式二计算的结果准确度更高。     

甲壳素类液晶高分子的研究Ⅳ.脱乙酰度对壳聚糖液晶性的影响

通过壳聚糖乙酰化法制备了不同脱乙酰度的壳聚糖 ,并研究了脱乙酰度这一结构因素对壳聚糖溶致液晶性的影响 .观察到脱乙酰度为 5 0 %左右时 ,壳聚糖在水中和二氯乙酸中的溶致液晶临界浓度最高 .壳聚糖在水中的溶致液晶临界浓度远低于在二氯乙酸中的临界浓度 .     

Influence of molecular parameters on the degradation of chitosan by a commercial enzyme

The degradative activities of neutral protease against chitosan samples with different molecular parameters were characterized. The effects of the degree of deacetylation (DD) and molecular weight (MW) of chitosan on its susceptibility to degradation were investigated. The DD and MW of the chitosans were determined using potentiometric titration and viscometry, respectively. The molecular weight distribution of initial and degraded commercial chitosan was investigated by gel permeation chromatography. Initial degradation rates (r) were determined from the plots of viscosity decrease against time of degradation. The time courses of degradation of chitosans with neutral protease were non-linear and the enzymatic hydrolysis was an endo-action. Classical Michaelis-Menten kinetic parameters were measured by analyzing the amount of reducing sugars and Eadie-Hofstee plots established that hydrolysis of chitosan by neutral protease obeyed Michaelis-Menten kinetics. Michaelis-Menten parameters and initial degradation rates were calculated and compared to determine the influences of DD and MW on hydrolysis. The results showed that higher DD and higher MW chitosans possessed a lower affinity for the enzyme and a slower degradation rate. Those samples with a lower DD and lower MW were more susceptible substrates.     

Degradation of chitosan with self-resonating cavitation

For the degradation of chitosan, a novel physical method of self-resonating cavitation with strong cavitation effects was investigated in this paper. The effects of initial concentration, pH, temperature, inlet pressure and cavitation time on the degradation efficiency of chitosan were evaluated. It was found that the degradation efficiency was positively correlated with temperature and cavitation time, but was negatively correlated with the solution concentration. The degradation efficiency was maximized at pH of 4.4 and inlet pressure of 0.4 MPa. Under the experimental conditions, the intrinsic viscosity of chitosan solution was reduced by 92.2%, which was twice as high as the degradation efficiency where a Venturi tube cavitator was used. The viscosity-average molecular weights of initial and degraded chitosan were 651 and 104 kD, respectively. The deacetylation degree of chitosan slightly decreased from 89.34% to 88.05%. Structures and polydispersity of initial and degraded chitosan were measured by Fourier-transform infrared spectroscopy (FT-IR), nuclear magnetic resonance hydrogen spectroscopy (¹H NMR), X-ray diffraction (XRD) and gel permeation chromatography (GPC). The results showed that the degradation process did not change the natural structure of chitosan. XRD peaks of the original chitosan were observed at 2θ of 9.59° and 20.00°, and the one at 2θ of 20.00° was obviously weakened after the degradation process, which indicated that the crystallinity of chitosan decreased significantly after the degradation. The polydispersity index of chitosan samples decreased from 3.17 to 2.75, indicating that the molecular-weight distribution of products after the degradation was more concentrated. The results proved that self-resonating cavitation prompted the degradation of chitosan and could reduce the polydispersity of the products for the production of oligochitosan with homogeneous molecular weights.     

LOW MOLECULAR WEIGHT O-CARBOXYMETHYLATED CHITOSANS DERIVED FROM IRRADIATED CHITOSAN AND THEIR ANTIBACTERIAL ACTIVITY

INTRODUCTIONChitin is the second most naturally abundant biopolymer and is found in a variety of organisms, including fungalcell walls, the exoskeleton of crustaceans, skeletal tissue of mollusks and the integument of insects.When treated with alkali, chitin can be deactylated and turned into chitosan, which is a linear binaryheteropolysaccharide composed of (1-4) linked 2-acetamido-2-deoxy-β-D-glucopyranose and 2-amino-2-deoxy-β-D-glucopyranose residues. Chitosan has a wide variety of …     

蘑菇中甲壳素的提取及结构研究

以蘑菇为原料提取甲壳素,并制备壳聚糖。通过滴定法测定由蘑菇制备的壳聚糖的脱乙酰度,用乌氏黏度计测定了比浓黏度,并研究了制备工艺中加热温度和碱处理时间对它们的影响,计算了其产率;对以蘑菇为原料制取的甲壳素、壳聚糖的结构通过红外光谱进行表征。结果表明,在碱处理时间为24h、加热温度为100℃的条件下有较高的脱乙酰度;比浓黏度随着碱处理时间的延长、加热温度的增加都呈下降的趋势;壳聚糖产率为1．69％。制取的甲壳素、壳聚糖的红外光谱图表明,甲壳素在蘑菇中主要是以α-构型存在,α-构型甲壳素在浓碱中经过脱乙酰后生成β-构型的壳聚糖。     

Gelation time and degradation rate of chitosan-based injectable hydrogel

Gelation time and degradation rate of thermally-sensitive aqueous solutions of chitosan/Gp (glycerophosphate disodium salt) have been studied. The effects of different parameters such as Gp salt concentration, solution temperature, degree of deacetylation of chitosan (DDA) and drug loading on the gelation time have been investigated. Gravimetric analysis, gel permeation chromatography and FTIR spectrophotometry were used to investigate the influence of the DDA and concentration of chitosan solution on hydrogel degradation. The presented results indicated that gelation time decreases by increasing Gp salt concentration, temperature, concentration and DDA of chitosan solutions, while drug loading has no significant effect on gelation time. Slower degradation profile was recorded for hydrogel with the higher DDA and concentration of chitosan in the primary solution. FTIR studies indicated that the chemical structure of chitosan macromolecules does not change significantly during the degradation. It could be concluded that biodegradation of chitosan hydrogel occurred via its surfaces.     

水溶性甲壳素及其膜的制备与表征

通过对高脱乙酰度壳聚糖进行均相乙酰化反应，制得脱乙酰度为51．7％的水溶性甲壳素，产物有很好的水溶性．通过红外光谱、X-射线衍射、差热分析等测试表征了水溶性甲壳素结构．结果表明，水溶性甲壳素的结构发生了较大变化，结晶性显著下降是导致水溶性增加的主要原因．     

EDTA脱钙法制备甲壳素

甲壳素的制备一般采用HCl脱钙,NaOH脱蛋白,两种化学品对甲壳素的分子链都有损害,而且能耗高,废弃物对环境污染较严重。本文用。EDTA替代HCl制备甲壳素,同等条件下所制得的甲壳素分子量高得多,其它各项综合性能指标均优,EDTA可回收,可减少环境污染。     

Mouse Acidic Chitinase Effectively Degrades Random-Type Chitosan to Chitooligosaccharides of Variable Lengths under Stomach and Lung Tissue pH Conditions

Chitooligosaccharides exhibit several biomedical activities, such as inflammation and tumorigenesis reduction in mammals. The mechanism of the chitooligosaccharides’ formation in vivo has been, however, poorly understood. Here we report that mouse acidic chitinase (Chia), which is widely expressed in mouse tissues, can produce chitooligosaccharides from deacetylated chitin (chitosan) at pH levels corresponding to stomach and lung tissues. Chia degraded chitin to produce N-acetyl-d-glucosamine (GlcNAc) dimers. The block-type chitosan (heterogenous deacetylation) is soluble at pH 2.0 (optimal condition for mouse Chia) and was degraded into chitooligosaccharides with various sizes ranging from di- to nonamers. The random-type chitosan (homogenous deacetylation) is soluble in water that enables us to examine its degradation at pH 2.0, 5.0, and 7.0. Incubation of these substrates with Chia resulted in the more efficient production of chitooligosaccharides with more variable sizes was from random-type chitosan than from the block-type form of the molecule. The data presented here indicate that Chia digests chitosan acquired by homogenous deacetylation of chitin in vitro and in vivo. The degradation products may then influence different physiological or pathological processes. Our results also suggest that bioactive chitooligosaccharides can be obtained conveniently using homogenously deacetylated chitosan and Chia for various biomedical applications.