新型复合壳聚糖水凝胶伤口敷料的制备及性能评价

发布时间：2018-05-01 03:09

本文选题：伤口敷料 + 壳聚糖　；参考：《安徽大学》2017年博士论文

【摘要】：伤口敷料是用于伤口护理的重要医药产品,新型伤口敷料可通过吸收伤口多余渗液或者持有水分从而保持创面的湿润,从而促进伤口愈合。壳聚糖及其衍生物为原料所制备的生物医学材料,因其生物相容性优异,能够止血并具有一定的抗菌性能,在促进伤口愈合方面的作用被广泛认可。本工作选择壳聚糖作为起始原料,以化学交联为主要技术手段,制备了三种壳聚糖复合交联薄膜/海绵及一种酸溶性竣甲基壳聚糖原位凝胶伤口敷料,并通过核磁共振(NMR)、傅里叶变换红外光谱(FTIR)、X-射线衍射(XRD)、热重分析(TGA)、扫描电子显微镜(SEM)等手段表征其结构;通过溶胀率检测、水蒸气及氧气透过率检测、细胞毒性实验、血液相容性实验、动物实验等方法测试其性能,并评价它们的应用前景。本论文的主要创新点在于:(1)制备了一种纤维素衍生物,丁二酰羟丙基甲基纤维素(HPMCS),并在EDC/NHS催化作用下将其与壳聚糖交联形成复合交联凝胶膜,对凝胶膜进行了结构表征并评估了其作为伤口敷料材料的性能;(2)研究了羧甲基魔芋葡甘聚糖(CMKGM)的制备条件,制备了高溶胀性CMKGM/CS复合交联海绵伤口敷料;(3)不使用任何有毒的交联剂或引发剂,采用PEG衍生物(SC-PEG-SC)与壳聚糖在溶液中直接反应生成凝胶,再干燥成膜。并通过小鼠全层皮肤伤口模型评估了其促进伤口愈合的效果;(4)制备了酸溶性羧甲基壳聚糖原位凝胶并研究了其伤口止血密封性能及在稀醋酸中的溶解性;合成的PEG-原酸酯活性酯(SC-OE-PEG-OE-SC)具有酸敏感原酸酯键,可在稀酸作用下降解。SC-OE-PEG-OE-SC溶液和羧甲基壳聚糖溶液混合即可生成原位凝胶;此原位凝胶不仅可以在紧急情况下用于止血并封闭伤口,还可以被稀醋酸溶解去除,便于伤口的进一步外科处理,防止伤口部位的二次伤害。本论文的主要研究内容和结论如下:(1)将羟丙甲纤维素(HPMC)经丁二酸酐改性生成具有游离羧基的丁二酰羟丙基甲基纤维素(HPMCS),然后用EDC/NHS作为催化剂,交联HPMCS和壳聚糖(CS),制备出三种不同配比的HPMCS/CS复合交联水凝胶薄膜;测试了该薄膜的拉伸性能、溶胀比、水蒸汽透过率和氧渗透性等理化性质;采用NIH3T3细胞培养和MTT法测定了其细胞毒性,采用兔血红细胞测定了其溶血情况;以庆大霉素为模型药物测试了交联膜的体外药物释放和杀菌性能,结果表明,HPMCS/CS复合交联水凝胶薄膜具有良好的抗拉强度与断裂伸长率,适宜的气体透过性,生物相容性好,对细胞无毒,无溶血性,载药凝胶薄膜能有效抑制大肠杆菌和金黄色葡萄球菌的生长。(2)采用异丙醇为反应介质,用氯乙酸在NaOH催化下使KGM上的部分羟基被羧甲基取代,得到羧甲基魔芋葡甘聚糖(CMKGM);在EDC/NHS催化作用下使CMKGM与壳聚糖交联,经冷冻干燥制备了不同比例的CMKGM/CS复合交联海绵;使用FTIR,XRD,SEM,TGA等方法对复合海绵的理化结构、热稳定性等性质进行了表征,测定了 CMKGM/CS复合交联海绵的溶胀性,孔隙率,水蒸气透过率,最后对其生物相容性和血液相容性进行了测定。结果表明,CMKGM/CS复合交联海绵具有良好的吸水溶胀性能及热稳定性,能有效减少水分蒸发损失,对细胞无毒,无溶血性。(3)以乙腈为溶剂,DMAP为催化剂,PEG和二琥珀酰亚胺碳酸酯(DSC)反应,得到两端含活性酯的PEG衍生物(二琥珀酰亚胺聚乙二醇酯,SC-PEG-SC),将SC-PEG-SC溶液与壳聚糖溶液混合,SC-PEG-SC两端的活性酯与壳聚糖的氨基反应,形成凝胶,经干燥后制得PEG活性酯/壳聚糖薄膜;采用NMR,FTIR,XRD,TGA等方法对复合交联膜的结构、热稳定性等性质进行了表征,并通过拉伸性能检测、水蒸气及氧气透过率检测研究了不同比例原料形成凝胶薄膜的性能差异;最后通过ICR小鼠全层皮肤伤口模型测试了 PEG活性酯/壳聚糖水凝胶膜作为伤口敷料的性能。结果表明,此复合交联膜热稳定性及吸水性能良好,拉伸强度和断裂延伸率都有明显提高,能有效减少水分蒸发损失,对细胞无毒,无溶血性,动物实验结果表明这种复合交联膜能促进ICR小鼠伤口愈合。(4)设计了一种用于紧急情况下处理伤口的酸溶性羧甲基壳聚糖原位水凝胶密封剂;先采用五步反应合成PEG-原酸酯活性酯(SC-OE-PEG-OE-SC),并与羧甲基壳聚糖溶液混合,使二者在伤口部位发生交联反应生成SC-OE-PEG-OE-SC凝胶并粘附于组织、封闭伤口;由于原酸酯键对酸敏感,采用稀醋酸溶液即可将凝胶溶解;对SC-OE-PEG-OE-SC的结构进行了 1HNMR检测,并测定了其细胞毒性,降解产物细胞毒性,聚合物及降解产物血液相容性;将SC-OE-PEG-OE-SC溶液和羧甲基壳聚糖溶液按不同的体积比制成复合凝胶后,研究了原料浓度对凝胶化时间的影响,凝胶的皮肤粘附性能、溶解性能,最后进行了封闭伤口动物实验;结果显示,SC-OE-PEG-OE-SC合成成功,聚合物及降解产物对细胞均无毒,无溶血反应;从原料浓度对SC-OE-PEG-OE-SC/CMC复合凝胶凝固时间的影响结果可见,SC-OE-PEG-OE-SC和CMC浓度越大,凝固时间越短,原位凝胶可牢固粘附于组织表面并随组织形变而扭曲,能紧密贴合于大鼠伤口表面,形成凝胶并将伤口止血封闭,用5%醋酸溶液脱脂棉覆盖可完全溶解凝胶。综上所述,本工作得到了既有抗菌性的透明薄膜敷料材料,也有高吸收性海绵敷料材料,还有用于紧急伤口护理的酸溶性原位凝胶材料,不仅功能齐全,可满足不同类型的伤口护理之需求,而且安全无毒,制备方法简便,因此具有较强的理论价值和实际应用前景。
[Abstract]:Wound dressing is an important medical product for wound care. The new wound dressing can keep the wound healed by absorbing excess seepage or holding water to keep the wound healed. The biomedical materials prepared by chitosan and its derivatives as raw materials can stop bleeding and have a certain degree of hemostasis because of its excellent biocompatibility. Antibacterial properties have been widely recognized in promoting wound healing. In this work, three chitosan composite crosslinked films / sponges and an acid soluble chitosan in situ gel wound dressing were prepared with chitosan as the starting material and chemical crosslinking as the main technical means. Infrared spectroscopy (FTIR), X- ray diffraction (XRD), thermogravimetric analysis (TGA), scanning electron microscope (SEM) and other means were used to characterize their structures. The properties of the water vapor and oxygen transmittance detection, cytotoxicity test, blood compatibility test and animal experiment were tested by means of swelling rate detection, and their application prospects were evaluated. The main innovations in this paper The points are: (1) a cellulose derivative, butylene hydroxypropyl methyl cellulose (HPMCS) was prepared and crosslinked with chitosan to form a composite crosslinked gel membrane under the catalysis of EDC/NHS. The structure of the gel membrane was characterized and its performance as a wound dressing material was evaluated. (2) the carboxymethyl konjac glucomannan (CMKGM) was studied. High swelling CMKGM/CS composite crosslinked sponge wound dressing was prepared. (3) without any toxic crosslinking agent or initiator, PEG derivative (SC-PEG-SC) and chitosan were used to react directly to the gel to form the gel and then dry into the film. The effect of the wound healing was evaluated by the whole layer skin wound model of mice. (4) The acid soluble carboxymethyl chitosan in situ gel was prepared and its wound sealing performance and solubility in dilute acetic acid were studied. The synthesized PEG- active ester (SC-OE-PEG-OE-SC) had acid sensitive ester bond, which could be used to degrade.SC-OE-PEG-OE-SC solution and carboxymethyl chitosan solution under dilute acid to produce in-situ coagulation. The gel can not only be used to stop bleeding and close the wound in an emergency, but also can be dissolved by dilute acetic acid to facilitate further surgical treatment of the wound and prevent the two injury of the wound site. The main contents and conclusions of this paper are as follows: (1) the hydroxypropyl methylene cellulose (HPMC) is modified by butyl two anhydride to be free. The carboxyl group of Ding Erxian hydroxypropyl methyl cellulose (HPMCS), then EDC/NHS as a catalyst, crosslinked HPMCS and chitosan (CS), was used to prepare three different HPMCS/CS composite crosslinked hydrogel films. The tensile properties, swelling ratio, water vapor transmittance and oxygen permeability of the film were tested, and NIH3T3 cell culture and MTT were used. The results showed that the HPMCS/CS composite crosslinked hydrogel film had good tensile strength and elongation at break, suitable gas permeability and good biocompatibility, and the results showed that the composite crosslinked hydrogel film had good tensile strength and elongation at break. The cells are nontoxic and non hemolytic. The film of drug carrier gel can effectively inhibit the growth of Escherichia coli and Staphylococcus aureus. (2) using isopropanol as the reaction medium, chloroacetic acid is used as a catalyst to replace the hydroxyl group on KGM by NaOH, and the carboxymethyl konjac glucomannan (CMKGM) is obtained; and CMKGM and chitosan are made under the catalysis of EDC/NHS. Different proportions of CMKGM/CS composite crosslinked sponges were prepared by freeze-drying. The physicochemical structure and thermal stability of the composite sponge were characterized by FTIR, XRD, SEM, TGA and other properties. The swelling, porosity and vapor permeability of the CMKGM/CS composite sponge were measured. Finally, the biocompatibility and blood compatibility of the sponges were obtained. The results showed that the CMKGM/CS composite crosslinked sponge had good swelling property and thermal stability, and could effectively reduce the loss of evaporation of water, and was nontoxic and hemolytic to cells. (3) the reaction of acetonitrile as a solvent, DMAP as a catalyst, PEG and two succinimide carbonate (DSC), obtained the PEG derivatives (two amber) containing active esters at both ends (two amber) SC-PEG-SC solution is mixed with chitosan solution, the active ester of the SC-PEG-SC at both ends of SC-PEG-SC is reacted with the amino group of chitosan to form the gel and the PEG active ester / chitosan film is prepared after drying. The structure and thermal stability of the composite crosslinked membrane are characterized by NMR, FTIR, XRD, TGA and so on. The properties of the gel films formed by different proportion of raw materials were studied by the test of tensile properties and the permeability of water vapor and oxygen. Finally, the performance of the PEG active ester / chitosan hydrogel membrane was tested by the full layer skin wound model of ICR mice as wound dressing. The results showed that the thermal stability and water absorption of the composite crosslinked membrane were good. The tensile strength and elongation at break were improved obviously, and the loss of water evaporation was reduced, the cells were nontoxic and non hemolytic. The results of animal experiments showed that the composite crosslinking membrane could promote the healing of wound in ICR mice. (4) an acid soluble carboxymethyl chitosan in situ hydrogel sealant used in emergency treatment was designed. A five step reaction was used to synthesize PEG- active ester (SC-OE-PEG-OE-SC), and mixed with carboxymethyl chitosan solution to make the two groups crosslinked at the wound site to produce SC-OE-PEG-OE-SC gel and adhere to the tissue and seal the wound. The cytotoxicity, cytotoxicity of degradation products, and the blood compatibility of polymer and degradation products were measured, and the effect of the concentration of SC-OE-PEG-OE-SC and carboxymethyl chitosan on the gelation time, the skin adhesion and solubility of the gel were studied. The 1HNMR and the carboxymethyl chitosan solution were prepared by different volume ratio. Finally, the closed wound animal experiment was carried out. The results showed that the SC-OE-PEG-OE-SC synthesis was successful. The polymer and the degradation products were non-toxic and no hemolytic reaction. The effect of the concentration of raw materials on the coagulation time of SC-OE-PEG-OE-SC/CMC composite gel showed that the greater the concentration of SC-OE-PEG-OE-SC and CMC, the shorter the solidification time, the gel in situ could be secured. It adheres to the surface of the tissue and distorts with the deformation of the tissue. It can close to the surface of the wound of the rat, form the gel and seal the wound, and cover the gel with the 5% acetic acid solution. To sum up, we have obtained the transparent film dressing materials with both antibacterial properties and high absorbability sponge dressing. The acid soluble in situ gel material used for emergency wound care is not only fully functional, but also meets the needs of different types of wound care. It is safe and non-toxic, and the preparation method is simple. Therefore, it has strong theoretical value and practical application prospects.

【学位授予单位】：安徽大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：TQ427.26;R318.08

【相似文献】