切换至 "中华医学电子期刊资源库"

中华损伤与修复杂志(电子版) ›› 2019, Vol. 14 ›› Issue (04) : 256 -262. doi: 10.3877/cma.j.issn.1673-9450.2019.04.004

所属专题: 文献

论著

含银介孔二氧化硅-壳聚糖复合材料的制备与性能研究
夏德萌1, 王胥人2, 王元辰1, 熊文韬3, 许硕贵1, 周潘宇1,()   
  1. 1. 200433 上海,海军军医大学第一附属医院急诊科
    2. 200433 上海,海军军医大学护理学院
    3. 200433 上海,海军军医大学学员三大队
  • 收稿日期:2019-07-02 出版日期:2019-08-01
  • 通信作者: 周潘宇
  • 基金资助:
    国家自然科学基金(81571887); 上海市青年科技"启明星"(18QA1405400); 海军军医大学校级军事医学研究课题(2017JS15); 海军军医大学海洋军事医学重点攻关项目(2015HJ06)

Preparation and properties of silver-containing mesoporous silica-chitosan composites

Demeng Xia1, Xuren Wang2, Yuanchen Wang1, Wentao Xiong3, Shuogui Xu1, Panyu Zhou1,()   

  1. 1. Department of Emergency, First Affiliated Hospital of Naval Medical University, Shanghai 200433, China
    2. Department of Nursing, Naval Medical University, Shanghai 200433, China
    3. Naval Medical University Students Third Battalion, Shanghai 200433, China
  • Received:2019-07-02 Published:2019-08-01
  • Corresponding author: Panyu Zhou
  • About author:
    Corresponding author: Zhou panyu, Email:
引用本文:

夏德萌, 王胥人, 王元辰, 熊文韬, 许硕贵, 周潘宇. 含银介孔二氧化硅-壳聚糖复合材料的制备与性能研究[J/OL]. 中华损伤与修复杂志(电子版), 2019, 14(04): 256-262.

Demeng Xia, Xuren Wang, Yuanchen Wang, Wentao Xiong, Shuogui Xu, Panyu Zhou. Preparation and properties of silver-containing mesoporous silica-chitosan composites[J/OL]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2019, 14(04): 256-262.

目的

探讨含银介孔二氧化硅-壳聚糖复合材料(Ag/MSN-Chi)的制备方法及其微观表征、细胞毒性、吸水性能、抗菌性能及止血性能。

方法

以正硅酸乙酯为前驱体,十六烷基三甲基溴化铵为致孔剂,采用离子交换法在介孔二氧化硅纳米粒子(MSN)中引入银离子,制备出具有抗菌作用的新型有序的含银介孔二氧化硅纳米粒子(Ag/MSN)材料。再利用烷基化壳聚糖负载Ag/MSN,制备出Ag/MSN-Chi。根据所用材料不同将实验分为实验组和空白对照组,实验组又分为3个亚组:MSN组、Ag/MSN组、Ag/MSN-Chi组,空白组为不加任何材料的阳性对照。计算MSN和Ag/MSN的比表面积、孔容、孔径和Ag/MSN与Ag/MSN-Chi的电荷。并通过吸水实验、体外凝血实验、抗菌实验对MSN、Ag/MSN和Ag/MSN-Chi的细胞毒性、吸水性能、止血性能及抗菌性能进行评价,计算细胞相对存活率、吸水率、凝血酶原时间(PT)、凝血活酶时间(APTT)及抑菌率。取健康成年新西兰大白兔18只,随机分成3组:对照组(采用医用纱布处理)、Ag/MSN组(采用Ag/MSN处理)、Ag/MSN-Chi组(采用Ag/MSN-Chi处理),每组6只,建立肝创伤出血模型,计算止血时间。数据比较采用方差分析和t检验。

结果

MSN的比表面积为(523.8±12.4) m2/g、孔容为(1.2±0.4) m3/g、孔径为(3.5±0.9) nm;Ag/MSN的比表面积为(521.6±11.7) m2/g、孔容为(1.15±0.5) m3/g、孔径为(3.6±0.7) nm,2种材料的比表面积、孔容、孔径比较差异均无统计学意义(t=0.224、0.135、0.015,P值均大于0.05)。经测量,Ag/MSN的Zeta电位为-19.7 mV,Ag/MSN-Chi的Zeta电位为10.27 mV,表明Ag/MSN表面电荷从负值变为正值。Ag/MSN-Chi组、Ag/MSN组和MSN组与小鼠成肌细胞共培养1、4、7 d的细胞相对存活率比较,差异均无统计学意义(F=2.61、4.72、3.52, P值均大于0.05)。Ag/MSN组吸水率分别与MSN组和Ag/MSN-Chi组比较,差异均无统计学意义(t=0.482、1.159,P值均大于0.05)。经检测,Ag/MSN-Chi组、Ag/MSN组、MSN组和空白对照组的PT比较,差异无统计学意义(F=10.28,P>0.05);Ag/MSN-Chi组、Ag/MSN组、MSN组和空白对照组APTT分别为(20.9±2.1)、(28.5±3.4)、(31.4±2.6)、(38.7±2.5) s,4组比较差异有统计学意义(F=8.70,P<0.05);Ag/MSN-Chi组、Ag/MSN组、MSN组APTT分别与空白对照组比较,差异均有统计学意义(t=9.443、4.186、3.506,P值均小于0.05);Ag/MSN-Chi组APTT与Ag/MSN组比较,差异有统计学意义(t=3.294,P<0.05)。MSN组在培养0.5、2、4、6、24 h 5个时间点抑菌率比较差异无统计学意义(F=5.437,P>0.05);培养0.5 h,Ag/MSN组和Ag/MSN-Chi组抑菌率分别为(99.7±5.2)%、(97.1±5.4)%,与培养0.5 h MSN组抑菌率(11.2±5.8)%比较,差异均有统计学意义(t=19.678、18.775, P值均小于0.05);培养24 h,Ag/MSN组和Ag/MSN-Chi组抑菌率分别为(73.2±5.1)%和(72.9±6.9)%,与MSN组(11.8±5.7)%比较,差异均有统计学意义(t=13.904、11.825, P值均小于0.05)。Ag/MSN-Chi组、Ag/MSN组和对照组止血时间分别为(12.3±1.5)、(17.2±3.4)、(28.1±3.8) s,3组比较差异有统计学意义(F=5.892,P<0.05);Ag/MSN-Chi组和Ag/MSN组止血时间分别与对照组比较,差异均有统计学意义(t=9.473、5.236, P值均小于0.05);且Ag/MSN-Chi组与Ag/MSN组止血时间比较,差异有统计学意义(t=3.230,P<0.05)。

结论

Ag/MSN-Chi在不增加细胞毒性的基础上具有有较好的吸水性能、止血性能及抗菌性能。

Objective

To investigate the preparation method of silver-containing mesoporous silica nanoparticle-chitosan(Ag/MSN-Chi) composites and their microscopic representation, cytotoxicity, water absorption, rapid hemostatic properties and antibacterial properties.

Methods

Using ethyl orthosilicate as precursor and cetyltrimethylammonium bromide as porogen, silver ions were introduced into mesoporous silica nanoparticles (MSN) by ion-exchange method to prepare novel ordered silver-containing MSN (Ag/MSN) material with antibacterial action. Ag/MSN-Chi was prepared by using alkylated chitosan loaded with Ag/MSN. The experiment were divided into experimental group and blank control group according to the materials used. The experimental group was further divided into three subgroups: MSN group, Ag/MSN group and Ag/MSN-Chi group. The blank group was a positive control without material. The specific surface area, pore volume, pore size of MSN and Ag/MSN, the charge of Ag/MSN and Ag/MSN-Chi were calculated. The cytotoxicity, water absorption performance, hemostatic and antibacterial properties of MSN, Ag/MSN and Ag/MSN-Chi were evaluated by water absorption test, in vitro coagulation test and antibacterial test to calculate the relative cell survival rate, water absorption rate and prothrombin time (PT), activated partial thromboplastin time (APTT), and bacteriostatic rate. Eighteen healthy adult New Zealand white rabbits were randomly divided into three groups: control group (treated with medical gauze), Ag/MSN group (treated with Ag/MSN), and Ag/MSN-Chi group (treated with Ag/MSN-Chi), 6 rats in each group. A hepatic trauma hemorrhage model was established and the hemostasis time was calculated. Data comparisons were analyzed by analysis of variance and t test.

Results

The specific surface area of MSN was (523.8±12.4) m2/g, the pore volume was (1.2±0.4) m3/g, and the pore diameter was (3.5±0.9) nm; the specific surface area of Ag/MSN was (521.6±11.7) m2/g, the pore volume was (1.15±0.5) m3/g, and the pore diameter was (3.6±0.7) nm, there was no significant difference in specific surface area, pore volume and pore size between the two materials (t=0.224, 0.135, 0.015; with P values above 0.05). The Zeta potential of Ag/MSN was measured to be -19.7 mV, and the Zeta potential of Ag/MSN-Chi was 10.27 mV, indicating that the surface charge of Ag/MSN changed from a negative value to a positive value. The relative survival rates of Ag/MSN-Chi group, Ag/MSN group and MSN group at 1, 4, and 7 days after co-culture with mouse myoblasts were not statistically significant (F=2.61, 4.72, 3.52; with P values above 0.05). The water absorption of Ag/MSN group was significantly different from that of MSN group and Ag/MSN-Chi group (t=0.482, 1.159; with P values above 0.05). There was no significant difference in PT between Ag/MSN-Chi group, Ag/MSN group, MSN group and blank control group (F=10.28, P>0.05). The APTT of Ag/MSN-Chi group, Ag/MSN group, MSN group and blank control group were (20.9±2.1), (28.5±3.4), (31.4±2.6), (38.7±2.5) s, respectively, the difference of the 4 groups was statistically significant (F=8.70, P<0.05); the APTT of Ag/MSN-Chi group, Ag/MSN group and MSN group were significantly different from the blank control group, the differences were statistically significant (t=9.443, 4.186, 3.506; with P values below 0.05); the difference between Ag/MSN group and Ag/MSN-Chi group of APTT was statistically significant (t=3.294, P<0.05). There was no significant difference in the inhibition rate between the MSN group at 0.5, 2, 4, 6 and 24 h (F=5.437, P>0.05); after incubation for 0.5 h, the inhibition rates of Ag/MSN group and Ag/MSN-Chi group were (99.7±5.2)% and (97.1±5.4)%, respectively, compared with the incubation rate of 0.5 h in MSN group (11.2±5.8)%, the differences were statistically significant (t=19.678, 18.775; with P values below 0.05). After 24 h of culture, the inhibition rates of Ag/MSN group and Ag/MSN-Chi group were (73.2±5.1)% and (72.9±6.9)%, respectively, compared with MSN group (11.8±5.7)%, the differences were statistically significant significance (t=13.904, 11.825; with P values below 0.05). The hemostasis time of Ag/MSN-Chi group, Ag/MSN group and control group were (12.3±1.5), (17.2±3.4), (28.1±3.8) s, respectively, the difference between the three groups was statistically significant (F=5.892, P<0.05); the Ag/MSN-Chi group and the Ag/MSN group were significantly different from the control group (t=9.473, 5.236; with P values below 0.05); and the Ag/MSN-Chi group was compared with the Ag/MSN group, the difference was statistically significant (t=3.230, P<0.05).

Conclusion

Ag/MSN-Chi has better water absorption performance, hemostatic property and antibacterial property without increasing cytotoxicity.

表1 Ag/MSN-Chi组、Ag/MSN组和MSN组小鼠成肌细胞相对存活率比较(%, ±s)
表2 Ag/MSN-Chi组、Ag/MSN组、MSN组和空白对照组APTT与PT比较(s, ±s)
图1 Ag/MSN-Chi组、Ag/MSN组和MSN组不同培养时间对大肠杆菌的抑菌率变化。Ag/MSN-Chi为含银介孔二氧化硅纳米粒子-壳聚糖;Ag/MSN为含银介孔二氧化硅纳米粒子;MSN为介孔二氧化硅纳米粒子
[1]
Björses K,Holst J. Various local hemostatic agents with different modes of action; an in vivo comparative randomized vascular surgical experimental study[J]. Eur J Vasc Endovasc Surg, 2007, 33(3): 363-370.
[2]
王胥人, 桂莉, 王毅欣. 美军战术战伤救护止血进展及启示[J]. 中国急救复苏与灾害医学杂志,2017, 12(8): 793-796.
[3]
Sohn VY,Eckert MJ,Martin MJ, et al. Efficacy of three topical hemostatic agents applied by medics in a lethal groin injury model[J]. J Surg Res, 2009, 154(2): 258-261.
[4]
邓乐君, 樊鸿浩, 李伟达,等. 可吸收止血材料的生物相容性研究进展[J]. 中国生物医学工程学报,2016, 35(2): 241-246.
[5]
李大为, 何进, 何凤利,等. 丝素蛋白/壳聚糖复合材料在组织工程中应用的研究进展[J]. 中国生物工程杂志,2017, 37(10): 111-117.
[6]
刘琳娜, 李学拥, 赵聪颖,等. 壳聚糖护创敷料用于深Ⅱ度烧伤创面的疗效和安全性:一项前瞻性、随机、单盲、阳性对照临床试验[J]. 中国组织工程研究,2017, 21(14): 2222-2226.
[7]
李正超, 刘璐, 张娟,等. 局部止血材料研究新进展[J]. 医疗卫生装备,2015, 36(10): 109-112.
[8]
胡何晶, 段军超. 纳米二氧化硅对凝血系统的影响及研究进展[J]. 毒理学杂志,2016, 30(5): 385-389.
[9]
李晓生, 刘昌胜, 袁媛,等. 介孔硅基干凝胶的制备及其止血性能的研究[J]. 无机材料学报,2008, 23(2): 327-331.
[10]
Dai C,Liu C,Wei J, et al. Molecular imprinted macroporous chitosan coated mesoporous silica xerogels for hemorrhage control[J]. Biomaterials, 2010, 31(30): 7620-7630.
[11]
Wang Y,Zhao Q,Han N, et al. Mesoporous silica nanoparticles in drug delivery and biomedical applications[J]. Nanomedicine, 2015, 11(2): 313-327.
[12]
Silver S. Bacterial silver resistance: molecular biology and uses and misuses of silver compounds[J]. FEMS Microbiol Rev, 2003, 27(2/3): 341-353.
[13]
Dai C,Yuan Y,Liu C, et al. Degradable, antibacterial silver exchanged mesoporous silica spheres for hemorrhage control[J]. Biomaterials, 2009, 30(29): 5364-5375.
[14]
Jalal M,Ansari MA,Ali SG, et al. Anticandidal activity of bioinspired ZnO NPs: effect on growth, cell morphology and key virulence attributes of species[J]. Artif Cells Nanomed Biotechnol, 2018, 46(sup1): 912-925.
[15]
王美娜, 李苑新, 蔡兴. 羧甲基壳聚糖的性能及应用概况[J]. 中国实验方剂学杂志,2015, 21(1): 228-232.
[16]
刘璐, 王淑芳, 李灵芝,等. 壳聚糖/聚谷氨酸聚电解质海绵的制备及止血作用研究[J]. 中国急救复苏与灾害医学杂志,2014, 9(5): 406-410.
[17]
叶梓, 王世杰, 李辉,等. 血管内皮细胞中的凝血物质:理论研究新进展[J]. 中国组织工程研究,2017, 21(4): 627-632.
[18]
Cox JM,Rall JM. Evaluation of XSTAT® and QuickClot® Combat Gauze® in a Swine Model of Lethal Junctional Hemorrhage in Coagulopathic Swine[J]. J Spec Oper Med, 2017, 17(3): 64-67.
[19]
Johnson D,Bates S,Nukalo S, et al. The effects of QuikClot Combat Gauze on hemorrhage control in the presence of hemodilution and hypothermia[J]. Ann Med Surg(Lond), 2014, 3(2): 21-25.
[20]
李继城, 李东东, 温莎莎,等. 季铵化壳聚糖止血海绵制备工艺的研究[J]. 化学推进剂与高分子材料,2017, 15(3): 68-71, 80.
[21]
Ahn SJ,Lee SJ,Kook JK, et al. Experimental antimicrobial orthodontic adhesives using nanofillers and silver nanoparticles[J]. Dent Mater, 2009, 25(2): 206-213.
[22]
余文珺, 胡滨, 张富强. 纳米载银无机抗菌剂对义齿基托树脂抗菌性能的影响[J]. 上海交通大学学报(医学版), 2006, 26(10): 1096-1098.
[23]
刘桐, 唐慧琴, 张学华,等. 镀银碳纳米管的抗菌性研究[J]. 功能材料,2007, 38(5): 802-805.
[24]
石凉, 汪涛, 吴大洋. 壳聚糖止血材料及最新研究进展[J]. 蚕业科学,2009, 35(4): 929-934.
[25]
卢斌, 钱利强, 章志量. 壳聚糖止血机理及应用进展研究[J]. 健康研究,2010, 30(1): 55-58.
[26]
尹刚, 侯春林, 刘万顺,等. 新型壳聚糖止血粉止血作用的实验研究[J]. 中国修复重建外科杂志,2009, 23(11): 1347-1349.
[27]
Grissom TE,Fang R. Topical hemostatic agents and dressings in the prehospital setting[J]. Curr Opin Anaesthesiol, 2015, 28(2): 210-216.
[28]
赵一璠, 赵京禹, 付小兵. 战创伤用局部快速止血材料的研究进展[J]. 中华创伤杂志,2018, 34(6): 534-539.
[1] 马桥桥, 张传开, 郭开今, 蒋涛, 王子豪, 刘勇, 郝亮. 可降解止血粉减少初次全膝关节置换术失血量的研究[J/OL]. 中华关节外科杂志(电子版), 2024, 18(05): 585-589.
[2] 赵锦伟, 李晓军, 刘桂宇, 荣晓玲, 黄明利, 宋文静, 高广凌, 鞠昌军. 固定平台假体膝关节单髁置换术中止血带应用策略研究[J/OL]. 中华关节外科杂志(电子版), 2024, 18(01): 39-47.
[3] 费扬, 赵晗希, 孙丽琴, 楼琴华, 胡骏程. 银杏叶提取物对糖尿病肾病患者的疗效及其对尿液外泌体miR-342-3p的干预研究[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(03): 219-224.
[4] 杨丽, 李文颖, 田园, 周林熙, 王伟. 脂质水胶体硫酸银敷料和蔗糖八硫酸盐敷料在糖尿病足溃疡中的应用进展[J/OL]. 中华损伤与修复杂志(电子版), 2024, 19(03): 271-275.
[5] 冯蓉琴, 王鹏, 李煜, 陆翮, 白晓智, 韩军涛. 抗菌肽在糖尿病创面愈合中作用的研究进展[J/OL]. 中华损伤与修复杂志(电子版), 2024, 19(01): 78-82.
[6] 李可心, 鲁瑞, 田燕, 王文俊, 李维, 万月强, 翟嵩, 刘拉羊. 阿达木单抗治疗银屑病致播散性肺结核一例并文献复习[J/OL]. 中华实验和临床感染病杂志(电子版), 2024, 18(04): 245-249.
[7] 王哲, 陈杨, 徐京杭, 李六亿, 樊亚楠, 纪童童, 胡可, 高慧, 于岩岩. 1 349例肠道门诊患者院前抗菌药物使用情况调查[J/OL]. 中华实验和临床感染病杂志(电子版), 2024, 18(01): 19-26.
[8] 王阅, 杨园梦, 何德亿, 孟雯, 陈昕煜, 李飞, 卢展民, 陆海霞. 基于口腔微生态的龋病防治研究进展[J/OL]. 中华口腔医学研究杂志(电子版), 2024, 18(06): 391-396.
[9] 王刚, 李涛, 刘玉芳. 胃癌根治手术后行抗菌药物治疗对患者肠道细菌移位及肠黏膜屏障功能的影响[J/OL]. 中华普外科手术学杂志(电子版), 2024, 18(02): 141-145.
[10] 李彦浇, 梁雷, 金钫, 王智伟. 银杏内酯B通过调控miR-24-3p对人牙周膜干细胞增殖、成骨分化的影响[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(04): 229-235.
[11] 王冬澳, 林超, 鄂长勇. 胰腺术后出血原因及诊治策略[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(04): 472-480.
[12] 韦小霞, 陈管洁, 李雪珠, 李晓青, 钱淑媛. 机械通气患者抗菌药物雾化吸入的临床实施[J/OL]. 中华重症医学电子杂志, 2024, 10(04): 334-337.
[13] 蒋心怡, 顾丹丹, 叶艳, 缪佳蓉. RNA测序研究抗菌肽KT2治疗溃疡性结肠炎的作用机制[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(01): 8-15.
[14] 陈慧, 邹祖鹏, 周田田, 张艺丹, 张海萍. 皮肤镜对头皮红斑性皮肤病辅助鉴别诊断的研究进展[J/OL]. 中华临床医师杂志(电子版), 2024, 18(07): 692-698.
[15] 向林, 江云颂, 程吕佳, 关炳生, 杨景哥. 生长素释放肽在肥胖治疗中的潜在应用价值研究进展[J/OL]. 中华肥胖与代谢病电子杂志, 2024, 10(04): 235-241.
阅读次数
全文


摘要