CN107261195B - 一种运动创伤康复敷料用抗菌生物质凝胶网的制备方法与用途 - Google Patents
一种运动创伤康复敷料用抗菌生物质凝胶网的制备方法与用途 Download PDFInfo
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Abstract
本发明属于生物材料技术领域,具体涉及一种运动创伤康复敷料用抗菌生物质凝胶的制备方法与用途。本发明以乳酸羟基乙酸共聚物、羟丙基壳聚糖、聚己内酯和医用几丁糖为原料通过静电纺丝制备出静电纺丝网,然后对静电纺丝网负载胶原蛋白制备出抗菌生物质凝胶网;本发明所用材料均为绿色环保材料,具有一定的生物相容性和可降解性,医用几丁糖可促进创伤组织生理性修复,抑制疤痕形成;制备的运动创伤康复敷料用抗菌生物质凝胶网不仅具有优异的降解性,而且降解速率恒定,属于线性降解,后期可以利用这一降解特性用于控制药物释放,到达控释的目的。
Description
技术领域
本发明属于生物材料技术领域,具体涉及一种运动创伤康复敷料用抗菌生物质凝胶的制备方法与用途。
背景技术
运动损伤的恢复是长期困扰运动员的难题,直接影响运动训练和运动技术的正常发挥。根据近年来的科学和统计报导,全球创伤市场的需求越来越大,特别是在外科手术类的创伤;全球创伤市场每年总人数已经突破1亿人次,并呈逐年增长的趋势。每年约有2千万人次因意外事故造成创伤与撕裂伤,而烧烫伤数量则约每年1千万人次;此外每年因慢性疾病、糖尿病、衰老退化所造成的溃疡性伤口则已经超过了3千万人次。
外科手术由于治疗的需求以及内窥镜手术的风行,手术风险已经大幅降低,但随之而来的是对于较佳的伤口照护及预防术后疤痕的需求。目前已经出现借助各种先进敷料进行伤口照护的方法,藉以缩短伤口愈合时间且同时消除疤痕。
传统的敷料是由天然植物纤维或动物毛发类物质所构成,如纱布、棉垫、羊毛、各类油纱布等,这类敷料仅为暂时性的覆盖材料,均需在一定的时间内加以更换。现有创伤敷料的研究让人们对于创伤敷料有了科学上的认识,且研究显示,较佳的创伤敷料是能够保持创伤处的良好细胞生长愈合环境,以及能够控制和吸收渗出物;透气、透湿,能阻止细菌侵入;能紧密贴敷于创伤面;可载设、释放药物;此外应具有良好的组织及血液兼容性,在从创伤面揭取时不发生粘连和脱屑;还应当具有较佳的机械性能和抗拉强度,以及使用方便的性质。在现有技术中,如2008年10月29日公开的中国专利申请号200810122438.1提供了一种医用水凝胶床上敷料的制备方法,再例如2005年1月12日公开的公开号为CN1562382A名称为含有聚氨酯水乳液的水凝胶型创伤敷料及其制备方法的专利申请,均为关于以水凝胶为基体的医用敷料。
敷料必须有适当的力学强度和机械性能,其次敷料最好有一定的生物相容性和可降解性,避免在创伤伤口处愈合过程中导致伤口与敷料黏连、生物排斥等现象。
发明内容
本发明的目的是提供一种运动创伤康复敷料用抗菌生物质凝胶网的制备方法,本发明以乳酸羟基乙酸共聚物、羟丙基壳聚糖、聚己内酯和医用几丁糖为原料通过静电纺丝制备出静电纺丝网,然后对静电纺丝网负载胶原蛋白制备出抗菌生物质凝胶网;本发明所用材料均为绿色环保材料,具有一定的生物相容性和可降解性,以具有生物降解性的羟丙基壳聚糖为抗菌剂替代传统的苯扎氯铵等抗菌剂,副作用大大降低,而且医用几丁糖可促进创伤组织生理性修复,抑制疤痕形成,减少组织粘连。
本发明是通过以下技术方案实现上述目的的,一种运动创伤康复敷料用抗菌生物质凝胶网的制备方法,包括以下步骤:
1)静电纺丝网的制备:将聚乳酸羟基乙酸共聚物、羟丙基壳聚糖、聚己内酯和医用几丁糖溶于四氢呋喃和氮氮二甲基甲酰胺的混合溶液中搅拌溶解,搅拌均匀后加入2%wt(以混合溶液重量为基准)的双甲基丙烯酸二缩三乙二醇酯和4%wt(以混合溶液重量为基准)的2-羟基-2-甲基-1-苯基丙酮得静电纺丝液;将静电纺丝液采用静电纺丝设备在进行静电纺丝,然后将静电纺丝编织成网,在光强度为210mW/cm3的紫外光源下照射20分钟,进行交联得静电纺丝网;
2)抗菌生物质凝胶网的制备:将I型胶原蛋白粉溶于0.1mol/L醋酸的乙醇溶液中,配制成2wt%的I型胶原蛋白溶液,然后加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和N-羟基琥珀酰亚胺得胶原蛋白溶液,将步骤1)制备的静电纺丝网置于胶原蛋白溶液中室温超声浸渍2h,浸渍结束后取出带有I型胶原蛋白的生物质凝胶网-70℃下冻干即得运动创伤康复敷料用抗菌生物质凝胶网;
优选的,步骤1)所述混合溶液中四氢呋喃与氮氮二甲基甲酰胺的体积比为1:2,实验过程中采用体积比为1:2的四氢呋喃与氮氮二甲基甲酰胺作为静电纺丝的溶液是因为可以对聚乳酸羟基乙酸共聚物、羟丙基壳聚糖、聚己内酯和医用几丁糖完全溶解,单纯采用一种或者其余配比(THF:DMF体积比尝试过10:1-1:10)溶解度均不是较好,只有THF:DMF=1:2具有意料不到的溶解效果;
优选的,步骤1)所述混合溶液中聚乳酸羟基乙酸共聚物的浓度为12-14 wt %,羟丙基壳聚糖的浓度为4-6wt%,聚己内酯的浓度为5-8wt%,医用几丁糖的浓度为4-6%wt;
优选的,步骤1)所述聚乳酸羟基乙酸共聚物中聚乳酸(PLA)与聚乙醇酸(PGA)的聚合比例为60:40;
优选的,步骤1)所述静电纺丝设备的电场电压为25KV;
优选的,步骤2)所述的胶原蛋白溶液中1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐的浓度为0.003%wt;N-羟基琥珀酰亚胺的浓度为0.002%wt;
聚己内酯在体内与生物细胞相容性很好,细胞可在其基架上正常生长,并可降解成CO2和H2O。
医用几丁糖是由蟹壳提纯的高分子化合物几丁质(chitin),经脱N—乙酰基再深加工后制成的一种聚氨基葡萄糖,是一种具有良好生物相容性、生物可降解性及生物学活性的医用高分子多糖类物质。医用几丁糖具有选择性促进上皮细胞、内皮细胞生长而抑制成纤维细胞生长的生物特性,从而促进组织生理性修复,抑制疤痕形成,减少组织粘连。
羟丙基壳聚糖是由壳聚糖改性而成的一种水溶性良好的壳聚糖衍生物,它既能溶于水又能溶于乙醇等有机溶剂,具有良好的乳化性、抗菌性、吸湿保湿性和表面活性,其吸湿保湿能力与透明质酸(HA)相当,乳化性和泡沫性与非离子表面活性剂Tween60相当。
由于本发明的运动创伤康复敷料用抗菌生物质凝胶网具有恒定的生物降解速率,所以本发明制备的运动创伤康复敷料用抗菌生物质凝胶网可用于药物的负载,制备成缓释药物系统。
与现有技术相比,本发明具有如下优点:
1)本发明提供了一种全新的运动创伤康复敷料用抗菌生物质凝胶网,该凝胶网具有优异的生物相容性和可降解性,医用几丁糖可促进创伤组织生理性修复,抑制疤痕形成,减少组织粘连;
2)本发明制备的运动创伤康复敷料用抗菌生物质凝胶网不仅具有优异的降解性,而且降解速率恒定,属于线性降解,后期可以利用这一降解特性用于控制药物释放,到达控释的目的。
附图说明
图1为本发明实施例1制备的静电纺丝网的扫描电镜图;
图2为本发明实施例1制备的运动创伤康复敷料用抗菌生物质凝胶网的扫描电镜图。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚明了,下面结合具体实施方式,对本发明进一步详细说明。应该理解,这些描述只是示例性的,而并非要限制本发明的范围。
I型胶原蛋白粉为牛跟腱胶原蛋白,来源于西格玛奥德里奇(上海)贸易有限公司,产品编号为C9879,CAS号为9007-34-5;
聚乳酸羟基乙酸共聚物来源于西格玛奥德里奇(上海)贸易有限公司,产品编号为P2066,共聚物中聚乳酸(PLA)与聚乙醇酸(PGA)的聚合比例为60:40。
实施例1
1)静电纺丝网的制备:将聚乳酸羟基乙酸共聚物、羟丙基壳聚糖、聚己内酯和医用几丁糖溶于四氢呋喃和氮氮二甲基甲酰胺的混合溶液中(体积比THF:DMF=1:2)搅拌溶解,搅拌均匀后加入2%wt(以混合溶液重量为基准)的双甲基丙烯酸二缩三乙二醇酯和4%wt(以混合溶液重量为基准)的2-羟基-2-甲基-1-苯基丙酮得静电纺丝液;将静电纺丝液采用静电纺丝设备(电场电压为25KV)在进行静电纺丝,然后将静电纺丝编织成网,在光强度为210mW/cm3的紫外光源下照射20分钟,进行交联得静电纺丝网;混合溶液中聚乳酸羟基乙酸共聚物的浓度为13 wt %,羟丙基壳聚糖的浓度为5wt%,聚己内酯的浓度为7wt%,医用几丁糖的浓度为5%wt;
对制备出的静电纺丝网采用扫描电镜进行表征,结果如图1所示:
2)抗菌生物质凝胶网的制备:将I型胶原蛋白粉溶于0.1mol/L醋酸的乙醇溶液中,配制成2wt%的I型胶原蛋白溶液,然后加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(在胶原蛋白溶液中的浓度为0.003%wt)和N-羟基琥珀酰亚胺(在胶原蛋白溶液中的浓度为0.002%wt)得胶原蛋白溶液,将步骤1)制备的静电纺丝网置于胶原蛋白溶液中室温超声浸渍2h,浸渍结束后取出带有I型胶原蛋白的生物质凝胶网-70℃下冻干即得运动创伤康复敷料用抗菌生物质凝胶网,其扫描电镜图如图2所示。
实施例2
1)静电纺丝网的制备:将聚乳酸羟基乙酸共聚物、羟丙基壳聚糖、聚己内酯和医用几丁糖溶于四氢呋喃和氮氮二甲基甲酰胺的混合溶液中(体积比THF:DMF=1:2)搅拌溶解,搅拌均匀后加入2%wt(以混合溶液重量为基准)的双甲基丙烯酸二缩三乙二醇酯和4%wt(以混合溶液重量为基准)的2-羟基-2-甲基-1-苯基丙酮得静电纺丝液;将静电纺丝液采用静电纺丝设备(电场电压为25KV)在进行静电纺丝,然后将静电纺丝编织成网,在光强度为210mW/cm3的紫外光源下照射20分钟,进行交联得静电纺丝网;混合溶液中聚乳酸羟基乙酸共聚物的浓度为14 wt %,羟丙基壳聚糖的浓度为6wt%,聚己内酯的浓度为8wt%,医用几丁糖的浓度为6%wt;
2)抗菌生物质凝胶网的制备:将I型胶原蛋白粉溶于0.1mol/L醋酸的乙醇溶液中,配制成2wt%的I型胶原蛋白溶液,然后加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(在胶原蛋白溶液中的浓度为0.003%wt)和N-羟基琥珀酰亚胺(在胶原蛋白溶液中的浓度为0.002%wt)得胶原蛋白溶液,将步骤1)制备的静电纺丝网置于胶原蛋白溶液中室温超声浸渍2h,浸渍结束后取出带有I型胶原蛋白的生物质凝胶网-70℃下冻干即得运动创伤康复敷料用抗菌生物质凝胶网。
孔隙率测量:取实施例1制备出的运动创伤康复敷料用抗菌生物质凝胶网切成2mmX2mm的样品,在10 mL的离心管中,倒满无水乙醇称重M1,取适量样品M2浸入无水乙醇中,超声震荡至完全脱除海绵中的气泡,使乙醇完全充于样品结构中,向离心管中添加无水乙醇至充满,称重为M3。取出吸满无水乙醇的样品,称量剩余的质量为M4,计算孔隙率为θ,重复5次测定,求孔隙率的平均值;
实施例1制备所得的孔隙率为92.3%,孔隙率高可以提高更大的空间用来吸收创伤处溶出的体液,同时吸附并激活血小板释放凝血因子,对创面止血,可作为止血材料。
降解速率测试:取实施例1制备的样品质量为W1的浸没入磷酸盐缓冲溶液(PH=7.4)溶液中,放置于培养箱中(37℃培养),分别于2、4、6、8、10周取出,干燥后称重W2,同时,更换PBS,材料称重后再次浸没入液体,放置于培养箱中,直至下一测试时间点,按照下述公式计算质量损失:
表1为降解率随时间的变化曲线
表1中的降解率结果表明,本发明制备的运动创伤康复敷料用抗菌生物质凝胶网在pH=7.4的磷酸盐缓冲溶液中降解速率基本为线性恒定。
为了验证本发明运动创伤康复敷料用抗菌生物质凝胶网可用于制备控释药物,在实施例1制备步骤2)中加入I型胶原蛋白粉的同时加入克拉霉素,制备出具有抗菌药物的生物质凝胶网,置于pH=7.4的磷酸盐缓冲水溶液中,分不同时间段采用HPLC(检测方法按照现行药典中克拉霉素的方法进行检测)测量水溶液中的浓度,浓度随时间的结果如表2所示:
表2药物浓度随时间的变化
试验结果表明,药物在生物质凝胶网中的释放速度前期基本是恒定的,在15天以后基本达到最大药物浓度,所以根据这一特性本发明制备的运动创伤康复敷料用抗菌生物质凝胶网完全可以作为一种载体对药物进行负载,达到缓控释药物的效果。
尽管已经详细描述了本发明的实施方式,但是应该理解的是,在不偏离本发明的精神和范围的情况下,可以对本发明的实施方式做出各种改变、替换和变更。
Claims (6)
1.一种运动创伤康复敷料用抗菌生物质凝胶网的制备方法,包括以下步骤:
1)静电纺丝网的制备:将聚乳酸羟基乙酸共聚物、羟丙基壳聚糖、聚己内酯和医用几丁糖溶于四氢呋喃和氮氮二甲基甲酰胺的混合溶液中搅拌溶解,搅拌均匀后加入2wt %的双甲基丙烯酸二缩三乙二醇酯和4wt %的2-羟基-2-甲基-1-苯基丙酮得静电纺丝液;将静电纺丝液采用静电纺丝设备进行静电纺丝,然后将静电纺丝编织成网,在光强度为210mW/cm2的紫外光源下照射20分钟,进行交联得静电纺丝网;所述混合溶液中聚乳酸羟基乙酸共聚物的浓度为12-14 wt %,羟丙基壳聚糖的浓度为4-6wt%,聚己内酯的浓度为5-8wt%,医用几丁糖的浓度为4-6wt %;
2)抗菌生物质凝胶网的制备:将I型胶原蛋白粉溶于0.1mol/L醋酸的乙醇溶液中,配制成2wt%的I型胶原蛋白溶液,然后加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和N-羟基琥珀酰亚胺得胶原蛋白溶液,将步骤1)制备的静电纺丝网置于胶原蛋白溶液中室温超声浸渍2h,浸渍结束后取出带有I型胶原蛋白的生物质凝胶网-70℃下冻干即得运动创伤康复敷料用抗菌生物质凝胶网。
2.根据权利要求1所述的制备方法,其特征在于:步骤1)所述混合溶液中四氢呋喃与氮氮二甲基甲酰胺的体积比为1:2。
3.根据权利要求1所述的制备方法,其特征在于:步骤1)所述混合溶液中聚乳酸羟基乙酸共聚物的浓度为13wt %,羟丙基壳聚糖的浓度为5wt%,聚己内酯的浓度为7wt%,医用几丁糖的浓度为5wt %。
4.根据权利要求1所述的制备方法,其特征在于:步骤1)所述聚乳酸羟基乙酸共聚物中聚乳酸与聚乙醇酸的聚合比例为60:40。
5.根据权利要求1所述的制备方法,其特征在于:步骤2)所述的胶原蛋白溶液中1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐的浓度为0.003wt %;N-羟基琥珀酰亚胺的浓度为0.002wt %。
6.一种运动创伤康复敷料用抗菌生物质凝胶网的用途,所述运动创伤康复敷料用抗菌生物质凝胶网由权利要求1-5任一所述方法制备,其特征在于:用于负载药物制备成缓释药物系统。
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