CN113717431A - 一种负载牛磺酸分子的胶原基支架材料及其制备方法 - Google Patents
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Abstract
一种负载牛磺酸分子的胶原基支架材料及其制备方法,包括以下步骤:(1)在37℃下,配制胶原溶液,将溶于酸的胶原溶液浇铸到模具中;(2)将浇铸的胶原溶液在‑20℃下预冻2小时,然后在‑80℃条件下预冻24小时以上,然后真空冷冻干燥48小时以上,获得结构均匀的胶原海绵支架;(3)分别配制牛磺酸溶液、碳化二亚胺和N‑羟基丁二酰溶液;(4)将胶原海绵支架浸泡到牛磺酸溶液中,同时加入碳化二亚胺和N‑羟基丁二酰溶液进行交联反应;(5)将交联水凝胶冷冻干燥后,即得到一种负载牛磺酸分子的胶原基支架材料。本材料具有较好的理化性能和生物相容性,且成型可控,可用于临床上眼科创面的修复和治疗,其制备工艺简单,设备简易。
Description
技术领域
本发明涉及一种负载牛磺酸分子的胶原基支架材料及其制备方法,具体涉及到一种制备具有良好理化性能和生物相容性的功能化胶原基角膜修复材料的方法。本发明所得的材料可应用于临床上眼科创面的修复和治疗。
背景技术
角膜疾病是一种十分常见的眼科病,约有三分之一的失明患者是由于角膜疾病而导致的。对于角膜病患者而言,需要进行角膜移植手术,采用供体的健康角膜来替代其损伤角膜,这就导致了对同种异体角膜的极大需求。但是目前在临床上,健康的角膜供体数量远少于角膜移植的需求量,所以人工角膜的研发就显得十分必要。
人工角膜发展最快的就是其材料的发展,目前制备人工角膜修复材料的方法有很多,大多数都集中在玻璃,聚甲基丙烯酸甲酯,水凝胶,硅凝胶,丙烯酸酯,多孔聚四氟乙烯等材料上。胶原(Collagen)作为人体组织细胞外最重要的不溶性纤维蛋白,在动物细胞中扮演结合组织的角色,是构成细胞外基质的骨架,胶原在细胞外基质中形成半结晶的纤维,给细胞提供抗张力和弹性,并在细胞的迁移和发育中起作用。鉴于此,胶原基材料在角膜修复领域中具有广阔的应用前景。牛磺酸(Taurine)又称α-氨基乙磺酸,是一种从牛磺酸中分离出来的非蛋白氨基酸,在体内以游离状态存在,与人体内胱氨酸、半胱氨酸的代谢密切相关。牛磺酸能够维持视网膜的正常功能,从而有效提高神经传导和视觉机能。此外,牛磺酸还具有调节晶体渗透压和抗氧化等重要作用,可抑制白内障的发生发展。本研究采用化学方法对胶原基材料进行牛磺酸负载,获得了一种具有良好理化性能和生物相容性的功能化胶原基支架材料。本专利获得的这种牛磺酸改性胶原基材料具有广阔的市场前景。
发明内容
本发明的目的在于克服现有技术的缺点,提供一种负载牛磺酸分子的胶原基支架材料及其制备方法。
本发明的目的通过以下技术方案实现:
一种负载牛磺酸分子的胶原基支架材料的制备方法,包括以下步骤:
(1)在37℃下,用酸溶液配制一定浓度的胶原溶液,将溶于酸的胶原溶液浇铸到模具中;
(2)将步骤(1)浇铸到模具中的胶原溶液在-20℃下预冻2小时,然后在-80℃条件下预冻24小时以上,然后真空冷冻干燥48小时以上,获得结构均匀的胶原海绵支架;
(3)分别配制一定浓度的牛磺酸溶液、碳化二亚胺和N-羟基丁二酰溶液;
(4)将步骤(2)得到的胶原海绵支架浸泡到步骤(3)得到的牛磺酸溶液中,同时加入碳化二亚胺和N-羟基丁二酰溶液进行交联反应,得到交联水凝胶;
(5)将步骤(4)得到的交联水凝胶冷冻干燥后,即得到一种负载牛磺酸分子的胶原基支架材料。
进一步地,步骤(1)和(2)中所述的胶原原料来源于牛筋,用0.01~0.1mol/L的乙酸或盐酸溶液配制胶原溶液。
进一步地,步骤(1)和(2)中所述胶原溶液的浓度为4.5~9.5mg/mL。
进一步地,步骤(2)中,真空冷冻干燥时真空度低于10Pa。
进一步地,步骤(3)中所述的牛磺酸溶液中,其质量分数为2.5~15.0%。
进一步地,步骤(4)中牛磺酸占胶原的质量比为0.5~3.5%,碳化二亚胺与胶原和牛磺酸的总质量比为1:(5.5~6.5),碳化二亚胺与N-羟基丁二酰的质量比为(1~1.2):(0.8~1.2)。
本发明在传统的胶原基角膜修复支架材料基础上,引入了能促进视网膜正常功能、并能有效提高视神经传导和视觉机能的牛磺酸分子。同时,牛磺酸还具有调节晶体渗透压和抗氧化等重要作用,可抑制白内障的发生。在这种负载牛磺酸分子的胶原基支架材料缓慢降解过程中,材料内物理吸附和化学接枝的牛磺酸分子可以先后缓释到眼部。本材料能够在实现角膜修复的同时促进眼部组织功能恢复。
具体实施方式
为了更好的理解本发明,下面结合实施例对本发明做进一步地说明,但本发明要求保护的范围并不局限于提到的实施例表示的范围。
实施例1
以胶原和牛磺酸作为原料,制备一种负载牛磺酸分子的胶原基支架材料。这种改性胶原基支架材料的制备步骤如下:
(1)在37℃下,配制6.5mg/mL的胶原溶液,将溶于醋酸的胶原溶液浇铸到模具中;
(2)将步骤(1)所浇铸的胶原溶液在-20℃下预冻2小时,然后在-80℃条件下预冻24小时以上,然后真空冷冻干燥48小时以上(真空度为8Pa),获得结构均匀的胶原海绵支架;
(3)分别配制质量分数为7.5%的牛磺酸溶液;配制浓度为15mg/mL的碳化二亚胺和N-羟基丁二酰溶液;
(4)将步骤(2)得到的胶原海绵支架浸泡到(3)得到的牛磺酸溶液中,牛磺酸占胶原的质量比为2.5%,同时加入碳化二亚胺(EDC)和N-羟基丁二酰(NHS)溶液进行交联反应;其中EDC:NHS=1:1,EDC:胶原和牛磺酸质量=1:6。
(5)将步骤(4)得到的交联水凝胶冷冻干燥后,即得到一种牛磺酸改性胶原基支架材料。
实施例2
以胶原和牛磺酸作为原料,制备一种负载牛磺酸分子的胶原基支架材料。这种改性胶原基支架材料的制备步骤如下:
(1)在37℃下,配制7.5mg/mL的胶原溶液,将溶于醋酸的胶原溶液浇铸到模具中;
(2)将步骤(1)所浇铸的胶原溶液在-20℃下预冻2小时,然后在-80℃条件下预冻24小时以上,然后真空冷冻干燥48小时以上(真空度为5Pa),获得结构均匀的胶原海绵支架;
(3)分别配制质量分数为8.5%的牛磺酸溶液;配制浓度为15mg/mL的碳化二亚胺和N-羟基丁二酰溶液;
(4)将步骤(2)得到的胶原海绵支架浸泡到(3)得到的牛磺酸溶液中,牛磺酸占胶原的质量比为3.0%,同时加入碳化二亚胺(EDC)和N-羟基丁二酰(NHS)溶液进行交联反应;其中EDC:NHS=1:1,EDC:胶原和牛磺酸质量=1:6。
(5)将步骤(4)得到的交联水凝胶冷冻干燥后,即得到一种负载牛磺酸分子的胶原基支架材料。
Claims (7)
1.一种负载牛磺酸分子的胶原基支架材料的制备方法,其特征在于,包括以下步骤:
(1)在37℃下,用酸溶液配制一定浓度的胶原溶液,将溶于酸的胶原溶液浇铸到模具中;
(2)将步骤(1)浇铸到模具中的胶原溶液在-20℃下预冻2小时,然后在-80℃条件下预冻24小时以上,然后真空冷冻干燥48小时以上,获得结构均匀的胶原海绵支架;
(3)分别配制一定浓度的牛磺酸溶液、碳化二亚胺和N-羟基丁二酰溶液;
(4)将步骤(2)得到的胶原海绵支架浸泡到步骤(3)得到的牛磺酸溶液中,同时加入碳化二亚胺和N-羟基丁二酰溶液进行交联反应,得到交联水凝胶;
(5)将步骤(4)得到的交联水凝胶冷冻干燥后,即得到一种负载牛磺酸分子的胶原基支架材料。
2.根据权利要求1所述的制备方法,其特征在于,步骤(1)和(2)中所述的胶原原料来源于牛筋,用0.01~0.1mol/L的乙酸或盐酸溶液配制胶原溶液。
3.根据权利要求1所述的制备方法,其特征在于,步骤(1)和(2)中所述胶原溶液的浓度为4.5~9.5mg/mL。
4.根据权利要求1所述的制备方法,其特征在于,步骤(2)中,真空冷冻干燥时真空度低于10Pa。
5.根据权利要求1所述的制备方法,其特征在于,步骤(3)中所述的牛磺酸溶液中,其质量分数为2.5~15.0%。
6.按照权利要求1所述的制备方法,其特征在于,步骤(4)中牛磺酸占胶原的质量比为0.5~3.5%,碳化二亚胺与胶原和牛磺酸的总质量比为1:(5.5~6.5),碳化二亚胺与N-羟基丁二酰的质量比为(1~1.2):(0.8~1.2)。
7.权利要求1~6任一一项方法制备的一种负载牛磺酸分子的胶原基支架材料。
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