CN112618777A - 海藻酸生物膜及其制备方法 - Google Patents
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Abstract
本发明属于生物医疗材料技术领域,尤其涉及海藻酸生物膜及其制备方法。本发明提供的海藻酸生物膜的制备方法,证实了用注浆法制备海藻酸生物新型膜的可行性,并弥补了海藻酸单独作为制膜原料的缺陷,并在制膜过程中添加了起到纳米过滤器作用的无机填料,可控制生物膜的膨胀率,改变并控制了药物释放、细胞附着和表面增殖的速度,多西环素抗生素也被装载在其中膜作为模型药物,以及用于预防在植入过程中可能发生的潜在细菌感染。本发明提供的海藻酸生物膜具有较强的溶胀性、较高的细胞相容性和细胞活性,并且有优异的抗菌性,适合作为细胞附着和生长的基质,从而在软组织再生中具有广阔的应用前景。
Description
技术领域
本发明属于生物医疗材料技术领域,尤其涉及海藻酸生物膜及其制备方法。
背景技术
世界各地有数百万人遭受严重、轻微和长期无法治愈的创伤。正常生活方式导致的各种伤口,如皮肤感染、烧伤和由糖尿病引起的溃疡,是造成慢性和不可治愈伤口的最常见原因。大多数皮肤损伤对传统伤口愈合疗法的反应极小,这就产生了全球对开发替代和先进的伤口愈合疗法/疗法的需求,以帮助慢性和无反应的伤口更快愈合和最小疤痕形成。目前,主要采用多种化学或物理交联方法制备了海藻酸盐水凝胶,用于伤口愈合、生物活性物质的传递以及损伤组织的再生。海藻酸盐的主要缺点是普遍缺乏较强的机械性能,细胞粘附性差,在哺乳动物中缺乏降解性。然而,通过将海藻酸盐与其他生物高聚物(如琼脂糖和壳聚糖)结合,或者将藻酸盐与高硫酸氢钠等分子直接氧化,这样提高了藻酸盐的机械性能和降解性,为基于藻酸盐的生物材料的研究带来了巨大的希望。最近报道了海藻酸-生物聚合物混合物薄膜的双重释放,可作为潜在的损伤护理材料或某些药品的无菌包装。现有的海藻酸-生物聚合物膜仍然存在溶胀性较弱,与细胞相容性较低,在植入过程中具有细菌感染的风险。
发明内容
针对上述现有技术的不足,本发明提供了海藻酸生物膜的制备方法,目的是为了解决现有的海藻酸-生物聚合物膜仍然存在溶胀性较弱,与细胞相容性较低,在植入过程中具有细菌感染的风险的技术问题。
本发明提供了海藻酸生物膜的制备方法,具体技术方案如下:
海藻酸生物膜的制备方法,包括如下步骤:
S1,将甲基纤维素溶于蒸馏水后,转入冰浴中继续搅拌,加热和冰浴反复多次,直到得到清晰的溶液,即为甲基纤维素溶液;
S2,将海藻酸溶于蒸馏水中,制备海藻酸溶液,将所述海藻酸溶液与步骤S1中的甲基纤维素溶液搅拌混合,获得混合液;
S3,将无机填料原位浸渍到步骤S2中的混合液中,制备聚合药物;
S4,将步骤S3中的聚合药物按照1:1-1:2的比例原位加载多西环素抗生素后,浇注在矩形盘子中过夜,形成共混膜,使溶剂完全蒸发,获得干燥膜;
S5,将步骤S4中的干燥膜在氯化钙溶液中浸泡交联后,洗涤消除氯化钙残留,并进行干燥处理,获得海藻酸生物膜。
在某些实施方式中,所述甲基纤维素溶液(MC)的浓度是1-10W/V%。MC在无水乙醇、乙醚、丙酮中几乎不溶。在70~90>℃的热水中迅速分散、溶胀,降温后迅速溶解,水溶液在常温下相当稳定,高温时能凝胶,并且此凝胶能随温度的高低与溶液互相转变。具有优良的润湿性、分散性、粘接性、增稠性、乳化性、保水性和成膜性,以及对油脂的不透性。所成膜具有优良的韧性、柔曲性和透明度。
在某些实施方式中,在步骤S2中,所述海藻酸(Alg)溶液的浓度为1-10W/V%;搅拌混合的时间为30-60min。海藻酸具有微溶于热水的特性,所以所需蒸馏水温度达到70℃即可,从中海藻中提取的海藻酸钠具有增稠、悬浮、乳化、稳定、形成凝胶、形成薄膜和纺制纤维等特性。非常适合用于医用敷料。
在某些实施方式中,在步骤S1和S2中,所述蒸馏水的温度为70-90℃。
在某些实施方式中,在步骤S3中,所述无机填料包括SrTiO3、5mol%铋掺杂SrTiO3以及2.5mol%铋和2.5mol%铜共掺杂SrTiO3。钛酸锶(SrTiO3)具有高介电常数,良好的绝缘性质,优良的物理化学稳定性和在可见光范围内的优异的透明度等优点,是一种重要的无机功能材料。
在某些实施方式中,在步骤S3中,所述原位浸渍的温度为20-30℃,时间为5-10天。
在某些实施方式中,在步骤S5中,所述氯化钙的浓度为5-10W/V%;所述浸泡的时间为1小时;所述干燥的温度为40℃,所述干燥的时间为12-24小时。氯化钙溶液具有导电、冰点比水低等特点,具有遇水散热的特征,并且有较好的吸附功能。
本发明还提供了根据上述方法制备的海藻酸生物膜。
本发明具有以下有益效果:本发明提供的海藻酸生物膜的制备方法,证实了用注浆法制备海藻酸生物新型膜的可行性,并弥补了海藻酸单独作为制膜原料的缺陷,并在制膜过程中添加了起到纳米过滤器作用的无机填料,可控制生物膜的膨胀率,改变并控制了药物释放、细胞附着和表面增殖的速度,多西环素抗生素也被装载在其中膜作为模型药物,以及用于预防在植入过程中可能发生的潜在细菌感染。本发明提供的海藻酸生物膜具有较强的溶胀性、较高的细胞相容性和细胞活性,并且有优异的抗菌性,适合作为细胞附着和生长的基质,从而在软组织再生中具有广阔的应用前景。
附图说明
图1是本发明提供的海藻酸生物膜的制备方法的流程图。
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白,以下结合具体实施例,并参照附图1,对本发明进一步详细说明。
实施例1
本实施例提供的海藻酸生物膜的制备方法,具体技术方案如下:
海藻酸生物膜的制备方法,包括如下步骤:
S1,将甲基纤维素溶于90℃的蒸馏水后,转入冰浴中继续搅拌,加热和冰浴反复多次,直到得到清晰的溶液,即为甲基纤维素溶液,甲基纤维素溶液的浓度是5W/V%。
S2,将海藻酸溶于90℃的蒸馏水中,制备浓度为5W/V%的海藻酸溶液,将所述海藻酸溶液与步骤S1中的甲基纤维素溶液搅拌混合45min,获得混合液。
S3,将无机填料SrTiO3原位浸渍(温度为20℃,时间为10天)到步骤S2中的混合液中,制备聚合药物。
S4,将步骤S3中的聚合药物按照1:2的比例原位加载多西环素抗生素后,浇注在矩形盘子中过夜,形成共混膜,使溶剂完全蒸发,获得干燥膜;
S5,将步骤S4中的干燥膜在浓度为8W/V%的氯化钙溶液中浸泡1小时,交联后,洗涤消除氯化钙残留,并在40℃下干燥处理24小时,获得海藻酸生物膜。
本实施例还提供了根据上述方法制备的海藻酸生物膜。
实施例2
海藻酸生物膜的制备方法,包括如下步骤:
S1,将甲基纤维素溶于70℃的蒸馏水后,转入冰浴中继续搅拌,加热和冰浴反复4次,直到得到清晰的溶液,即为甲基纤维素溶液,甲基纤维素溶液的浓度是10W/V%。
S2,将海藻酸溶于70℃的蒸馏水中,制备浓度为10W/V%的海藻酸溶液,将所述海藻酸溶液与步骤S1中的甲基纤维素溶液搅拌混合60min,获得混合液。
S3,将无机填料SrTiO3原位浸渍(温度为30℃,时间为5天)到步骤S2中的混合液中,制备聚合药物。
S4,将步骤S3中的聚合药物按照1:1的比例原位加载多西环素抗生素后,浇注在矩形盘子中过夜,形成共混膜,使溶剂完全蒸发,获得干燥膜;
S5,将步骤S4中的干燥膜在浓度为10W/V%的氯化钙溶液中浸泡1小时,交联后,洗涤消除氯化钙残留,并在40℃下干燥处理18小时,获得海藻酸生物膜。
本实施例还提供了根据上述方法制备的海藻酸生物膜。
实施例3
海藻酸生物膜的制备方法,包括如下步骤:
S1,40ml蒸馏水(80℃)中配制MC溶液(1%W/V)30min,转入冰浴中继续搅拌,加热和冰浴反复3次,直到得到清晰的溶液,即为甲基纤维素溶液。
S2,将海藻酸溶于70-90℃的蒸馏水中,制备浓度为1W/V%的海藻酸溶液,将所述海藻酸溶液与步骤S1中的甲基纤维素溶液在25℃磁搅拌器中混合30min,获得混合液。
S3,将无机填料SrTiO3原位浸渍(温度为25℃,时间为7天)到步骤S2中的混合液中,制备聚合药物。
S4,将步骤S3中的聚合药物按照1:1.3的比例原位加载多西环素抗生素后,浇注在矩形盘子中过夜,形成共混膜,使溶剂完全蒸发,获得干燥膜;
S5,将步骤S4中的干燥膜在浓度为5W/V%的氯化钙溶液中浸泡1小时,交联后,洗涤消除氯化钙残留,并在40℃下干燥处理12小时,获得海藻酸生物膜。
本实施例还提供了根据上述方法制备的海藻酸生物膜。
先进的多功能膜在大分子传递方面的应用早有报道。特别是乙基纤维素作为纤维素的一种衍生物,通过改进三轴静电纺丝技术,以一种新型的核-壳纳米纤维的形式来传递酮洛芬。这些乙基纤维素纳米纤维载体展示了一种理想的持续药物输送,这反映了纤维素衍生物在药物输送系统中令人印象深刻的特性。由于甲基纤维素的有益特性,包括生态友好性(因为它是一种天然聚合物)、较高的溶胀能力和良好的细胞亲和力,以及在非常低的浓度下具有较高的粘度,在材料学领域应用较多。此外,甲基纤维素在保留种植体形状方面也是有效的,这在创面的长期治疗中非常重要。因此,海藻酸钠和甲基纤维素的结合将克服海藻酸盐的局限性,提供一个可持续的伤口恢复。
此外,有报道称,不同浓度的Sr和/或Ca的掺入促进了含磷酸盐玻璃的骨诱导性能。此外,Ti的掺入延长了玻璃的生物降解时间。含17.5mol%Sr的玻璃的碱性磷酸酶活性比对照高3.6倍。一些膜在组织再生中的成功应用的潜在适用性与包括Ca和Sr离子在内的几种离子的释放密切相关。同时,研究表明Cu2+在组织血管化、刺激人内皮细胞增殖和间充质干细胞(MSC)分化方面具有令人印象深刻的作用。因此,Cu2+在生物活性药物中的加入会增加(HIF)-1(缺氧诱导因子)和VEGF(血管内皮生长因子)的表达。用BMSC(骨髓间充质干细胞)和HDMEC(人真皮微血管内皮细胞)培养试验了掺铜的衍生支架,观察到VEGF表达增加,这种支架对骨髓间充质干细胞无毒性。利用铜来重新放置受损组织的吸引力不仅在于它是几种催化剂的基本辅因子,而在于它的促血管生成作用,还在于铜具有普遍的抗菌效果。特别是在植入治疗过程中,感染污染的危险给患者带来了极大的不便,这类疾病很难用抗感染治疗。
综上所述,本发明提供的海藻酸生物膜的制备方法,证实了用注浆法制备海藻酸生物新型膜的可行性,并弥补了海藻酸单独作为制膜原料的缺陷,并在制膜过程中添加了起到纳米过滤器作用的无机填料,可控制生物膜的膨胀率,改变并控制了药物释放、细胞附着和表面增殖的速度,多西环素抗生素也被装载在其中膜作为模型药物,以及用于预防在植入过程中可能发生的潜在细菌感染。本发明提供的海藻酸生物膜具有较强的溶胀性、较高的细胞相容性和细胞活性,并且有优异的抗菌性,适合作为细胞附着和生长的基质,从而在软组织再生中具有广阔的应用前景。
上述仅本发明较佳可行实施例,并非是对本发明的限制,本发明也并不限于上述举例,本技术领域的技术人员,在本发明的实质范围内,所作出的变化、改型、添加或替换,也应属于本发明的保护范围。
Claims (8)
1.海藻酸生物膜的制备方法,其特征在于,包括如下步骤:
S1,将甲基纤维素溶于蒸馏水后,转入冰浴中继续搅拌,加热和冰浴反复多次,直到得到清晰的溶液,即为甲基纤维素溶液;
S2,将海藻酸溶于蒸馏水中,制备海藻酸溶液,将所述海藻酸溶液与步骤S1中的甲基纤维素溶液搅拌混合,获得混合液;
S3,将无机填料原位浸渍到步骤S2中的混合液中,制备聚合药物;
S4,将步骤S3中的聚合药物按照1:1-1:2的比例原位加载多西环素抗生素后,浇注在矩形盘子中过夜,形成共混膜,使溶剂完全蒸发,获得干燥膜;
S5,将步骤S4中的干燥膜在氯化钙溶液中浸泡交联后,洗涤消除氯化钙残留,并进行干燥处理,获得海藻酸生物膜。
2.根据权利要求1所述的海藻酸生物膜的制备方法,其特征在于,在步骤S1中,所述甲基纤维素溶液的浓度是1-10W/V%。
3.根据权利要求1所述的海藻酸生物膜的制备方法,其特征在于,在步骤S2中,所述海藻酸溶液的浓度为1-10W/V%;搅拌混合的时间为30-60min。
4.根据权利要求1所述的海藻酸生物膜的制备方法,其特征在于,在步骤S1和S2中,所述蒸馏水的温度为70-90℃。
5.根据权利要求1所述的海藻酸生物膜的制备方法,其特征在于,在步骤S3中,所述无机填料包括SrTiO3、5mol%铋掺杂SrTiO3以及2.5mol%铋和2.5mol%铜共掺杂SrTiO3。
6.根据权利要求1所述的海藻酸生物膜的制备方法,其特征在于,在步骤S3中,所述原位浸渍的温度为20-30℃,时间为5-10天。
7.根据权利要求1所述的海藻酸生物膜的制备方法,其特征在于,在步骤S5中,所述氯化钙的浓度为5-10W/V%;所述浸泡的时间为1小时;所述干燥的温度为40℃,所述干燥的时间为12-24小时。
8.海藻酸生物膜,其特征在于,根据权利要求1-7任一项所述的方法制备的海藻酸生物膜。
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