CN104998294B - 一种杀菌止血的医用敷料的制备方法 - Google Patents

一种杀菌止血的医用敷料的制备方法 Download PDF

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CN104998294B
CN104998294B CN201510468248.5A CN201510468248A CN104998294B CN 104998294 B CN104998294 B CN 104998294B CN 201510468248 A CN201510468248 A CN 201510468248A CN 104998294 B CN104998294 B CN 104998294B
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陈启旺
潘俊
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Abstract

本发明涉及一种杀菌止血的医用敷料的制备方法,采用分阶段的真空冷冻干燥技术处理羊膜并浸渍中药提取物,制备出一种有效的杀菌止血的医用敷料,可以广泛应用于各种不同程度的创面的治疗,与现有技术相比,本发明制备得到的医用敷料止血止痛、促进愈合,并且对人体无副作用,使用安全。本发明的制备方法简单易行,成本低,无污染。

Description

一种杀菌止血的医用敷料的制备方法
技术领域
本发明属于医用材料领域,具体涉及一种杀菌止血的医用敷料的制备方法。
背景技术
在人们日常生活中,烧伤、烫伤时常发生,造成各种深度不同的创面,并极易造成感染,导致各种并发症,因此用于该领域的有效的医用敷料的研制是急需解决的问题。羊膜由于自身独有的特性受到国内外学者关注,羊膜从细胞滋养层衍化而来,是胚胎双层膜的内层,由上皮细胞层,基底膜和无血管基质组成,羊膜中包含多种胶原蛋白以及层粘连蛋白、纤连蛋白、糖胺聚糖、透明质酸等。正是这些活性物质的存在,可以促进上皮细胞的黏附移行,诱导上皮分化,防止上皮凋亡,使得羊膜充当一种“可移植的基底膜”来促进上皮化。此外,羊膜基质中还包含有很多生长因子,比如神经生长因子(NGF),肝细胞生长因子(HGF),角质细胞生长因子(KGF),转化生长因子-α(TGF-α),转化生长因子-β1(TGF-β1),表皮生长因子(EGF),碱性成纤维细胞生长因子(bFGF)等等,能够促进眼表上皮化、减轻炎性反应、抑制纤维组织增生和新生血管形成。因此,近年来羊膜在眼表重建中的应用备受瞩目。
发明内容
本发明的目的在于利用羊膜与特定中草药结合共同制备出一种有效的杀菌止血的医用敷料,以解决现有技术中存在的上述问题。
本发明采用如下技术方案:一种杀菌止血的医用敷料的制备方法,包括以下步骤:
(1)消毒杀菌:用生理盐水清洗羊膜后,浸泡于过氧化氢水溶液中2-3h,然后用水清洗干净;将消毒后的羊膜浸泡于含有抗生素的水溶液2~3h;
(2)分阶段真空冷冻干燥:将步骤(1)得到的羊膜置于5~10g/L的海藻酸钠和1~5g/L透明质酸溶液中静置10~15h后,进行真空冷冻干燥,先在2~5℃下预冻2~3h,然后降温到-10~-15℃,冷冻3~5h,在继续降温到-20~-25℃,冷冻3~5h,在继续降温到-40~-50℃,冷冻7~10h,控制羊膜的水分含量在5~10%;
(3)浸渍中药:将步骤(2)得到的羊膜浸渍在中药提取物中,加热使浸泡温度在30~50℃,浸泡时间为10~15h;其中所述的中药提取物通过以下方法制备得到:按以下质量份取中药药材,大青叶8-10份、蛇床子3-5份、三七3-5份、黄芩1-5份、蒲公英1-3份、野菊花2-3份、金银花1-3份、骨丹3-5份、七里单6-8份、乳香2-3份、没药5-10份、丁香2-5份、红花1-3份、大黄3-5份、甘草3-5份、苍术1-2份,加水煎煮1-2小时,得到的煎煮液经超滤和纳滤后,得到中药精制液,然后加入医用酒精和水混合,调节溶液中酒精浓度为65-70%,并加热至60-70℃得到所述的中药提取物;
(4)真空干燥:将步骤(3)处理后的羊膜进行真空干燥处理,温度为60~70℃,干燥时间2-4小时;
(5)封装灭菌:经过真空干燥后的羊膜进行包装、灭菌即可在常温下保存。
在本发明的优选的实施方案中,所述过氧化氢水溶液的浓度为2~4g/L。
在本发明的优选的实施方案中,所述的抗生素选自青霉素、链霉素、庆大霉素中的一种或是两种,其用量分别为50μg/mL、30μg/mL、10μg/mL;
在本发明的优选的实施方案中,所述步骤(4)的真空度为0.01-0.05MPa。
本发明制备的医用敷料可以广泛应用于各种不同程度的创面的治疗,与现有技术相比,本发明具备以下有益效果:
(1)经过长期试验研究,发现采用分阶段的真空冷冻干燥技术,将羊膜在逐级降低的温度下冷冻干燥,阶梯式降温,能够最大程度的保留了其胶原和活性因子成分,使得羊膜的组成没有受到破坏,经检测,所制得的医用敷料含有Ⅰ型胶原290.1±1.0μg/mg、Ⅲ型胶原356.8±2.0μg/mg,Ⅳ型胶原47.1±1.0μg/mg,肝细胞生长因子71.2±1.0μg/mg,碱性成纤维细胞生长因子56.1±1.0μg/mg;而新鲜羊膜中含Ⅰ型胶原303.5±5.3μg/mg,Ⅲ型胶原384.2±4.5μg/mg,Ⅳ型胶原53.6±1.7μg/mg,肝细胞生长因子89.9±3.2μg/mg,碱性成纤维细胞生长因子71.1±1.0μg/mg。
(2)通过浸渍本发明的特定中药配方,使得得到的羊膜敷料能够更有效的止血止痛、促进愈合,并且对人体无副作用,使用安全。
(3)本发明的制备方法简单易行,成本低,无污染。
具体实施方式
以下结合实例对本发明技术方案及其效果作进一步的说明。
实施例1
(1)消毒杀菌:用生理盐水清洗羊膜后,浸泡于2g/L过氧化氢水溶液中2h,然后用水清洗干净;将消毒后的羊膜浸泡于含有50μg/mL青霉素的水溶液3h;
(2)分阶段真空冷冻干燥:将步骤(1)得到的羊膜置于8g/L的海藻酸钠和5g/L透明质酸溶液中静置10h后,进行真空冷冻干燥,先在3℃下预冻3h,然后降温到-10℃,冷冻5h,在继续降温到-20℃,冷冻5h,在继续降温到-50℃,冷冻8h,控制羊膜的水分含量在7%;
(3)浸渍中药:将步骤(2)得到的羊膜浸渍在中药提取物中,加热使浸泡温度在30℃,浸泡时间为15h;其中所述的中药提取物通过以下方法制备得到:按以下质量份取中药药材,大青叶10份、蛇床子3份、三七4份、黄芩2份、蒲公英2份、野菊花2份、金银花2份、骨丹3份、七里单6份、乳香2份、没药6份、丁香3份、红花2份、大黄3份、甘草3份、苍术1份,加水煎煮2小时,得到的煎煮液经超滤和纳滤后,得到中药精制液,然后加入医用酒精和水混合,调节溶液中酒精浓度为70%,并加热至60℃得到所述的中药提取物;
(4)真空干燥:将步骤(3)处理后的羊膜进行真空干燥处理,真空度为0.02MPa,温度为70℃,干燥时间2小时;
(5)封装灭菌:经过真空干燥后的羊膜进行包装、灭菌即可在常温下保存。
经检测,所制得的医用敷料含有Ⅰ型胶原290.1μg/mg、Ⅲ型胶原356.8μg/mg,Ⅳ型胶原47.1μg/mg,肝细胞生长因子71.2μg/mg,碱性成纤维细胞生长因子56.1μg/mg,与新鲜羊膜的成分较为接近,说明上述制备方法基本没有对羊膜的成分造成破坏,极大程度的保持了其原貌。
将制得的医用敷料用于创面,测试其对创面愈合的影响(率,时间)。选用动物的深II度皮肤烫伤模型作为评价敷料对创面愈合影响的动物模型。
将雄性Wistar大鼠随机分组,包括自然愈合组、市售常规医用敷料组、本发明医用敷料组,各组观察时相点为伤后3、7、14、21d,每组每时相点10只动物。将大鼠麻醉成功后用80℃水浴烫15秒造成大鼠背部10%深II度皮肤烫伤(病检切片证实),伤后经腹腔给予5ml生理盐水。除自然愈合组外,各组分别用相应敷料覆盖创面。伤后各确定时相点测创面愈合率,先将创面描记在半透明纸上,再以此为模板,将质地均匀的硬纸片剪成同样大小,然后用天平称重,以硬纸片重量间接地表示创面面积大小。按下式计算创面愈合率:创面愈合率(%)=(原始创面面积一未愈合创面面积)/原始创面面积。病理检查,了解创面愈合质量,并根据创面愈合情况确定愈合时间。试验结果显示,本发明医用敷料组,在创面恢复时间上和创面愈合率两方面都优于自然愈合组、市售常规医用敷料组。
本发明并非限定于所述特定的实施例及说明,并且不脱离在权利要求范围中所请求的本发明的要点,本发明所属的技术领域具有通常知识的人员均可进行各种变形,并且所述变形所属于本发明的保护范围。

Claims (1)

1.一种杀菌止血的医用敷料的制备方法,其特征在于,包括以下步骤:
(1)消毒杀菌:用生理盐水清洗羊膜后,浸泡于过氧化氢水溶液中2-3h,然后用水清洗干净;将消毒后的羊膜浸泡于含有抗生素的水溶液2~3h;
(2)分阶段真空冷冻干燥:将步骤(1)得到的羊膜置于5~10g/L的海藻酸钠和1~5g/L透明质酸溶液中静置10~15h后,进行真空冷冻干燥,先在2~5℃下预冻2~3h,然后降温到-10~-15℃,冷冻3~5h,在继续降温到-20~-25℃,冷冻3~5h,在继续降温到-40~-50℃,冷冻7~10h,控制羊膜的水分含量在5~10%;
(3)浸渍中药:将步骤(2)得到的羊膜浸渍在中药提取物中,加热使浸泡温度在30~50℃,浸泡时间为10~15h;其中所述的中药提取物通过以下方法制备得到:按以下质量份取中药药材,大青叶8-10份、蛇床子3-5份、三七3-5份、黄芩1-5份、蒲公英1-3份、野菊花2-3份、金银花1-3份、骨丹3-5份、七里单6-8份、乳香2-3份、没药5-10份、丁香2-5份、红花1-3份、大黄3-5份、甘草3-5份、苍术1-2份,加水煎煮1-2小时,得到的煎煮液经超滤和纳滤后,得到中药精制液,然后加入医用酒精和水混合,调节溶液中酒精浓度为65-70%,并加热至60-70℃得到所述的中药提取物;
(4)真空干燥:将步骤(3)处理后的羊膜进行真空干燥处理,温度为60~70℃,干燥时间2-4小时;
(5)封装灭菌:经过真空干燥后的羊膜进行包装、灭菌即可在常温下保存。
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