CN107184961A - 一种活性细胞修复剂及其制备方法 - Google Patents
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- CN107184961A CN107184961A CN201710392295.5A CN201710392295A CN107184961A CN 107184961 A CN107184961 A CN 107184961A CN 201710392295 A CN201710392295 A CN 201710392295A CN 107184961 A CN107184961 A CN 107184961A
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Abstract
本发明属于生物工程技术领域,具体涉及一种活性细胞修复剂及其制备方法。本发明活性细胞修复剂包含以下组分及其质量百分比:芦荟多糖8‑13%、壳寡糖6‑8%、壳聚糖4‑6%、透明质酸4‑5%、维生素B5 3‑5%、胶原蛋白2.5‑5%、谷胱甘肽2‑3%、苹果多酚1‑3%、十八醇4‑8%、白凡士林2‑5%、单硬脂酸甘油酯3‑5%、甘油6‑10%、苯氧乙醇0.2‑0.7%和水23.3‑54.3%。本发明的技术优势在于制备得到的活性细胞修复剂在皮肤外伤伤口愈合、减少瘢痕形成、术后粘连等皮肤修复方面有很好的效果,并且无毒副作用,有较大的临床应用价值。
Description
技术领域
本发明属于生物工程技术领域,具体涉及一种活性细胞修复剂及其制备方法。
背景技术
皮肤是人体的重要器官,起着控制体温、防止感染及体液流失、免疫及传感的作用。由于创伤、擦伤、皮肤溃烂和烧伤等原因,可能导致皮肤的大范围伤害。皮肤的损伤很容易造成细菌感染和体液流失,并引起各种临床并发症。现有技术中对于医治创伤所采用的敷料按照材质和用途可划为传统敷料、动物类敷料、合成敷料、药物敷料和固定用敷料,现有技术中的敷料存在以下缺点:不能很好的吸收创面渗液,维护创面温度,为创面愈合提供良好的局部环境;不能维护伤口适量的血液流通和养分,一致血管内皮肿胀而阻碍了血管生长。
中国专利CN100502953C公开了一种创面活性修复材料,主要由以下成分组成:透明质酸、二氧化硅、氧化钠、氧化钙和五氧化二磷,所述创面修复材料可以刺激表皮细胞和形成纤维细胞的移行、降低新生修复血管的循环阻力,阻碍粒细胞的沉积、抑制血管内皮肿胀,减少细胞碎片在血管内的堆积,该专利采用物理混合技术制备的多物质混合粉料,其晶体结构依旧是独立单晶混合料,但在创面渗液吸收、促血管生长、改善微循环、促进创面愈合、减少瘢痕形成等方面效果有待改进。
中国专利申请CN101879307A公开了一种活性细胞修复制剂,所述制剂由壳聚糖、壳寡糖、胶原蛋白为活性成分与药用辅料组成,该发明虽然在术后/外伤伤口愈合、减少瘢痕形成有很好的的效果,但其药用辅料涉及亚硫酸钠,而亚硫酸钠对眼睛、皮肤、粘膜有刺激作用,因而人体存在潜在的威胁性。
发明内容
为解决现有技术中活性细胞修复剂修复效果不佳以及某些产品含有对人体刺激性组分等问题,本发明提供一种活性细胞修复剂及其制备方法。
本发明一种活性细胞修复剂,包含以下组分及其质量百分比:
芦荟多糖8-13%、壳寡糖6-8%、壳聚糖4-6%、透明质酸4-5%、维生素B53-5%、胶原蛋白2.5-5%、谷胱甘肽2-3%、苹果多酚1-3%、十八醇4-8%、白凡士林2-5%、单硬脂酸甘油酯3-5%、甘油6-10%、苯氧乙醇0.2-0.7%和水23.3-54.3%。
优选的所述的活性细胞修复剂,由以下组分及其质量百分比组成:
芦荟多糖10.5%、壳寡糖7%、壳聚糖5%、透明质酸4.5%、维生素B54%、胶原蛋白3.75%、谷胱甘肽2.5%、苹果多酚2%、十八醇6%、白凡士林3.5%、单硬脂酸甘油酯4%、甘油8%、苯氧乙醇0.45%和水38.8%。
再优选的,所述的芦荟多糖的提取方法包括以下步骤:
S1、取芦荟鲜叶2-5kg和水5-10L,将芦荟鲜叶在水中浸泡20-40min,然后切去表皮,将内层凝胶置于水中浸洗6-10h,1000-1200r/min转速、15-20℃温度下离心10-20min,抽滤并将所得的芦荟提取液减压浓缩即得芦荟凝胶浓缩汁,向芦荟凝胶浓缩汁中加入乙酸调节pH至5-6;
S2、取S1得到的芦荟凝胶浓缩汁20-100g和无水乙醇500-1000mL,将芦荟凝胶浓缩汁加入无水乙醇中超声萃取处理15-30min,然后于800-1000r/min、15-20℃下离心5-10min过滤即得。
另外,本发明还提供了一种活性细胞修复剂的制备方法,包括以下步骤:
S1、备料:按质量比称取相应原料;
S2、取白凡士林和十八醇置于烧杯中,水浴温度70-80℃,使其熔化得到油液;将芦荟多糖、壳寡糖、壳聚糖、透明质酸、维生素B5、苹果多酚和水置于另一烧杯中水浴温度70-80℃,使其溶解得到水液;将水液加入油液中,在混合液中加入胶原蛋白、谷胱甘肽、单硬脂酸甘油酯、甘油和苯氧乙醇,搅拌均匀;
S3、用紫外线灯进行杀菌处理,降至室温即得。
优选的,所述的活性细胞修复剂的制备方法步骤S2中制备水液和油液的水浴温度为75℃。
壳寡糖CAS号:148411-57-8;壳聚糖CAS号:9012-76-4;透明质酸CAS号:9004-61-9;维生素B5CAS号:137-08-6;胶原蛋白CAS号:9064-67-9;谷胱甘肽CAS号:70-18-8;苹果多酚CAS号:85251-63-4;十八醇CAS号:112-92-5;白凡士林CAS号:8009-03-8;单硬脂酸甘油酯CAS号:123-94-4;苯氧乙醇CAS号:122-99-6。
芦荟多糖主要存在于芦荟叶的凝胶部位,即由叶皮所包围的透明黏状部分,目前已被检出的有乙酰化葡聚糖、葡甘聚糖、阿拉伯-半乳聚糖等,作为芦荟凝胶中的主要活性成分之一,具有免疫、促进细胞分裂和伤口愈合的功能,对多种由于免疫力下降引起的疾病都有显著的疗效。
壳寡糖具有保湿、活化机体细胞、阻止皮肤粗糙和老化,抑制皮肤表面有害菌滋生、抑菌抗皮肤病和吸收紫外线功能等功效,且有促进伤口愈合的功能,可用于动物外商或骨折的辅助治疗,可以促进皮肤修复,提高机体免疫力和抑菌药理作用。
壳聚糖具有止痛、止血、促进伤口愈合、减小疤痕的作用,且有良好的生物相溶性、生物降解性以及良好的成膜性、抗凝血性等功能。
透明质酸是人皮肤表皮及真皮的主要基质成分之一,其生理功能是能使水分进入细胞间隙,并与蛋白质结合而形成蛋白凝胶,将细胞粘在一起,发挥正常的细胞代谢作用,起到保持细胞水分,保护细胞不受病原菌的侵害,加快恢复皮肤组织,提高创口愈合再生能力,减少疤痕,增强免疫力等作用。在医药方面,透明质酸用于非甾体消炎药,关节治疗、眼科,心外科手术的辅助药品,在治疗烫伤、烧伤、冻伤、人造皮肤方面有着独到作用。
维生素B5是一种优异的皮肤保护剂,能刺激细胞的分裂增殖,增进纤维芽细胞的增生,有协助修复皮肤组织的功能,具消炎及愈合表皮创伤之作用,并能减轻配方中其他组分对皮肤的刺激与损伤,有助于伤口痊愈可制造抗体抗传染病,治疗手术后休克,防止疲劳,缓解多种抗生素的毒副作用。
胶原蛋白参与细胞的迁移、分化和增殖,使骨、腱、软骨和皮肤具有一定的机械强度。胶原蛋白因具有弱的抗原性和良好的生物相溶性目前在烧伤、创伤、眼角膜疾病、美容矫形、硬组织修复、创面止血等领域得到广泛的应用。
谷胱甘肽是一种含γ-酰胺键和疏基的三肽,由谷氨酸、半胱氨酸及甘氨酸组成。谷胱甘肽存在于几乎身体的每一个细胞。能帮助保持正常的免疫系统的功能,并具有抗氧化作用和整合解毒作用。
苹果多酚是苹果中所含多元酚类物质的通称,是一类具有独特生理活性和化学活性的天然产物,它能够维护胶原的合成,具有极强的抗氧化能力,能够有效清除人体内多余的自由基,延缓衰老,并具有防止维生素损失、抗过敏、抗肿瘤、抗突变、抗辐射、防龋齿、增强肌力,改善皮肤弹性、减少皱纹产生、保持皮肤细腻的作用。
白凡士林具防水性,是一种非常好的保湿用品,用于配制医药的药膏及皮肤保护油膏用的原料,白凡士林的化学惰性使得它对任何类型的皮肤都没有刺激性;单硬脂酸甘油酯具有优良的乳化性能,且对皮肤刺激性小、生物降解性好。
本发明各组分相互配合、协同作用,对皮肤术后/外伤伤口愈合、减少瘢痕形成、术后粘连等皮肤修复方面有很好的效果。
与现有技术相比,本发明的技术优势在于:采用水提醇沉方法对芦荟叶中的芦荟多糖进行提取分离;制备得到的活性细胞修复剂安全无毒、无刺激,在皮肤外伤伤口愈合、减少瘢痕形成、术后粘连等皮肤修复方面有很好的效果,并且无毒副作用,有较大的临床应用价值。
具体实施方式
下面通过具体实施例对本发明做进一步详细说明,这些实施例仅用于例证的目的,绝不限制本发明的保护范围。
实施例1-3,对比例1-6
表1列出了本发明所提供的实施例1-3和对比例1-6的活性细胞修复剂的组成及原料配比。
表1实施例和对比例活性细胞修复剂的组成及原料配比
对比例1与实施例2的区别在于:不含维生素B5,增加壳寡糖的含量。
对比例2与实施例2的区别在于:不含苹果多酚,增加胶原蛋白的含量。
对比例3与实施例2的区别在于:不含壳聚糖,增加芦荟多糖的含量。
对比例4与实施例2的区别在于:不含透明质酸,增加胶原蛋白的含量。
对比例5与实施例2的区别在于:不含谷胱甘肽,增加苹果多酚的含量。
对比例6与实施例2的区别在于:不含壳寡糖,增加壳聚糖的含量。
实施例1活性细胞修复剂的制备方法为:
S1、备料:按质量比称取相应原料;
S2、取白凡士林和十八醇置于烧杯中,水浴温度70℃,使其熔化得到油液;将芦荟多糖、壳寡糖、壳聚糖、透明质酸、维生素B5、苹果多酚和水置于另一烧杯中水浴温度70℃,使其溶解得到水液;将水液加入油液中,在混合液中加入胶原蛋白、谷胱甘肽、单硬脂酸甘油酯、甘油和苯氧乙醇,搅拌均匀;
S3、用紫外线灯进行杀菌处理,降至室温即得。
实施例2活性细胞修复剂的制备方法为:
S1、备料:按质量比称取相应原料;
S2、取白凡士林和十八醇置于烧杯中,水浴温度75℃,使其熔化得到油液;将芦荟多糖、壳寡糖、壳聚糖、透明质酸、维生素B5、苹果多酚和水置于另一烧杯中水浴温度75℃,使其溶解得到水液;将水液加入油液中,在混合液中加入胶原蛋白、谷胱甘肽、单硬脂酸甘油酯、甘油和苯氧乙醇,搅拌均匀;
S3、用紫外线灯进行杀菌处理,降至室温即得。
实施例3活性细胞修复剂的制备方法为:
S1、备料:按质量比称取相应原料;
S2、取白凡士林和十八醇置于烧杯中,水浴温度80℃,使其熔化得到油液;将芦荟多糖、壳寡糖、壳聚糖、透明质酸、维生素B5、苹果多酚和水置于另一烧杯中水浴温度80℃,使其溶解得到水液;将水液加入油液中,在混合液中加入胶原蛋白、谷胱甘肽、单硬脂酸甘油酯、甘油和苯氧乙醇,搅拌均匀;
S3、用紫外线灯进行杀菌处理,降至室温即得。
对比例1-6的制备方法与实施例2类似。
产品性能评价测试
试验例1:急性毒理评价
试验材料:体重18-22g的昆明种小鼠200只雌雄各半、实施例1-3和对比例1-6活性细胞修复剂。
用药方法:采用雌雄分别随机均衡分组法,共分为20组,每组10只雌雄各半,其中2组为空白对照组,其他组分别对应实施例1-3和对比例1-6的细胞修复剂的进行一次性灌胃给药(按10.0g/kg的剂量给药)组和尾静脉注射试验(按2.0g/kg的剂量给药)组。
评价方法:给药连续观察7个小时以上,随后每天上、下午各观察一次,连续观察14天,详细记录各项观察指标。
急性毒理评价:未出现中毒和死亡,小鼠的外观、行为行动,精神状态正常,体重增长与对照组无差别。
受试物在本试验中灌胃给药剂量达10.0g/kg,注射给药剂量达2.0g/kg,而且没有死亡,因此本受试物可视为实际无毒。
试验例2:疗效评价
试验材料:采用新西兰大白兔20只,体重2.25-2.50kg,距脊柱两侧2.0cm处切割直径为2.0cm的对称创伤口10个(每侧5个),深度从皮肤到肌层,伤口的厚度为0.3cm,用生理盐水冲洗伤口,20只大白兔处于同一位置的伤口作为同一组,备用。
用药方法:第1组伤口不作处理,用凡士林医用纱布覆盖,处理后的伤口用无菌纱布包扎好(空白对照组);第2组伤口用实施例1配比制备的活性细胞修复剂涂抹于伤口表面(实验组1),隔日换药;第3组伤口用实施例2制备的活性细胞修复剂涂抹于伤口表面(实验组2),隔日换药;第4组伤口按本发明实施例3配比制备的活性细胞修复剂涂抹于伤口表面(实验组3),隔日换药;第5组伤口按本发明对比例1配比制备的活性细胞修复剂涂抹于伤口表面(实验组4),隔日换药;第6组伤口按本发明对比例2配比制备的活性细胞修复剂涂抹于伤口表面(实验组5),隔日换药;第7组伤口按本发明对比例3配比制备的活性细胞修复剂涂抹于伤口表面(实验组6),隔日换药;第8组伤口按本发明对比例4配比制备的活性细胞修复剂涂抹于伤口表面(实验组7);第9组伤口按本发明对比例5配比制备的活性细胞修复剂涂抹于伤口表面(实验组8);第10组伤口按本发明对比例6配比制备的活性细胞修复剂涂抹于伤口表面(实验组9)。
评价方法:分别于上药后3、5、7、10天,肉眼观察表皮、肉芽组织的生长情况,此外考察各实验组分泌物、创缘皮下有无积液、创面皮缘有无坏死。同时测量创面面积:伤后即刻及3、5、7、10和14天,用透明膜覆盖创面画膜,剪膜,称重换算成面积。数据以均数计算,疗效评价结果如表2所示。
表2实施例和对比例细胞修复剂疗效评价
从表2可以看出,各组采用不同方法处理伤口在上皮组织和伤口收缩作用上有明显的差别。伤后3天:空白组情况较差,但实验组1-3创面收缩明显,优于实验组4-9及空白对照组;伤后14天:实验组1-3创面已基本愈合、无明显疤痕,而实验组4-9虽基本愈合但是形成疤痕明显,空白组大部分伤口仍湿润,仍有渗出物,伤口收缩不明显。结果表明本发明各组分相互配合协同作用,对皮肤术后/外伤伤口愈合、减少瘢痕形成、术后粘连等皮肤修复方面有很好的效果。
以上仅是本发明的优选实施方式,应当指出的是,上述优选的实施方式不应视为对本发明的限制,本发明的保护范围应当以权利要求所限定的范围为准。对于本技术领域的普通技术人员来说,在不脱离本发明的精神和范围内,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
Claims (5)
1.一种活性细胞修复剂,其特征在于,包含以下组分及其质量百分比:
芦荟多糖8-13%、壳寡糖6-8%、壳聚糖4-6%、透明质酸4-5%、维生素B53-5%、胶原蛋白2.5-5%、谷胱甘肽2-3%、苹果多酚1-3%、十八醇4-8%、白凡士林2-5%、单硬脂酸甘油酯3-5%、甘油6-10%、苯氧乙醇0.2-0.7%和水23.3-54.3%。
2.根据权利要求1所述的活性细胞修复剂,其特征在于,由以下组分及其质量百分比组成:芦荟多糖10.5%、壳寡糖7%、壳聚糖5%、透明质酸4.5%、维生素B54%、胶原蛋白3.75%、谷胱甘肽2.5%、苹果多酚2%、十八醇6%、白凡士林3.5%、单硬脂酸甘油酯4%、甘油8%、苯氧乙醇0.45%和水38.8%。
3.根据权利要求1或2所述的活性细胞修复剂,其特征在于,所述的芦荟多糖的提取方法包括以下步骤:
S1、取芦荟鲜叶2-5kg和水5-10L,将芦荟鲜叶在水中浸泡20-40min,然后切去表皮,将内层凝胶置于水中浸洗6-10h,1000-1200r/min转速、15-20℃温度下离心10-20min,抽滤并将所得的芦荟提取液减压浓缩即得芦荟凝胶浓缩汁,向芦荟凝胶浓缩汁中加入乙酸调节pH至5-6;
S2、取S1得到的芦荟凝胶浓缩汁20-100g和无水乙醇500-1000mL,将芦荟凝胶浓缩汁加入无水乙醇中超声萃取处理15-30min,然后于800-1000r/min、15-20℃下离心5-10min过滤即得。
4.根据权利要求1-3任一项所述的活性细胞修复剂的制备方法,其特征在于,包括以下步骤:
S1、备料:按质量比称取相应原料;
S2、取白凡士林和十八醇置于烧杯中,水浴温度70-80℃,使其熔化得到油液;将芦荟多糖、壳寡糖、壳聚糖、透明质酸、维生素B5、苹果多酚和水置于另一烧杯中水浴温度70-80℃,使其溶解得到水液;将水液加入油液中,在混合液中加入胶原蛋白、谷胱甘肽、单硬脂酸甘油酯、甘油和苯氧乙醇,搅拌均匀;
S3、用紫外线灯进行杀菌处理,降至室温即得。
5.根据权利要求4所述的活性细胞修复剂的制备方法,其特征在于,步骤S2中制备水液和油液的水浴温度为75℃。
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