CN107714609A - 一种羊膜基膜、基于羊膜基膜的羊膜面膜及其制备方法 - Google Patents
一种羊膜基膜、基于羊膜基膜的羊膜面膜及其制备方法 Download PDFInfo
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- CN107714609A CN107714609A CN201711088597.XA CN201711088597A CN107714609A CN 107714609 A CN107714609 A CN 107714609A CN 201711088597 A CN201711088597 A CN 201711088597A CN 107714609 A CN107714609 A CN 107714609A
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- amnion
- basement membrane
- facial mask
- placenta
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Abstract
本发明涉及基于羊膜基膜的羊膜面膜及其制备方法,羊膜基膜为去除了纤维细胞层、海绵层的动物胎盘羊膜,羊膜基膜采用健康动物胎盘的羊膜,经分离、加工,滋养、消毒等工序制备而成,将制备好的羊膜基膜与面膜载帖物直接帖合制成羊膜面膜;本发明使用动物胎盘之羊膜制备面膜,面膜与皮肤的帖合度更好,对皮肤的剌激性更小;同时羊膜作为一种天然、有活性的生物材料,具有以下特点:首先,天然羊膜中富含营养和生物活性物质,可显著改善面膜产品的功能性成份,从而丰富面膜内容,增加面膜功效;其次,羊膜基膜可通过交联、吸附、结合等作用添加各种外源性营养与功能成份,增加了面膜的产品种类。
Description
技术领域
本发明涉及美容护肤品及美容护肤品制备方法的技术领域,尤其涉及一种羊膜基膜、基于羊膜基膜的羊膜面膜及其制备方法。
背景技术
面(帖)膜作为一种常见的美容产品,因其使用便捷、效果好而广受消费者喜爱,自2013年以来,面膜已成为大众化妆品消费的第一选择;目前市场上售卖的面膜按功效可分为营养面膜、清洁面膜、补水(保湿)面膜、美白面膜、滋养面膜、抗皱面膜、祛斑面膜、修复面膜8大类;按面膜的材质分类,有无纺布、蚕丝、生物纤维、泥膜等;国内各种面膜的品种达数百种之多。
根据AC尼尔森一份市场调查报告显示,2012年,中国面膜市场销售规模高达78.5亿元,同比增长33%,与此同时,面膜在中国现有市场渗透率为17.7%,而化妆品市场渗透率83.3%。贴式面膜(面帖膜)产品在中国,将逐步壮大成为一个独立、庞大的市场。
面膜作为快速、有效改变肌肤状况的护肤类产品,近几年的发展呈现出飞速增长趋势;其关注点是:(1)提倡发展生物面膜产品,减少或消除化学成分面膜。目前生物纤维面膜是国内仅有的生物类面膜产品,其优势之处在于:①生物面膜具有良好的亲水性、透氧性、粘附性;②与皮肤有良好的帖合度;③对皮肤无刺激、无损伤。2007年,第一个隐形面膜诞生,是将原用于医学界处理烫伤专用之仿生真皮——“隐形薄膜”演变成当今美容行业面膜的顶级材质;隐形面膜符合人体脸部工学设计,超强贴合度可以最大限度地保留并补充肌肤所需的水分和养分的面膜,隐形面膜是仿造人体皮肤表皮组织的天然材料,承托高浓度精华液,可贴于整个面部,与肌肤完全贴合,且具有良好的通透性,保证肌肤的正常呼吸,阻隔空气中的污染物。(2)开发天然、绿色无刺激性的面膜材料与面膜产品。生物纤维面膜是由木醋杆菌自然发酵制成的纤维体,具有类似皮肤的功能;该面膜质地非常柔软,弹性好,韧性强,无刺激无异味,不易生长细菌、不含荧光物质,容易清洗,是纯天然健康环保用品;由于具有不被人体排斥的特性,在医学界被用来作为心血管修复与人造皮肤使用;鉴于上述特点,生物纤维素面膜在市场广受青睐。(3)多功能、多用途性。面膜所承载的功能众多:营养、美白、清洁、补水、美白、滋养、抗皱、修复等,所以,一款产品所具备的功能越多越好,这就要求面膜产品具有多功能性和多用途性,从而满足广大消费者日益增长的要求。
羊膜是众多哺乳动物胎盘的一种重要结构,为一种透明、有韧性,无神经、血管、淋巴管分布的天然生物膜,其厚度约0.02~0.05mm。羊膜组织的抗原性极低,不引起异物反应,这是因为羊膜的上皮细胞层未表达白细胞抗原(HLA-II),导致其缺乏免疫原性。
羊膜取材方便,来源广泛,成本低,在医学临床作为一种生物学重建材料而广泛应用,羊膜具有抑制炎症、减少纤维化、降低疼痛等生物学作用,而大量的现代研究阐明,羊膜中含有数十种生物活性物质,产生广泛的生物学功能。在羊膜的基质成分中含有多种胶原(Collagen)、整合素(Integrin)、层粘连蛋白(LN)和板层体(LB)等,它们具有维持表皮完整性,防止上皮细胞凋亡,促进上皮细胞移行与分化作用。羊膜中含有多种生长因子如:表皮生长因子(EGF)、转化生长因子(TGF)、角质细胞生长因子(KGF)、肝细胞生长因子(HGF)、和碱性成纤维细胞生长因子(bFGF)等,概况起来,这些细胞因子的综合作用可归纳为3类:⑴促进上皮细胞增殖与分化,众多的细胞因子和生物酶即促进了上皮增殖,同时也改善上皮组织代谢,从而维持上皮的完整与稳定;⑵促进胶原沉积和组织修复,多向性细胞因子具有刺激各种靶细胞生长的作用,还能诱导上皮细胞形成单层集落,促进组织修复;⑶加速伤口愈合、抑制纤维化,产生抗瘢痕作用,其机理是某些细胞因子通过抑制转化生长因子β(TGF-β)的合成,从而抑制纤维母细胞的活性,减少瘢痕的形成。此外,羊膜中还含有多种酶类,已揭示的主要有三类:一类是基质金属蛋白酶(MMP)和金属蛋白酶组织抑制因子(TIMP),有报道认为这类酶在抗新生血管化和间质纤维化过程中发挥重要作用;另一类是多种蛋白酶抑制因子(如:α-1抗胰蛋白酶、α-2抗糜蛋白酶等)、抗炎因子(如:白细胞介素-1受体拮抗剂IL-1Ra)和抗血管形成因子(如:内皮抑素前体蛋白EIPP、血小板反应蛋白-1TSP-1)等,这些因子通过抑制相应的蛋白酶而发挥抗炎作用,抑制血管新生。第三类是神经营养因子(如:神经生长营养因子NGF、脑源性神经生长因子BDNF等),具有很强的神经保护作用。
羊膜是机体内最厚的膜结构,通常由5层组织结构:上皮细胞层、基底膜、致密层、纤维细胞层及海绵层。上皮细胞层含有羊膜上皮细胞和由羊膜上皮细胞分泌的各种生物活性成份,上皮细胞合成并分泌基质成份,维持膜的稳定性。基底膜以网状纤维为主,富含具硫酸肝素的蛋白多糖,是羊膜中发挥屏障作用的组织结构。致密层薄而致密,起支撑和滤过作用。
此外,天然羊膜还具有一定的抗感染能力,有人(Uberti等,2011)报道,巨噬细胞杀灭大肠杆菌的能力与羊膜源性细胞因子溶液的浓度呈正相关关系。
羊膜的种种特性表明,把动物(绵羊、山羊、鹿)羊膜用作人类面膜在技术上是可行的,并克服了使用人类羊膜而产生的伦理和道德问题:首先,动物(绵羊、山羊、鹿)羊膜来源广泛,成本低廉;其次不存在因种间差异而产生的排异反应,且种间差异越大其生物学效应越好;第三,国内外大量的羊胎盘产品(如各种胎盘素、冻干粉、胶囊、口服液及外用产品等)通过注射、口服、外用的方式用于人类,其安全性、有效性毋容质疑,然而目前没有现有技术采用羊膜作为贴式面膜的制备原料。
发明内容
本发明的目的在于针对上述问题,提供一种羊膜基膜、基于羊膜基膜的羊膜面膜及其制备方法,以解决现有的化学成分面膜刺激和损害皮肤,生物面膜品种单一、功效单一,羊膜用途较少等技术问题。
本发明所解决的技术问题可以采取以下方案来实现:一种基于羊膜基膜的羊膜面膜,其特征在于:所述羊膜面膜包括羊膜基膜及面膜载贴物,所述羊膜基膜由动物胎盘羊膜经去除海绵层和纤维细胞层并保留上皮细胞层、基底膜及致密层而制成,所述面膜载贴物贴合于所述羊膜基膜的致密层表面。
上述的基于羊膜基膜的羊膜面膜,所述羊膜基膜由动物胎盘羊膜经去除海绵层和纤维细胞层并保留上皮细胞层、基底膜及致密层之后,先经滋养液滋养、再经消毒液消毒而制成。
上述的基于羊膜基膜的羊膜面膜,所述滋养液为在生理盐水中添加维生素C及硫酸软骨素,所述消毒液为在蒸馏水中添加质量分数为15%~20%的过氧乙酸。
上述的基于羊膜基膜的羊膜面膜,每1000ml所述生理盐水中添加8~12g的所述维生素C以及8~12g的所述硫酸软骨素,每1000ml所述蒸馏水中添加10~15ml的所述过氧乙酸。
上述的基于羊膜基膜的羊膜面膜,在进行滋养时将羊膜置于所述滋养液中,在18~23℃下浸渍30分钟以上,在进行消毒时将羊膜用所述消毒液在18~23℃的温度条件下浸泡4小时以上。
上述的基于羊膜基膜的羊膜面膜,所述面膜载贴物为聚丙烯无纺布或硝酸纤维素滤膜。
上述的基于羊膜基膜的羊膜面膜,提供所述动物胎盘的动物为山羊或绵羊或鹿。
一种用于制备上述的基于羊膜基膜的羊膜面膜的方法,其特征在于:所述方法包括如下步骤:(1)制备羊膜基膜:具体包括如下制备步骤:(a)动物健康检查:检查、评估胎盘供体活体的健康状况,选择健康供体;(b)胎盘收取与清洗:收取所选择的健康供体的胎盘,并对收取的胎盘进行清洗;(c)羊膜分离与加工:将羊膜从经清洗后的胎盘上分离,分离后的羊膜首先剥离子叶,之后去除所述海绵层和纤维细胞层;(d)滋养与消毒:将去除了海绵层和纤维细胞层的羊膜置于滋养液中进行滋养,之后使用消毒液对经滋养后的羊膜进行消毒,消毒后的羊膜经清洗后形成羊膜基膜;(2)制备羊膜面膜:将所述面膜载贴物贴于经步骤(1)制备好的羊膜基膜的致密层表面。
上述的用于制备基于羊膜基膜的羊膜面膜的方法,所述滋养液为在无菌生理盐水中添加维生素C及硫酸软骨素,所述消毒液为在蒸馏水中添加质量分数为15%~20%的过氧乙酸。
上述的用于制备基于羊膜基膜的羊膜面膜的方法,每1000ml所述无菌生理盐水中添加8~12g的维生素C以及8~12g的硫酸软骨素,每1000ml所述蒸馏水中添加10~15ml的所述过氧乙酸
上述的用于制备基于羊膜基膜的羊膜面膜的方法,在所述步骤(d)中将羊膜置于所述滋养液中,在18~23℃的温度条件下浸渍30分钟以上,将羊膜用所述消毒液在18~23℃的温度条件下浸泡4小时以上。
上述的用于制备基于羊膜基膜的羊膜面膜的方法,在所述步骤(d)之后还包括步骤(e)羊膜基膜的保存:将经消毒后的羊膜基膜在常温下放置15分钟以上,之后放入甘油与DMEM混合液中密封,在低于零下80摄氏度的环境下进行保存。
上述的用于制备基于羊膜基膜的羊膜面膜的方法,所述甘油与所述DMEM的体积比为1:1,所述DMEM包含如下组分:无水氯化钙、硝酸铁、氯化钾、无水硫酸镁、氯化钠、无水磷酸二氢钠、丁二酸、丁二酸钠、L-盐酸精氨酸、L-盐酸胱氨酸、甘氨酸、L-盐酸组氨酸、L-异亮氨酸、L-亮氨酸、L-盐酸赖氨酸、L-甲硫氨酸、L-苯丙氨酸、L-丝氨酸、L-苏氨酸、L-色氨酸、L-酪氨酸、L-缬氨酸、D-泛酸钙、酒石酸胆碱、叶酸、肌醇、烟酰胺、核黄素、盐酸硫胺、盐酸吡哆辛、葡萄糖、丙酮酸钠、酚红。
一种用于制备面膜的羊膜基膜,其特征在于:所述羊膜基膜由动物胎盘羊膜经去除海绵层和纤维细胞层并保留上皮细胞层、基底膜及致密层之后,先经滋养液滋养、再经消毒液消毒而制成。
上述的用于制备面膜的羊膜基膜,所述滋养液为在生理盐水中添加维生素C及硫酸软骨素,所述消毒液为在蒸馏水中添加质量分数为15%~20%的过氧乙酸。
上述的用于制备面膜的羊膜基膜,每1000ml所述生理盐水中添加8~12g的维生素C以及8~12g的硫酸软骨素,每1000ml所述蒸馏水中添加10~15ml的所述过氧乙酸。
上述的用于制备面膜的羊膜基膜,在进行滋养时将羊膜置于所述滋养液中,在18~23℃的温度条件下浸渍30分钟以上,在进行消毒时将羊膜用所述消毒液在18~23℃的温度条件下浸泡4小时以上。
上述的用于制备面膜的羊膜基膜,提供所述动物胎盘的动物为山羊或绵羊或鹿。
本发明使用动物胎盘之羊膜制备贴式面膜,羊膜具有独特的生物学构造与活性,是制备贴式面膜理想的天然材料,作为一种高分子生物膜材料,具有良好的亲水性、透氧性和粘附性,把天然羊膜用作面膜不仅对皮肤无刺激性,还具有良好的帖合度;更为突出的是,天然羊膜具备了显著的生物学美容功能:①促进上皮分化,维持正常上皮表型;②改善皮肤营养,产生显著的皮肤保键、营养、美容和清洁等功效;③抑制纤维化,产生抗瘢痕作用;天然羊膜不仅可直接制备面帖膜,也可通过改变其活性成份制备各种功能性面膜,产生全面的营养与美容功能。
本发明促进了现有贴式面膜产品的升级换代,改善产品质量,提高产品功效,并减少产品对皮肤的刺激和损害,本发明使用动物胎盘之羊膜制备面帖膜,使面膜与皮肤的帖合度更好,对皮肤的剌激性更小。同时羊膜作为一种天然、有活性的生物材料,具有以下特点:首先,天然羊膜中富含营养和生物活性物质,可显著改善面膜产品的功能性成份,从而丰富了面膜内容,增加了面膜功效;其次,以羊膜基膜制备的面膜载帖物,可通过交联、吸附、结合等作用添加各种外源性营养与功能成份,极大地增加了面膜的产品种类;由于羊膜具有完整的组织结构和上皮细胞层、基底膜、致密层,上皮细胞的粘附作用和其它组织中胶原纤维构成的网状结构,为外源性营养、功能性成份提供了交联、吸(粘)附、结合的空间结构与组织基础,不仅能有效添加,还能保持其生物学活性,这是人工面膜纸、无纺布及其它面膜载帖物无法比拟的优势。
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地展示,下面将对本发明实施方式作进一步地详细描述。
实施例
本发明羊膜基膜的制备方法为:动物(绵羊、山羊、鹿均可)健康检查→胎盘收取与清洗→羊膜分离与加工→羊膜滋养与消毒→羊膜基膜;羊膜基膜贴式面膜的制备方法为:羊膜基膜→载帖物帖合→面帖膜。以绵羊、山羊和鹿胎盘获取羊膜,具有来源广泛、成本低廉的特点,我国每年经屠宰场集中淘汰的怀羔母羊超过600万只,以每只胎盘收取25×25cm羊膜为计,理论上全年可产羊膜375×107cm2。
以下以羊胎盘为例说明:
⑴动物健康检查:即屠宰前检查,为取得健康羊胎盘,对供体羊群的健康状况进行准确评估,方法是依据当地的“畜群健康标准”并结合个体抽检进行评估,确认供体羊健康。
①畜群健康标准主要评价标准有以下5条:
--畜群主要生产性能(胴体重、产羔率、羔羊成活率、出栏率)不低于所在地平均水平(-10%);
--病死率、淘汰率和流产率不高于当地平均水平(+10%);
--有明确的免疫程序和免疫标识制度,接种密度达到100%;
--有完整的畜群健康管理程序;
--有良好的兽医保障体系。
②个体抽检
根据羊群大小随机抽取5-10%待宰羊只,按以下项目逐项检查:被毛有无光泽、可视粘膜是否正常,精神状态、呼吸是否均匀,皮肤有无结痂、鼻镜干湿状况、有无分泌物,食欲是否正常、营养状况,体温、体表淋巴结等是否正常,排便是否正常。
③判定标准
畜群健康标准5条符合,个体抽检包括体温在内的90%的指标合格,即判为合格,认定羊群健康。
⑵胎盘收取与清洗
①通常于有资质的屠宰场收取胎盘,在屠宰车间收取集中宰杀的健康母羊胎盘。
②胎盘成熟度与收取
母羊怀羔时间90~150天,离体胎盘的收取时间不超过2个小时,胎盘收取、清洗的全过程不超过2小时。
③胎盘收取与清洗
洁净条件下收取胎盘、除去胎儿、羊水、污物、血水、血块等,用清水冲洗3次,再用无菌生理盐水洗2次。
④胎盘的检验
A,胎盘质量检验
按胎盘质量标准,依次检验胎盘外观、子叶饱满度、子叶分离数、重量,应符合要求。检验合格的胎盘-20℃速冻保存,保存时间以不超过6个月为宜。胎盘的质量标准为:新鲜胎盘淡红色,无异味、赘生物,无创伤、变色或腐败,手感柔软无结节。子叶饱满,直径≥1.0cm,母体子叶和胎儿子叶结合完好,分离子叶数不超过子叶总数的10%。单只胎盘的重量应为500~1000g。
B,胎盘病原微生物检验通过实验室检查排除羊布鲁氏病、结核病、口蹄疫,支原体、衣原体感染等病,确认胎盘合格。
布鲁氏病:应用豚鼠接种法检验,具体方法为:3批供试胎盘各取样10g,剪成块状(1×1cm),加等量灭菌生理盐水分别浸泡30min后,取出胎盘组织,所留液体摇匀,各取1ml混匀。豚鼠3只皮下注射1.0ml/只,3~4周后剖杀豚鼠,如脾脏、肝脏与肾脏出现炎性坏死小病灶,并在脾脏、淋巴结涂片中找到病原体,可判为阳性。
结核病:应用P氏固体培养基法检验,具体方法为:3批供试胎盘各取样10g,剪成块状(1×1cm),加等量灭菌生理盐水分别浸泡30min后,取出胎盘组织,所留液体摇匀,各取1ml混匀。接种于培养基上,室温(培养箱)培养至少20日。若无菌生长判为阴性。若有呈黄白色颗粒状、干燥、革兰氏染色阳性的菌落生长,判为阳性。
口蹄疫:应用乳鼠法检验,具体方法为:3批供试胎盘各取样10g,剪成块状(1×1cm),加等量灭菌生理盐水分别浸泡30min后,取出胎盘组织,所留液体摇匀,各取1ml混匀。取2~3日龄的乳鼠4~5只(不少于4只),颈部皮下接种0.1ml/只,连续观察7日,不出现以下症状并全部健活为阴性。出现下述症状者可判为阳性:后腿运动障碍、进而麻痹,头部不能抬起,呼吸困难,终因心肌麻痹死亡。
支原体:按现行《中国兽药典》附录进行检验,不得检出。
衣原体:应用小白鼠腹腔接种法检验,具体方法为:3批供试胎盘各取样10g,剪成块状(1×1cm),加等量灭菌生理盐水分别浸泡30min后,取出胎盘组织,所留液体摇匀,各取1ml混匀。接种于小白鼠腹腔内(0.5ml/只),如全部健活可判为阴性;若接种动物在7~10日内死亡,取脾、肺、肝制涂片检查,检出衣原体和嗜硷性包涵体可判为阳性。
⑶羊膜分离与加工
①羊膜分离取胎盘,自羊膜和绒毛膜之间的空隙处人工钝性分离。冷冻保存的羊膜应在完全解冻后分离羊膜。
②羊膜加工分离后的羊膜首先剥离子叶,再上皮细胞层朝下平铺于乙酸纤维薄膜纸上,用细胞刮子刮除海绵层和纤维细胞层,后用无菌生理盐水洗3次除去纤维细胞层和海绵层残余。除去纤维细胞层和海绵层的羊膜按需要裁剪、成形。
⑷羊膜基膜滋养与消毒
①羊膜基膜滋养经加工成形的羊膜基膜置于滋养液中,在18~23℃的温度条件下浸渍30分钟后取出用无菌生理盐水清洗。所述羊膜基膜滋养液为:维生素C 10g、硫酸软骨素10g、无菌生理盐水加至1000ml。
本发明中的羊膜滋养是改善、提高羊膜生物活性的重要环节,硫酸软骨素作为一种高分子物质,其大分子基团带有负电荷,可与羊膜上皮共价结合,从而在羊膜上皮形成一个保护层。实验证明,这一保护层不仅对羊膜上皮产生保护作用,还显著提高了羊膜上皮和基底膜的生物学活性,维生素C的抗氧化和保护胶原蛋白的作用进一步巩固了硫酸软骨素的羊膜保护功能,二者协同对羊膜基膜产生良好的滋养作用(引字中国专利公开号CN102132697A)。
②羊膜基膜消毒处理过的羊膜基膜用专用消毒液浸泡4小时(18~23℃)后取出,以无菌生理盐水充分清洗2次。所述羊膜基膜消毒液为:质量分数为15%的过氧乙酸15ml、蒸馏水加至1000ml。
③羊膜基膜保存暂不用于羊膜面帖膜制备的羊膜基膜按下法保存:于18~23℃下放置15分钟,使羊膜上的水分充分挥发,放入甘油与DMEM(1:1,V/V)混合液中密封,-80℃保存。保存时间可达6个月。所述DMEM的组分及含量如下表所示:
⑸羊膜面帖膜的制备
①载帖物帖合将羊膜基膜上皮面朝上,直接帖于载帖物(聚丙烯无纺布、硝酸纤维素滤膜)之上,使羊膜基膜与载帖物之间帖合完好即为羊膜基膜贴式面膜。
②羊膜基膜的再加工低温保存的羊膜基膜于灭菌生理盐水中复水30分钟后,即可用于载帖物帖合。
③羊膜面帖膜的保存对制备好的羊膜面贴膜进行真空密封包装、在2~8℃下保存,保存期为6个月。
以上所述,仅为本发明的具体实施方式,但本发明的保护范围并不局限于此,任何熟悉本发明的技术人员在本发明揭露的技术范围内,可轻易想到变化或替换,都应涵盖在本发明的保护范围之内。因此,本发明的保护范围应以所述权利要求的保护范围为准。
Claims (10)
1.一种基于羊膜基膜的羊膜面膜,其特征在于:所述羊膜面膜包括羊膜基膜及面膜载贴物,所述羊膜基膜由动物胎盘羊膜经去除海绵层和纤维细胞层并保留上皮细胞层、基底膜及致密层而制成,所述面膜载贴物贴合于所述羊膜基膜的致密层表面。
2.根据权利要求1所述的基于羊膜基膜的羊膜面膜,其特征在于:所述羊膜基膜由动物胎盘羊膜经去除海绵层和纤维细胞层并保留上皮细胞层、基底膜及致密层之后,先经滋养液滋养、再经消毒液消毒而制成。
3.根据权利要求2所述的基于羊膜基膜的羊膜面膜,其特征在于:所述滋养液为在生理盐水中添加维生素C及硫酸软骨素,所述消毒液为在蒸馏水中添加质量分数为15%~20%的过氧乙酸。
4.根据权利要求1~3中任意一项所述的基于羊膜基膜的羊膜面膜,其特征在于:提供所述动物胎盘的动物为山羊或绵羊或鹿。
5.一种用于制备如权利要求1~4中任意一项所述的基于羊膜基膜的羊膜面膜的方法,其特征在于:所述方法包括如下步骤:
(1)制备羊膜基膜:具体包括如下制备步骤:(a)动物健康检查:检查、评估胎盘供体活体的健康状况,选择健康供体;(b)胎盘收取与清洗:收取所选择的健康供体的胎盘,并对收取的胎盘进行清洗;(c)羊膜分离与加工:将羊膜从经清洗后的胎盘上分离,分离后的羊膜首先剥离子叶,之后去除所述海绵层和纤维细胞层;(d)滋养与消毒:将去除了海绵层和纤维细胞层的羊膜置于滋养液中进行滋养,之后使用消毒液对经滋养后的羊膜进行消毒,消毒后的羊膜经清洗后形成羊膜基膜;
(2)制备羊膜面膜:将所述面膜载贴物贴于经步骤(1)制备好的羊膜基膜的致密层表面。
6.根据权利要求5所述的用于制备基于羊膜基膜的羊膜面膜的方法,其特征在于:所述滋养液为在无菌生理盐水中添加维生素C及硫酸软骨素,所述消毒液为在蒸馏水中添加质量分数为15%~20%的过氧乙酸。
7.根据权利要求6所述的用于制备基于羊膜基膜的羊膜面膜的方法,其特征在于:每1000ml所述无菌生理盐水中添加8~12g的维生素C以及8~12g的硫酸软骨素,每1000ml所述蒸馏水中添加10~15ml的所述过氧乙酸
8.根据权利要求7所述的用于制备基于羊膜基膜的羊膜面膜的方法,其特征在于:在所述步骤(d)中将羊膜置于所述滋养液中,在18~23℃的温度条件下浸渍30分钟以上,将羊膜用所述消毒液在18~23℃的温度条件下浸泡4小时以上。
9.根据权利要求5~8中任意一项所述的用于制备基于羊膜基膜的羊膜面膜的方法,其特征在于:在所述步骤(d)之后还包括步骤(e)羊膜基膜的保存:将经消毒后的羊膜基膜在常温下放置15分钟以上,之后放入甘油与DMEM混合液中密封,在低于零下80摄氏度的环境下进行保存。
10.根据权利要求9所述的用于制备基于羊膜基膜的羊膜面膜的方法,其特征在于:所述甘油与所述DMEM的体积比为1:1,所述DMEM包含如下组分:无水氯化钙、硝酸铁、氯化钾、无水硫酸镁、氯化钠、无水磷酸二氢钠、丁二酸、丁二酸钠、L-盐酸精氨酸、L-盐酸胱氨酸、甘氨酸、L-盐酸组氨酸、L-异亮氨酸、L-亮氨酸、L-盐酸赖氨酸、L-甲硫氨酸、L-苯丙氨酸、L-丝氨酸、L-苏氨酸、L-色氨酸、L-酪氨酸、L-缬氨酸、D-泛酸钙、酒石酸胆碱、叶酸、肌醇、烟酰胺、核黄素、盐酸硫胺、盐酸吡哆辛、葡萄糖、丙酮酸钠、酚红。
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