CN110179679A - 含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华、面膜及其制备方法 - Google Patents

含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华、面膜及其制备方法 Download PDF

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CN110179679A
CN110179679A CN201910553939.3A CN201910553939A CN110179679A CN 110179679 A CN110179679 A CN 110179679A CN 201910553939 A CN201910553939 A CN 201910553939A CN 110179679 A CN110179679 A CN 110179679A
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umbilical cord
mesenchymal stem
stem cells
cord mesenchymal
culture
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许键炜
吕宇
何志旭
周谊霞
秦臻
王品权
舒莉萍
熊英
刘忠山
吴西军
张丽娟
汪显耀
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Chenggong Kunming District People's Hospital
Guizhou Medical University
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Chenggong Kunming District People's Hospital
Guizhou Medical University
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Abstract

本发明涉及一种含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华、面膜及其制备方法，属于面膜技术领域。本发明的生物养颜面膜以与人体组织有高度相容性的丝素蛋白为基料，脐带来源的间充质干细胞无血清培养浓缩液为主料，自可以改变皮肤创面微环境，从而加快皮肤修复速率；干细胞还可以调节皮肤的稳态，抑制炎性因子分泌，促进抗炎介质释放，促进新生血管生成，加快皮肤的修复重建，改善皮肤损害状况。面膜还辅以多种植物活性成分以及人工合成护肤分子，保湿补水，润泽清新的水感瞬间渗透，滋养修护，缓解肌肤干渴，促进保湿成分渗透，加倍提升洁面力度，为肌肤补充营养，发挥面膜最大功效。

Description

含脐带间充质干细胞培养液提取物的生物养颜修护面膜精 华、面膜及其制备方法
技术领域
本发明属于面膜技术领域，具体涉及一种含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华、面膜及其制备方法。
背景技术
间充质干细胞(mesenchymal stem cells，MSCs)因其具有自我复制、多向分化潜能、造血支持和促进干细胞植入、免疫调控、异体移植不易发生免疫排斥，在体内或体外特定的诱导条件下，可分化为骨、肌肉、神经、肝、皮肤等多种组织细胞。而且MSCs可以在低温下长期保存，特性也较为稳定。这些特点使得MSCs在再生医学方面具有极强的应用价值而备受关注。MSCs同时也可以分泌一些重要的生物活性因子，包括具促进组织再生的因子如肝细胞生长因子(Hepatocyte growth factor,HGF)、血管内皮生长因子(vascularendothelial growth factor,VEGF)等，可以促进受损皮肤再生，修护老化皮肤。而脐带来源的间充质干细胞(human umbilical cord mesenchymal stem cells，HUCMSCs)除具有其他来源的MSCs同样的优点外，更原始、增殖能力更强，长期传代不易分化，不会产生肿瘤细胞，病毒和其他病原微生物感染及传播几率也相对较低，且取材方便，无伦理学争议，在新兴的转化医学中展现出广阔的应用前景。
我们在前期的研究中，用Elisa法检测了脐带来源的间充质干细胞无血清培养上清液中，能够促进组织再生修复的一些主要细胞因子，发现随着培养时间的延长，其分泌的量逐渐增加。详见下表1：
表1能够促进组织再生修复的一些主要细胞因子
目前市场上传统的同类产品主要是通过外源性物质对肌肤产生短暂治疗的目的，而本发明产品利用其所含的丰富的细胞活性因子，通过肌肤渗透，进入组织细胞内，激活皮肤内的干细胞，促进其增殖分化，使衰老细胞获得修复或更新，皮肤再生，达到长期治疗的目的。传统产品需要坚持每天使用，且传统产品功能相对单一，主要是美白、补水等功能，使用后需要立即清洗面部，因此对生物养颜修护面膜进行研究就显得尤为重要。
发明内容
本发明的目的是为了解决现有技术的不足，而提供一种含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华、面膜及其制备方法，本发明制得的面膜通过一个疗程10天左右的使用，可使皮肤获得长久修护，且本发明产品可以蚕丝膜布为载体，对皮肤无刺激，使用后可不必立即清洗面部，同时本发明产品同时兼具修复、美白、补水、嫩肤、润肤等多功能。
本发明采用如下技术方案：
含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华，以30份所述生物养颜修护面膜精华的重量为基础，包括以下重量份成分：水16份、脐带间充质干细胞培养浓缩液9份、卵磷脂0.20份、PE份-12聚二甲基硅烷交联聚物0.22份、野大豆(GLYCINE SOJA)0.26份、燕麦(AVENA SATIVA)肽0.18份、虎杖(POLYGONUM CUSPIDATUM)提取物0.13份、黄芩(SCUTELLARIA BAICALENSIS)提取物0.24份、母菊(CHAMOMILLA RECUTITA)花/叶提取物0.26份、苹果(MALUS PUMILA)提取物0.23份、茶(CAMELLIA SINENSIS)叶提取物0.20份、积雪草(CENTELLA ASIATICA)根提取物0.25份、迷迭香(ROSMARINUS OFFICINALIS)提取物0.09份、甘油聚全-26 0.44份、聚甘油-10 0.39份、双丙甘醇0.21份、EDTA二钠0.05份、丁二醇0.17份、α熊果苷0.23份、乙酰半胱氨酸0.12份、精氨酸阿魏酸盐0.13份、1-甲基乙内酰脲-2-酰亚胺0.08份、聚山梨醇酯-80 0.09份、植物甾醇类0.22份、甘油辛酸酯0.18份、辛酰羟肟酸0.08份、对羟基苯乙酮0.11份、卡波姆0.12份、三乙醇胺0.10份、香精0.02份。
更进一步地，所述脐带间充质干细胞培养浓缩液由L-DMEM细胞培养基和无血清培养基培养间充质干细胞48小时后浓缩获得。
本发明还提供所述含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华的制作方法，步骤如下：脐带间充质干细胞培养浓缩液的制作：选择增殖能力强、状态好的第3-5代人脐带间充质干细胞，用无血清培养基培养48h后，收集培养上清液30mL，1000×g离心15min除去细胞碎片，再经0.45μm无菌滤膜过滤到20mL规格的超滤离心管中，-30℃冻干机内冻干4小时，获得脐带间充质干细胞培养浓缩液；将其余成分加入至所得脐带间充质干细胞培养浓缩液中，即得所述脐带间充质干细胞培养液提取物的生物养颜修护面膜精华。
本发明还提供一种含脐带间充质干细胞培养液提取物的生物养颜修护面膜，起包含权利要求上述含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华。
本发明还提供一种所述含脐带间充质干细胞培养液提取物的生物养颜修护面膜的使用方法，步骤如下：用温水清洁额面部肌肤后，将面膜完整覆盖于额面部，用手指按压抚平面膜，使其完整敷贴于额面部，保留20～30分钟后揭下，将剩余的精华按摩至完全吸收，每天一片，连续使用10天为一疗程。
本发明与现有技术相比，其有益效果为：
第一：本发明的生物养颜面膜以与人体组织有高度相容性的丝素蛋白为基料，脐带来源的间充质干细胞无血清培养浓缩液为主料，富含干细胞因子(SCF)、内皮细胞生长因子(VEGF)、表皮细胞生长因子(EGF)、肿瘤坏死因子(TNF)、肝细胞生长因子(HGF)、神经生长因子(NGF)、纤维母细胞生长因子(FGF)、胰岛素样生长因子(IGF)、粒细胞集落刺激因子(G－CSF)、白细胞介素(IL)、胶原蛋白、溶菌酶等多种活性因子。自我更新快，具有较强的增殖能力和多向分化的潜能，并且能表达、合成、分泌或调节生长因子或者直接提供修复细胞来促进受损皮肤恢复；还可以改变皮肤创面微环境，从而加快皮肤修复速率；干细胞还可以调节皮肤的稳态，抑制炎性因子分泌，促进抗炎介质释放，促进新生血管生成，加快皮肤的修复重建，改善皮肤损害状况。面膜还辅以多种植物活性成分以及人工合成护肤分子，保湿补水，润泽清新的水感瞬间渗透，滋养修护，缓解肌肤干渴，促进保湿成分渗透，加倍提升洁面力度，为肌肤补充营养，发挥面膜最大功效。
第二：本发明制得的面膜通过一个疗程10天左右的使用，可使皮肤获得长久修护，且本发明产品可以蚕丝膜布为载体，对皮肤无刺激，使用后可不必立即清洗面部，同时本发明产品同时兼具修复、美白、补水、嫩肤、润肤等多功能。
说明书附图
图1为HE染色含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华对烧伤小鼠皮肤的治疗作用。
具体实施方式
下面结合实施例对本发明作进一步的详细描述。
以下为实施例所用原料的说明：
实施例
含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华，以30份所述生物养颜修护面膜精华的重量为基础，包括以下重量份成分：水16份、脐带间充质干细胞培养浓缩液9份、卵磷脂0.20份、PE份-12聚二甲基硅烷交联聚物0.22份、野大豆(GLYCINE SOJA)0.26份、燕麦(AVENA SATIVA)肽0.18份、虎杖(POLYGONUM CUSPIDATUM)提取物0.13份、黄芩(SCUTELLARIA BAICALENSIS)提取物0.24份、母菊(CHAMOMILLA RECUTITA)花/叶提取物0.26份、苹果(MALUS PUMILA)提取物0.23份、茶(CAMELLIA SINENSIS)叶提取物0.20份、积雪草(CENTELLA ASIATICA)根提取物0.25份、迷迭香(ROSMARINUS OFFICINALIS)提取物0.09份、甘油聚全-26 0.44份、聚甘油-10 0.39份、双丙甘醇0.21份、EDTA二钠0.05份、丁二醇0.17份、α熊果苷0.23份、乙酰半胱氨酸0.12份、精氨酸阿魏酸盐0.13份、1-甲基乙内酰脲-2-酰亚胺0.08份、聚山梨醇酯-80 0.09份、植物甾醇类0.22份、甘油辛酸酯0.18份、辛酰羟肟酸0.08份、对羟基苯乙酮0.11份、卡波姆0.12份、三乙醇胺0.10份、香精0.02份。
其中，所述脐带间充质干细胞培养浓缩液由L-DMEM细胞培养基和无血清培养基培养间充质干细胞48小时后浓缩获得，具体组成如下表2所示：
表2 L-DMEM细胞培养基(粉末型)成分
(2)无血清培养基成分：
表3所含15种氨基酸：
名称 含量(mg/l)
亮氨酸 60.00
脯氨酸 70.00
丝氨酸 25.00
色氨酸 30.00
苏氨酸 180.00
半胱氨酸 30.00
缬氨酸 50.00
蛋氨酸 50.00
异亮氨酸 50.00
酪氨酸 40.00
谷氨酸 40.00
苯丙氨酸 60.00
赖氨酸 60.00
组氨酸 30.00
精氨酸 84.00
表4所含其他成分：
所述含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华的制作方法，步骤如下：脐带间充质干细胞培养浓缩液的制作：选择增殖能力强、状态好的第3-5代人脐带间充质干细胞，用无血清培养基培养48h后，收集培养上清液30mL，1000×g离心15min除去细胞碎片，再经0.45μm无菌滤膜过滤到20mL规格的超滤离心管中，-30℃冻干机内冻干4小时，获得脐带间充质干细胞培养浓缩液；将其余成分加入至所得脐带间充质干细胞培养浓缩液中，即得所述脐带间充质干细胞培养液提取物的生物养颜修护面膜精华。
将本发明的含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华用于烧伤小鼠的皮肤愈合，具体使用方法：将小鼠固定于老鼠固定器内并暴露烧伤皮肤，面膜剪成小块平敷于小鼠烧伤部位，保留30分钟后揭掉面膜，待皮肤的面膜精华液完全吸收。每天一次，持续至小鼠烧伤皮肤完全愈合。由图1可知，烧伤模型组和HUCMSCs治疗组的烧伤皮肤创面面积均减小，烧伤模型组皮肤创面愈合时间为20-25天，含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华治疗组烧伤皮肤创面愈合时间为14-18天。含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华治疗组的烧伤皮肤创面愈合时间较明显小于烧伤模型组。
治疗后，烧伤模型组和含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华治疗组的烧伤皮肤创面愈合后都有瘢痕，烧伤模型组皮肤创面愈合后瘢痕厚度约为500μm，含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华治疗组皮肤创面愈合后瘢痕厚度约30-50μm，含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华治疗组的烧伤皮肤创面愈合后瘢痕大小明显小于单纯烧伤组，且治疗14天组疗效优于治疗7天组，HE染色结果显示治疗组皮肤各层结构与正常组类似。
将本发明的含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华，以蚕丝膜布为载体，制作成面膜成品，面膜的使用方法如下：用温水清洁额面部肌肤后，将面膜完整覆盖于额面部，用手指按压抚平面膜，使其完整敷贴于额面部，保留20～30分钟后揭下，将剩余的精华按摩至完全吸收，每天一片，连续使用10天为一疗程。
本发明的含脐带间充质干细胞培养液提取物的生物养颜修护面膜在使用时的注意事项：
1.本品为一次性使用；
2.为避免面膜内活性成分失活，应将面膜储存于4～10℃低温环境；
3.在寒冷天气使用时，使用前可将未拆封面膜置于40℃温水中浸泡5分钟预热后使用。
上述实施例对本发明的实施方式作了详细说明，但是本发明并不限于上述实施方式，在本领域普通技术人员所具备的知识范围内，还可以在不脱离本发明宗旨的前提下做出各种变化。

Claims (5)

1.含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华，其特征在于，以30份所述生物养颜修护面膜精华的重量为基础，包括以下重量份成分：水16份、脐带间充质干细胞培养浓缩液9份、卵磷脂0.20份、PE份-12聚二甲基硅烷交联聚物0.22份、野大豆（0.26份、燕麦肽0.18份、虎杖提取物0.13份、黄芩提取物0.24份、母菊花/叶提取物0.26份、苹果提取物0.23份、茶叶提取物0.20份、积雪草根提取物0.25份、迷迭香提取物0.09份、甘油聚全-260.44份、聚甘油-10 0.39份、双丙甘醇0.21份、EDTA二钠0.05份、丁二醇0.17份、α熊果苷0.23份、乙酰半胱氨酸0.12份、精氨酸阿魏酸盐0.13份、1-甲基乙内酰脲-2-酰亚胺0.08份、聚山梨醇酯-80 0.09份、植物甾醇类0.22份、甘油辛酸酯0.18份、辛酰羟肟酸0.08份、对羟基苯乙酮0.11份、卡波姆0.12份、三乙醇胺0.10份、香精0.02份。
2.根据权利要求1所述的含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华，其特征在于，所述脐带间充质干细胞培养浓缩液由L-DMEM细胞培养基和无血清培养基培养间充质干细胞48小时后浓缩获得。
3.权利要求1所述的含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华的制作方法，其特征在于，步骤如下：脐带间充质干细胞培养浓缩液的制作：选择增殖能力强、状态好的第3-5代人脐带间充质干细胞，用无血清培养基培养48 h后，收集培养上清液30 mL，1000×g离心15 min除去细胞碎片，再经0.45 μm无菌滤膜过滤到20 mL规格的超滤离心管中，-30℃冻干机内冻干4小时，获得脐带间充质干细胞培养浓缩液；将其余成分加入至所得脐带间充质干细胞培养浓缩液中，即得所述脐带间充质干细胞培养液提取物的生物养颜修护面膜精华。
4.含脐带间充质干细胞培养液提取物的生物养颜修护面膜，其特征在于，包含权利要求1所述的含脐带间充质干细胞培养液提取物的生物养颜修护面膜精华。
5.权利要求4所述的含脐带间充质干细胞培养液提取物的生物养颜修护面膜的使用方法，其特征在于，步骤如下：用温水清洁额面部肌肤后，将面膜完整覆盖于额面部，用手指按压抚平面膜，使其完整敷贴于额面部，保留20～30分钟后揭下，将剩余的精华按摩至完全吸收，每天一片，连续使用10天为一疗程。
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Application publication date: 20190830