CN108113960A - 一种用于红血丝皮肤的组合物及其制备方法 - Google Patents
一种用于红血丝皮肤的组合物及其制备方法 Download PDFInfo
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Abstract
本发明提供了一种用于红血丝皮肤的组合物，包括皮肤干细胞培养上清液、根皮素、植物甾醇、松花粉、珍珠粉、白藜芦醇、芦荟提取物、紫草素、磷酸腺苷、谷氨酰胺；所述皮肤干细胞培养基中添加神经干细胞裂解物。本发明有效清除皮肤红血丝，不仅皮肤红血丝症状得以大大缓解，并且皮肤敏感状况得到改善；同时治疗效果可长效保持，不会反复。
Description
技术领域
本发明属于美容化妆领域，主要涉及用于红血丝皮肤的组合物。
背景技术
红血丝是一种面部问题肌肤，呈现红血丝一丝丝纵横交错，如蜘蛛网般，分散性分布，严重者会连成片状，变成红脸。红血丝皮肤敏感，角质层薄弱导致毛细血管位置更容易接触和感知到外界环境变化，紫外线、过冷过热、情绪激动、温度突然变化等都会导致脸色更红；严重者还会形成沉积性色斑。
红血丝的形成多是后天由于各种损伤因素作用形成的。皮肤的表皮层是没有毛细血管的，表皮层的细胞营养主要来源棘层，当棘层细胞受损，自身供给营养能力下降。皮肤自身具备修复能力，此时真皮层的毛细血管自主向上扩张，帮助棘层细胞提供营养，从而形成红血丝。表皮层的细胞破坏越严重、受损时间越长、毛细血管增生移行越重，红血丝就越严重。当皮肤受损30%，是呈点状的红血丝；皮肤受损50%，是呈线状的红血丝；皮肤受损70%，是呈网状的红血丝；皮肤受损90%，是呈片状的红血丝。
改善红血丝的方法常采用化妆品涂覆于面部，然而大多只起到遮挡作用，而一些外用产品虽然可以抑制红血丝，但同时在皮肤又遗留一些有害毒素，致使出现斑点，皮肤变质，皮肤更加敏感脆弱，而且并不能从根本清除红血丝。而物理疗法，如冷冻治疗、激光治疗和同位素放射治疗等，虽然当时有效，但容易造成皮肤毁损、形成溃疡、瘢痕或放射性坏死等后遗症，美容效果并不理想。
CN105596249、CN106563089、CN107397876等采用中药组合物治疗红血丝，然而其制备、使用不方便，不适合日常护肤。
发明内容
本发明的目的在于提供一种用于红血丝皮肤的组合物，对皮肤无刺激，并能有效改善皮肤的红血丝状态，同时还能改善皮肤敏感状况。
本发明的目的是通过以下措施实现的：
一种用于红血丝皮肤的组合物，包括皮肤干细胞培养上清液、根皮素、植物甾醇、松花粉、珍珠粉、白藜芦醇、芦荟提取物、紫草素、磷酸腺苷、谷氨酰胺；所述皮肤干细胞培养基中添加神经干细胞裂解物。该组合物被命名为抗红肌液。
优选的，各组分用量配比为：皮肤干细胞培养上清液800-1500份、根皮素0.5-5份、植物甾醇5-15份、松花粉1-10份、珍珠粉5-20份、白藜芦醇0.005-0.05份、芦荟提取物3-8份、谷氨酰胺0.05-0.5份、紫草素0.5-5份，磷酸腺苷0.005-0.05份，以质量份数计。
所述神经干细胞裂解物是指神经干细胞破坏或溶解后细胞及细胞内物质的释放后所获得的混合物。包括胞质部分、细胞壁部分和代谢产物等。优选的，所述细胞裂解采用冻融法；具体地，将神经干细胞置于液氮中冷却，35-40℃融解，重复3次以上；膜过滤收集过膜物，即得。所述膜采用300-500nm孔径滤膜。
皮肤干细胞培养上清液是对皮肤干细胞培养3天以上收集得到的上清液。在培养24-48h后，在培养基中添加上述神经干细胞裂解物。兼顾生产效率和营养，以3-5天为佳。采用的培养基是：基础培养基为DMEM/F12、25-30μg/L VEGF、30-40μg/L EGF、5-10μg/L的NGF、5～50mg/L的人血清蛋白、20-30μg/L维生素E、柠檬酸铁1-3mmol/L、谷氨酰胺1-2mmol/L、庆大霉素 10-100g/L、腐胺0.8-2.0mg/L。该培养基培养24-48h后将神经干细胞裂解物加入培养基，用量为培养基的0.5-2wt%。
本发明还提供了一种护肤品，其含有上述能够用于敏感肌肤的组合物及常规化妆品级助剂，其中不含酒精、香精或防腐剂。
本发明还提供了上述用于敏感肌肤的组合物在制备敏感肌肤用护肤品中的应用。
本发明所述组合可以与其他皮肤活性成分一起用于各种产品制剂，例如，爽肤水、洁面乳、乳剂如乳液、面霜、面膜、凝胶等。
本发明所述组合物可以被用作保留型或洗去型组合物。例如，保留型组合物可以是被局部地施用至皮肤并在皮肤上保留一段时间( 比如5-30min或1-24h）；洗去型组合物可以是施用至皮肤然后在一段时间如小于5min内从皮肤去除或洗去的产品。洗去型组合物可以是皮肤清洁剂、洗发水、护发素或皂。保留型组合物的实例子可以是皮肤保湿剂、防晒剂、面膜、晚霜或日霜。
本发明所述芦荟提取物、是指水提、醇提、榨汁等方式得到的提取物。
有益效果
1. 本发明有效清除皮肤红血丝，扭转目前红血丝治疗饮鸩止渴的局面，不仅皮肤红血丝症状得以大大缓解，而且无任何致敏状况或加重皮肤敏感状况，反而使得皮肤敏感状况得到改善；同时治疗效果可长效保持，不会反复。本发明各种有效成分彼此协同增效，增强皮肤屏障功能，提高受损皮肤的免疫力，祛除皮肤红血丝、改善皮肤敏感性等多种症状。
2. 本发明安全、无刺激、无毒副作用，长期使用不会造成皮肤累计伤害。
3. 本发明不添加防腐剂、乳化剂、醇醚类、抗氧化剂、稳定剂、色素、香精等有毒害物质即可具有优异的修复敏感肌肤效果。
附图说明
图1 细胞增殖实验结果。
具体实施方式
下面通过实施例对本发明进行具体的描述，有必要在此指出的是本实施例只能用于对本发明进行进一步说明，不能理解为对本发明保护范围的限制，该领域的技术熟练人员可以根据上述本发明的内容作出一些非本质的改进和调整。本发明所述抗红肌液符合《化妆品安全技术规范》。
实施例1
一种用于红血丝皮肤的组合物，包括以下步骤：
1. 神经干细胞裂解物的制备
将人神经干细胞置于液氮中冷却30min，37℃细胞培养箱中融解，重复3次；250nm孔径膜过滤收集过膜物，即得。放入37℃保温备用。
2. 皮肤干细胞培养上清液的制备：
配置培养基：基础培养基为DMEM/F12、25μg/LVEGF、35μg/LEGF、8μg/L的NGF、20mg/L的人血清蛋白、25μg/L维生素E、柠檬酸铁2mmol/L、谷氨酰胺2mmol/L、庆大霉素 50g/L、腐胺1.6mg/L。将人皮肤干细胞在该培养基中，37℃培养24h后，在培养基中加入1wt%步骤（1）制得的神经干细胞裂解物，再培养24h，然后收集细胞液，离心，取上清，500nm孔径滤膜过滤，即得。
3. 肌液的制备
芦荟提取物是将芦荟茎去皮后的凝胶用水清洗后，进行破碎榨汁，去渣后即得。
将根皮素0.2份、植物甾醇8份、松花粉5份、珍珠粉10份、白藜芦醇0.01份、芦荟提取物5份、谷氨酰胺0.2份、紫草素3份，磷酸腺苷0.02份加入步骤（1）所得皮肤干细胞培养上清液（1200份）中充分混合；在50℃保温2小时，置于4℃冷冻箱内保存。
如无特别说明，本发明所述份数均以质量份数计。
实施例2
一种用于红血丝皮肤的组合物，包括以下步骤：
1. 神经干细胞裂解物的制备
将人神经干细胞置于液氮中冷却30min，37℃细胞培养箱中融解，重复5次；250nm孔径膜过滤收集过膜物，即得。放入37℃保温备用。
2. 皮肤干细胞培养上清液的制备：
配置培养基：基础培养基为DMEM/F12、30μg/LVEGF、40μg/LEGF、10μg/L的NGF、45mg/L的人血清蛋白、30μg/L维生素E、柠檬酸铁3mmol/L、谷氨酰胺2mmol/L、庆大霉素 80g/L、腐胺2.0mg/L。将人皮肤干细胞在该培养基中，37℃培养15h后，在培养基中加入1.5wt%步骤（1）制得的神经干细胞裂解物，再培养60h，然后收集细胞液，离心，取上清，500nm孔径滤膜过滤，即得。
3. 肌液的制备
将根皮素4份、植物甾醇12份、松花粉8份、珍珠粉12份、白藜芦醇0.02份、芦荟提取物8份、谷氨酰胺0.3份、紫草素4份，磷酸腺苷0.03份加入步骤（1）所得皮肤干细胞培养上清液（1500份）中充分混合；在50℃保温2小时，置于4℃冷冻箱内保存。
实施例3
一种用于红血丝皮肤的组合物，包括以下步骤：
1. 神经干细胞裂解物的制备
将人神经干细胞置于液氮中冷却30min，37℃细胞培养箱中融解，重复3次；250nm孔径膜过滤收集过膜物，即得。放入37℃保温备用。
2. 皮肤干细胞培养上清液的制备：
配置培养基：基础培养基为DMEM/F12、25μg/LVEGF、30μg/LEGF、5μg/L的NGF、10mg/L的人血清蛋白、20μg/L维生素E、柠檬酸铁1mmol/L、谷氨酰胺1mmol/L、庆大霉素 30g/L、腐胺1.0mg/L。将人皮肤干细胞在该培养基中，37℃培养30h后，在培养基中加入0.6wt%步骤（1）制得的神经干细胞裂解物，再培养80h，然后收集细胞液，离心，取上清，500nm孔径滤膜过滤，即得。
3. 肌液的制备
将根皮素0.5份、植物甾醇5份、松花粉1份、珍珠粉5份、白藜芦醇0.005份、芦荟提取物3份、谷氨酰胺0.05份、紫草素0.5份，磷酸腺苷0.005份加入步骤（1）所得皮肤干细胞培养上清液（1000份）中充分混合；在50℃保温2小时，混合均匀，置于4℃冷冻箱内保存。
实施例4
一．抗红肌液对细胞增殖的影响
1. 将人角质形成细胞HaCAT、神经细胞SHSY5Y、人微血管内皮细胞HMEC-1，置于37℃、5%CO2的培养箱中培养，取对数生长期细胞用于实验；
2. 将细胞接种于96孔板，每种细胞分为以下几组，每组设6个平行孔：
（1）100ul，DMEM/F12（含2%FBS）：空白对照组；
（2）100ul，DMEM/F12（含10%FBS）；细胞；标准对照组
（3）100ul，DMEM/F12（含2%FBS+1.0μl抗红肌液）；细胞；
（4）100ul，DMEM/F12（含2%FBS+2.0μl抗红肌液）；细胞；
（5）100ul，DMEM/F12（含2%FBS+4.0μl抗红肌液）；细胞；
（6）100ul，DMEM/F12（含2%FBS+8.0μl抗红肌液）；细胞；
（7）100ul，DMEM/F12（含2%FBS+16.0μl抗红肌液）；细胞；
（8）100ul，DMEM/F12（含2%FBS+20.0μl抗红肌液）；细胞。
3. MTT检测细胞增殖情况，如图1所示。
实验结果显示不同剂量抗红肌液对人角质形成细胞、神经细胞、人微血管内皮细胞均有增殖作用。与对照组相比，1.0-20uL/mL剂量的抗红肌液均有明显的细胞增殖作用，并表现剂量依赖性。
二. 长效实验
（1）受试物：本抗红肌液。
（2）动物：白化豚鼠。
（3）试验方法：试验前约24h，将动物背部脊柱两侧毛剪掉，去毛范围左、右各约3cmx 3cm。取受试物0.5ml 涂在一侧皮肤上，另一侧涂抹清水作为对照，两侧在涂抹后以两层纱布和一层玻璃纸覆盖，再以无刺激性胶布封闭固定6小时，然后用温水清洗以除去残留受试物。每天1 次，连续12个月。
（4）按照《化妆品安全技术规范》规定的皮肤反应来测试红斑和水肿情况。每天记录一次，并未发现任何红斑和焦痂形成，无任何水肿形成。
每个月进行血液检查、尿检、临床化学检查，未发现实验组和对照组有差异。
12个月后进行肉眼剖检和组织病理检查，为发现实验组和对照组有差异。
本发明长期使用安全可靠，能持续改善皮肤敏感问题。
三. 敏感性红血丝肌肤改善测试
1.受试人群：
红血丝人群：脸部皮肤可见粗细不等的毛细血管扩张、颜色淡红、鲜红或紫红色，压之不褪色，对称或不对称，局限或泛发。按严重程度进行评分：4分为有众多明显的红血丝，集结成团或成片，颜色深浅不一，皮肤发暗，有灼痛及瘙痒感者；3分为有明显红血丝，颜色发红，有刺痛感；2分为部分略有轻微红血丝、颜色浅者；1分为部分外表与正常皮肤稍有不同，基本与周围皮肤齐平，颜色略红或略白；0分为表面皮肤光滑，无红血丝、无痛痒、弹性良好、皮肤颜色基本正常。2-4分的个体认为其具有红血丝皮肤，作为受试人群，共筛选到55名。
2.实验方法：早晚各一次，日常清洁面部后用导入仪导入该抗红肌液，连续使用12周，实验期间未使用其它保湿、抗衰老等护肤品。
3. 效果检测：
（1）受试前和受试后（受试12周时）进行对比后按照1中评分标准进行评分。
（2）皮肤生物学实验：受试第一天当天在受试前检测皮肤红斑、皮肤角质层水含量；12周后再次检测皮肤红色素、皮肤角质层水含量、皮肤弹性情况。由皮肤黑色素和血红素测试仪（德国CK公司）测定皮肤红色素情况；SCH电学测量仪（Skicon-200）测定皮肤角质层水含量；皮肤弹性测试仪（德国CK公司）测试皮肤弹性。
（3）受试结束后3个月回访，观察面部红血丝状况。
4. 实验结果：（1）评分所得分数均值从治疗前3.2下降到治疗后1.4。
（2）红色素测试前后下降了32%以上、皮肤角质层水含量测试前后上升了20%以上，皮肤弹性测量前后增加了8%以上。
（3）受试结束后3个月回访，面部红血丝状况均保持或优于受试时候结束时的状态。
受试人群受试前均具有红血丝皮肤的特征性表现，经过12周治疗后，受试者的红血丝大幅下降；皮肤红色素下降，含水量上升，皮肤弹性增加，不仅皮肤红血丝症状得以大大缓解，而且本产品无任何致敏状况或加重皮肤敏感状况，反而治疗前后皮肤敏感状况得到改善；同时治疗效果可长效保持，不会反复。
Claims (9)
1.一种用于红血丝皮肤的组合物，包括皮肤干细胞培养上清液、根皮素、植物甾醇、松花粉、珍珠粉、白藜芦醇、芦荟提取物、紫草素、磷酸腺苷、谷氨酰胺；所述皮肤干细胞培养基中添加神经干细胞裂解物。
2.如权利要求1所述的用于敏感肌肤的组合物，皮肤干细胞培养上清液800-1500份、根皮素0.5-5份、植物甾醇5-15份、松花粉1-10份、珍珠粉5-20份、白藜芦醇0.005-0.05份、芦荟提取物3-8份、谷氨酰胺0.05-0.5份、紫草素0.5-5份，磷酸腺苷0.005-0.05份，以质量份数计。
3.如权利要求1或2所述的用于敏感肌肤的组合物，所述神经干细胞裂解物是将神经干细胞置于液氮中冷却，35-40℃融解，重复3次以上；膜过滤收集过膜物。
4.如权利要求3所述的用于敏感肌肤的组合物，所述膜采用300-500nm孔径滤膜。
5.如权利要求1所述的用于敏感肌肤的组合物，皮肤干细胞培养上清液是对皮肤干细胞培养3天以上收集得到的上清液。
6.如权利要求5所述的用于敏感肌肤的组合物，皮肤干细胞培养所采用的培养基为：基础培养基为DMEM/F12、25-30μg/L VEGF、30-40μg/L EGF、5-10μg/L的NGF、5～50mg/L的人血清蛋白、20-30μg/L维生素E、柠檬酸铁1-3mmol/L、谷氨酰胺1-2mmol/L、庆大霉素 10-100g/L、腐胺0.8-2.0mg/L；采用所述培养基培养24-48h后，培养基中加入神经干细胞裂解物，添加量为培养基的0.5-2wt%。
7.一种护肤品，其含有权利要求1-6任一项所述的能够改善敏感肌肤的组合物及常规化妆品级助剂，其中不含酒精、香精或防腐剂。
8.权利要求7所述的护肤品，其制成以下形式之一：爽肤水、洁面乳、乳剂如乳液、面霜、面膜、凝胶、皮肤清洁剂、洗发水、护发素、皂、皮肤保湿剂、防晒剂、面膜、晚霜或日霜。
9.权利要求1-5任一项所述的用于敏感肌肤的组合物在制备敏感肌肤用护肤品中的应用。
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| CN105055285A (zh) * | 2015-08-26 | 2015-11-18 | 浙江奥瑞健生物技术有限公司 | 一种皮肤营养液及其制备方法 |
| CN105125467A (zh) * | 2015-09-30 | 2015-12-09 | 青岛海之源智能技术有限公司 | 一种祛红血丝乳液及其制备方法 |
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| CN105055285A (zh) * | 2015-08-26 | 2015-11-18 | 浙江奥瑞健生物技术有限公司 | 一种皮肤营养液及其制备方法 |
| CN105125467A (zh) * | 2015-09-30 | 2015-12-09 | 青岛海之源智能技术有限公司 | 一种祛红血丝乳液及其制备方法 |
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