CN111956787A - 一种毛发增长剂和育毛剂及其应用 - Google Patents
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Abstract
本发明公开了一种毛发增长剂和育毛剂及其应用，所述毛发增长剂和育毛剂由胎盘组织和脐带组织中的干细胞中分离提取得到，包括高活性多肽分子、复合细胞生长因子、细胞活化因子以及外泌体。本发明中育毛剂的原材料由胎盘和脐带中的干细胞分离提取得到，不含有有害化学成分、基因重组的生长因子、生长激素和重金属等；所述育毛剂中的生物活性成分既能有效改善头皮组织新陈代谢，也可阻止和逆转皮肤衰老修复成纤维细胞损伤以促进毛囊细胞的再生，对毛发生长毛囊生物制药有重要价值。
Description
技术领域
本发明涉及增毛剂技术领域，尤其涉及一种毛发增长剂和育毛剂及其应用。
背景技术
毛发的生长往往受到营养、神经内分泌和自身免疫的影响，临床上常见的脱发性疾病有斑秃、雄激素性脱发及生理性脱发。目前市场上的防脱发产品多种多样，对于脱发只能提供短期的改善作用，停止用药后，患者容易出现再次脱发的现象。另外，现有防脱发产品对于毛发的再生作用效果微乎其微，大大降低了防脱发产品的使用效果。
发明内容
本发明的目的在于：为了解决现有技术存在的问题，而提出一种毛发增长剂和育毛剂及其应用。本发明提供的毛发增长剂和育毛剂的生物活性既能有效改善头皮组织新陈代谢，也可阻止和逆转皮肤衰老修复成纤维细胞损伤以促进毛囊细胞的再生，调节并平衡头皮组织新陈代谢，修复受伤后的皮肤，从而改善局部微环境降低毛囊重建障碍，提高头皮组织毛发的再生能力。
为了实现上述目的，本发明采用了如下技术方案：一种毛发增长剂和育毛剂，所述毛发增长剂和育毛剂由胎盘组织和脐带组织中的干细胞中分离提取得到，包括高活性多肽分子、复合细胞生长因子、细胞活化因子以及外泌体。
作为对上述方案的进一步描述：所述高活性多肽分子的总含量为1pg/ml-500ug/ml，所述复合细胞生长因子的总合量为10pg/ml-1000ug/ml，所述细胞活化因子的总含量为3pg/ml-1700ug/ml，所述外泌体的总含量为1pg/ml-2000ug/ml。
作为对上述方案的进一步描述：所述复合细胞生长因子中主要包括表皮生长因子EGF、成纤维细胞生长因子FGF、神经生长因子NGF、肝细胞生长因子HGF、胎盘生长因子PLGF、角化细胞生长因子KGF、血管内皮生长因子VEGF和胰岛素样生长因子IGF。
作为对上述方案的进一步描述：所述外泌体包含CD9、CD63、CD81和PLAP中的一种或几种，可以诱导毛囊外根鞘细胞或毛囊组织样的结构形成。
作为对上述方案的进一步描述：所述细胞活化因子为T细胞活化因子，也具有促进毛囊干细胞生长的活性。
作为对上述方案的进一步描述：所述高活性多肽分子的分子量为500-1000道尔顿。
作为对上述方案的进一步描述：所述胎盘组织和脐带组织中的干细胞选自脐带血干细胞、脐带华氏胶干细胞、羊膜干细胞和胎盘干细胞中的一种或几种。
作为对上述方案的进一步描述：所述毛发增长剂和育毛剂由以下方法分离提取：
S1、将胎盘组织和脐带组织中的干细胞进行组织分离工程；
S2、将分离出的干细胞进行培养，然后进行磁珠破碎工程；
S3、将破碎的细胞进行过筛和分子筛精制，得到所需高活性多肽分子、复合细胞生长因子、细胞活化因子和外泌体，配置成所述毛发增长剂和育毛剂。
本发明另一方面，还提供所述毛发增长剂和育毛剂在促进毛囊毛发增长中的应用，毛囊的发育是表皮细胞和其下的间质相互作用诱导形成，重建一个毛囊生长模型，对阐明毛囊生物学的许多生理特征有重要价值。所述增毛剂可诱导免疫缺陷和裸鼠的毛囊形成，也可阻止和逆转皮肤衰老修复成纤维细胞损伤以促进毛囊细胞的再生，增毛剂对人的毛囊毛发的增长也有促进作用特别是对受伤后再生新毛囊，新产生的毛囊建立形成干细胞群，表达已知毛囊分化分子标记，产生毛杆，并且进入毛囊生长的各个阶段。
综上所述，由于采用了上述技术方案，本发明的有益效果是：本发明中育毛剂的原材料由胎盘和脐带组织中的干细胞分离提取得到，不含有有害化学成分、基因重组的生长因子、生长激素和重金属等；所述育毛剂中的生物活性成分既能有效改善头皮组织新陈代谢，也可阻止和逆转皮肤衰老修复成纤维细胞损伤以促进毛囊细胞的再生，对毛发生长毛囊生物制药有重要价值。
附图说明
图1为本发明提出的毛发增长剂和育毛剂裸鼠局部使用实验图；
图2小鼠皮肤内导入鉴定实验图；
图3为本发明提出的毛发增长剂和育毛剂的裸鼠全身使用实验图；
图4为本发明提出的毛发增长剂和育毛剂在脱发人群使用的效果对比图。
具体实施方式
下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其它实施例，都属于本发明保护的范围。
实施例1
胎盘干细胞的分离
取新鲜胎盘组织进行无菌和病毒检测，然后将胎盘组织上羊膜、绒毛膜等剥离，胎盘组织分离后，切割为边长为2cm左右的小块，加入与糊状组织体积相等的生理盐水，然后进行细胞破碎机工程或磁珠破碎工程。
脐带血干细胞的分离培养
(1)脐带血全血细胞分离和血浆进行预处理；
(2)细胞培养皿包被抗CD34抗体和抗CD44抗体；
(3)向细胞培养皿中加入步骤(1)预处理后的脐带血全血细胞，分离培养有核细胞。
羊膜和脐带华氏胶干细胞的分离培养
将羊膜组织和华氏胶组织分别进行切割为边长为小于1cm左右的小块，进行细胞培养和继代，收集P3-P5代细胞，加入与糊状组织体积相等的生理盐水进行细胞破碎机工程或磁珠破碎工程，然后研磨细胞，过筛，细胞筛为100um，过滤液分离提取检测，得到复合细胞生长因子及其他有效成分。
将获得的细胞因子、多肽分子、细胞因子及外泌体混合物采用无菌生理盐水，林格氏液配置成毛发增长剂和育毛剂，增毛剂中高活性多肽分子分子量为500-550道尔顿，多肽分子的含量为1-50pg/ml，复合细胞生长因子的总含量为70ng/ml-600ng/ml，细胞活化因子的总含量为3-300pg/ml，外泌体的总含量为25-530ng/ml，复合细胞生长因子中主要成分的含量如下：胰岛素样生长因子IGF含量为50-230ng/ml，血管内皮生长因子VEGF含量为30-260pg/ml，纤维细胞生长因子-5 FGF-5含量为20-500pg/ml，表皮生长因子EGF含量为20-250pg/ml，神经生长因子NGF含量为20-350ng/ml，肝细胞生长因子HGF含量为50-1000pg/ml；胎盘生长因子PLGF含量为10-200pg/ml，角化细胞生长因子KGF含量为10-300pg/ml。
外泌体中CD9含量为1-30ng/ml，CD63 TSPAN30含量为10-100pg/ml，CD81含量为25-500ng/ml。
实施例2裸鼠增毛全身和局部实验
试剂：采用实施例1制得的育毛剂；
仪器：压缩空气式雾化器(雾化颗粒＜5μm，雾化量0.2ml/min)；
小鼠：成年裸鼠各4只+对照2只不干预；
进行裸鼠局部喷雾(口鼻)和全身喷雾试验，每次雾化进行3min每周5次持续3周，每次使用量1ml，使用3周后评价结果。
实验结果：
局部增毛结果：2周后2只出现发毛，3周后发毛显著，发毛率为100％。
全身增毛结果：有3周后出现发毛4周后发毛显著，发毛率100％。
对照组无发毛0％，如图1所示。
现有技术中不用培养技术无法使裸鼠毛发增长。本发明克服了基因缺陷，给药2周后裸鼠局部表面观察到毛发生长，促进毛囊的再生，对毛发生长毛囊生物制药有重要价值。
实施例3增毛剂的小鼠皮肤内导入鉴定
裸鼠皮肤标记实验将实施例1中的育毛液加入细胞核标记物1∶1000；喷雾3日后解剖检测冷冻切片，结果裸鼠皮肤毛囊有细胞核的评价荧光标记UV390nm，细胞核染色，病理表示毛囊的细胞核有显示，如图2所示。
实施例4小鼠损伤后皮肤修复和毛发增长的实验
4周小鼠2只，背部皮肤剃毛，其中一只进行皮肤10mm的表面创伤。对照组剃毛无创伤。次日将2只小鼠每日进行全身喷雾，每次3min；4日后皮肤表面创伤的小鼠皮肤不仅修复其毛发、毛发增长覆盖全身、与无损伤的小鼠对比有差别，见图3a；每日喷雾后2周无损伤的小鼠生毛和覆盖全身，见图3b。
实施例5
所述育毛剂对人的毛囊毛发的增长也有促进作用，特别是对受伤后再生新毛囊。新产生的毛囊建立了干细胞群，表达已知毛囊分化分子标记，产生毛杆，并且进入毛囊生长的各个阶段。采用实施例1制得的育毛剂对一个40岁左右的脱发实验对象进行喷雾实验，实验前如图4a所示，20天后，实验对象毛发长出，如图4b所示。
由上述实施例可知，毛囊细胞诱导且对免疫缺陷和裸鼠毛囊形成，也可阻止和逆转皮肤衰老修复成纤维细胞损伤以促进毛囊细胞的再生，对人的毛囊毛发的增长也有促进作用。毛发增长剂和育毛剂促进毛囊的表皮细胞发育也促进间质的生长期、退化期、休止期三个阶段的循环周期。再生重建一个毛囊生长模型，对毛囊再生具有生理特征的价值。
以上所述，仅为本发明较佳的具体实施方式，但本发明的保护范围并不局限于此，任何熟悉本技术领域的技术人员在本发明揭露的技术范围内，根据本发明的技术方案及其发明构思加以等同替换或改变，都应涵盖在本发明的保护范围之内。
Claims (9)
1.一种毛发增长剂和育毛剂，其特征在于，所述毛发增长剂和育毛剂由胎盘组织和脐带组织中的干细胞中分离提取得到，包括高活性多肽分子、复合细胞生长因子、细胞活化因子以及外泌体。
2.根据权利要求1所述的毛发增长剂和育毛剂，其特征在于，所述高活性多肽分子的总含量为1pg/ml-500ug/ml，所述复合细胞生长因子的总含量为10pg/ml-1000ug/ml，所述细胞活化因子的总含量为3pg/ml-1700ug/ml，所述外泌体的总含量为1pg/ml-2000ug/ml。
3.根据权利要求1所述的毛发增长剂和育毛剂，其特征在于，所述复合细胞生长因子中主要包括表皮生长因子EGF、成纤维细胞生长因子FGF、神经生长因子NGF、肝细胞生长因子HGF、胎盘生长因子PLGF、角化细胞生长因子KGF、血管内皮生长因子VEGF和胰岛素样生长因子IGF。
4.根据权利要求1所述的毛发增长剂和育毛剂，其特征在于，所述外泌体包含CD9、CD63、CD81和PLAP中的一种或几种。
5.根据权利要求1所述的毛发增长剂和育毛剂，其特征在于，所述细胞活化因子为T细胞活化因子。
6.根据权利要求1所述的毛发增长剂和育毛剂，其特征在于，所述高活性多肽分子的分子量为500-1000道尔顿。
7.根据权利要求1所述的毛发增长剂和育毛剂，其特征在于，所述胎盘组织和脐带组织中的干细胞选自脐带血干细胞、脐带华氏胶干细胞、羊膜干细胞和胎盘干细胞中的一种或几种。
8.根据权利要求1所述的毛发增长剂和育毛剂，其特征在于，所述毛发增长剂和育毛剂由以下方法分离提取：
S1、将胎盘组织和脐带组织中的干细胞进行组织分离工程；
S2、将分离出的干细胞进行培养，然后进行磁珠破碎工程；
S3、将破碎的细胞进行过筛和分子筛精制，得到所需高活性多肽分子、复合细胞生长因子、细胞活化因子和外泌体，配置成所述毛发增长剂和育毛剂。
9.一种如权利要求1～8任一所述的毛发增长剂和育毛剂在促进毛囊毛发增长中的应用。
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