CN114983857A - 一种太平洋鳕鱼皮明胶肽微针的制备方法及其应用 - Google Patents
一种太平洋鳕鱼皮明胶肽微针的制备方法及其应用 Download PDFInfo
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Abstract
本发明提出一种太平洋鳕鱼皮明胶肽微针的制备方法及其应用,将太平洋鳕鱼皮明胶肽与先进的微针技术相结合,将太平洋鳕鱼皮明胶肽与透明质酸的混合物覆盖于微针模板表面,脱模后获得了一种含有太平洋鳕鱼皮明胶肽的微针贴片,能够改善明胶肽皮肤渗透性差,释放速率慢等问题。
Description
技术领域
本发明属于美容微整形技术领域,具体涉及一种太平洋鳕鱼皮明胶肽微针的制备方法及其应用。
背景技术
随着社会的发展和生活质量的提升,人们开始更加关注自己的形象,同时也对美提出了更高的要求。在社会日益发展的今天,医学所要承担的责任已不仅仅只是治病救人。美容就是建立在医疗基础之上的,是医学的一部分。
面部皮肤因常常暴露于紫外线、烟草烟雾和污染等有害的环境因素下,其脂质氧化和衰老进程更快于身体其他部位。因此抗氧化抗衰老是现代医学美容技术的重要组成部分,但是传统的抗氧化物质在美容行业的具体应用中,都存在一定的问题,如:易溶解、稳定性差、渗透性差等。研究表明,水解胶原蛋白能够产生胶原蛋白低聚肽,这些肽类能够发挥美容和抗氧化衰老的作用。
传统的美容手段包括涂抹美容、激光美容和口服,肽作为小分子药物最常见的为口服给药。但蛋白质和肽类药物经口服后,肠胃道上皮会充当蛋白质吸收的物理和生化屏障,导致其利用度极低通常小于1-2%。其次口服美容因其作用机制,无法在短期内出现明显效果,往往需要长达几个月的时间。
鱼胶原蛋白经酶解后制的胶原蛋白肽,因其低聚性,易于被人体吸收利用,胶原蛋白低聚肽还具有良好的生物相容性和渗透性,且对人体无刺激性,同时胶原蛋白低聚肽可以为机体合成胶原蛋白提供原料,因此常被应用于美容和抗衰老等领域。
微针用于美容已经开展的较为成熟,微针可以通过微针滚轮上的许多微小镇头刺激皮肤,并且能够定位、定层、定量的将活性成分直接导入深层皮肤细胞,从而达到各种美容效果。可溶性微针还具有使用方便、卫生,且不必担心微针在皮内断裂等优点。
发明内容
本发明为解决上述技术问题,将肽与微针结合,提高肽的利用度。
第一方面,提供一种太平洋鳕鱼皮明胶肽微针的制备方法,包括:
S10:获取太平洋鳕鱼皮明胶肽和深海鱼皮明胶并混合,将混合物覆盖在微针模板表面;
S20:加热浓缩后加入透明质酸和PVA溶液脱模,得到含有太平洋鳕鱼皮明胶肽的可溶性微针。
本发明将太平洋鳕鱼皮明胶肽与先进的微针技术相结合,将太平洋鳕鱼皮明胶肽与透明质酸的混合物覆盖于微针模板表面,脱模后获得了一种含有太平洋鳕鱼皮明胶肽的微针贴片,能够改善明胶肽皮肤渗透性差,释放速率慢等问题。
优选的,所述步骤S10包括获取太平洋鳕鱼皮明胶肽的步骤:
S11:采用碱液浸泡太平洋鳕鱼皮,除去胶原蛋白外的杂质;
S12:用水清洗碱处理过的太平洋鳕鱼皮并浸泡,搅拌,提取明胶;
S13:采用离心设备,对提取的明胶离心处理并收集上清液,将所述上清液冷冻干燥处理,获取干物质明胶粉;
S14:将所述明胶粉溶于甘氨酸缓冲溶液中;
S15:采用胃蛋白酶、胰蛋白酶和α-糜蛋白酶水解明胶,并不断搅拌;
S16:煮沸明胶后制成冻干备用。
优选的,所述S11中,碱液浸泡太平洋鳕鱼皮前,需将太平洋鳕鱼皮清洗,以去除表面粘液并剪成小块,例如尺寸为1cm*2cm;碱液为0.5%-1.5%的Ca(OH)2溶液;浸泡时间为20h-28h;浸泡期间更换一次到五次的碱液。
优选的,所述S12中,清洗过的太平洋鳕鱼皮采用pH为5.5到6.5的水浸泡28h-32h,水温为55℃-65℃,并持续搅拌。
优选的,所述S13中,离心处理时,根据需求,转速2800rmp-3200rmp,离心时间8min-12min。
优选的,所述S14中,将所述明胶粉溶解于0.01M的甘氨酸缓冲溶液中,调节pH=2。
优选的,所述S15中,依次使用胃蛋白酶、胰蛋白酶和α-糜蛋白酶各水解3.5h-4.5h,E/S=1:100,期间不断搅拌。
优选的,所述S16中,煮沸明胶8min-12min后,制成冻干存于-75℃--85℃备用。
优选的,所述S10中,将80mg/mL的太平洋鳕鱼明胶肽和深海鱼皮明胶1:1混合,并将混合物覆盖在微针模板表面;所述S20中,加热浓缩后加入含水量33%-37%的透明质酸;所述微针侧开口结构,针长300μm,间距800μm,直径270μm。
第二方面,提供一种太平洋鳕鱼皮明胶肽微针,采用第一方面所述的方法获取。
由上述对本发明的描述可知,本发明具有以下有益效果:
1、本发明首次将太平洋鳕鱼皮明胶肽和透明质酸混合作为基质材料制作成可溶性微针;
2、本发明使用方便、卫生、安全系数高;
3、本发明能够是明胶肽快速进入皮肤并缓慢释放药物发挥功效;
4、本发明缩短了明胶肽的疗效时间;
5、本发明具有优秀的除皱保湿效果。
附图说明
此处所说明的附图用来提供对本发明的进一步理解,构成本发明的一部分,本发明的示意性实施例及其说明用于解释本发明,并不构成对本发明的不当限定。
其中:
图1是本发明太平洋鳕鱼皮可溶性微针的形貌表征图;
图2是本发明实施例二中微针力学性能测试图;
图3是本发明实施例二中体外微针刺入皮肤试验结果图;
图4是本发明实施例二中微针刺入皮肤溶解试验结果图;
图5是本发明实施例二中微针刺入皮肤后愈合情况结果图。
具体实施方式
为了使本发明所要解决的技术问题、技术方案及有益效果更加清楚、明白,以下结合附图和实施例,对本发明进行进一步详细说明。应当理解,此处所描述的具体实施例仅用以解释本发明,并不用于限定本发明。
实施例一:
获取太平洋鳕鱼明胶肽,并将80mg/mL的太平洋鳕鱼明胶肽和深海鱼皮明胶1:1混合,并将混合物覆盖在微针模板表面,负压除泡,并吸取含气泡的溶液,加热浓缩,二次加入含水量35%透明质酸,后再次加热浓缩,经过光固化滴加PVA溶液,加热烘干脱模后,得到含太平洋鳕鱼皮明胶肽的可溶性微针。
在一实施例中,获取太平洋鳕鱼明胶肽的步骤包括:将太平洋鳕鱼皮清洗干净,除去表面的粘液并剪成小块(1cm×2cm),以1:4的比例浸泡在1%的Ca(OH)2溶液中,浸泡24h,期间更换三次1%的Ca(OH)2溶液,以去除胶原蛋白外的其他杂质,用水彻底清洗碱处理的太平洋鳕鱼皮后,将太平洋鳕鱼皮碎片至于pH=6的60℃水中浸泡30h,并持续搅拌提取明胶,将混合物至于离心机中,3000rmp,离心10min,收集上清液冷冻干燥,得到干物质——明胶粉,将明胶溶解于0.01M的甘氨酸缓冲溶液中,调节pH=2,依次使用胃蛋白酶、胰蛋白酶和α-糜蛋白酶各水解4h,E/S=1:100,期间不断搅拌,煮沸10min,冻干,存于-80℃备用。
实施例二:
根据实施例一提供的方法获取一种太平洋鳕鱼皮明胶肽微针,通过光学显微镜进行表征观察,测得微针侧开口结构,针长300μm,间距800μm,直径270μm,如图1。
利用物性分析仪来表征太平洋鳕鱼皮明胶肽可溶性微针的机械强度,如图2。
将健康SD雌性大鼠通过脱颈椎处死并脱毛后进行皮肤剥离,擦干水分后角质层侧朝上并固定在泡沫板上,用自制的300μm微针贴片对皮肤进行垂直按压给药30s,用1%盼蓝对穿刺后皮肤样本进行染色,如图3,对染色前和染色后皮肤进行拍照记录考察微针刺入皮肤的性能,使得微针在皮下停留不同时间间隔0s,1min,5min,10min,30min,60min,用显微镜观察可以观察在不同扎针时间间隔内微针的溶解情况,60min后微针的剩余高度最低,如图4,对微针刺入前和微针刺入后0s、30s、60s、2min、3min、4min、5min的大鼠皮肤进行拍照,观察微针刺入后5min内皮肤逐渐愈合,如图5。
上面结合附图对本发明进行了示例性描述,显然本发明具体实现并不受上述方式的限制,只要采用了本发明的方法构思和技术方案进行的各种非实质性的改进,或未经改进将本发明的构思和技术方案直接应用于其它场合的,均在本发明的保护范围之内。
Claims (10)
1.一种太平洋鳕鱼皮明胶肽微针的制备方法,其特征在于,包括:
S10:获取太平洋鳕鱼皮明胶肽和深海鱼皮明胶并混合,将混合物覆盖在微针模板表面;
S20:加热浓缩后加入透明质酸和PVA溶液脱模,得到含有太平洋鳕鱼皮明胶肽的可溶性微针。
2.根据权利要求1所述的一种太平洋鳕鱼皮明胶肽微针的制备方法,其特征在于,所述步骤S10包括:
S11:采用碱液浸泡太平洋鳕鱼皮,除去胶原蛋白外的杂质;
S12:用水清洗碱处理过的太平洋鳕鱼皮并浸泡,搅拌,提取明胶;
S13:离心处理并收集上清液,将所述上清液冷冻干燥处理,获取干物质明胶粉;
S14:将所述明胶粉溶于甘氨酸缓冲溶液中;
S15:采用胃蛋白酶、胰蛋白酶和α-糜蛋白酶水解明胶,并不断搅拌;
S16:煮沸明胶后制成冻干备用。
3.根据权利要求2所述的一种太平洋鳕鱼皮明胶肽微针的制备方法,其特征在于,所述S11中,碱液浸泡太平洋鳕鱼皮前,将太平洋鳕鱼皮清洗,去除表面粘液并剪成小块;碱液为0.5%-1.5%的Ca(OH)2溶液;浸泡时间为20h-28h;浸泡期间更换一次到五次的碱液。
4.根据权利要求2所述的一种太平洋鳕鱼皮明胶肽微针的制备方法,其特征在于,所述S12中,清洗过的太平洋鳕鱼皮采用pH为5.5到6.5的水浸泡28h-32h,水温为55℃-65℃,并持续搅拌。
5.根据权利要求2所述的一种太平洋鳕鱼皮明胶肽微针的制备方法,其特征在于,所述S13中,离心处理时,转速2800rmp-3200rmp,离心时间8min-12min。
6.根据权利要求2所述的一种太平洋鳕鱼皮明胶肽微针的制备方法,其特征在于,所述S14中,将所述明胶粉溶解于0.01M的甘氨酸缓冲溶液中,调节pH=2。
7.根据权利要求2所述的一种太平洋鳕鱼皮明胶肽微针的制备方法,其特征在于,所述S15中,依次使用胃蛋白酶、胰蛋白酶和α-糜蛋白酶各水解3.5h-4.5h,E/S=1:100,期间不断搅拌。
8.根据权利要求2所述的一种太平洋鳕鱼皮明胶肽微针的制备方法,其特征在于,所述S16中,煮沸明胶8min-12min后,制成冻干存于-75℃--85℃备用。
9.根据权利要求1所述的一种太平洋鳕鱼皮明胶肽微针的制备方法,其特征在于,所述S10中,将80mg/mL的太平洋鳕鱼明胶肽和深海鱼皮明胶1:1混合,并将混合物覆盖在微针模板表面;所述S20中,加热浓缩后加入含水量33%-37%的透明质酸;所述微针侧开口结构,针长300μm,间距800μm,直径270μm。
10.一种太平洋鳕鱼皮明胶肽微针,其特征在于,采用权利要求1到9任一项所述的方法获取。
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