CN111544361A - 一种抑制牙斑菌菌膜活性的漱口水 - Google Patents
一种抑制牙斑菌菌膜活性的漱口水 Download PDFInfo
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Abstract
本发明涉及医药配方技术领域,尤其是一种抑制牙斑菌菌膜活性的漱口水。该漱口水包括以下重量份计的原料:蜂胶醇提物、倍他环糊精、茶多酚、薄荷香精、纳米银粒子、金纳米棒吸附物、维生素D、苹果酸、酒石酸、硫酸多糖、明胶、表面活性剂、双氯苯双胍己烷、苦参超微粉和纯化水。本发明针对牙斑菌中数量最多的变异链球菌表面蛋白样参与粘附形成生物膜,通过干扰变异链球菌细胞膜功能蛋白的合成,促进药物对生物膜的渗透,减少菌体对双氯苯双胍己烷的耐药性,显著降低牙斑菌生物膜活性。
Description
技术领域
本发明涉及医药配方技术领域,尤其是一种抑制牙斑菌菌膜活性的漱口水。
背景技术
漱口水是人们口常生活中不可缺少的口腔清洁卫生用品,其基本功能在于清洁牙齿,清新口气,达到保持口腔清洁的目的。近年来,由于消费者保健意识不断增强,对漱口水提出了更多的要求,除了洁齿还增加了一些辅助预防口腔疾病,对口腔有护理保健作用的功能,因此,国内外漱口水中具有各种功能型的产品成为市场主流,又由于中药植物活性成分具有天然、安全等西药无法比拟的优点,故市场上以中药活性成分的功能型漱口水成为当今发展的重点。近年来,随着中药现代化技术的发展,对中药活性成分研究利用的方法也得到了很好发展,这为漱口水利用中药活性成分来辅助预防口腔疾病提供了很好的技术支持。但是现有技术中的漱口水缺乏针对牙斑菌特性的专门配方。如专利号为CN201810118780.8的一种含三七稀有皂苷的漱口水及其制备方法,其成分对于在形成较好生物膜的牙斑菌抑制中效果有限,对于口腔环境的深层次清洁效果并不理想;再如专利号为CN201910046403.2的一种含酸性杀菌剂的漱口水及其制备方法,其杀菌主要成分为次氯酸水溶液,稳定性不高在保存使用中操作性不高,同时杀菌方式较为单一,对于菌群复杂的口腔,要达到长效抑菌的目的有一定的难度;又如专利号为CN201710120533.7的一种含有薁磺酸钠的漱口散剂及其制备方法,其杀菌方式常规,对于口腔菌群的针对性不强,同时其采用散剂,配制时溶液的均一性稳定性难以保证,杀菌效果也会受到影响。
发明内容
为了解决现有技术中存在的上述技术问题,本发明提供一种抑制牙斑菌菌膜活性的漱口水,以重量份计,包括以下原料:蜂胶醇提物60-70份、倍他环糊精10-20份、茶多酚40-50份、薄荷香精40-50份、纳米银粒子3-5份、金纳米棒吸附物5-8份、维生素D20-28份、苹果酸20-26份、酒石酸10-20份、硫酸多糖10-18份、明胶5-8份、表面活性剂5-10份、双氯苯双胍己烷5-10份、苦参超微粉5-15份和纯化水200-350份。
进一步的,所述蜂胶醇提物的制作方法为:将蜂胶与醇溶液按照质量比1:5混合,加热到50-55℃,搅拌20-25min,收集上清液备用;在残留物中加入其质量3-5倍的醇溶液,加热至60-65℃,搅拌20-30min,取上清液;将两次上清液合并,在0.1-0.3个标准大气压、45-48℃下干燥即得所述蜂胶醇提物;所述醇溶液由质量比为5:1:3的乙醇、丙醇、水混合而得,利用不同极性溶剂的特性,提取出种类更为丰富的蜂胶醇提物。
进一步的,所述纳米银粒子尺寸为3-55nm,利用较小粒径的纳米银粒子,提高杀菌效果,保证溶液体系的稳定性。
进一步的,所述金纳米棒吸附物的制作方法为:将黄连、藿香、牛膝、艾叶按照质量比1:3:2:5混合,粉碎为200-300目的粉末,将粉末与其质量10-15倍的纯净水混合浸泡30-35min,用微波加热到40-45℃,保温5-10min,将混合溶液经400目滤布过滤,所得滤液喷雾干燥后得到提取物;将提取物在30-35℃下和纯净水配制成饱和溶液备用;在饱和溶液中加入其质量10-15%的金纳米棒,在0.3-0.4个标准大气压下加热沸腾3-5min即可;通过微波处理,破坏细胞壁,提高提取效果,利用低压下的沸腾,促进金纳米棒吸附溶液中的溶质,同时低压降低沸点,避免功能组分的分解。
进一步的,所述表面活性剂由质量比为3:1的卵磷脂、蔗糖酯混合而成。
进一步的,所述苦参超微粉的制作方法为:将苦参粉碎为80-100目的粉末,在30-35℃下经微波处理2-3min,然后将粉末置于超微粉碎机中加工成苦参超微粉,通过微波改变苦参植物细胞构型,提高超微粉碎效果。
本发明所述的抑制牙斑菌菌膜活性的漱口水,制备方法如下:
(1)将蜂胶醇提物、倍他环糊精、纯化水混合,在35-38℃下搅拌10-20min,使得蜂胶醇提物和倍他环糊精由充分的时间进行包合;
(2)将维生素D、苹果酸、酒石酸、硫酸多糖、明胶、表面活性剂、双氯苯双胍己烷、苦参超微粉依次加入步骤(1)的溶液中混合,搅拌均匀,促进溶液形成稳定体系,减少纳米粒子的聚合;
(3)将茶多酚、薄荷香精、纳米银粒子、金纳米棒吸附物依次加入步骤(2)的溶液中搅拌均匀,将溶液在30-35℃下经300-400W的超声波处理10-15min即可,通过超声波使得溶液中的各种成分可以均匀分布,提高溶液体系的稳定性。
有益效果
本发明通过双氯苯双胍己烷、苦参超微粉、纳米粒子组成杀菌广谱性的药物,利用纳米银粒子的渗透性,深入生物膜内部杀灭细菌,同时,采用金纳米棒的高比表面特性,吸附药物,使得药物通过金纳米棒深入口腔各部位,更好的粘附在菌斑上,增加药物与口腔细菌的接触时间提高杀菌效率。同时,本发明漱口液在吸附在菌斑上,使得药效不会因为时间的延长而衰减,通过纳米粒子吸附形成的多孔体系,减少杀菌药物的分解,通过纳米粒子在菌斑膜表面的渗透,有效降低菌斑生物膜的生物活性,达到杀菌的同时破坏口腔细菌的生长环境。
而且,本发明针对牙斑菌中数量最多的变异链球菌表面蛋白附属结构作为粘结素完成粘附作用,特别加强药物对变异链球菌细胞膜的干扰,影响其功能蛋白的合成。具体来说,通过纳米银粒子、金纳米棒吸附物、苦参超微粉与双氯苯双胍己烷的协同作用,利用纳米粒子具有的表面效应、小尺寸效应,纳米银粒子渗透扩散引起牙斑菌的菌体细胞膜畸形,导致菌体细胞膜表面出现凹陷,增大菌体细胞膜与药物的接触面积,提高菌体与药物中的金纳米棒吸附物、双氯苯双胍己烷、苦参超微粉等抑菌成分的接触,更大限度的发挥出药物对菌体的作用;并且,药物中的金纳米棒吸附物含有黄连、藿香、牛膝、艾叶的提取成分,纳米银粒子造成的菌体细胞膜表面形变增加了金纳米棒吸附物的附着时间,提高药物的持续作用;再通过苦参超微粉和黄连、藿香、牛膝、艾叶的提取成分改变菌体功能蛋白的表达,降低菌体的生理活性,使得菌体缢缩变形,导致生物膜结构空隙增大,利于药物进一步进入生物膜内部;通过促进牙斑菌菌体细胞的膜的异形变化,协同中药成分彻底杀灭牙斑菌,降低牙斑菌对双氯苯双胍己烷产生耐药性的概率。
本发明使用的药物可溶性高,纳米粒子细度合适,便于均匀分散在溶液中,再通过蜂胶醇提物、倍他环糊精、苹果酸、酒石酸、硫酸多糖、明胶、表面活性剂的融合,提高溶液的离子均衡度和溶液稳定性,还具有丰富口感,促进牙周炎症伤口的愈合。
具体实施方式
下面结合具体的实施方式来对本发明的技术方案做进一步的限定,但要求保护的范围不仅局限于所作的描述。
实施例1
一种抑制牙斑菌菌膜活性的漱口水,包括以下份计的原料:蜂胶醇提物60份、倍他环糊精10份、茶多酚40份、薄荷香精40份、纳米银粒子3份、金纳米棒吸附物5份、维生素D20份、苹果酸20份、酒石酸10份、硫酸多糖10份、明胶5份、表面活性剂5份、双氯苯双胍己烷5份、苦参超微粉5份和纯化水200份。所述蜂胶醇提物的制作方法为:将蜂胶与醇溶液按照质量比1:5混合,加热到50℃,搅拌20min,收集上清液备用;在残留物中加入其质量3倍的醇溶液,加热至60℃,搅拌20min,取上清液;将两次上清液合并,在0.1个标准大气压、45℃下干燥即得所述蜂胶醇提物;所述醇溶液由质量比为5:1:3的乙醇、丙醇、水混合而得。所述纳米银粒子尺寸为3nm。所述金纳米棒吸附物的制作方法为:将黄连、藿香、牛膝、艾叶按照质量比1:3:2:5混合,粉碎为200目的粉末,将粉末与其质量10倍的纯净水混合浸泡30min,用微波加热到40℃,保温5min,将混合溶液经400目滤布过滤,所得滤液喷雾干燥后得到提取物;将提取物在30℃下和纯净水配制成饱和溶液备用;在饱和溶液中加入其质量10%的金纳米棒,在0.3个标准大气压下加热沸腾3min即可。所述表面活性剂由质量比为3:1的卵磷脂、蔗糖酯混合而成。所述苦参超微粉的制作方法为:将苦参粉碎为80目的粉末,在30℃下经微波处理2min,然后将粉末置于超微粉碎机中加工成苦参超微粉。
本实施例的抑制牙斑菌菌膜活性的漱口水,制备方法如下:
(1)将蜂胶醇提物、倍他环糊精、纯化水混合,在35℃下搅拌10min;
(2)将维生素D、苹果酸、酒石酸、硫酸多糖、明胶、表面活性剂、双氯苯双胍己烷、苦参超微粉依次加入步骤(1)的溶液中混合,搅拌均匀;
(3)将茶多酚、薄荷香精、纳米银粒子、金纳米棒吸附物依次加入步骤(2)的溶液中搅拌均匀,将溶液在30℃下经300W的超声波处理10min即可。
实施例2
一种抑制牙斑菌菌膜活性的漱口水,包括以下份计的原料:蜂胶醇提物70份、倍他环糊精20份、茶多酚50份、薄荷香精50份、纳米银粒子5份、金纳米棒吸附物8份、维生素D28份、苹果酸26份、酒石酸20份、硫酸多糖18份、明胶8份、表面活性剂10份、双氯苯双胍己烷10份、苦参超微粉15份和纯化水350份。所述蜂胶醇提物的制作方法为:将蜂胶与醇溶液按照质量比1:5混合,加热到55℃,搅拌25min,收集上清液备用;在残留物中加入其质量5倍的醇溶液,加热至65℃,搅拌30min,取上清液;将两次上清液合并,在0.3个标准大气压、48℃下干燥即得所述蜂胶醇提物;所述醇溶液由质量比为5:1:3的乙醇、丙醇、水混合而得。所述纳米银粒子尺寸为55nm。所述金纳米棒吸附物的制作方法为:将黄连、藿香、牛膝、艾叶按照质量比1:3:2:5混合,粉碎为300目的粉末,将粉末与其质量15倍的纯净水混合浸泡35min,用微波加热到45℃,保温10min,将混合溶液经400目滤布过滤,所得滤液喷雾干燥后得到提取物;将提取物在35℃下和纯净水配制成饱和溶液备用;在饱和溶液中加入其质量15%的金纳米棒,在0.4个标准大气压下加热沸腾5min即可。所述表面活性剂由质量比为3:1的卵磷脂、蔗糖酯混合而成。所述苦参超微粉的制作方法为:将苦参粉碎为100目的粉末,在35℃下经微波处理3min,然后将粉末置于超微粉碎机中加工成苦参超微粉。
本实施例的抑制牙斑菌菌膜活性的漱口水,制备方法如下:
(1)将蜂胶醇提物、倍他环糊精、纯化水混合,在38℃下搅拌20min;
(2)将维生素D、苹果酸、酒石酸、硫酸多糖、明胶、表面活性剂、双氯苯双胍己烷、苦参超微粉依次加入步骤(1)的溶液中混合,搅拌均匀;
(3)将茶多酚、薄荷香精、纳米银粒子、金纳米棒吸附物依次加入步骤(2)的溶液中搅拌均匀,将溶液在35℃下经400W的超声波处理15min即可。
实施例3
一种抑制牙斑菌菌膜活性的漱口水,包括以下份计的原料:蜂胶醇提物67份、倍他环糊精17份、茶多酚44份、薄荷香精46份、纳米银粒子4份、金纳米棒吸附物6份、维生素D26份、苹果酸23份、酒石酸15份、硫酸多糖13份、明胶7份、表面活性剂8份、双氯苯双胍己烷8份、苦参超微粉12份和纯化水280份。所述蜂胶醇提物的制作方法为:将蜂胶与醇溶液按照质量比1:5混合,加热到53℃,搅拌23min,收集上清液备用;在残留物中加入其质量5倍的醇溶液,加热至60℃,搅拌30min,取上清液;将两次上清液合并,在0.1个标准大气压、48℃下干燥即得所述蜂胶醇提物;所述醇溶液由质量比为5:1:3的乙醇、丙醇、水混合而得。所述纳米银粒子尺寸为3nm。所述金纳米棒吸附物的制作方法为:将黄连、藿香、牛膝、艾叶按照质量比1:3:2:5混合,粉碎为300目的粉末,将粉末与其质量10倍的纯净水混合浸泡35min,用微波加热到40℃,保温10min,将混合溶液经400目滤布过滤,所得滤液喷雾干燥后得到提取物;将提取物在35℃下和纯净水配制成饱和溶液备用;在饱和溶液中加入其质量10%的金纳米棒,在0.4个标准大气压下加热沸腾3min即可。所述表面活性剂由质量比为3:1的卵磷脂、蔗糖酯混合而成。所述苦参超微粉的制作方法为:将苦参粉碎为100目的粉末,在30℃下经微波处理3min,然后将粉末置于超微粉碎机中加工成苦参超微粉。
本实施例的抑制牙斑菌菌膜活性的漱口水,制备方法如下:
(1)将蜂胶醇提物、倍他环糊精、纯化水混合,在36℃下搅拌20min;
(2)将维生素D、苹果酸、酒石酸、硫酸多糖、明胶、表面活性剂、双氯苯双胍己烷、苦参超微粉依次加入步骤(1)的溶液中混合,搅拌均匀;
(3)将茶多酚、薄荷香精、纳米银粒子、金纳米棒吸附物依次加入步骤(2)的溶液中搅拌均匀,将溶液在33℃下经350W的超声波处理13min即可。
对比例设置如下表:
试验例1
为了验证本发明方案的漱口水的质量,按照实施例1-3分别制作漱口水,标记为实验组1、实验组2、实验组3;按照对比例1-7操作,分别标记为对照组1-7。选择无全身系统性疾病,近期未服用抗生素的龋齿患者110名,男女各半,患者年龄在8-35岁之间,其平均年龄为24.15岁。将患者随机分为11组,分别对应实验1-3组,对照1-8组;患者取对应组漱口水每天含漱2次,早晚餐后各1次,每次10m1,含漱60s,含漱后30min内不得进食,连续使用1个月。实验期结束后,在患者2h以上未进食、漱口及刷牙,局部隔湿条件下,取患者龋齿菌斑打碎,连续倍比稀释至10-5,选择各稀释度的菌斑液0.1ml,均匀涂布在MSB培养基上,37℃培养48h。体视显微镜下分别计数菌斑内总菌落数CFU/ml。同时,分别将各组漱口水180ml盛装在200ml烧杯中,用保鲜膜封口3/4,在35℃的室内放置4个月后,观察漱口水的状态。
实验结果
由表可以看出,实验组1-3的总菌落数明显低于对照组1-7,且对照组1菌落数高于对照组2-6,所以本发明中苦参超微粉,纳米银粒子、金纳米棒吸附物协同效果明显。可见在本发明的方案之中,兼顾了漱口水的杀菌效果和稳定性,对照组1-3的配比变化中降低了溶液的稳定性,由于溶液稳定性降低进一步加剧了杀菌效果的衰减。所以本发明漱口水杀菌效果显著,并且溶液稳定,适合长时间的保存。
实验例2
选取年龄35-55之间,身体正常无其他疾病,牙菌斑明显的患者88名,将患者随机分为11组,分别对应实验1-3组,对照1-8组;实验组使用实施例1-3的漱口水,对照组使用对比例1-8的漱口水;患者取对应组漱口水每天含漱2次,早晚餐后各1次,每次10m1,含漱60s,含漱后30min内不得进食,连续使用1个月。实验前和实验后取患者牙间形成生物膜的菌斑,将菌斑倒置在滴有一滴生理盐水的盖玻片上,立即在CLSM下观察,在488nm波长激光激发下,活菌呈绿色、死菌呈红色。以图像分析软件(ZeissLSMImageBrowser,Germany)分析扫描图像,定量分析绿色及红色像素,菌斑生物膜活性(BV%)=绿荧光量/(绿荧光量+红荧光量)×100%。
实验结果:
实验前生物膜活性 | 实验后生物膜活性 | |
实验组1 | 43.92% | 8.65% |
实验组2 | 51.59% | 11.27% |
实验组3 | 33.87% | 7.80% |
对照组1 | 59.35% | 36.66% |
对照组2 | 57.90% | 31.54% |
对照组3 | 55.53% | 32.00% |
对照组4 | 47.84% | 30.42% |
对照组5 | 62.52% | 31.18% |
对照组6 | 53.84% | 32.74% |
对照组7 | 65.14% | 39.81% |
对照组8 | 55.21% | 36.42% |
由表可以看出,本发明方案中,实验组1-3的生物膜活性降低显著,试验后菌斑的生物膜活性降低显著。所以本发明漱口水对于口腔的环境的净化效果明显。
Claims (7)
1.一种抑制牙斑菌菌膜活性的漱口水,其特征在于,以重量份计,包括以下原料:蜂胶醇提物60-70份、倍他环糊精10-20份、茶多酚40-50份、薄荷香精40-50份、纳米银粒子3-5份、金纳米棒吸附物5-8份、维生素D20-28份、苹果酸20-26份、酒石酸10-20份、硫酸多糖10-18份、明胶5-8份、表面活性剂5-10份、双氯苯双胍己烷5-10份、苦参超微粉5-15份和纯化水200-350份。
2.根据权利要求1所述的抑制牙斑菌菌膜活性的漱口水,其特征在于,所述蜂胶醇提物的制作方法为:将蜂胶与醇溶液按照质量比1:5混合,加热到50-55℃,搅拌20-25min,收集上清液备用;在残留物中加入其质量3-5倍的醇溶液,加热至60-65℃,搅拌20-30min,取上清液;将两次上清液合并,在0.1-0.3个标准大气压、45-48℃下干燥即得所述蜂胶醇提物;所述醇溶液由质量比为5:1:3的乙醇、丙醇、水混合而得。
3.根据权利要求1所述的抑制牙斑菌菌膜活性的漱口水,其特征在于,所述纳米银粒子尺寸为3-55nm。
4.根据权利要求1所述的抑制牙斑菌菌膜活性的漱口水,其特征在于,所述金纳米棒吸附物的制作方法为:将黄连、藿香、牛膝、艾叶按照质量比1:3:2:5混合,粉碎为200-300目的粉末,将粉末与其质量10-15倍的纯净水混合浸泡30-35min,用微波加热到40-45℃,保温5-10min,将混合溶液经400目滤布过滤,所得滤液喷雾干燥后得到提取物;将提取物在30-35℃下和纯净水配制成饱和溶液备用;在饱和溶液中加入其质量10-15%的金纳米棒,在0.3-0.4个标准大气压下加热沸腾3-5min即可。
5.根据权利要求1所述的抑制牙斑菌菌膜活性的漱口水,其特征在于,所述表面活性剂由质量比为3:1的卵磷脂、蔗糖酯混合而成。
6.根据权利要求1所述的抑制牙斑菌菌膜活性的漱口水,其特征在于,所述苦参超微粉的制作方法为:将苦参粉碎为80-100目的粉末,在30-35℃下经微波处理2-3min,然后将粉末置于超微粉碎机中加工成苦参超微粉。
7.根据权利要求1所述的抑制牙斑菌菌膜活性的漱口水,其特征在于,制备方法如下:
(1)将蜂胶醇提物、倍他环糊精、纯化水混合,在35-38℃下搅拌10-20min;
(2)将维生素D、苹果酸、酒石酸、硫酸多糖、明胶、表面活性剂、双氯苯双胍己烷、苦参超微粉依次加入步骤(1)的溶液中混合,搅拌均匀;
(3)将茶多酚、薄荷香精、纳米银粒子、金纳米棒吸附物依次加入步骤(2)的溶液中搅拌均匀,将溶液在30-35℃下经300-400W的超声波处理10-15min即可。
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