CN117462428A - 精氨酸与含氟生物活性玻璃的组合物以及防龋口腔护理产品 - Google Patents
精氨酸与含氟生物活性玻璃的组合物以及防龋口腔护理产品 Download PDFInfo
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- CN117462428A CN117462428A CN202311473795.3A CN202311473795A CN117462428A CN 117462428 A CN117462428 A CN 117462428A CN 202311473795 A CN202311473795 A CN 202311473795A CN 117462428 A CN117462428 A CN 117462428A
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- Prior art keywords
- fluorine
- arginine
- bioactive glass
- containing bioactive
- oral care
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Abstract
本发明公开了精氨酸与含氟生物活性玻璃的组合物以及防龋口腔护理产品,精氨酸与含氟生物活性玻璃的组合提高了抑制釉质在酸性条件脱矿的能力,促进了再矿化,同时增强了对致龋菌如变形链球菌的抑制效果,表现出了协同增强的致龋效果,并且降低了材料剂量,增加了生物安全性。本发明具有以下优势:精氨酸促进了含氟生物活性玻璃里氟的可控释放及利用;具备更好的抗菌效果及抑制脱矿效果;节约成本、降低剂量、增加生物安全性;优化使用口感;达到长效缓释可控防龋。
Description
技术领域
本发明涉及一种口腔护理产品,具体涉及精氨酸与含氟生物活性玻璃的组合物以及防龋口腔护理产品。属于口腔护理技术领域。
背景技术
全世界龋病高发,我国5岁儿童患龋率高达70%,龋病是在以细菌为主的多种因素作用下,牙体硬组织发生的慢性、进行性破坏的一种疾病。牙体组织的脱矿及再矿化一直伴随在龋病的发生发展过程中,龋病的发生是脱矿与再矿化失去平衡所致,当脱矿占据主导时,龋病形成。因此,一种好的防龋材料应能抑制致龋菌、抑制釉质脱矿。
以往的研究大多用氟化物进行防龋,含氟牙膏含有1450ppm的可溶性氟(氟化钠或单氟磷酸钠)能降低患龋率,可溶性氟化物的浓度通过唾液稀释,迅速降低。通常情况下,其浓度每分钟会减半,所以在90分钟左右,氟的浓度就无法达到治疗浓度。常用的口腔护理产品面临可溶性氟产品氟化物释放快,无法达到可控缓释,低浓度的氟化物防龋效果较差,高浓度的氟可能引起氟斑牙等慢性氟中毒。
L-精氨酸是一种天然存在的氨基酸,已知其参与人体内若干代谢过程,唾液中也能检测到精氨酸的存在。不仅作为蛋白质合成的前提,还用作尿素、多胺、谷氨酸、肌酸和胍丁胺的合成前体。精氨酸可作为底物参与细菌代谢过程,从而提高口腔微环境的pH值,抑制致龋菌生长,有利于防止龋病发生。但精氨酸释放较快,无法实现长效防龋的效果。
生物活性玻璃具备良好的生物安全性及生物活性,可在口内降解形成羟基磷灰石,将氟添加到生物活性玻璃结构中形成的含氟生物活性玻璃可降解,可控释放离子,可形成氟磷灰石,氟磷灰石相对于羟基磷灰石来说具备更好的耐酸度,在酸性条件下可降低牙齿釉质表面的溶解度,氟磷灰石的耐酸侵蚀能力是羟基磷灰石的10倍。氟的可控释放是随着微小的玻璃颗粒与唾液接触、溶解而进行的。而传统的口腔护理产品为游离的氟离子或生物活性玻璃产品里添加游离的氟离子,释放较快,作用时间短。含氟生物活性玻璃在口腔中可发生降解,提高口腔pH值,可控释放氟、钙和磷离子、进而形成氟磷灰石,增加釉质显微硬度,具备矿化特性。但现有的含氟生物活性玻璃抑制致龋菌效果较差,需要较高浓度才能有抑菌效果,而较高浓度的含氟生物活性玻璃可能产生慢性氟中毒等副作用,且口腔护理产品中使用的生物活性玻璃粒径常大于38μm,口腔护理产品使用过程中沙砾感较明显,影响口感。
发明内容
本发明的目的是为克服上述现有技术的不足,提供精氨酸与含氟生物活性玻璃的组合物以及防龋口腔护理产品。
为实现上述目的,本发明采用下述技术方案:
1、精氨酸与含氟生物活性玻璃的组合物,两者的质量比为1:2~2:1,所述精氨酸为L-精氨酸,所述含氟生物活性玻璃的粒径小于38μm。
优选的,精氨酸与含氟生物活性玻璃的质量比为1:1。
优选的,所述含氟生物活性玻璃是以CaF2、SiO2、P2O5、CaCO3为原料,采用高温熔融法制备得到。
进一步优选的,以摩尔百分占比计,各原料组成如下:CaF2 0.5~8%,SiO2 20~50%,P2O5 2~10%,余量为CaCO3。
进一步优选的,具体制备方法如下:将各原料充分混匀后,加热至1400~1500℃,熔融1小时,倒入去离子水中冷却,收集玻璃熔块,干燥,研磨,过38μm筛网,即得所述的含氟生物活性玻璃。
2、前述组合物在制备防龋口腔护理产品中的应用。
3、一种防龋口腔护理产品,含有前述组合物。
优选的,在所述口腔护理产品中,精氨酸、含氟生物活性玻璃的质量含量分别为0.05~12wt%、0.05~12wt%。
优选的,所述口腔护理产品还包含甘油、羟乙基纤维素、山梨醇、月桂酸硫酸酯钠、水。
优选的,所述口腔护理产品还包含载体,所述载体包括乳化剂、增稠剂、调味剂、填充剂等。
进一步优选的,所述乳化剂包括月桂醇硫酸酯钠、月桂酰肌氨酸钠等。
进一步优选的,所述增稠剂可包含一种或多种多糖。多糖的一些非限制示例包括:淀粉、琼脂、明胶、果胶、羧甲基纤维素、羟乙基纤维素、羟丙基纤维素等纤维素化合物。
进一步优选的,所述调味剂选自:薄荷醇、肉桂、水杨酸甲酯、桂皮、香兰素等。
进一步优选的,所述填充剂包括湿润剂、防腐剂等,其中,所述湿润剂选自甘油、水、聚乙二醇、木糖醇、山梨醇、乳糖醇等;所述防腐剂选自对羟基苯甲酸甲酯、苯甲酸和苯甲酸钠等。
优选的,以质量百分比计,所述口腔护理产品是由以下组分混合制成的:精氨酸0.05~12wt%,含氟生物活性玻璃0.05~12wt%,乳化剂1~4wt%,增稠剂0.5~4wt%,调味剂0.5~4wt%,余量为填充剂。
例如:以所述口腔护理组合物总重量计,所述精氨酸或含氟生物活性玻璃可以为0.05、0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1.0、1.1、1.2、1.3、1.4、1.5、1.6、1.7、1.8、1.9、2.0、2.1、2.2、2.3、2.4、2.5、2.6、2.7、2.8、2.9、3.0、3.1、3.2、3.3、3.4、3.5、3.6、3.7、3.8、3.9、4.0、4.1、4.2、4.3、4.4、4.5、4.6、4.7、4.8、4.9、5.0、5.1、5.2、5.3、5.4、5.5、5.6、5.7、5.8、5.9、6.0、6.1、6.2、6.3、6.4、6.5、6.6、6.7、6.8、6.9、7.0、7.1、7.2、7.3、7.4、7.5、7.6、7.7、7.8、7.9、8.0、8.1、8.2、8.3、8.4、8.5、8.6、8.7、8.8、8.9、9.0、9.1、9.2、9.3、9.4、9.5、9.6、9.7、9.8、9.9、10.0、10.1、10.2、10.3、10.4、10.5、10.6、10.7、10.8、10.9、11.0、11.1、11.2、11.3、11.4、11.5、11.6、11.7、11.8、11.9、12.0wt%或其之间的任意范围。
优选的,所述口腔护理产品包括但不限于:牙膏,漱口水,牙贴,凝胶,胶囊。
本发明的有益效果:
本发明公开了精氨酸与含氟生物活性玻璃的组合物,两者的组合提高了抑制釉质在酸性条件脱矿的能力,促进了再矿化,同时增强了对致龋菌如变形链球菌的抑制效果,表现出了协同增强的致龋效果,并且降低了材料剂量,增加了生物安全性。本发明具有以下优势:精氨酸促进了含氟生物活性玻璃里氟的可控释放及利用;具备更好的抗菌效果及抑制脱矿效果;节约成本、降低剂量、增加生物安全性;优化使用口感;达到长效缓释可控防龋。
本发明的组合物可进一步制备获得防龋口腔护理产品,用于:
①减少或抑制龋齿的形成;
②减少、修复或抑制牙釉质的损伤;
③减少或抑制牙齿的脱矿化并促进牙齿的再矿化;
④减少牙齿的超敏性;
⑤降低产酸细菌的水平;
⑥减少或抑制口腔中微生物生物膜形成;
⑦促进全身健康;
⑧清洁牙齿和口腔。
本发明的工作原理如下:
精氨酸的正电荷基团与含氟生物活性玻璃的氟离子键合,促进含氟生物活性玻璃里氟的释放,提高氟在釉质表面的摄取,抑制变形链球菌等致龋菌的生长及生物膜的形成,促进钙磷离子在釉质表面沉积,形成氟磷灰石,氟磷灰石与羟基磷灰石相比,具备更低的溶解度,增强釉质表面的抗酸能力,氟磷灰石的耐酸侵蚀能力是羟基磷灰石的10倍。氟的可控释放是随着微小的玻璃颗粒与唾液接触、溶解而进行的,作用时间可长达约10小时。所以能进一步抑制釉质脱矿,促进脱矿釉质的再矿化。本发明避免了其他钙磷再矿化技术中钙、磷离子与氟离子联合使用时溶解度过低的难题,可产生较好的协同作用。同时在较低浓度时就可产生较好的抑制致龋菌和抑制脱矿的效果。
本发明采用的粒径<38μm的含氟生物活性玻璃与精氨酸进行联合,有较好的生物安全性及使用口感体验,达到长效缓释可控防龋,可为儿童龋病预防提供一个新的思路,可形成牙膏、漱口水、牙贴、凝胶和胶囊多种载体形式。
附图说明
图1为各治疗组样品失重率(注:***代表其他治疗组与对照组相比有显著统计学差异,(p<0.0001),A-G组组间无统计学差异)。
图2为各治疗组SEM观察釉质表面形貌(10000×,Normal代表正常釉质块抛光后)。
图3为各治疗组溶液里氟离子浓度(注:a-e代表各组间有显著统计学差异,p<0.05)。
图4为各治疗组对变形链球菌4h抑制效果。
图5为各治疗组对变形链球菌2h抑制效果,其中,A为精氨酸1.5wt%,B为精氨酸:含氟生物活性玻璃=3:1(精氨酸1.125wt%+含氟生物活性玻璃0.375wt%),C为精氨酸:含氟生物活性玻璃=1:1(精氨酸0.75wt%+含氟生物活性玻璃0.75wt%),D为精氨酸:含氟生物活性玻璃=1:3(精氨酸0.375wt%+含氟生物活性玻璃1.125wt%),E为含氟生物活性玻璃1.5wt%。
图6为各治疗组对变形链球菌24h抑制效果。
图7为SEM观察各治疗组对变形链球菌生物膜的影响,其中,A为对照组,B为精氨酸1.5wt%,C为含氟生物活性玻璃1.5wt%,D为精氨酸1.5wt%+含氟生物活性玻璃1.5wt%,E为精氨酸0.75wt%+含氟生物活性玻璃0.75wt%。
具体实施方式
下面结合附图和实施例对本发明进行进一步的阐述,应该说明的是,下述说明仅是为了解释本发明,并不对其内容进行限定。
除非特别规定,否则本文及本说明书中其他处表达的所有百分比和量均应理解为重量百分比,给定的量是基于材料的有效重量计算。
参考实施例
一种含氟生物活性玻璃,采用高温熔融法应用马弗炉(Carbolite.Gero,英国)进行烧制,包含6mol%的CaF2(Alfa Aesar,中国),35.9mol%的SiO2(上海国药,中国),5.9mol%的P2O5(Sigma,美国),其余为CaCO3(Sigma,美国)。按照配方比,称取分析级SiO2、P2O5、CaCO3、CaF2,充分混匀后倒入铂铑坩埚中,在马弗炉中加热至1500℃,熔融1小时后,迅速将熔融的玻璃倒入去离子水中冷却,将收集到的玻璃熔块在60℃的恒温鼓风干燥机中干燥过夜。称取100g玻璃于行星球研磨仪中研磨2个7分钟(转速设置为350rmp),随后使用过滤筛震动过滤20分钟后,收集直径小于38μm的含氟生物活性玻璃微粒用于后续应用及研究。
实施例1:
(1)样本收集
取口腔颌面外科门诊因阻生或正畸新鲜拔除(1个月内)的第三磨牙一百颗(获得伦理审批且取得患者同意),离体牙纳入标准:牙面无龋坏、无裂纹、无形态及色泽异常、无色素沉着,将离体牙放进体积浓度75%酒精中4℃保存备用。
(2)实验设计
建立牙釉质脱矿模型:牙体用低速切割机在流动水冷却下制备小釉质块(釉质厚度约1mm,长宽约3-4mm),另制备大釉质块包埋于树脂中,抛光釉质呈镜面,超声冲洗,去离子水漂洗后储存于4℃冰箱备用。将釉质块浸泡于脱矿液(乙酸溶液pH=4.0)24h得到脱矿离体牙模型。
(3)实验分组及处理
将牙釉质块随机分成8组,试验组:以采用上述精氨酸+含氟生物活性玻璃质量比1:1制得的组合物+脱矿液(乙酸溶液pH=4.0)处理;阴性对照组为采用脱矿液(乙酸溶液pH=4.0)处理;阳性对照组采用单纯精氨酸组合物(不含含氟生物活性玻璃)和单纯含氟生物活性玻璃组合物(不含精氨酸)+脱矿液(乙酸溶液pH=4.0)处理。脱矿24h后将釉质样品用含去离子水冲洗后浸泡于上述处理液中96h。具体实例实施见表1。
表1.实施例1实例实施
处理及测试方法:小釉质块在脱矿前、脱矿后、含治疗溶液处理后分别用天平称取釉质块重量,大釉质块在处理完毕后冲洗、干燥、喷金,使用扫描电镜SEM观察釉质表面形貌,并用氟离子选择电极测量溶液里氟含量。小釉质块重量损失如图1所示:
根据上述结果可以看出,D、G组氟离子浓度分别比C、F组高(图3),说明精氨酸可以促进组合物里氟离子释放,从而提高氟的利用度,促进氟磷灰石的形成,提高釉质表面抗酸能力;釉质重量经A、D、G组处理后重量没有进一步损失,且得到了部分恢复(图1),说明促进了釉质再矿化,A组在较低的浓度就取得了与其他组一样的抑制脱矿效果;A、D、G组处理后牙釉质表面形貌变平坦,可见大量晶体沉积,牙釉质表面相对均匀、光滑、平坦,而B、C、E、F组牙釉质表面相对较松散,H组釉质表面呈蜂窝状(图2),表明A、D、G组相对于单独精氨酸及含氟生物活性玻璃组具备更优的抑制脱矿效果。
实施例2:
研究组合物对致龋菌变形链球菌的抑制效果,试验组:以采用上述精氨酸与含氟生物活性玻璃质量比1:1制得的组合物处理;阴性对照组为BHI培养基溶液(脑心浸液,Oxoid,美国)处理;阳性对照组采用单纯精氨酸组合物(不含氟生物活性玻璃)和单纯含氟生物活性玻璃组合物(不含精氨酸)处理。
处理方法:分别采取Alamarblue评估组合物抑制变形链球菌2h、4h的效果,涂布平板评估抑制变形链球菌24h的效果,通过扫描电镜SEM观察组合物对变形链球菌24h生物膜形成的影响。
变形链球菌(ATCC 25175),购买自中国普通微生物菌种保藏管理中心(CGMCC)。
Alamarblue评估组合物抑制变形链球菌2h、4h的效果方法如下:将变形链球菌对数期菌液稀释至106CFU/mL备用,在96孔板每孔中加入100μL菌液,随后加入100μL材料悬浊液同时设置空白对照组(菌液+BHI培养基),在厌氧条件下((85% N2、10% H2、5%CO2)37℃培养2h及4h后,每孔加入20μL Alamarblue,避光孵育30min。4500rpm下离心5min,每孔吸取100μL的上清液转移至新的96孔板,测量在570nm和600nm波长下的吸光度。与空白对照组比较,计算得到细菌生长抑菌率。
涂布平板评估抑制变形链球菌24h的效果方法如下:将106CFΜ/mL的对数期变形链球菌菌液与材料充分混匀后,置于37℃厌氧箱中的摇床上(100rpm/min)共培养24h,同时设置空白对照组(未加材料的菌液)。共培养24h结束时,将所有样品重复吸取100μL转移到另一个含有900μL BHI培养基的EP管中,制备10倍梯度的连续稀释的样品。取100μL的菌悬液用玻璃三角棒均匀涂布于BHI琼脂平板上,在37℃的厌氧培养箱中培养。培养48h后取出平板拍照,通过菌落计数仪统计平板上的菌落数量计算得到原始菌液浓度。
扫描电镜SEM观察组合物对变形链球菌24h生物膜形成的影响方法:将对数期变形链球菌菌液0.2ml加入至含细胞爬片的12孔板中,加入1ml材料悬浊液,并补充0.8mlBHI培养基,同时设置空白对照组(未加材料的菌液)。在厌氧条件下37℃静置培养24h。培养后用磷酸盐缓冲溶液缓慢冲洗2次,再向每孔中加入2ml2.5%戊二醛过夜固定。分别用一系列乙醇(50%、70%、80%、90%和100%)梯度分级脱水,每次脱水时间为15min。真空冷冻干燥沉淀24h,放入离子喷射仪中喷金,并使用扫描电子显微镜观察。
精氨酸组能在4h、24h起到较好的抑菌效果,而含氟生物活性玻璃抑菌效果较差(图4、图6),精氨酸+含氟生物活性玻璃在抑制变形链球菌时起到了最佳效果,说明二者协同改善抑菌效果。
由图5可知,不同比例的精氨酸与含氟生物活性玻璃可产生不同的抑菌效果,其中精氨酸与含氟生物活性玻璃质量比为1:1时在2h可产生对变形链球菌最佳的抑菌效果。
由图7可知,在变形链球菌生物膜形成方面,与对照组(图7中A)相比,精氨酸(图7中B)无明显效果,含氟生物活性玻璃(图7中C)可以抑制生物膜形成,二者联合可以明显降低变形链球菌生物膜形成,生物膜形成的量最少(图7中D、E),表明精氨酸与生物活性玻璃联合组相对于单独精氨酸及含氟生物活性玻璃组具备更优的抗菌效果。
上述结果表明精氨酸与生物活性玻璃联合具备更优的防龋功效。
上述虽然结合附图对本发明的具体实施方式进行了描述,但并非对本发明保护范围的限制,在本发明的技术方案的基础上,本领域技术人员不需要付出创造性劳动即可做出的各种修改或变形仍在本发明的保护范围以内。
Claims (10)
1.精氨酸与含氟生物活性玻璃的组合物,其特征在于,两者的质量比为1:2~2:1,所述精氨酸为L-精氨酸,所述含氟生物活性玻璃的粒径小于38μm。
2.根据权利要求1所述的组合物,其特征在于,所述含氟生物活性玻璃是以CaF2、SiO2、P2O5、CaCO3为原料,采用高温熔融法制备得到。
3.根据权利要求2所述的组合物,其特征在于,以摩尔百分占比计,各原料组成如下:CaF2 0.5~8%,SiO2 20~50%,P2O5 2~10%,余量为CaCO3。
4.根据权利要求2所述的组合物,其特征在于,具体制备方法如下:将各原料充分混匀后,加热至1400~1500℃,熔融1小时,倒入去离子水中冷却,收集玻璃熔块,干燥,研磨,过38μm筛网,即得所述的含氟生物活性玻璃。
5.权利要求1所述组合物在制备防龋口腔护理产品中的应用。
6.一种防龋口腔护理产品,其特征在于,含有权利要求1所述组合物。
7.根据权利要求6所述的一种防龋口腔护理产品,其特征在于,在所述口腔护理产品中,精氨酸、含氟生物活性玻璃的质量含量分别为0.05~12wt%、0.05~12wt%。
8.根据权利要求6所述的一种防龋口腔护理产品,其特征在于,所述口腔护理产品还包含甘油、羟乙基纤维素、山梨醇、月桂酸硫酸酯钠、水。
9.根据权利要求6所述的一种防龋口腔护理产品,其特征在于,所述口腔护理产品还包含载体,所述载体包括乳化剂、增稠剂、调味剂、填充剂。
10.根据权利要求6所述的一种防龋口腔护理产品,其特征在于,所述口腔护理产品包括但不限于:牙膏,漱口水,牙贴,凝胶,胶囊。
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