CN114588059A - 一种口腔防蛀膏的制备方法及其应用 - Google Patents
一种口腔防蛀膏的制备方法及其应用 Download PDFInfo
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Abstract
本发明公开了一种口腔防蛀膏的制备方法及其应用，该防蛀膏由中药单体组合物和膏体基质制成。该中药单体组合物中含有厚朴酚、和厚朴酚、甘草酸、甘草甜素、甘草酸二钾、氯化两面针碱等活性成分。本发明制备所得的口腔防蛀膏质地细腻，活性成分清晰明确，中药单体组合物多成分多靶点多通路协同增效发挥作用，抑菌作用强，能够有效减少牙菌斑生物膜对牙齿的粘附，具有防蛀、抑菌、清热、消肿止痛的功效，能够封闭牙齿窝沟点隙，阻止细菌进入，有效改善牙龈出血、蛀牙等多种口腔疾病。
Description
技术领域
本发明涉及日用化学技术领域，具体涉及一种口腔防蛀膏的制备方法及其应用。
背景技术
龋齿俗称蛀牙，是一种常见的口腔疾病，发病因素与口腔卫生密切相关，细菌的存在是发病的主要原因。口腔中存在的细菌主要有葡萄球菌、链球菌、奈瑟氏菌、乳酸杆菌、痰螺旋菌、色柔氏螺旋体等。其中，龋齿的主要致病菌是乳酸杆菌和链球菌，它们可以使人体口腔中残留的碳水化合物发酵产酸。同时，细菌在牙齿表面形成一层粘附性的膜，该膜能使乳酸长时间与牙齿表面接触，最终使牙釉质中的羟基磷灰石逐渐溶解。随着牙釉质的不断损失，牙齿硬组织将不断软化、缺损，最终龋洞形成，不断扩大至牙髓，危及患者健康。目前，龋齿的治疗方法主要是非创伤性充填手术，使用器械清除牙齿龋坏组织后用离子材料填充龋洞。龋齿的预防手段主要是使用含氟牙膏，抑制菌斑生成从而预防龋齿，但含氟牙膏中未添加明确的活性成分，治疗效果不明显，临床应用上有一定的局限性。
中国医学史上对龋齿的防治早有记载，齐国时期已有大夫用针灸和汤剂治疗龋齿。据研究表明，多种中药都具有抑菌杀虫功效，中药复方能够协同发挥作用，增强药效。中药在治疗疾病上有多成分多靶点多通路协同发挥的特点，与传统的中药相比，中药单体都具有确切的活性成分，药理作用清晰，作用靶点与药效机理明确，对口腔细菌都有良好的抑制效果，毒性少，不会产生耐药性，能够长期使用，防治龋齿。
发明内容
本发明的目的是为了克服现有技术的不足，提供一种口腔防蛀膏的制备方法。本发明的另一个目的在于提供一种口腔防蛀膏在治疗龋齿中的应用。本发明的创新之处在于该防蛀膏加入了中药单体组合物，活性成分清晰明确，多组分多靶点协同发挥作用，抑菌作用强。该防蛀膏涂敷在牙齿表面能够封闭牙齿窝沟点隙，阻止细菌进入，使用方法简单快捷，用后不需漱口，产品安全有效。
为了实现上述目的，本发明是通过以下技术方案予以实现的：
一种口腔防蛀膏，按重量百分比计算，中药单体组合物1％-3％，膏体基质97％-99％。
优选的，按重量份计算，中药单体组合物包括厚朴酚3-7份、和厚朴酚4-6份、甘草酸4-7份、甘草甜素3-8份、甘草酸二钾2-6份、氯化两面针碱3-7份。
优选的，该防蛀膏还含有保湿剂、甜味剂、粘合剂、氟化护齿剂、摩擦剂、芳香剂、防腐剂。
优选的，保湿剂为甘油和丙二醇，甜味剂为木糖醇，粘合剂为羧甲基纤维素钠和卡波姆，氟化护齿剂为氟化钠，摩擦剂为二氧化硅，芳香剂为薄荷醇，防腐剂为羟苯甲酯，溶剂纯化水。
优选的，按重量百分比计算，膏体基质包括:甘油15％-30％，丙二醇20％-40％，木糖醇0.1％-0.4％，羧甲基纤维素钠3％-7％，卡波姆2％-8％，氟化钠1％-3％，二氧化硅15％-30％，薄荷醇0.5％-1％，羟苯甲酯0.3％-0.7％，余量为纯化水。
该防蛀膏的制备包括以下步骤：
(1)将中药单体组合物、甘油、丙二醇、木糖醇、氟化钠、羟苯甲酯加入到纯化水中，搅拌均匀得到混合溶液。
(2)将羧甲基纤维素钠、卡波姆、二氧化硅混合均匀得到混合物料。
(3)将混合溶液与混合物料加入真空搅拌机中，搅拌均匀至无颗粒状得到膏体。
(4)将薄荷醇加入膏体中，继续搅拌至均匀，密封分装即得口腔防蛀膏。
本发明制备的防蛀膏加入了厚朴酚、和厚朴酚、甘草酸、甘草甜素、甘草酸二钾、氯化两面针碱，活性成分清晰明确，能够预防龋齿，封闭牙齿窝沟点隙，阻断细菌进入，提高牙齿釉质的耐酸腐蚀性。将该防蛀膏涂抹到牙齿表面，能够有效抑制变异链球菌的生长，降低各细菌产酸能力，改变菌斑膜的性质，最终减少龋齿致病菌的粘附，有效防治龋齿。
厚朴酚是一种具有联苯二酚结构的木质素类化合物，具有抑菌、抗病毒、抗氧化、抗癌、镇痛抗炎、抗过敏、杀虫等多种药理活性。厚朴酚对龋齿的主要致病菌——变异链球菌，有极佳的抑菌效果，对常见真菌、耐酸菌、革兰氏阳性菌、酵母菌等也有良好的抑菌效果。厚朴酚还能通过抑制淋巴细胞芽生、抑制β-葡糖苷酸酶和溶菌酶的释放从而达到抗炎镇痛的效果。
和厚朴酚有良好的抗菌抗炎作用，对牙龈卟啉单胞菌、幽门螺杆菌、大肠埃希菌、沙门菌、产气肠杆菌等都有良好的抑菌作用，可以抑制杆菌的群体感应系统中的Luxs基因表达使AI-2分泌减少，最终降低mqsR、flhDC、flic基因表达，最终抑制生物被膜形成，产生抗菌作用；通过抑制iNOS、COX-2、白三烯的合成与释放，产生广谱抗炎作用。
甘草酸具有抗病毒、抗肿瘤、抗菌等作用，对牙龈卟啉单细胞有显著抑制作用。甘草酸能抑制细胞内钙离子浓度，改变p-ERK的表达，恢复线粒体的功能达到抗病毒作用；通过抑制NF-κB的表达诱导细胞凋亡产生抗肿瘤作用；通过激活巨噬细胞的吞噬能力达到抗菌作用，对沙门氏菌也有显著的抑制作用。
甘草甜素有良好的抑菌、提高免疫力的作用，可以通过抑制葡糖基转移酶活性影响变异链球菌生物膜的形成达到抗菌作用；通过参与细胞免疫应答反应提高人体抵御念珠菌感染的能力。
甘草酸二钾有抑菌、抗炎镇痛、抗过敏等作用，甘草酸二钾对白色葡萄球菌、金黄色葡萄球菌、大肠埃希氏菌等细菌均由抑菌作用；甘草酸二钾能影响一氧化氮、白细胞介素6、前列腺素的释放达到抗炎镇痛作用；影响HMGB1信号通路，负性调控HMGB1的生成与释放从而达到治疗疾病的效果。
氯化两面针碱有良好的抗菌、抗肿瘤、消炎止痛作用。氯化两面针碱对耐甲氧西林葡萄球菌、芽孢杆菌、化脓性链球菌等都有抑菌活性；通过调控STAT3、ERK等信号通路抑制蛋白基因表达从而使癌细胞凋亡；通过抑制MAPK磷酸化和P65的转录减少炎症因子的释放从而达到抗炎镇痛作用。
附图说明
图1为SD大鼠上下颌骨标本图。
图2为大鼠磨牙纵磨片RhB染色后荧光显微镜观察图。
图3为抑菌环对比图。
具体实施方式
下面结合具体实施例对本发明作出进一步地详细阐述，所述实施例只用于解释本发明，并非用于限定本发明的范围。下述实施例中所使用的试验方法如无特殊说明，均为常规方法；所使用的材料、试剂等，如无特殊说明，为可从商业途径得到的试剂和材料。下列各实施例中，所涉及百分比均是质量百分比计算。
实施例1
中药单体组合物3％，甘油15％，丙二醇20％，木糖醇0.2％，羧甲基纤维素钠5％，卡波姆6％，氟化钠2％，二氧化硅20％，薄荷醇0.5％，羟苯甲酯0.3％，余量为纯化水。所述中药单体组合物包括：厚朴酚3份、和厚朴酚4份、甘草酸4份、甘草甜素3份、甘草酸二钾2份、氯化两面针碱3份。
制备方法：
(1)将中药单体组合物、甘油、丙二醇、木糖醇、氟化钠、羟苯甲酯加入到纯化水中，搅拌均匀得到混合溶液。
(2)将羧甲基纤维素钠、卡波姆、二氧化硅混合均匀得到混合物料。
(3)将混合溶液与混合物料加入真空搅拌机中，搅拌均匀至无颗粒状得到膏体。
(4)将薄荷醇加入膏体中，继续搅拌至均匀，密封分装即得口腔防蛀膏。
实施例2
中药单体组合物3％，甘油20％，丙二醇25％，木糖醇0.3％，羧甲基纤维素钠6％，卡波姆7％，氟化钠2％，二氧化硅25％，薄荷醇0.8％，羟苯甲酯0.4％，余量为纯化水。所述中药单体组合物包括：厚朴酚4份、和厚朴酚4份、甘草酸5份、甘草甜素5份、甘草酸二钾5份、氯化两面针碱6份。
制备方法：
(1)将中药单体组合物、甘油、丙二醇、木糖醇、氟化钠、羟苯甲酯加入到纯化水中，搅拌均匀得到混合溶液。
(2)将羧甲基纤维素钠、卡波姆、二氧化硅混合均匀得到混合物料。
(3)将混合溶液与混合物料加入真空搅拌机中，搅拌均匀至无颗粒状得到膏体。
(4)将薄荷醇加入膏体中，继续搅拌至均匀，密封分装即得口腔防蛀膏。
实施例3
中药单体组合物2％，甘油25％，丙二醇30％，木糖醇0.3％，羧甲基纤维素钠6％，卡波姆7％，氟化钠2％，二氧化硅25％，薄荷醇0.8％，羟苯甲酯0.4％，余量为纯化水。所述中药单体组合物包括：厚朴酚5份、和厚朴酚6份、甘草酸7份、甘草甜素7份、甘草酸二钾4份、氯化两面针碱4份。
制备方法：
(1)将中药单体组合物、甘油、丙二醇、木糖醇、氟化钠、羟苯甲酯加入到纯化水中，搅拌均匀得到混合溶液。
(2)将羧甲基纤维素钠、卡波姆、二氧化硅混合均匀得到混合物料。
(3)将混合溶液与混合物料加入真空搅拌机中，搅拌均匀至无颗粒状得到膏体。
(4)将薄荷醇加入膏体中，继续搅拌至均匀，密封分装即得口腔防蛀膏。
实施例4
中药单体组合物3％，甘油30％，丙二醇20％，木糖醇0.4％，羧甲基纤维素钠7％，卡波姆3％，氟化钠1％，二氧化硅15％，薄荷醇0.6％，羟苯甲酯0.3％，余量为纯化水。所述中药单体组合物包括：厚朴酚7份、和厚朴酚6份、甘草酸7份、甘草甜素8份、甘草酸二钾5份、氯化两面针碱5份。
制备方法：
(1)将中药单体组合物、甘油、丙二醇、木糖醇、氟化钠、羟苯甲酯加入到纯化水中，搅拌均匀得到混合溶液。
(2)将羧甲基纤维素钠、卡波姆、二氧化硅混合均匀得到混合物料。
(3)将混合溶液与混合物料加入真空搅拌机中，搅拌均匀至无颗粒状得到膏体。
(4)将薄荷醇加入膏体中，继续搅拌至均匀，密封分装即得口腔防蛀膏。
实施例5
中药单体组合物2％，甘油20％，丙二醇40％，木糖醇0.2％，羧甲基纤维素钠7％，卡波姆7％，氟化钠3％，二氧化硅25％，薄荷醇0.8％，羟苯甲酯0.3％，余量为纯化水。所述中药单体组合物包括：厚朴酚4份、和厚朴酚5份、甘草酸4份、甘草甜素4份、甘草酸二钾5份、氯化两面针碱4份。
制备方法：
(1)将中药单体组合物、甘油、丙二醇、木糖醇、氟化钠、羟苯甲酯加入到纯化水中，搅拌均匀得到混合溶液。
(2)将羧甲基纤维素钠、卡波姆、二氧化硅混合均匀得到混合物料。
(3)将混合溶液与混合物料加入真空搅拌机中，搅拌均匀至无颗粒状得到膏体。
(4)将薄荷醇加入膏体中，继续搅拌至均匀，密封分装即得口腔防蛀膏。
实施例6
中药单体组合物2％，甘油23％，丙二醇26％，木糖醇0.2％，羧甲基纤维素钠7％，卡波姆7％，氟化钠3％，二氧化硅17％，薄荷醇0.8％，羟苯甲酯0.2％，余量为纯化水。所述中药单体组合物包括：厚朴酚5份、和厚朴酚5份、甘草酸4份、甘草甜素8份、甘草酸二钾5份、氯化两面针碱7份。
制备方法：
(1)将中药单体组合物、甘油、丙二醇、木糖醇、氟化钠、羟苯甲酯加入到纯化水中，搅拌均匀得到混合溶液。
(2)将羧甲基纤维素钠、卡波姆、二氧化硅混合均匀得到混合物料。
(3)将混合溶液与混合物料加入真空搅拌机中，搅拌均匀至无颗粒状得到膏体。
(4)将薄荷醇加入膏体中，继续搅拌至均匀，密封分装即得口腔防蛀膏。
实施例7
中药单体组合物2％，甘油20％，丙二醇30％，木糖醇0.2％，羧甲基纤维素钠5％，卡波姆6％，氟化钠3％，二氧化硅20％，薄荷醇0.8％，羟苯甲酯0.3％，余量为纯化水。所述中药单体组合物包括：厚朴酚4份、和厚朴酚5份、甘草酸4份、甘草甜素4份、甘草酸二钾5份、氯化两面针碱4份。
制备方法：
(1)将中药单体组合物、甘油、丙二醇、木糖醇、氟化钠、羟苯甲酯加入到纯化水中，搅拌均匀得到混合溶液。
(2)将羧甲基纤维素钠、卡波姆、二氧化硅混合均匀得到混合物料。
(3)将混合溶液与混合物料加入真空搅拌机中，搅拌均匀至无颗粒状得到膏体。
(4)将薄荷醇加入膏体中，继续搅拌至均匀，密封分装即得口腔防蛀膏。
实施例8
中药单体组合物2％，甘油17％，丙二醇28％，木糖醇0.2％，羧甲基纤维素钠6％，卡波姆8％，氟化钠2％，二氧化硅19％，薄荷醇0.8％，羟苯甲酯0.3％，余量为纯化水。所述中药单体组合物包括：厚朴酚7份、和厚朴酚4份、甘草酸4份、甘草甜素4份、甘草酸二钾3份、氯化两面针碱7份。
制备方法：
(1)将中药单体组合物、甘油、丙二醇、木糖醇、氟化钠、羟苯甲酯加入到纯化水中，搅拌均匀得到混合溶液。
(2)将羧甲基纤维素钠、卡波姆、二氧化硅混合均匀得到混合物料。
(3)将混合溶液与混合物料加入真空搅拌机中，搅拌均匀至无颗粒状得到膏体。
(4)将薄荷醇加入膏体中，继续搅拌至均匀，密封分装即得口腔防蛀膏。
为了验证本产品的性能，以具体实施例1中所制的口腔防蛀膏为例，进行如下实验测试。
一、防蛀膏对牙本质抗酸性能实验
1.实验材料
SD大鼠30只、口腔防蛀膏、体式显微镜、高速涡轮手机、电子天平、超声波清洗机、微量滴定管、冰箱、恒温水浴锅。
2.实验原理
酸性物质可以导致牙齿脱釉，导致龋齿的生成，钙离子含量是评价牙齿抗酸性能的重要指标，用EDTA络合滴定法测定牙齿中钙离子含量评价该防蛀膏对牙本质的抗酸性能。
3.实验步骤
(1)样本制备：收集大鼠磨牙，清洗干净后用高速涡轮手机磨除前磨牙近中面的牙釉质，暴露牙本质面，划分2×2mm实验开窗区，开窗区外用指甲油涂布。
(2)样本处理：将样本随机分成2组，每组60个，实验组在实验开窗区涂敷口腔防蛀膏5分钟后用去离子水清洗。对照组：在实验开窗区涂敷去离子水5分钟。
(3)钙离子浓度测定：取10个20ml的离心管，灭菌后加入10ml的脱矿液，将样品放入离心管中，分别在8h、16h、24h、32h、40h从离心管中取1ml脱矿液，用EDTA络合滴定法测定脱矿液中钙离子浓度。
(4)数据处理：用SPSS17.0统计学软件对数据进行分析。
4.实验结果
统计分析如表1所示，不同组溶出的钙离子浓度有显著性的差异(p＜0.05)。每组溶出钙离子浓度随着时间的延长而增加。与对照组相比，实验组钙离子浓度更低，说明本防蛀膏能够有效延缓酸性物质对牙齿的腐蚀。
表1不同时间点在酸蚀液中各组牙本质钙离子浓度(x-±s，n＝30)
注：*与空白对照组比较，p＜0.05
二.防蛀膏防蛀效果实验
1.实验材料
口腔防蛀膏、高露洁全面防蛀清新薄荷牙膏、SD大鼠30只、变异链球菌、BHI培养基、MSB培养基。
2.实验原理
制备大鼠龋齿模型，在体视显微镜下根据keyes评分法对大鼠磨牙龋损程度进行记分，评价该防蛀膏的防蛀效果。
3.实验步骤
(1)分组与处理：SD大鼠30只，随机编号，在大鼠口腔内接种变异链球菌，细菌定植成功后将大鼠随机分成3组，A组为实验组，B组为阳性药物对照组，C组为空白对照组。用水合氯醛麻醉大鼠后，A组每日在磨牙牙合面涂防蛀膏3分钟后用去离子水清洗，B组每日在磨牙牙合面用高露洁全面防蛀清新薄荷牙膏刷牙3分钟后用去离子水清洗，C组每日在牙合面涂去离子水，持续3个月。在此期间，每周在大鼠口腔内接种1次变异链球菌。
(2)数据测量：3个月后处死所有大鼠，截取大鼠上下颌骨，清洗晾干，在体视显微镜下根据keyes评分法对大鼠磨牙龋齿龋损程度进行记分。
4.实验结果
用spss17.0统计软件进行检测，p＜0.05为显著性差异。表2为各组大鼠磨牙平画面和窝沟龋Keyes记分表，图1为大鼠上下颌骨标本，图2为大鼠磨牙纵磨片RhB染色后荧光显微镜观察图。A组和B组均采取了防龋措施，磨损程度均远远低于C组。A组和B组相比，A组窝沟龋全层和平滑面龋釉质记分更低，说明A组的防蛀效果更佳，表明本发明制备的口腔防蛀膏防蛀效果优良。
表2各组大鼠磨牙平滑面和窝沟龋keyes记分的比较
注：E：龋损仅累及釉质；Ds：龋损累及釉质和牙本质外层1/4以内；Dm：龋损累及牙本质厚度的1/4～3/4；Dx：龋损累及深度超过牙本质厚度的3/4
三.口腔防蛀膏抑菌效果实验
1.实验材料
口腔防蛀膏、高露洁全面防蛀清新薄荷牙膏、变异链球菌、脑心浸出液肉汤培养基、脑心浸出液琼脂培养基、脱纤维羊血、生化培养箱、游标卡尺。
2.实验原理
变异链球菌是龋病发病的主要致病菌，本实验通过抑菌环直径测量法评价该防蛀膏的抑菌效果。
3.实验步骤
(1)菌种复苏与增菌：取出干菌种管，加入少量含脱纤维羊血的脑心浸出液肉汤培养基至菌种融化分散为止。取10ml含脱纤维羊血的脑心浸出液肉汤培养基于试管中，滴加菌液，放入生化培养箱中培养72h。
(2)制备血琼脂培养基：取24g脑心浸出液琼脂固体、0.2g半胖氨酸和500ml蒸馏水于锥形瓶中，121℃高压蒸汽灭菌20min，冷却后加20ml脱纤维羊血，50℃水浴备用。
(3)制备细菌混悬液：在生化培养箱中取出变异链球菌，加入PBS浊度管中，用移液枪逐步稀释，在浊度仪上调节菌液浓度，直至细菌混悬液浓度为1.5×108CFU/ml。
(4)药液的制备：分别取50mg口腔防蛀膏、50mg高露洁全面防蛀清新薄荷牙膏于锥形瓶中，加入10ml去离子水稀释，溶解完全后得到药液。
(5)制备实验平板：取出血琼脂培养基，在血琼脂平板上打3个直径为6mm的小孔，分别加入1ml口腔防蛀膏药液、1ml高露洁全面防蛀清新薄荷牙膏药液，1ml去离子水，重复3个平板。将平板放入生化培养箱中培养，72h后用游标卡尺测量抑菌圈直径。1号小孔添加口腔防蛀膏药液作为实验组，2号小孔添加高露洁全面防蛀清新薄荷牙膏药液作为阳性药物对照组，3号小孔添加去离子水作为空白对照组。
4.实验结果
采用SPSS21.0统计学软件处理数据，计量资料用x-±s表示，用t检验，p＜0.05为差异具有统计学意义。抑菌圈直径大于7mm均可视为有抑菌效果，如表3与图3抑菌环对比图所示，口腔防蛀膏与高露洁全面防蛀清新薄荷牙膏对变异链球菌均有明显的抑菌效果，口腔防蛀膏的抑菌效果更加优良。
表3各药液对变异链球菌抑菌环直径分析(x-±s，mm)
注：与阳性药物对照组相比较，*P＜0.01
上述实施例为本发明较佳的实施方式，但本发明的实施方式并不受上述实施例的限制，其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本发明的保护范围之内。
Claims (5)
1.一种口腔防蛀膏的制备方法及其应用，其特征在于，所述口腔防蛀膏按重量百分比计算，由中药单体组合物1％-3％和膏体基质97％-99％组成。
2.如权利要求1所述的一种口腔防蛀膏的制备方法及其应用，其特征在于，所述中药单体组合物由以下重量份的原料组成：厚朴酚3-7份、和厚朴酚4-6份、甘草酸4-7份、甘草甜素3-8份、甘草酸二钾2-6份、氯化两面针碱3-7份。
3.如权利要求1所述的一种口腔防蛀膏的制备方法及其应用，其特征在于，所述膏体基质由以下重量百分比的原料组成：甘油15％-30％，丙二醇20％-40％，木糖醇0.1％-0.4％，羧甲基纤维素钠3％-7％，卡波姆2％-8％，氟化钠1％-3％，二氧化硅15％-30％，薄荷醇0.5％-1％，羟苯甲酯0.3％-0.7％，余量为纯化水。
4.如权利要求1所述的一种口腔防蛀膏的制备方法及其应用，其特征在于，该防蛀膏的制备包括以下步骤：
(1)将中药单体组合物、甘油、丙二醇、木糖醇、氟化钠、羟苯甲酯加入到纯化水中，搅拌均匀得到混合溶液。
(2)将羧甲基纤维素钠、卡波姆、二氧化硅混合均匀得到混合物料。
(3)将混合溶液与混合物料加入真空搅拌机中，搅拌均匀至无颗粒状得到膏体。
(4)将薄荷醇加入膏体中，继续搅拌至均匀，密封分装即得口腔防蛀膏。
5.如权利要求1所述的一种口腔防蛀膏在防治蛀牙以及牙龈炎症中的应用。
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