CN115518005B - 一种口腔护理凝胶及其制备方法 - Google Patents
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Abstract
本申请涉及口腔护理的技术领域，具体公开了一种口腔护理凝胶及其制备方法。一种口腔护理凝胶，包括以下重量百分比的原料：巴戟天多糖提取物2‑5%、凝胶0.5‑1%、奥硝唑1‑3%；其制备方法为：将巴戟天多糖提取物、凝胶、奥硝唑、水按配比混合均匀，包装，得到口腔护理凝胶。本申请的口腔护理凝胶可用于口腔牙周炎的治疗与预防，其具有较好的抗炎症效果，并且可以减少牙槽骨吸收，促进牙槽骨生长。
Description
技术领域
本申请涉及口腔护理的技术领域，更具体地说，它涉及一种口腔护理凝胶及其制备方法。
背景技术
口腔清理不规范，龈牙结合部堆积的牙菌斑及其中的有害物质长期作用于牙龈，引起炎症，而如果炎症未能及时治疗，炎症可由牙龈向深层扩散到牙周膜、牙槽骨和牙骨质而发展为牙周炎，由于炎症的扩展，牙周膜被破坏，牙槽骨逐渐吸收，牙龈与牙根分离，使龈沟加深而形成牙周袋，牙周组织被破坏，特别是牙槽骨吸收加重时，支持牙齿力量不足，出现牙齿松动、移位等现象。因此口腔护理凝胶尤为重要。
目前，市面上的口腔护理凝胶多通过使用添加如硝唑类、咪唑类衍生物、四环素类等抗生素药物来达到抗菌消炎作用，但是市面上的口腔护理凝胶均没有减少牙槽骨吸收、促进牙槽骨再生的作用，即使杀灭牙周炎细菌，牙槽骨的损伤也不能修复。
发明内容
为了减少牙槽骨吸收、促进牙槽骨再生，本申请提供一种口腔护理凝胶及其制备方法。
第一方面，本申请提供一种口腔护理凝胶，采用如下的技术方案：
一种口腔护理凝胶，包括以下重量百分比的原料：巴戟天多糖提取物2-5％、凝胶0.5-1％、奥硝唑1-3％。
通过采用上述技术方案，巴戟天是由茜草科植物巴戟天的干燥根炮制而成，性味甘、辛、微温，归肝、肾经，为我国“四大南药”之一，其有抗炎镇痛、抗肿瘤、抗抑郁、增强免疫、抗骨质疏松等作用。
多糖为巴戟天的重要有效成分之一，巴戟天多糖在体外能够通过诱导NO和炎性因子的表达，下调免疫抑制性细胞因子发挥免疫刺激作用；在体内则通过调节脾脏免疫细胞数量，特别是增加干扰素-γ(Interferon-γ，IFN-γ)阳性的自然杀伤细胞(Naturalkiller cells，NK)数量诱导免疫刺激，激活先天性和适应性免疫应答。尤其在免疫应答不足的情况下，巴戟天能发挥有益的免疫调节作用。然而在病原微生物感染情况下，巴戟天抑制LPS诱导的THP-1巨噬细胞分泌TNF-α，IL-1β等炎性因子、降低感染部位的炎性浸润、以及通过协调辅助型T细胞Th1、Th17、Th22等介导的细胞因子抑制免疫反应，恢复免疫平衡，减轻过度免疫应答造成的组织损伤。在口腔护理凝胶中加入巴戟天多糖提取物，对牙菌斑所导致的口腔慢性炎症及自身免疫性牙槽骨吸收具有预防和治疗作用，原材料价廉易取得、无毒副作用，是一种理想的纯中药制剂的口腔护理凝胶。
优选的，所述巴戟天多糖提取物的重量百分比为3-4％。
通过采用上述技术方案，进一步优化巴戟天多糖提取物在口腔护理凝胶中的配比，优化口腔护理凝胶的性能。
优选的，所述巴戟天多糖提取物的制备方法为：
1)预处理
将巴戟天用乙醇回流浸提2-3，过滤，滤渣干燥备用；
2)热水浸提
将步骤1)得到的滤渣用80-90℃的水回流提取2-3次，提取结束后，抽滤、离心，取上清，即为巴戟天多糖粗提液；
3)醇沉
将步骤2)所得多糖粗提液减压浓缩成黏稠浸膏状，然后加入乙醇使溶液含醇量为60-70％，混合均匀，在4-5℃下醇沉12-14h，离心，收集沉淀物，冷冻干燥得到巴戟天多糖。
通过采用上述技术方案，先用醇提对巴戟天进行预处理，然后再用热水进行浸提，最后再进行醇沉，醇提、热水浸提、醇沉相互配合，提高巴戟天多糖的提取率。
优选的，所述步骤2)中滤渣与水的配比为：1g滤渣对应10-15ml水。
通过采用上述技术方案，滤渣与水的料液比会影响巴戟天多糖的提取率，在本申请限定的料液比内巴戟天多糖提取物的提取率达到最大水平，料液比过高或过低均会导致提取率降低。
优选的，所述凝胶为海藻酸钠水凝胶，其制备方法为：
将10-15重量份海藻酸钠加入氯化钠溶液中，混合均匀，然后在45-50℃下处理30-40min，然后静置10-12h，得到混合液；向混合液中加入丙烯酰胺，混合均匀，并加入0.1-0.2重量份过硫酸铵、0.003-0.005重量份四甲基乙二胺、0.09-0.15重量份N，N-亚甲基双丙烯酰胺，然后在0-3℃下搅拌15min；离心、干燥，得到海藻酸钠水凝胶。
通过采用上述技术方案，制备的海藻酸钠凝胶，具有多孔结构，且空隙大小不一，它允许营养物质的扩散和细胞内生长，以促进牙周组织再生中的血管生成和成骨，同时良好的空隙也可以容纳药物颗粒镶嵌其中，维持药物的释放。另外，制得的海藻酸钠凝胶还可以抵消奥硝唑对细胞生长的抑制作用，促进细胞增殖，促进牙槽骨再生。
优选的，所述海藻酸钠水凝胶的制备方法为：
将10-15重量份海藻酸钠加入氯化钠溶液中，混合均匀，然后在45-50℃下处理30-40min，然后静置10-12h，得到混合液；向混合液中加入丙烯酰胺，混合均匀，并加入0.1-0.2重量份过硫酸铵、0.003-0.005重量份四甲基乙二胺、0.09-0.15重量份N，N-亚甲基双丙烯酰胺，然后在0-3℃下搅拌15min；然后再加入0.1-0.2重量份过氧化钙粒子，混合均匀，然后离心、干燥，得到海藻酸钠水凝胶。
口腔中，血管网络是组织内细胞的唯一氧来源，而组织内氧弥散的距离短，口腔凝胶用于口腔内伤患处时，会阻止氧弥散，导致口腔内部分区域严重缺氧，缺氧会导致细胞死亡、组织坏死，进而影响骨缺损的愈合或再生。通过采用上述技术方案，在口腔凝胶中添加过氧化钙颗粒，过氧化钙是常用的固体释氧材料之一，相比于过氧化氢，过氧化钙释氧速度缓慢且持久，因此不易造成局部活性氧的积累而破坏氧化还原平衡，维持氧的有效浓度，提高干细胞的存活率，减少缺氧诱导的细胞凋亡，促进牙槽骨再生。
优选的，在加入所述过氧化钙粒子时，随过氧化钙粒子加入0.15-0.3重量份抗坏血酸。
过氧化钙遇水反应生成过氧化氢及氢氧化钙，前者为强氧化剂，继续分解为氧气和水；后者是强碱。因此，如果二者产生过多会破坏周围细胞的氧化还原平衡及酸碱平衡，导致细胞死亡，影响骨再生。通过采用上述技术方案，添加抗坏血酸，以便克服过氧化钙遇水释放氧气过程中产生过多的活性氧及碱性物质。保证细胞的正常生长，促进牙槽骨再生。
优选的，所述过氧化钙粒子的粒径为10-20nm。
通过采用上述技术方案限定的过氧化钙粒径，使得过氧化钙颗粒不易凝结，反应速度快；若过氧化钙颗粒过大，遇水容易凝结成块，反应速度过于缓慢；若过氧化钙颗粒过大，则容易团聚。
优选的，其还包括0.4-1％的纤连蛋白。
通过采用上述技术方案，纤连蛋白存在于多种动物细胞表面的大分子细胞外膜蛋白，是细胞外基质和基底膜中的主要非胶原性糖蛋白，是在人体所有组织中发现的进化上保守的糖蛋白，可以充当三维支架，指导其他细胞外基质组件的组装，并通过整联蛋白结合域和生长因子结合域调节细胞行为，促进下游反应，促进细胞募集，增殖和分化，口腔凝胶中的纤连蛋白可以促进牙槽骨细胞的增殖、再生，从而促进牙槽骨的再生。
第二方面，本申请提供一种口腔护理凝胶的制备方法，采用如下的技术方案：
一种口腔护理凝胶的制备方法，包括以下步骤：
将巴戟天多糖提取物、凝胶、奥硝唑、水按配比混合均匀，包装，得到口腔护理凝胶。
通过采用上述技术方案，制备方法简单易操作，对生产设备没有特殊需求，适合工业化生产。
综上所述，本申请具有以下有益效果：
1、由于本申请采用在口腔凝胶中加入巴戟天多糖提取物，对IL-6炎症因子的抑制率可以达到88.26-95.72％，在大鼠实验中，新骨面积可以达到0.378-0.407mm2，牙槽嵴吸收率可以达到13.83-20.87％，有较好的抗炎症效果，并且减少牙槽骨吸收，促进牙槽骨生长。
2、本申请中优选采用在凝胶中加入过氧化钙与抗坏血酸，进一步提高口腔凝胶对牙槽骨再生的促进作用。
具体实施方式
以下结合实施例对本申请作进一步详细说明。
原料和中间体的制备例
原料
本申请实施例原料均可通过市售获得：
乙醇为95％乙醇。
制备例
制备例I-1
一种巴戟天多糖提取物，其提取方法为：
1)预处理
将巴戟天洗净，烘干，破碎过40目筛，称取100g巴戟天干粉，用10倍体积的乙醇回流浸提2次，冷却后过滤，滤渣干燥备用；
2)热水浸提
将步骤1)得到的滤渣用90℃的水回流提取2次，滤渣与水的配比为：1g滤渣对应10ml水，提取时间为2h，提取结束后，抽滤、离心，取上清，即为巴戟天多糖粗提液；
3)醇沉
将步骤2)所得多糖粗提液减压浓缩成黏稠浸膏状，然后加入乙醇使溶液含醇量为65％，混合均匀，在4℃下醇沉14h，离心，收集沉淀物，冷冻干燥得到巴戟天多糖。
制备例I-2
与制备例1不同的是，制备例2中步骤2)中热水浸提的次数为3次。
制备例I-3
与制备例1不同的是，制备例3中步骤2)中滤渣与水的配比为：1g滤渣对应15ml水。
制备例I-4
与制备例1不同的是，制备例4中步骤2)中滤渣与水的配比为：1g滤渣对应8ml水。
制备例I-5
与制备例1不同的是，制备例5中步骤2)中滤渣与水的配比为：1g滤渣对应20ml水。
制备例I-6
一种巴戟天多糖提取物，其提取方法为：
1)预处理
将巴戟天洗净，烘干，破碎过40目筛，称取100g巴戟天干粉，用10倍体积的水回流浸提2次，冷却后过滤，滤渣干燥备用；
2)热水浸提
将步骤1)得到的滤渣用90℃的水回流提取2次，滤渣与水的配比为：1g滤渣对应10ml水，提取时间为2h，提取结束后，抽滤、离心，取上清，即为巴戟天多糖粗提液；
3)醇沉
将步骤2)所得多糖粗提液减压浓缩成黏稠浸膏状，然后加入乙醇使溶液含醇量为65％，混合均匀，在4℃下醇沉14h，离心，收集沉淀物，冷冻干燥得到巴戟天多糖。
制备例II-1-II-3
一种海藻酸钠水凝胶，其制备方法为：
按照表1的原料配比，将氯化钠溶于水中，配置氯化钠溶液，将海藻酸钠分3次加入氯化钠溶液中，在25℃下持续搅拌4h，然后在45℃环境中搅拌混合35min，然后在25℃中静置10h，得到混合液；向混合液中加入丙烯酰胺，混合均匀，并加入过硫酸铵、四甲基乙二胺、N，N-亚甲基双丙烯酰胺，然后在2℃下搅拌15min；离心、干燥，得到海藻酸钠水凝胶。
表1制备例II-1-制备例II-3原料配比表(g)
制备例II-4-II-6
与制备例II-2不同的是，制备例II--4-II-6中还分别含有0.1g、0.2g、0.3g过氧化钙粒子，过氧化钙粒子的粒径为10-20nm，其制备方法为：
将氯化钠溶于水中，配置氯化钠溶液，将海藻酸钠分3次加入氯化钠溶液中，在25℃下持续搅拌4h，然后在45℃环境中搅拌混合35min，然后在25℃中静置10h，得到混合液；向混合液中加入丙烯酰胺，混合均匀，并加入过硫酸铵、四甲基乙二胺、N，N-亚甲基双丙烯酰胺，然后在2℃下搅拌15min；然后再加入过氧化钙粒子，混合均匀，然后离心、干燥，得到海藻酸钠水凝胶。
制备例II-7-II-9
与实施例II-5不同的是，实施例II-7-II-9中还分别含有0.15g、0.30g、0.45g抗坏血酸，抗环血酸随过氧化钙粒子一起加入。
制备例II-10
与制备例II-8不同的是，制备例II-10中过氧化钙粒子的粒径为5-10nm。
实施例
实施例1-5
一种口腔护理凝胶，其制备方法为：
按照表2中的原料配比表，将巴戟天多糖提取物、凝胶、奥硝唑、水按配比混合均匀，包装，得到口腔护理凝胶；
其中巴戟天多糖提取物来自于制备例I-1，凝胶来自于制备例II。
表2实施例1-5原料配比表(g)
实施例8-12
与实施例7不同的是，实施例8-12中的巴戟天多糖提取物分别来自于制备例I-2-I-6。
实施例13-21
与实施例9不同的是，实施例13-21中的巴戟天多糖提取物分别来自于制备例II-2-II-10。
对比例
对比例1
与实施例1不同的是，对比例1中不含巴戟天多糖提取物。
对比例2
与实施例1不同的是，对比例2中巴戟天多糖提取物的重量百分比为10％。
性能检测试验
检测方法/试验方法
对实施例1-21以及对比例1-2中制得的口腔护理凝胶进行以下性能检测，检测结果见表3：采用成纤维细胞的细胞实验，检测对IL-6炎症因子的抑制率；
以大鼠为实验对象，采用Micro-CT对各组大鼠牙槽骨进行扫描、分析，用Java(Image J-1.51r；NIH，USA)软件进行图像处理和分析，测量新骨面积与牙槽嵴吸收率，牙槽嵴吸收率以唇、舌牙槽嵴吸收率总和计。
表3性能检测结果
结合实施例1-21和对比例1-2，并结合表3可以看出，实施例1-21中的炎症因子抑制率高于对比例1-2，这说明本申请的口腔凝胶防治感染性口腔炎症的效果更好；实施例1-21中的牙槽嵴吸收率低于对比例1-2，这说明本申请的口腔凝胶可以减少唇舌对牙槽骨的吸收；实施例1-21中的新骨面积高于对比例1-2，这说明本申请的口腔凝胶可以促进牙槽骨的再生。
结合实施例1与对比例1-2，并结合表3可以看出，实施例1中的口腔凝胶的抗炎症效果、减少牙槽骨吸收的效果、促进牙槽骨再生的效果均优于对比例1-2，这可能是巴戟天多糖提取物通过对口腔内的免疫调节，对牙菌斑所导致的口腔慢性炎症及自身免疫性牙槽骨吸收具有预防和治疗作用，且巴戟天多糖提取物的添加量在本申请的限定范围内，效果更好。
结合实施例3与实施例6-7，并结合表3可以看出，实施例6-7中的口腔凝胶的抗炎症效果、减少牙槽骨吸收的效果、促进牙槽骨再生的效果均优于实施例3，这可能是纤连蛋白的加入可以提高牙槽骨的再生。
结合实施例3与实施例8-12，并结合表3可以看出，实施例9中的口腔凝胶在炎症抑制以及牙槽骨再生方面表现更优，这说明，巴戟天多糖提取物在炎症抑制以及牙槽骨再生方面有着较大的影响作用，而制备例I-3中提取巴戟天多糖提取物的工艺更优。
结合实施例13-17，并结合表3可以看出，实施例15-17中的口腔凝胶的在牙槽骨再生方面表现优于实施例13-14，这可能是因为过氧化钙可以维持口腔内有效氧的浓度，提高干细胞的存活率，减少缺氧诱导的细胞凋亡，促进牙槽骨再生。
结合实施例16-21，并结合表3可以看出，实施例17-21中的口腔凝胶的在牙槽骨再生方面表现优于实施例16，这可能是因为抗坏血酸的加入、可以克服过氧化钙加入的不足，保证细胞的正常生长，促进牙槽骨再生。
本具体实施例仅仅是对本申请的解释，其并不是对本申请的限制，本领域技术人员在阅读完本说明书后可以根据需要对本实施例做出没有创造性贡献的修改，但只要在本申请的权利要求范围内都受到专利法的保护。
Claims (9)
1.一种口腔护理凝胶，其特征在于，其包括以下重量百分比的原料：巴戟天多糖提取物2-5%、凝胶0.5-1%、奥硝唑1-3%；
所述凝胶为海藻酸钠水凝胶，其制备方法为：将10-15重量份海藻酸钠加入氯化钠溶液中，混合均匀，然后在45-50℃下处理30-40min，然后静置10-12h，得到混合液；向混合液中加入160-200重量份丙烯酰胺，混合均匀，并加入0.1-0.2重量份过硫酸铵、0.003-0.005重量份四甲基乙二胺、0.09-0.15重量份N，N-亚甲基双丙烯酰胺，然后在0-3℃下搅拌15min；离心、干燥，得到海藻酸钠水凝胶。
2.根据权利要求1所述的一种口腔护理凝胶，其特征在于：所述巴戟天多糖提取物的重量百分比为3-4%。
3.根据权利要求1所述的一种口腔护理凝胶，其特征在于：所述巴戟天多糖提取物的制备方法为：
1）预处理
将巴戟天用乙醇回流浸提2-3次，过滤，滤渣干燥备用；
2）热水浸提
将步骤1）得到的滤渣用80-90℃的水回流提取2-3次，提取结束后，抽滤、离心，取上清，即为巴戟天多糖粗提液；
3）醇沉
将步骤2）所得多糖粗提液减压浓缩成黏稠浸膏状，然后加入乙醇使溶液含醇量为60-70%，混合均匀，在4-5℃下醇沉12-14h，离心，收集沉淀物，冷冻干燥得到巴戟天多糖。
4.根据权利要求3所述的一种口腔护理凝胶，其特征在于：所述步骤2）中滤渣与水的配比为：1g滤渣对应10-15ml水。
5.根据权利要求1所述的一种口腔护理凝胶，其特征在于：所述海藻酸钠水凝胶的制备方法为：将10-15重量份海藻酸钠加入氯化钠溶液中，混合均匀，然后在45-50℃下处理30-40min，然后静置10-12h，得到混合液；向混合液中加入160-200重量份的丙烯酰胺，混合均匀，并加入0.1-0.2重量份过硫酸铵、0.003-0.005重量份四甲基乙二胺、0.09-0.15重量份N，N-亚甲基双丙烯酰胺，然后在0-3℃下搅拌15min；然后再加入0.1-0.2重量份过氧化钙粒子，混合均匀，然后离心、干燥，得到海藻酸钠水凝胶。
6.根据权利要求5所述的一种口腔护理凝胶，其特征在于：在加入所述过氧化钙粒子时，随过氧化钙粒子加入0.15-0.3重量份抗坏血酸。
7.根据权利要求5所述的一种口腔护理凝胶，其特征在于：所述过氧化钙粒子的粒径为10-20nm。
8.根据权利要求1所述的一种口腔护理凝胶，其特征在于：其还包括0.4-1%的纤连蛋白。
9.一种权利要求1-8任一所述的口腔护理凝胶的制备方法，其特征在于，包括以下步骤：
将巴戟天多糖提取物、凝胶、奥硝唑、水按配比混合均匀，包装，得到口腔护理凝胶。
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