CN109526940A - 一种新型离体活髓牙保存液及其制备方法 - Google Patents

一种新型离体活髓牙保存液及其制备方法 Download PDF

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CN109526940A
CN109526940A CN201811559823.2A CN201811559823A CN109526940A CN 109526940 A CN109526940 A CN 109526940A CN 201811559823 A CN201811559823 A CN 201811559823A CN 109526940 A CN109526940 A CN 109526940A
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liquid
tooth
teeth
alive
marrow
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王之发
韩冷
汪维健
段建民
李潇
吴补领
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Southern Hospital Southern Medical University
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02Preservation of living parts
    • A01N1/0205Chemical aspects
    • A01N1/021Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
    • A01N1/0226Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients

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Abstract

本发明公开了一种新型离体活髓牙保存液及其制备方法,属于医药技术领域,在HBSS液中添加青霉素200U/ml、链霉素100U/ml、地塞米松20μg/ml、左旋谷氨酰胺L‑glutamine,272μg/ml,再调其PH值在7.2~7.4之间,过滤消毒后得要制备的牙齿保存液。本发明的保存液经过实验发现,牙齿保存在保存液中24h以后,PDLC仍然保有良好的活力和增殖能力。

Description

一种新型离体活髓牙保存液及其制备方法
技术领域
本发明属于医学技术领域,涉及一种新型离体活髓牙保存液及其制备方法。
背景技术
根据世界卫生组织对牙外伤的分类,牙脱位(avulsion)是指牙齿完全从牙槽窝中分离。牙脱位常见于儿童的牙外伤,占恒牙外伤的16%和乳牙外伤的7%~21%。如果不能即刻再植,脱位牙齿必须保存于合适介质中以保持牙根表面牙周膜细胞(periodontalligament cell,PDLC)的活力,减少牙再植后牙根吸收的发生率。
脱位牙的再植成功率主要与牙齿离体时间和牙周膜细胞受损程度有关,此外还可能与牙位、牙根发育、牙萌出、年龄、牙根数目、牙槽窝炎症、局部创伤及牙根污染等因素有关。牙齿完全脱位后,附着于其上的牙周膜细胞在很短时间内即开始坏死,因而牙再植前的最关键因素是维持牙周膜细胞的活性,最大限度地保存牙周膜细胞,从而减少患牙发生牙根吸收,恢复牙齿的正常功能。学者们认为脱位牙应该在15~20min内尽快再植。然而,由于医疗知识缺乏和医疗条件的限制,脱位牙往往不能立即复位,牙再植的时间一般都在牙脱位后1~4h。在这种情况下,脱位牙应保存于合适的介质中以等待牙再植的时机。而且,牙齿的自体移植成功率也与移植牙牙根表面牙周膜细胞的活力有关。如果牙周膜受到较大损伤,移植后亦会发生牙根吸收。由此可见,牙脱位后应尽快再植;如不能即刻再植,也应将其保存在保存液内,以延缓牙根表面牙周膜细胞坏死的速度,为随后的牙再植增加成功的概率。
美国牙髓医师学会(American Association of Endodontists,AAE)推荐使用HBSS液(Hank’s balanced salt solution,HBSS)作为脱位牙保存的最佳介质,因为HBSS可以长期维持牙周膜细胞的活性。因而,HBSS一直被作为研究牙齿保存液的金标准。但是其作为牙齿保存液效果在不同报道中有所差异。而且有研究显示HBSS液保存PDLC活性的效果随其存放时间延长而变差。
而且,目前国外市场上已有商品化的牙齿保存液,如Dentosafe和EMT ToothSaver分别是德国和美国公司研发的牙齿保存液。这两种产品不但能维持幸存牙周膜细胞的活性,而且能溶解牙根表面坏死细胞,脱落牙在这类产品中可有效地保存超过24h,显著地提高牙再植成功率
我国在脱落牙保存再植方面研究较少,更是几乎没有牙齿保存液的相关研究报道。因此,有必要研发符合我国国情、可有效保存牙齿、显著延长牙齿离体保存时间的牙齿保存液,以提高牙再植成功率,减少并发症。
发明内容
本发明的目的在于克服上述技术存在的缺陷,提供一种新型离体活髓牙保存液及其制备方法。在传统HBSS液用于保存牙周膜细胞的基础上,研制了改良HBSS液,证实了其可以维持PDLC的体外存活能力和增殖能力,表明其可以用于体外脱落牙齿的保存。
其具体技术方案为:
一种新型离体活髓牙保存液,每ml HBSS液中各个组成成分的含量为:青霉素200U、链霉素100U、地塞米松20μg、左旋谷氨酰胺272μg,PH值在7.2~7.4之间。
一种本发明新型离体活髓牙保存液的制备方法,在HBSS液中添加青霉素200U/ml、链霉素100U/ml、地塞米松20μg/ml、左旋谷氨酰胺L-glutamine,272μg/ml,再调其PH值在7.2~7.4之间,过滤消毒后得要制备的牙齿保存液。
与现有技术相比,本发明的有益效果为:1.本保存液可以有效地保护离体牙牙周膜干细胞,延长其体外存活能力,并维持其具有良好的增殖能力;2.制备方便,易于保存和运输;3.产品可存放于幼儿园、公园等人口密集的场所,方便使用;4.具有一定的经济效益和社会健康保护意义。本发明的保存液经过实验发现,牙齿保存在保存液中24h以后,PDLC仍然保有良好的活力和增殖能力。
附图说明
图1是牙齿保存液中保持24h之后分离培养的PDLC光镜下观察;
图2是牙齿保存液中保持24h之后分离培养的PDLC的增殖能力。
具体实施方式
下面结合附图和具体实施例对本发明的技术方案作进一步详细地说明。
本发明在传统HBSS液用于保存牙周膜细胞的基础上,研制了改良HBSS液,证实了其可以维持PDLC的体外存活能力和增殖能力,表明其可以用于体外脱落牙齿的保存。改良HBSS液的配制:在HBSS液中添加青霉素(200U/ml)、链霉素(100U/ml)、地塞米松(20μg/ml)、左旋谷氨酰胺(L-glutamine,272μg/ml),再调其PH值在7.2~7.4之间,过滤消毒后即是我们要制备的牙齿保存液。然后我们将口腔科门诊拔除下来的年轻患者的正畸牙(第一或第二前磨牙)立即存放在保存液中,24h后,取其牙周膜进行牙周膜细胞的分离培养。实验发现,牙齿保存在保存液中24h以后,PDLC仍然保有良好的活力和增殖能力(图1和2)。
根据上述实验结果,我们认为改良的HBSS液可以作为一种新型的离体活髓牙保存液,特别是用于如脱落年轻恒牙等脱落牙的保存,从而保存离体牙牙周膜细胞的活性和增殖能力,为脱落牙再植后与牙槽骨发生生理性结合提供细胞学基础,有利于脱落牙再植后能够维持牙列的完整性和良好的咬合关系,并发挥正常的咀嚼功能,进而提高脱落牙再植的成功率。
以上所述,仅为本发明较佳的具体实施方式,本发明的保护范围不限于此,任何熟悉本技术领域的技术人员在本发明披露的技术范围内,可显而易见地得到的技术方案的简单变化或等效替换均落入本发明的保护范围内。

Claims (2)

1.一种新型离体活髓牙保存液,其特征在于,每ml HBSS液中各个组成成分的含量为:青霉素200U、链霉素100U、地塞米松20μg、左旋谷氨酰胺272μg,PH值在7.2~7.4之间。
2.一种本发明新型离体活髓牙保存液的制备方法,其特征在于,在HBSS液中添加青霉素200U/ml、链霉素100U/ml、地塞米松20μg/ml、左旋谷氨酰胺L-glutamine,272μg/ml,再调其PH值在7.2~7.4之间,过滤消毒后得要制备的牙齿保存液。
CN201811559823.2A 2018-12-12 2018-12-12 一种新型离体活髓牙保存液及其制备方法 Pending CN109526940A (zh)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009044943A1 (en) * 2007-10-01 2009-04-09 Seoul National University Industry Foundation Various human dental stem cells having a mineralization ability and the method for culturing them
CN104705289A (zh) * 2015-03-01 2015-06-17 安徽新生命干细胞科技有限公司 一种离体牙齿保存液

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009044943A1 (en) * 2007-10-01 2009-04-09 Seoul National University Industry Foundation Various human dental stem cells having a mineralization ability and the method for culturing them
CN104705289A (zh) * 2015-03-01 2015-06-17 安徽新生命干细胞科技有限公司 一种离体牙齿保存液

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李鑫等: "改良HBSS 液对体外培养人牙周膜细胞增殖活性的影响", 《实用口腔医学杂志》 *
王立晖等: "《生物活性多肽特性与营养学应用研究》", 30 November 2016, 天津大学出版社 *

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Application publication date: 20190329