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使用不同细胞培养物对含抗坏血酸和盐酸溶菌酶的药用口香糖的细胞毒性、增殖及微生物活性的研究

The Study of the Cytotoxicity, Proliferative and Microbiological Activity of the Medicated Chewing Gum with Ascorbic Acid and Lysozyme Hydrochloride Using Different Culture of Cells.

作者信息

Maslii Yuliia, Garmanchuk Liudmyla, Ruban Olena, Dovbynchuk Taisa, Herbina Nataliia, Kasparaviciene Giedre, Bernatoniene Jurga

机构信息

Department of Industrial Technology of Drugs, National University of Pharmacy, 61002 Kharkiv, Ukraine.

Department of Drug Technology and Social Pharmacy, Faculty of Pharmacy, Medical Academy, Lithuanian University of Health Sciences, LT-50161 Kaunas, Lithuania.

出版信息

Pharmaceutics. 2023 Jul 5;15(7):1894. doi: 10.3390/pharmaceutics15071894.

Abstract

Medicated chewing gum with lysozyme hydrochloride and ascorbic acid as active pharmaceutical ingredients was developed for application in dentistry. The aim of this research was to study the cytotoxicity, proliferative, and microbiological activities of the active ingredients in different types of cell cultures. The preclinical study of active pharmaceutical ingredients and their combinations was carried out using culture lines such as HepG2 (human hepatocarcinoma cells), Hek293 (human embryonic kidney cells), and MAEC (mouse aortic endothelial cells). MTT assays were used to analyse cytotoxicity and proliferative activity, while the state of antioxidant protection was assessed by the content of sulfhydryl groups and catalase activity. The determination of lipid peroxidation products was based on the level of TBA-active products. As a microbiological model for studying the effect of the developed dental medicine on the ability of the oral cavity microorganisms to form biofilms, the following strains were used: , , , , and . The optical density of the formed biofilm was evaluated by the intensity of the experimental sample's colour on a StatFax 303 Plus photometer at a wavelength of 630 nm. The combination of ascorbic acid and lysozyme hydrochloride in the established concentrations (20 mg and 10 mg per 1 gum, respectively) resulted in a slight stimulation of cell proliferation without any toxic effects and increased antioxidant protection, preventing the development of oxidative stress. It was found that, in contrast to the separately used active substances, the combination of lysozyme hydrochloride and ascorbic acid inhibits the biofilm formation of all studied microorganisms and shows the ability to destroy diurnal biofilms of and fungi of the genus , indicating potentiation and summation of the active pharmaceutical ingredients' composition effects in the developed dental medicine. Due to the observed positive pharmacological and microbiological action, the combination of lysozyme hydrochloride and ascorbic acid in the medicated chewing gum serves as a promising tool for the prevention and treatment of infectious and inflammatory diseases of the periodontium and mucous membranes and the prevention of caries.

摘要

以盐酸溶菌酶和抗坏血酸作为活性药物成分的药用口香糖被开发用于牙科领域。本研究的目的是研究不同类型细胞培养物中活性成分的细胞毒性、增殖和微生物活性。使用HepG2(人肝癌细胞)、Hek293(人胚胎肾细胞)和MAEC(小鼠主动脉内皮细胞)等细胞系对活性药物成分及其组合进行临床前研究。MTT法用于分析细胞毒性和增殖活性,而通过巯基含量和过氧化氢酶活性评估抗氧化保护状态。脂质过氧化产物的测定基于TBA活性产物的水平。作为研究开发的牙科药物对口腔微生物形成生物膜能力影响的微生物模型,使用了以下菌株:[具体菌株名称缺失]。通过在波长630nm下使用StatFax 303 Plus光度计测量实验样品的颜色强度来评估形成的生物膜的光密度。抗坏血酸和盐酸溶菌酶以既定浓度(每片口香糖分别为20mg和10mg)组合使用,导致细胞增殖略有刺激,无任何毒性作用,并增强了抗氧化保护,防止氧化应激的发生。结果发现,与单独使用的活性物质相比,盐酸溶菌酶和抗坏血酸的组合抑制了所有研究微生物的生物膜形成,并显示出破坏[具体微生物名称缺失]的日间生物膜和[具体真菌属名称缺失]真菌生物膜的能力,表明在开发的牙科药物中活性药物成分组合效应的增强和累加。由于观察到的积极药理和微生物作用,药用口香糖中盐酸溶菌酶和抗坏血酸的组合是预防和治疗牙周和粘膜感染性和炎症性疾病以及预防龋齿的有前途的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee18/10386584/d60382076de0/pharmaceutics-15-01894-g001.jpg

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