In Vitro Synergistic Antibacterial Activity Of Tea Tree Oil And KBN Thixotropic Gel

Objective:Anti-Candida albicans(KBN)thixotropic gel is a clinical experience for the treatment of Vulvovaginal Candidiasis(VVC).Its preparation consists of Coptis chinensis,Saururi chinensis,Isatis indigotica,Celosia cristata,Ewgewia caryophyllata,Borneolum synyheticum and other herbs extract and Avicin auxiliary.It is a new anti-fungal medicine,being continually studied by the task group,with good comparative advantages of large drug loading and antibacterial effect.Its prescription Were contained by"Directory of names of raw materials used in cosmetics" which has been published by the State Food and Drug Administration.So it can be convenient to develop Anti-bacterial and anti-inflammatory health products of care solution,shower gel,hand sanitizer and so on.Tea tree oil(TTO)is the excellent natural antibacterial agent,with pharmacological activity of broad-spectrum antibacterial,anti-virus,anti-inflammatory,anti-tumor,anti-oxidation,anti-inflammatory pain,cough and phlegm,killing mites,adjusting immunity and so on.In view of the strong pharmacological activity of TTO,it is widely used in preventive treatment and other areas of life.This paper discusses whether TTO and KBN thixotropic gel have synergistic antibacterial effect,so as to develop Makeup,Disinfection and other Daily health products for fungi and bacteria susceptible crowd and provide the basis of efficacy study.Methods:1.To prefer proportion of compatibility ratio of TTO and KBN thixotropic gel of combination of anti-Candida albicans in vitro.The minimal inhibitory-concentration(MIC)and minimal bactericidal concentration(MBC)of TTO and KBN thixotropic gel of combination of anti-Candida albicans in vitro will be determined by agar dilution method and microdilution method respectively.Then evaluating the two drugs combined antibacterial and sterilization efficacy,and prefering the best antimicrobial compatibility ratio.2.In vitro anti-Candida albicans of KBN preparation containing different TTO.To determine the MIC and MBC of anti-Candida albicans by agar dilution method and microdilution method and investigate the antibacterial activity effects of the Content of TTO in KBN Preparationstea.We will also verify the Best synergistic antimicrobial ratio.3.In vitro dynamic anti-Candida albicans of 2.91%TTO-KBN preparation.According to the optimal synergistic bactericidal ratio of TTO and KBN preparation,the effect of dynamic anti-Candida albicans of 2.91%TTO-KBN preparation will be investigated by colony counting method,So as to obtain the drug conc'entration-time-bactericidal rate of the relationship between the three.4.In vitro broad-spectrum antibacterial of 5.66%TTO-KBN preparation.According to the optimum synergistic antibacterial ratio of TTO and KBN preparation,the MIC and MBC of anting seven kinds of test bacteria of 5.66%TTO-KBN preparation,TTO and KBN preparation will be determined and compared by agar dilution method and microdilution method respectively.5.Electron Microscopic Study on the Mechanism of Anti-Candida albicans of TTO and KBN preparation.Candida albicans will be respectively treated with 2 times of MBC of TTO,KBN preparation and 2.91%TTO-KBN preparation.Then the ultrastructural effect of Candida albicans will be observed under transmission electron microscope and we will also explore its mechanism of antibacterial effect.ResuLts:1.When used alone,the MIC of TTO and KBN preparation were 1.91 mg/mL and 13.03 mg/mL and MBC were 7.64 mg/mL and 104.25 mg/mL,respectively.When TTO and KBN preparation were the best synergistic bacteriostasis ratio(6:100)and the best synergistic Sterilization ratio(3:100),the fractional Inhibitory Concentration(FIC)and the fractional bactericidal Concentration(FBC)were 0.5 and 0.25,respectively.Their MIC were 0.48 mg/mL and 3.26 mg/mL,respectively.And their MIBC were 0.96 mg/mL and 13.03 mg/mL,respectively.The MIC and MBC of Xianglian lotion were 31.25 mg/mL and 62.50 mg/mL,respectively.2.The MIC and MBC of KBN thixotropic gel containing 0.10%?9.09%TTO were 1.63?13.03 mg/mL and 3.26?52.13 mg/mL,respectively.At the same time,the bactericidal effect of 2.91%TTO-KBN preparation and 5.66%TTO-KBN preparation were consistent with the previous period.3.When 2.91%TTO-KBN preparation concentration was 52 mg/mL,its bactericidal effect was better than the separate drug group and Xianglian lotion at 3 min;In addition to KBN preparation group,the bactericidal effect of other groups were equivalent at 5 min;Each Group with complete bactericidal effect in more than 6 hours.And when 3%TTO-KBN preparation concentration was 26 mg/mL,its bactericidal effect was comparable to that of TTO,and better than Xianglian lotion at 3 and 5 min;The bactericidal effect of each group was similar in more than 6 and 8 hours.4.When TTO,KBN preparation,5.66%TTO-KBN preparation,and Xianglian lotion concentration were 7.64 mg/mL,104.25 mg/mL,26.06 mg/mL and 125.00 mg/ml respectively,they had bacteriostatic effect on seven test bacteria;In addition to TTO group had no bacteriostatic effect on hemolytic streptococcus,each group also had bacteriostatic effect on seven test bacteria.According to the FIC,5.66%TTO-KBN preparation had synergistic or additive effect to most test bacteria,and had a broad spectrum antibacterial effect.5.In the normal control group,Candida albicans were almost round,and their structures of cell membrane and cell wall were intact.In the positive control group,Candida albicans were "shell",and their structure of cell membrane and cell wall were damaged and incomplete.In the teat group,Candida albicans cell membrane were seriously absent,and were continuously damaged.Their structures of cell wall were loose,and there were cracks,and their cytoplasmic content had leakaged.ConcLusion:TTO and KBN thixotropic gel has synergistic antifungal effect in vitro.When 2.91%TTO-KBN preparation concentration is 26.06 mg/ml,as VVC susceptible crowd shower gel,or the need for drugs in the local long time retention of gels,suppositories and other daily products dose concentration development;The concentration of 5.66%TTO-KBN preparation is 26.06 mg/mL,as the development of dose concentration of broad-spectrum antibacterial daily chemical products.The two drugs may by changing the internal microstructure(cell membrane,cell wall,nucleus)and to achieve synergize antibacterial effect.The above research combined the research results of foreign natural botanical drugs with the theory and experience of traditional Chinese medicine.It fully reflects the characteristics and advantages of the two drugs combined with antibacterial,and provides the theoretical basis for the development of Makeup,disinfection and other daily health products,and efficacy demands to achieve better results,and it has a good market prospects.