Journal of Prevention and Treatment for Stomatological Diseases ›› 2021, Vol. 29 ›› Issue (7): 462-467.doi: 10.12016/j.issn.2096-1456.2021.07.005

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Inhibitory effect of baicalin on Streptococcus mutans UA159 in vitro

WU Ju(),WANG Ling,LIU Xingrong()   

  1. School of Stomatological, Southwest Medical University, Department of Children’s Stomatology, Affiliated Stomatological Hospital of Southwest Medical University, Luzhou 646000, China
  • Received:2020-10-26 Revised:2020-12-22 Online:2021-07-20 Published:2021-04-19
  • Contact: Xingrong LIU E-mail:302871518@qq.com;liuxingrong163@163.com
  • Supported by:
    Joint Scientific Research Project of Sichuan Provincial Department of Science and Technology Luzhou Municipal People’s Government and Luzhou Medical College(14JC0049-LY53)

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Abstract:

Objective To investigate the inhibitory effect of baicalin on Streptococcus mutans UA159 in vitro.Methods The minimum inhibitory concentration (MIC) of baicalin on Streptococcus mutans UA159 was determined by the liquid multiple dilution method combined with the OD600 value measured by microplate. The OD600 value of Streptococcus mutans UA159 in different concentrations of baicalin was measured by an enzyme mapping instrument. A growth curve was drawn, and the adhesion rate and adhesion inhibition rate were calculated. The effect of baicalin on the formation of Streptococcus mutans UA159 biofilms was observed by the crystal violet quantitative method and scanning electron microscopy. The effect of baicalin on the total number of Streptococcus mutans UA159 bacteria was observed by scanning electron microscopy.Results The MIC of baicalin on Streptococcus mutans UA159 was 12 mg/mL. With increasing baicalin concentration, the growth rate of Streptococcus mutans UA159 was slowed, the adhesion rate of Streptococcus mutans UA159 decreased and the adhesion inhibition rate increased（P < 0.05）. The results of crystal violet quantitative method showed that compared with the bacterial control group, the biofilm formation of Streptococcus mutans UA159 was significantly reduced after adding baicalin at 0 h, 6 h and 12 h (P < 0.001). Under a scanning electron microscope, the total number of bacteria decreased significantly after adding baicalin at 0 h, 6 h and 12 h.Conclusion Baicalin can inhibit the growth and reproduction of Streptococcus mutans UA159 in vitro as well as the adhesion and biofilm formation of Streptococcus mutans UA159.

Key words: baicalin, natural medicine, Streptococcus mutans UA159, caries, minimum inhibitory concentration, growth curve, adhesion rate, adhesion inhibition rate, biofilm formation, in vitro study

CLC Number: 

  • R78

Table 1

ΔOD600 value of Streptococcus mutans UA159 at different baicalin concentrations $\bar{x}\pm s$,n=3"

Tube numer Baicalin concentration
（mg/mL）		 ΔOD600值 P
1 24 0.046 ± 0.007 < 0.001
2 12 0.094 ± 0.001 < 0.001
3 6 0.182 ± 0.004 < 0.001
4 3 0.265 ± 0.002 < 0.001
5 1.5 0.377 ± 0.003 0.406
6 0.75 0.377 ± 0.005 0.544
7 0.375 0.378 ± 0.008 0.595
8 24 0.020 ± 0.005 < 0.001
9 0 0.377 ± 0.007 0.496
10 0 0.380 ± 0.005 -

Table 2

ΔOD600 value of Streptococcus mutans UA159 at different concentrations of baicalin at different time points $\bar{x}\pm s$,n=3"

Time（h） Baicalin concentration F P
12 mg/mL 6 mg/mL 3 mg/mL 1.5 mg/mL 0 mg/mL P1 P2 P3 P4
4 0.123 ± 0.002 0.152 ± 0.002 0.184 ± 0.004 0.192 ± 0.003 0.211 ± 0.004 349.182 < 0.001 < 0.001 < 0.001 < 0.001
8 0.174 ± 0.005 0.236 ± 0.008 0.311 ± 0.005 0.375 ± 0.003 0.380 ± 0.002 895.601 < 0.001 < 0.001 < 0.001 0.246
12 0.211 ± 0.005 0.315 ± 0.009 0.411 ± 0.003 0.458 ± 0.004 0.466 ± 0.003 1 180.943 < 0.001 < 0.001 < 0.001 0.103
24 0.277 ± 0.003 0.327 ± 0.003 0.452 ± 0.005 0.466 ± 0.003 0.473 ± 0.002 2 197.682 < 0.001 < 0.001 < 0.001 0.068

Figure 1

Growth curves of Streptococcus mutans UA159 at different concentrations of baicalin"

Table 3

Effects of different concentrations of baicalin on adhesion of Streptococcus mutans UA159 $\bar{x}\pm s$,n=3"

Baicalin concentration Adhesion rate（%） Adhesion inhibition rate（%）
12 mg/mL 28.00 ± 0.36* 33.83 ± 0.92△
6 mg/mL 31.13 ± 0.47* 25.77 ± 2.18△
3 mg/mL 33.06 ± 0.20* 22.18 ± 0.69△
1.5 mg/mL 35.47 ± 0.40* 16.18 ± 0.71△
0 mg/mL 42.32 ± 0.45 0

Table 4

Effects of baicalin on biofilm formation of Streptococcus mutans UA159 $\bar{x}\pm s$,n=3"

Group OD600 P
0 h 0.132 ± 0.003 < 0.000
6 h 0.247 ± 0.007 < 0.000
12 h 0.288 ± 0.006 < 0.000
Solvent control 0.376 ± 0.004 0.304
Bacteria control 0.381 ± 0.007 -

Figure 2

Effects of baicalin on the total number of Streptococcus mutans UA159 bacteria was observed under SEM a-b: bacteria control group, UA159 grew well; a: SEM (× 5 000); b: SEM (× 10 000); c-d: added 10 mg/mL baicalin at 0 h, UA159 grew in pairs; c: SEM (× 5 000); d: SEM (× 10 000); e-f: added 10 mg/ml baicalin at 6 h, the total number of UA159 decreased significantly; e: SEM (× 5 000); f: SEM (× 10 000); g-h: added 10 mg/mL baicalin at 12 h, the total number of UA159 decreased significantly; g: SEM (× 5 000); h: SEM (× 10 000); i-j: in the solvent control group, there were no significant differences compared with the bacteria control group; i: SEM (× 5 000); j: SEM (× 10 000); SEM: scanning electron microscope."

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