Int J Stomatol ›› 2021, Vol. 48 ›› Issue (1): 35-40.doi: 10.7518/gjkq.2021019

• Original Articles • Previous Articles     Next Articles

Antimicrobial effect of chlorhexidine on Candida albicans in vitro according to D2O-labeled single-cell Raman micro-spectroscopy

Li Fan1,2,Zhang Lijuan1,2,Tan Kaixuan2,3,Zhang Ying1,Lu Jie2,3,Li Shanshan1,Yang Fang1,2()   

  1. 1. School of Stomatology, Qingdao University, Qingdao 266003, China
    2. Stomatology Center, Qingdao Municipal Hos-pital, Qingdao 266071, China
    3. School of Stomatology, Dalian Medical University, Dalian 116044, China
  • Received:2020-04-05 Revised:2020-09-28 Online:2021-01-01 Published:2021-01-20
  • Contact: Fang Yang
  • Supported by:
    This study was supported by National Natural Science Foundation of China(81670979);This study was supported by National Natural Science Foundation of China(31600099)


Objective To evaluate the universality of D2O-labeled single-cell Raman micro-spectroscopy and the bacteriostatic effect of chlorhexidine (CHX) on Candida albicans (C. albicans) in vitro. Methods The universality of D2O-labeled single-cell Raman micro-spectroscopy was investigated by exploring the growth of C. albicans under various doses of D2O and the regularity of D2O intake by C. albicans on the basis of the temporal change in OD600 and C-D ratio, respectively. Furthermore, we determined the effects of inhibiting the growth and CHX metabolism on C. albicans according to minimum inhibitory concentration (MIC) and minimum inhibitory concentration based on metabolic activity (MIC-MA) values, using broth dilution test and D2O-labeled single-cell Raman micro-spectroscopy, respectively. Results The growth of C. albicans was not significantly affected at concentrations below or equal to 30% D2O, and C. albicans can actively metabolize D2O, as indicated by Raman micro-spectroscopy results. Moreover, the C-D ratio of C. albicans at the stationary phase was positively correlated with D2O concentration. The MIC and MIC-MA of CHX were 4 and 8 μg·mL -1, respectively. Under the MIC, the growth of C. albicans was completely inhibited, but metabolic activity was active. Metabolic activity can be inhibited only when the concentration of 2×MIC was reached.Conclusion D2O- labeled single-cell Raman micro-spectroscopy is suitable for evaluating the metabolism activity of C. albicans. The commonly used concentration CHX in clinical practice effectively inhibits the growth and even the metabolism of C. albicans.

Key words: Candida albicans, chlorhexidine, heavy water, single cell, Raman micro-spectroscopy

CLC Number: 

  • R780.2


Fig 1

Temporal change of OD600 for Candida albicans CICC32380 under various quality score D2O"

Fig 2

Change of single cell Raman spectrum for Candida albicans CICC32380 under various quality score D2O"

Fig 3

There was a linear relationship between the C-D ratio of Candida albicans CICC32380 at stable stage and D2O concentration followed by single cell Raman spectrum measurement"

Fig 4

Temporal change of single cell Raman spectrum for Candida albicans CICC32380 under 30% D2O"

Fig 5

Temporal change of C-D ratio curve and growth curve of Candida albicans CICC32380 under 30% D2O"

Tab 1

Measurement of MIC for Candida albicans CICC32380 under CHX"

CHX质量浓度/(μg·mL-1 ΔOD600
空白对照组 1.33×10-3±9.43×10-4
0(阴性对照组) 1.49×10-1±1.48×10-2
1 1.37×10-1±3.74×10-3
2 1.15×10-1±4.32×10-3
4 1.00×10-3±1.41×10-3
8 3.33×10-4±4.71×10-4
16 3.33×10-4±1.25×10-4
32 1.00×10-3±0.00

Tab 2

Measurement of MIC-MA for Candida albicans CICC32380 under CHX"

CHX质量浓度/(μg·mL-1 ΔC-D ratio
0(阴性对照组) 6.54×10-2±4.04×10-3
2 6.63×10-2±4.80×10-3
4 7.14×10-2±4.45×10-3
8 -8.90×10-4±5.67×10-4
12 -9.12×10-3±2.84×10-3

Fig 6

Measurement of MIC-MA under CHX for Candida albicans CICC32380"

Fig 7

Temporal dynamics of the C-D ratio under various quality score CHX"

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