AIMS Medical Science, 2015, 2(3): 150-161. doi: 10.3934/medsci.2015.3.150

Research article

Export file:


  • RIS(for EndNote,Reference Manager,ProCite)
  • BibTex
  • Text


  • Citation Only
  • Citation and Abstract

Mass Spectrometry Imaging of Chlorhexidine and Bacteria in a Model Wound

1 Department of Chemistry and Biochemistry, Montana State University, Bozeman MT 59717, USA;
2 BioScience Laboratories Inc., Bozeman MT 59718, USA;
3 Center for Biofilm Engineering, Montana State University, Bozeman MT 59717, USA

The ability to generate two-dimensional images of a wound that contains information about the distribution of bacteria overlaid with the distribution of drugs and metabolites could enhance our understanding of wound healing processes. Advances in technology are leading to a rapid expansion in mass spectrometry-based imaging. When combined with the ability of matrix assisted laser desorption ionization to ionize a wide range of molecules, imaging mass spectrometry is a powerful biomedical research tool. However, this technique has yet to be used to investigate bacterial colonization of wounds or the distribution of antimicrobial agents on tissue. To address this, distribution and persistence of the antimicrobial agent chlorhexidine on a model human tissue was investigated. The ability to detect and localize Staphylococcus aureus on the same tissue model was also addressed. Sub-millimeter resolution ion images from these experiments show the promise of using mass spectrometry imaging to investigate the growth and treatment of bacteria on skin. This methodology will be of value in the development of wound dressings with improved antimicrobial properties and a more careful analysis of the concentration of antimicrobial agents required to prevent biofilm formation and persistence.
  Article Metrics


1. James GA, Swogger E, Wolcott R, et al. (2007) Biofilms in chronic wounds. Wound Repair Regen 16: 37-44.

2. Brackman G, De Meyer L, Nelis HJ, et al. (2013) Biofilm inhibitory and eradicating activity of wound care products against Staphylococcus aureus and Staphylococcus epidermidis biofilms in an in vitro chronic wound model. J Appl Microbiol 114: 1833-1842.    

3. Demidova-Rice TN, Hamblin MR, Herman IM (2012) Acute and impaired wound healing: pathophysiology and current methods for drug delivery, part 1: normal and chronic wounds: biology, causes, and approaches to care. Adv Skin Wound Care 25: 304-314.    

4. Lazarus GS, Cooper DM, Knighton DR, et al. (1994) Definitions and guidelines for assessment of wounds and evaluation of healing. Wound Repair Regen 2: 165-170.    

5. Davies G, Francist J, Martin A, et al. (1954) 1:6-Di-4'-Chlorophenyldiguanidohexane (“Hibitane”*). Laboratory investigation of a new antibacterial agent of high potency. Brit J Pharm Chemoth 9: 192-196.

6. Ayliffe GAJ, Noy MF, Babb JR, et al. (1983) A comparison of pre-operative bathing with soap in the prevention of wound infection. J Hosp Infect 4: 237-244.    

7. Hayek LJ, Emerson JM, Gardner AMN (1987) A placebo-controlled trial of the effect of two preoperative baths or showers with chlorhexidine detergent on postoperative wound infection rates. J Hosp Infect 10: 165-172.    

8. Garibaldi RA (1988) Prevention with of intraoperative chlorhexidine wound contamination shower and scrub. J Hosp Infect 11: 5-9.    

9. Lynch W, Davey PG, Malek M, et al. (1992) Cost-effectiveness analysis of the use of chlorhexidine detergent in preoperative whole-body disinfection in wound infection prophylaxis. J Hosp Infect 21: 179-191.    

10. Jeansonne MJ, White RR (1994) A Comparison of 2.0% hlorhexidine gluconate and 5.25% sodium hypochlorite as antimicrobial endodontic irrigants. J Endodont 20: 276-278.

11. McBain AJ, Bartolo RG, Catrenich CE, et al. (2003) Effects of a chlorhexidine gluconate-containing mouthwash on the vitality and antimicrobial susceptibility of in vitro oral bacterial ecosystems. Appl Environ Microb 69: 4770-4776.    

12. Jones CG (1997) Chlorhexidine: is it still the gold standard? Periodontol 2000 15: 55-62.    

13. Zong Z, Kirsch LE (2012) Studies on the instability of chlorhexidine, Part I: kinetics. J Pharm Sci 101: 2417-2427.    

14. Hidalgo E, Dominguez C (2001) Mechanisms underlying chlorhexidine-induced cytotoxicity. Toxicol in Vitro 15: 271-276.    

15. Russell AD (1986) Chlorhexidine: antibacterial action and bacterial resistance. Infection 14: 212-215.    

16. Agarwal A, Nelson TB, Kierski PR, et al. (2012) Polymeric multilayers that localize the release of chlorhexidine from biologic wound dressings. Biomaterials 33: 6783-6792.    

17. Lavoine N, Tabary N, Desloges I, et al. (2014) Controlled release of chlorhexidine digluconate using β-cyclodextrin and microfibrillated cellulose. Colloids Surfaces B 121: 196-205.    

18. Caprioli RM, Farmer TB, Gile J (1997) Molecular imaging of biological samples: Localization of peptides and proteins using MALDI-TOF MS. Anal Chem 69: 4751-1760.    

19. Chughtai K, Heeren RMA (2010) Mass spectrometric imaging for biomedical tissue analysis. Chem Rev 110: 3237-3277.    

20. Hart PJ, Francese S, Claude E, et al. (2011) MALDI-MS imaging of lipids in ex vivo human skin. Anal Bioanal Chem 401: 115-125.    

21. Kaluzhny Y, Kandárová H, Hayden P, et al. (2011) Development of the EpiOcularTM eye irritation test for Hazard identification and labelling of eye irritating chemicals in response to the requirements of the EU. Altern Lab Anim 39: 339-364.

22. Ye H, Gemperline E, Venkateshwaran M, et al. (2013) MALDI mass spectrometry-assisted molecular imaging of metabolites during nitrogen fixation in the Medicago truncatula-Sinorhizobium meliloti symbiosis. Plant J 75: 130-145.    

23. Ammons MCB, Ward LS, James GA (2011) Anti-biofilm efficacy of a lactoferrin/xylitol wound hydrogel used in combination with silver wound dressings. Int Wound J 8: 268-273.    

24. Agostinho AM, Hartman A, Lipp C, et al. (2011) An in vitro model for the growth and analysis of chronic wound MRSA biofilms. J Appl Microbiol 111: 1275-1282.    

25. Knochenmuss R, Zenobi R (2003) MALDI ionization: The role of in-plume processes. Chem Rev 103: 441-452.    

26. Annesley T M (2003) Ion suppression in mass spectrometry. Clin Chem 49: 1041-1044.    

Copyright Info: © 2015, Brian Bothner, et al., licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (

Download full text in PDF

Export Citation

Article outline

Show full outline
Copyright © AIMS Press All Rights Reserved