Investigating the Growth of Pseudomonas aeruginosa and Its Influence on Osteolysis in Human Bone: An In Vitro Study
Ahmed Al Ghaithi, Atika Al Bimani, Sultan Al Maskari
Keywords :
Biofilm, Infection, Osteomyelitis
Citation Information :
Al Ghaithi A, Al Bimani A, Al Maskari S. Investigating the Growth of Pseudomonas aeruginosa and Its Influence on Osteolysis in Human Bone: An In Vitro Study. 2021; 16 (3):127-131.
Background: Isolation of the causal microorganisms in osteomyelitis presents a major challenge for treating clinicians. Several methods have been proposed to rapidly and accurately identify microorganisms. There has been an increasing interest in using Raman spectroscopy in the field of microbial detection and characterisation. This paper explores the use of Raman spectroscopy identification as one of the most difficult-to-isolate microorganisms causing osteomyelitis.
Methods and results: Fresh healthy human bone samples were collected from patients undergoing a total knee replacement. These samples were then inoculated with fresh overnight Pseudomonas aeruginosa (PAO) cultures. Bacteria growth and bone ultrastructural changes were monitored over a period of 6 weeks. The experiment demonstrated ultrastructural bony destruction caused by osteolytic PAO secretions. Raman-specific spectral signatures related to the cellular membranes of PAO structures were spotted indicating survival of bacteria on the bone surface.
Conclusion: This study showed the promising ability of Raman spectroscopy to identify the presence of bacteria on the surface of inoculated bone samples over time. It was able to detect the osteolytic activity of the bacteria as well as ultrastructure specific to PAO virulence. This method may have a role as an aid to existing diagnostic methods for fast and accurate bacterial identification in bone infection.
Franco-Duarte R, Černáková L, Kadam S, et al. Advances in chemical and biological methods to identify microorganisms—from past to present. Microorganisms 2019;7(5):130. DOI: 10.3390/microorganisms7050130.
Strola SA, Baritaux J-C, Schultz E, et al. Single bacteria identification by Raman spectroscopy. J Biomed Opt 2014;19(11):111610. DOI: 10.1117/1.JBO.19.11.111610.
Hamasha K, Mohaidat QI, Putnam RA, et al. Sensitive and specific discrimination of pathogenic and nonpathogenic Escherichia coli using Raman spectroscopy–a comparison of two multivariate analysis techniques. Biomed Opt Express 2013;4(4):481–489. DOI: 10.1364/BOE.4.000481.
Sandt C, Smith-Palmer T, Pink J, et al. Confocal Raman microspectroscopy as a tool for studying the chemical heterogeneities of biofilms in situ. J Appl Microbiol 2007;103(5):1808–1820. DOI: 10.1111/j.1365-2672.2007.03413.x.
Khalid M, Bora T, Ghaithi AA, et al. Raman Spectroscopy detects changes in bone mineral quality and collagen cross-linkage in Staphylococcus infected human bone. Sci Rep 2018;8(1):9417. DOI: 10.1038/s41598-018-27752-z.
Calhoun JH, Manring MM, Shirtliff M. Osteomyelitis of the long bones. Semin Plast Surg 2009;23(2):59–72. DOI: 10.1055/s-0029-1214158.
Laghmouche N, Compain F, Jannot A-S, et al. Successful treatment of Pseudomonas aeruginosa osteomyelitis with antibiotic monotherapy of limited duration. J Infect 2017;75(3):198–206. DOI: 10.1016/j.jinf.2017.06.006.
Pence I, Mahadevan-Jansen A. Clinical instrumentation and applications of Raman spectroscopy. Chem Soc Rev 2016;45(7): 1958–1979. DOI: 10.1039/c5cs00581g.
Morris MD, Mandair GS. Raman assessment of bone quality. Clin Orthop 2011;469(8):2160–2169. DOI: 10.1007/s11999-010-1692-y.
Zięba-Palus J, Michalska A. Photobleaching as a useful technique in reducing of fluorescence in Raman spectra of blue automobile paint samples. Vib Spectrosc 2014;74:6–12. DOI: 10.1016/j.vibspec.2014.06.007.
Shoulders MD, Raines RT. Collagen structure and stability. Annu Rev Biochem 2009;78:929–958. DOI: 10.1146/annurev.biochem.77.032207.120833.
Magalhães FL, Machado AMC, Paulino E, et al. Raman spectroscopy with a 1064-nm wavelength laser as a potential molecular tool for prostate cancer diagnosis: a pilot study. J Biomed Opt 2018;23(12):1–6. DOI: 10.1117/1.JBO.23.12.121613.
Keleştemur S, Avci E, Çulha M. Raman and surface-enhanced Raman scattering for biofilm characterization. Chemosensors 2018;6(1):5. DOI: 10.3390/chemosensors6010005.
Pseudomonas aeruginosa responds to exogenous polyunsaturated fatty acids (PUFAs) by modifying phospholipid composition, membrane permeability, and phenotypes associated with virulence. BMC Microbiol 2018;18:117. DOI: 10.1186/s12866-018-1259-8.
Barák I, Muchová K. The role of lipid domains in bacterial cell processes. Int J Mol Sci 2013;14(2):4050–4065. DOI: 10.3390/ijms14024050.
Abdel-Mawgoud AM, Lépine F, Déziel E. Rhamnolipids: diversity of structures, microbial origins and roles. Appl Microbiol Biotechnol 2010;86(5):1323–1336. DOI: 10.1007/s00253-010-2498-2.
Gelder JD, Gussem KD, Vandenabeele P, et al. Reference database of Raman spectra of biological molecules. J Raman Spectrosc 2007;38(9):1133–1147. DOI: 10.1002/jrs.1734.
Inoue T, Shingaki R, Fukui K. Inhibition of swarming motility of Pseudomonas aeruginosa by branched-chain fatty acids. FEMS Microbiol Lett 2008;281(1):81–86. DOI: 10.1111/j.1574-6968.2008.01089.x.
Inoue T, Kuroda T, Ohara N. 12-Methyltetradecanoic acid, a branched-chain fatty acid, represses the extracellular production of surfactants required for swarming motility in Pseudomonas aeruginosa PAO1. Jpn J Infect Dis 2012;65(2):126–131. PMID: 22446119.
Polisetti S, Baig NF, Morales-Soto N, et al. Spatial mapping of pyocyanin in Pseudomonas aeruginosa bacterial communities using surface enhanced Raman scattering. Appl Spectrosc 2017;71(2): 215–223. DOI: 10.1177/0003702816654167.
