CONFOCAL LASER SCANNING MICROSCOPE ANALYSIS ON POST-BIOFILM ASSESSMENT OF BIOFILM-PRODUCING OSTEOMYELITIC STAPHYLOCOCCUS AUREUS TREATED WITH NEW GENTAMICIN-NIGELLA SATIVA FUSION EMULSION (GNFE)

Yaakob KI, Mohd Shafri MA*, Mohd Yusof N, Mohamed F

  • Mohd Shafri MA

Abstract

CONFOCAL LASER SCANNING MICROSCOPE ANALYSIS ON POST-BIOFILM ASSESSMENT OF BIOFILM-PRODUCING OSTEOMYELITIC STAPHYLOCOCCUS AUREUS TREATED WITH NEW GENTAMICIN-NIGELLA SATIVA FUSION EMULSION (GNFE)

 

Yaakob KI1,5, Mohd Shafri MA*[1]1,4, Mohd Yusof N3, Mohamed F2,4,5

1Department of Biomedical Science, 2Department of Pharmaceutical Technology,

3Orthopaedic Department, Kulliyyah of Medicine, International Islamic University Malaysia IIUM, 25200 Kuantan, Malaysia. 4International Institute of Halal Research & Training (INHART), Ground Floor, Block E0, Kulliyyah of Engineering, IIUM, P.O.Box 10, 50728, Kuala Lumpur, Malaysia. 5IKOP Sdn. Bhd., Pilot Plant Pharmaceutical Manufacturing, International Islamic University Malaysia IIUM, 25200 Kuantan, Malaysia

 

Biofilm complicates osteomyelitis as there is antibiotic resistance and toxicity involved. In order to overcome the challenges of current treatment, gentamicin, the current antibiotic of choice for osteomyelitis, is fused with Nigella sativa oil to form an emulsion. Its efficacy as anti-biofilm agent is assessed using confocal laser scanning microscope (CLSM) against different strains of biofilm-producing Staphylococcus aureus. These strains of S.aureus were first allowed to express biofilm before being exposed for 24 hours to the emulsion with (0.1% (w/v) gentamicin; 40.2% (v/v) N.sativa. Later, the emulsion was removed and the biofilm was stained with fluorescence staining. The slides were viewed under CLSM at 100 times resolution. 3D images of biofilm were reconstructed, using Image J software, to measure the thickness of biofilm and viability of bacteria cells. Results revealed that the emulsion significantly reduced biofilm thickness compared to gentamicin and N.sativa alone in all strains of S.aureus (Tukey’s test p < 0.05). The emulsion was also able to produce more than 80% and 15% surface percentage (%) of non-viable (dead) bacteria in the sensitive and resistant strain, respectively, at a significant level when compared to  gentamicin and N.sativa (Tukey’s test p < 0.05). As a conclusion, this new fusion of gentamicin-N.sativa may be effective towards the biofilm of S.aureus, and can be developed further as a new promising anti-biofilm agent in osteomyelitis.

 

Keywords: biofilm, gentamicin-N.sativa, S.aureus, CLSM

 

* Corresponding author: Tel: +6 019 515 4964

E-mail: affendishafri@iium.edu.my

Published
2015-12-31
Section
Original Research Articles