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Improved Enzyme Protection Assay to Study Staphylococcus aureus Internalization and Intracellular Efficacy of Antimicrobial Compounds
Published on: September 8, 2021
Ultrasonically enhanced rifampin activity against internalized Staphylococcus aureus
Si-Feng Shi1, Xian-Long Zhang, Chen Zhu
1Department of Orthopaedic Surgery, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200233, P.R. China.
Abstract:
Staphylococcus aureus (S. aureus) is the principle causative agent of osteomyelitis, accounting for 80% of all human cases. S. aureus internalized in osteoblasts escapes immune response, including engulfment by phagocytes. It also escapes the action of a number of antibiotics. Ultrasound increases cell membrane permeability to a number of drugs. Following an internalization assay, we used low-frequency, low-power ultrasound combined with the antibiotic rifampin to target S. aureus internalized in human osteoblasts. Tryptic soy agar (TSA) was used to quantitate the antibacterial effect of rifampin combined with low-frequency ultrasound. A Cell Counting Kit-8 (CCK-8) assay was used to evaluate cell viability following exposure to ultrasound. Our data revealed that rifampin successfully penetrates into osteoblasts and kills internalized S. aureus in osteoblasts, while low-frequency ultrasound promotes this process. Ultrasound had a negative impact on the cell viability of osteoblasts; however, this damage was slight and reversible. Ultrasound-enhanced antibiotic efficiency to bacteria internalized in the osteoblasts may contribute to the control of chronic infection to reduce recurrence.
Insights
Low-frequency ultrasound enhances rifampin
Area of Science:
- Microbiology
- Biomedical Engineering
- Cell Biology
Background:
- Staphylococcus aureus (S. aureus) is the primary cause of osteomyelitis, leading to 80% of human cases.
- Intracellular S. aureus within osteoblasts evades immune responses and antibiotic treatments.
- Ultrasound technology can enhance cell membrane permeability, potentially improving drug delivery.
Purpose of the Study:
- To investigate the efficacy of combining low-frequency ultrasound with rifampin against S. aureus internalized in human osteoblasts.
- To assess the impact of ultrasound on osteoblast viability.
- To explore ultrasound-enhanced antibiotic delivery for treating intracellular bacterial infections.
Main Methods:
- Internalization assays were performed to quantify S. aureus within osteoblasts.
- Low-frequency, low-power ultrasound was applied in combination with rifampin.
- Antibacterial effects were measured using Tryptic Soy Agar (TSA), and cell viability was assessed with a Cell Counting Kit-8 (CCK-8) assay.
Main Results:
- Rifampin demonstrated effectiveness in penetrating osteoblasts and eliminating internalized S. aureus.
- Low-frequency ultrasound significantly enhanced the antibacterial activity of rifampin.
- Ultrasound exposure caused minimal and reversible damage to osteoblast cell viability.
Conclusions:
- Combining low-frequency ultrasound with rifampin is a promising strategy for targeting intracellular S. aureus in osteomyelitis.
- This approach enhances antibiotic efficacy against bacteria residing within osteoblasts.
- Ultrasound-enhanced antibiotic therapy may offer a novel method for controlling chronic S. aureus infections and reducing recurrence.
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