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Quantification of inflammatory cellular responses using real-time polymerase chain reaction
LeeAnn O Bailey1, Newell R Washburn, Carl G Simon
1Biomaterials Group, Polymers Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899-8545, USA. leeann.bailey@nist.gov
Journal of Biomedical Materials Research. Part A
|April 2, 2004
Summary
This study quantifies cellular inflammatory responses to biomaterials. Polymethylmethacrylate (PMMA) microspheres, combined with lipopolysaccharide (LPS), significantly elevated cytokine levels in macrophages, indicating a synergistic inflammatory reaction.
Area of Science:
- Biomaterials Science
- Immunology
- Tissue Engineering
Background:
- Biomaterials are crucial for tissue engineering but often trigger complex inflammatory responses.
- Understanding these responses is key to improving biomaterial biocompatibility.
Purpose of the Study:
- To develop improved methods for quantifying cellular inflammatory responses to biomaterials.
- To enhance the understanding of the body's reaction to implanted biomaterials.
Main Methods:
- Established an in vitro system to analyze and quantify cellular inflammatory responses.
- Incubated murine macrophages (RAW 264.7) with polymethylmethacrylate (PMMA) microspheres and lipopolysaccharide (LPS).
- Quantified genetic material using real-time reverse transcription polymerase chain reaction (RT-PCR) and assessed cellular viability via fluorescence microscopy.
Main Results:
- Both LPS and PMMA microspheres individually increased cytokine levels compared to controls.
- PMMA increased cytokine levels by 87.9-fold, while LPS caused a 258-fold increase at 8 hours.
- Combined LPS and PMMA exposure showed a synergistic effect, increasing cytokine expression 336-fold at 8 hours.
Conclusions:
- The developed in vitro system effectively quantifies inflammatory responses to biomaterials.
- PMMA microspheres elicit a significant inflammatory response in macrophages.
- A synergistic inflammatory effect occurs when macrophages are exposed to both PMMA and LPS.