Inflammatory Cytokine Quantification of Cell-SCK Interactions
via RT-PCR
Experimental Approach
Measurement of the acute inflammatory reaction
in response to peptide derivatized shell cross-linked (SCK)
nanoparticle exposure is a critical component in the assessment
of the biocompatibility. The inflammatory response to peptide
functionalized SCKs was found to be dependent, with respect
to functionality and concentration.
Results
Shell cross-linked (SCK) nanoparticles functionalized
with different amounts of the protein transduction domain (PTD)
of HIV-1 Tat causes statistically significant increases in IL-1?
production after 24 h of incubation that are both functionalization
and concentration dependent as quantified using RT-PCR. Error
bars are representative of one standard deviation from the mean
of triplicate samples, each harvested from three separate populations
of RAW 264.7 cells, and are the estimates of the standard uncertainties.
The measured increases of IL1-b production in response to
PTD, SCK, and PTD functionalized SCKs measured from 18.8 fold
to 483.5 fold increases over TCPS and 1 fold to 25.4 fold increase
over SCKs of similar concentration and show statistical dependence
with regard to both concentration and PTD functionalization.
The differences in TNF-a and IL1-b expression measured in this
series reflect different pathways of signal transduction with
regard to both the timeframe and the severity of the response
to the respective inflammatory stimuli.
NIST Contributors
Matthew L. Becker
Lee Ann O. Bailey
NIST Contributors
Professor Karen L. Wooley
Edward E. Remsen and Dipanjan Pan
Washington University
Biomaterials Group
Polymers Division
Materials Science and Engineering Laboratory
