Long-term subcutaneous microdialysis sampling and qRT-PCR of MCP-1, IL-6 and IL-10 in freely-moving rats

Abstract

Cytokines are important mediators of the wound healing response. However, sampling of cytokines from the interstitial fluid at a healing wound site in experimental animals is a challenge. Microdialysis sampling is an in vivo collection option for this purpose as it permits continuous sampling, while remaining contiguous with the wound microenvironment. The polymeric membrane of the microdialysis probe is a foreign material thus allowing a unique approach to sample cytokines generated during a foreign body response (FBR). The focus of these studies was to use microdialysis sampling to collect cytokines from a microdialysis probe implant site in a rat model of a FBR up to 6 days post implantation. Fluorescent bead-based immunoassays (Luminex™) were used to quantify monocyte chemoattractant protein-1 (MCP-1/CCL2), interleukin-6 (IL-6) and interleukin-10 (IL-10) in the dialysates. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to cross validate the protein measurements obtained via micorodialysis sampling. A histological examination of tissue was also performed to assess the progression in leukocyte extravasation and collagen deposition surrounding implanted probes. Our findings demonstrate that in vivomicrodialysis sampling can be used to collect temporal concentrations of cytokines which are consistent with wound healing and the development of a FBR.