Track D

MethodsEAC were decellularized by a detergent-based protocol for 40h (40h-dEAC) or 72h (72h-dEAC) and coated with or without 100ng/ml CCN1.Subsequently, THP-1 cells were seeded onto the scaffolds for 24 hours.For negative and positive controls THP-1 cells were seeded on tissue culture plastic stimulated with or without 100nM PMA.Expression of pro-inflammatory cytokines TNF-alpha, MIP-1alpha, MCP-1 and IL-1beta and anti-inflammatory cytokines IL1-ra and IL-10 were determined using quantitative reverse transcriptase PCR. ResultsCytokine expression in THP-1 cells seeded onto uncoated scaffolds showed for 40h-dEAC a mean 3.3-fold increase versus plastic significant for TNF-alpha, IL-1beta and IL-10 (p<0.05) and for 72h-dEAC a 15.9-fold increase significant for TNF-alpha, MIP, MCP-1 and IL-1beta (p<0.01).72h-dEAC induced an overall higher cytokine expression except for IL-1beta and IL-10.CCN1-coating strongly induced cytokine expression on both 40h-and 72h-dEAC.For 40h-dEAC, expression of TNF-alpha, MIP, MCP-1alpha, and IL-1beta showed a mean 113.9-fold increase (p<0.001) whereas on 72h-dEAC expression of all cytokines except IL10 showed a mean 166-fold increase (p<0.001).Absolute cytokine expression on CCN1-coated scaffolds showed no significant difference versus uncoated dEAC. ConclusionTHP-1 cells respond to CCN1-coated dEAC by overall higher cytokine expression.As this seem to be contradictory to the beneficial effects of CCN1 observed before, further research is needed to determine the effect of pro-and antiinflammatory cytokine up regulation on wound healing, fibrotic tissue formation and angiogenetic properties in vivo.