(Georgia Institute of Technology, 2010-11-15)
Rogers, Todd H.
The goal of the work presented was to further understand how both the body and dendritic cells (DCs) interact and respond to biomaterials through receptor-mediated mechanisms. The role of Toll-like receptor 4 (TLR4) was investigated in the host response to biomaterials, and it was found that TLR4-deficient mice (in comparison to wild-type) had a delayed acute inflammatory response as seen through an altered adherent leukocyte profile on implanted polymer discs. However, following a 2 week implantation, the response was resolved potentially through compensatory receptors. Therefore, TLR4 may aid in the initial response to a biomaterial through recognition of 'danger signal' molecules. An investigation into the role of TLR4 in the response of DCs to biomaterials was investigated using murine bone marrow-derived DCs (BMDC), and PLGA film or microparticle treatment of BMDCs resulted in TLR4-dependent signs of slight maturation in non/loosely adherent BMDCs. However, further investigation into BMDC populations within the culture system revealed that non/loosely adherent BMDCs took on an activated/mature phenotype while adherent BMDCs appeared to be less mature and more responsive to both LPS and biomaterial stimuli. Therefore, it was concluded that investigations into the responsiveness of BMDCs to stimuli in the future analyze both adherent and non/loosely adherent populations. Lastly, the role of integrin-mediated adhesion in biomaterial-induced DC maturation was investigated. Gene expression analysis revealed that PLGA treatment of human DCs increased adhesion molecule expression (including β1 and β2 integrin subunits), LPS treatment reduced adhesion molecule expression and agarose treatment did not alter their expression. Antibody blocking techniques pinpointed the role of β2 integrins (and not β1 integrins) in both the adhesion of DCs to TCPS or PLGA substrates and the regulation of a DC maturation marker (CD86). β2 (and not β1) was found co-localized with F-actin in podosomes of DCs adhering to PLGA, and the direct interaction of β2 (and not β1) to PLGA substrate was confirmed through crosslinking and immunofluorescence studies. Therefore, DCs utilized β2 integrins for both adhesion and maintenance of immunomodulatory status. This aids the field of tissue engineering and vaccine design by further developing the criteria for biomaterial-influenced immunomodulation.