Physical stress such as the surface roughness of the implants may activate the NFkB signalling pathway in macrophages. This activation is intimately related to the mechanism(s) by which the macrophage interacts with the surface through serum proteins and/or the formation of membrane rafts. This thesis examines the role of surface topography on activation of the NFkB signalling pathway in macrophages.
We examined the effect of implant surface topography on activating the NFkB signalling pathway in the RAW 264.7 macrophage cell line. We also examined the effect surface roughness had on the adhesion of the macrophages using the different media. To finish, we observed the effect the different media and the surface roughness had on the morphology of the macrophages by Scanning Electron Microscopy.
Activation of the NFkB pathway was surface topography dependent. The Smooth surface showed the highest level of activation followed by the Etched then the SLA. Addition of suboptimal concentrations of LPS mildly enhanced the response by signalling through the Toll receptor. Activation of NFKB occurred in the absence of fetal calf sera, although to a lesser extent. All three surfaces had very few cells with nuclear translocation at the 5 minutes time point with no significant statistical differences between the surfaces. After 30 minutes, translocation reached comparable levels to those surfaces tested with complete medium. Disruption of the lipid rafts affected the triggering and signalling of the NFkB pathway. This inhibitory effect was concentration and time dependent. Smooth surfaces bound more macrophages in the 30 minutes assay. Fetal calf serum appeared to be very critical for adhesion and spreading of the macrophages on the various surfaces examined. Removal of cholesterol did not affect adhesion or spreading on their respective surfaces.
We have clearly demonstrated that the lipid rafts along with surface topography play a role in the activation on NFKB. This in-vitro study has demonstrated that surface topography modulated activation of the NFKB signalling pathway in a time-dependent manner. However, at present, it is unclear through which receptor(s) / surface structure the signal pathway is initiated.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:BVAU./2436 |
| Date | 05 1900 |
| Creators | Ali, Tarek Adel |
| Publisher | University of British Columbia |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | 5240553 bytes, application/pdf |
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