Titanium (Ti) is a well known metallic biomaterial extensively used in dental, orthopaedic-, and occasionally also in blood contacting applications. It integrates well to bone and soft tissues, and is shown upon blood plasma contact to activate the intrinsic pathway of coagulation and bind complement factor 3b. The material properties depend largely on those of the nm-thick dense layer of TiO2 that becomes rapidly formed upon contact with air and water. The spontaneously formed amorphous Ti-oxide has a pzc similar to 5-6 and its water solubility is at the order of 1-2 micromolar. It is often subjected to chemical- and heat treatments in order to increase the anatase- and ruble crystallinity, to modify the surface topography and to decrease the water solubility. In this work, we prepared sol gel derived titanium and smooth PVD titanium surfaces, and analysed their oxide and protein deposition properties in human blood plasma before and after annealing at 100-500 degrees C or upon UVO-treatment for up to 96 hours. The blood plasma results show that complement deposition vanished irreversibly after heat treatment at 250-300 degrees C for 30 minutes or after UVO exposure for 24 hours or longer. XPS and infrared spectroscopy indicated change of surface water/hydroxyl binding upon the heat- and UVO treatments, and increased Ti oxidation. XRD analysis confirmed an increased crystallinity and both control (untreated) and annealed smooth titanium displayed low XRD-signals indicating some nanocrystallinity, with predominantly anatase phase. The current results show that the behaviour of titanium dioxide in blood contact can be controlled through relatively simple means, such as mild heating and illumination in UV-light, which both likely irreversibly change the stoichiometry and structure of the outmost layers of titanium dioxide and its OH/H2O binding characteristics.

In the present study the initial blood compatibility of glycerol monooleate (GMO)-coated surfaces was evaluated after deposition to surfaces and in bulk. The model surface was silica onto which multiple layers of fibrinogen or human serum albumin (HSA) was immobilized. The protein-coated surfaces were subsequently dip-coated in GMO in ethanol and used for blood plasma and whole blood experiments. The characterization methods included null ellipsometry, scanning electron microscopy, imaging of coagulation, hemolysis test and whole blood coagulation time by free oscillation rheometry.

The results showed a GMO film thickness of approximately 350 angstrom (similar to 4 mu g/cm(2)) upon dip-coating in ethanolic solution. A major part of the deposited layer detached in aqueous solutions, especially during shear conditions. The coagulation time on GMO was significantly prolonged compared to that on HSA coated silica. Whole blood tests showed that GMO is a very weak hemolytic agent. Deposited GMO detached easily from surfaces upon rinsing or shearing, although a stable layer with undefined phase structure and a thickness of 50-70 angstrom remained on HSA and fibrinogen precoated surfaces. This indicates that GMO has stronger adhesive forces to its substrate compared to the cohesive forces acting within the bulk GMO. The ability of GMO to detach from itself and tentatively form micelles or lipid bilayers when subjected to flowing blood may be of use in extravascular applications. It is concluded that GMO results in weak blood activation, and the material may in spite of this be suitable in selected biomaterial applications, especially as a biosealant and in colloidal dispersions.

Methodology for studies of effects of heating multilayers of human serum albumin (HSA) and anti-HSA is presented. Multilayers of anti-HSA were prepared on silicon substrates and studied with infrared spectroscopic ellipsometry equipped with a heat stage. The refractive index N = n + ik and the layer thickness are determined and the amide bands are analyzed. It is found that HSA/anti-HSA multilayers are stable for shorter times at temperatures above 100 °C, except for small thickness changes. Also pilot studies of effects of heating monolayers of proteins adsorbed on gold substrates is presented.

There is evidence of high matrix metalloproteinase (MMP) activity around sutures inserted into tendons. This probably results in tissue breakdown, allowing the suture to cut through the tendon, and thus contributes to repair-site elongation and gap formation. We therefore hypothesized that treatment with the MMP inhibitor doxycycline would improve the sutureholding capacity of tendon. Animals, methods and results In the first sub-study, rats received a suture in the Achilles tendon. One group was treated with systemic doxycycline and the other received no treatment. At 3, 5, and 7 days, suture-holding capacity was measured mechanically. The pull-out force and energy were reduced in all tendons, at 3 days compared to freshly inserted sutures, but no further reduction was detected at later time points. Doxycycline- treated tendons showed improved suture-holding capacity as measured by higher energy uptake than in untreated tendons. Force at failure showed a trend towards improvement. The effect was most evident on day 3. In the second sub-study, sutures were coated with doxycycline. At 3 days, local doxycycline treatment caused improved suture-holding capacity—as measured by higher force at failure and higher energy uptake.

C-reactive protein (CRP) interacts with phosphorylcholine (PC), Fcγ receptors, complement factor C1q and cell nuclear constituents, yet its biological roles are insufficiently understood. The aim was to characterize CRP-induced complement activation by ellipsometry. PC conjugated with keyhole limpet hemocyanin (PC-KLH) was immobilized to cross-linked fibrinogen. A low-CRP serum with different amounts of added CRP was exposed to the PC-surfaces. The total serum protein deposition was quantified and deposition of IgG, C1q, C3c, C4, factor H, and CRP detected with polyclonal antibodies. The binding of serum CRP to PC-KLH dose-dependently triggered activation of the classical pathway. Unexpectedly, the activation was efficiently down-regulated at CRP levels >150 mg/L. Using radial immunodiffusion, CRP–C1q interaction was observed in serum samples with high CRP concentrations. We propose that the underlying mechanism depends on fluid-phase interaction between C1q and CRP. This might constitute another level of complement regulation, which has implications for systemic lupus erythematosus where CRP is often low despite flare-ups.