Pentti Tengvall

Titanium in Medicine : material science, surface science, engineering, biological responses and medical applications

This comprehensive book provides state-of-the-art scientific and technical information in a clear format and consistent structure making it suitable for formal course work or self-instruction. T ...

A comparative study of protein adsorption on titanium oxide surfaces using in situ ellipsometry, optical waveguide lightmode spectroscopy, and quartz crystal microbalance/dissipation

Abstract The adsorption kinetics of three model proteins—human serum albumin, fibrinogen and hemoglobin—has been measured and compared using three different experimental techniques: optical waveguide lightmode spectroscopy (OWLS), ellipsometry (ELM) and quartz crystal microbalance (QCM-D). The studies were complemented by also monitoring the corresponding antibody interactions with the pre-adsorbed protein layer. All measurements were performed with identically prepared titanium oxide coated substrates. All three techniques are suitable to follow in-situ kinetics of protein–surface and protein–antibody interactions, and provide quantitative values of the adsorbed adlayer mass. The results have, however, different physical contents. The optical techniques OWLS and ELM provide in most cases consistent and comparable results, which can be straightforwardly converted to adsorbed protein molar (‘dry’) mass. QCM-D, on the other hand, produces measured values that are generally higher in terms of mass. This, in turn, provides valuable, complementary information in two respects: (i) the mass calculated from the resonance frequency shift includes both protein mass and water that binds or hydrodynamically couples to the protein adlayer; and (ii) analysis of the energy dissipation in the adlayer and its magnitude in relation to the frequency shift (c.f. adsorbed mass) provides insight about the mechanical/structural properties such as viscoelasticity.

Physico-chemical considerations of titanium as a biomaterial.

Physico-chemical properties of titanium are discussed. Special attention is paid to those of amorphous TiO 2 that contact tissues in vivo. In aqueous environments TiO 2. (aq) has low ion-formation tendency and low reactivity with macromolecules. This is accompanied by low toxicity. Titanium does not facilitate reactive oxygen radical generation during inflammatory conditions as observed in in-vitro experiments. The outermost layers of the oxide are in the Ti(IV) oxidation state, although using electron spin resonance (ESR) techniques, formation of Ti(III) is observed at atmospheric conditions. The impact of similarities between water and TiO 2 is speculated upon, and the physico-chemical properties of titanium are tentatively linked to some in-vivo consequences.

Titanium-hydrogen peroxide interaction: model studies of the influence of the inflammatory response on titanium implants.

In vitro studies of titanium and TiO2 as well as other metals were carried out to investigate the role of these metals in the inflammatory response through the Fenton reaction. The TiOOH matrix formed traps the superoxide radical, so that no or very small amounts of free hydroxyl radicals are produced. Ellipsometry and spin trapping with spectrophotometry and electron spin resonance (ESR) were used to study the interaction between Ti and H2O2. Spectrophotometry results indicated that Ti, Zr, Au and Al are low free OH-radical producers. We propose a new model for the titanium-tissue interface where the oxidized titanium surface is covered with a hydrated TiOOH matrix after the inflammatory reaction. This matrix is suggested to possess good ion exchange properties, and extracellular components may interact with the Ti(IV)-H2O2 compound before matrix formation. The TiOOH matrix is formed when the H2O2 coordinated to the Ti(IV)-H2O2 complex is decomposed to water and oxygen. Superoxide (O2-) may be bound therein. The oxide layer initially present may be partly reformed to a TiOOH matrix due to the interaction with hydrogen peroxide.

Titanium with different oxides: in vitro studies of protein adsorption and contact activation

Adsorption of albumin (HSA) and fibrinogen (Fib) from human blood plasma onto titanium surfaces with varying oxide properties was studied with an enzyme-linked immunosorbent assay. The intrinsic activation of blood coagulation (contact activation) was studied in vitro using a kallikrein-sensitive substrate. The sample surfaces were characterized with Fourier transform Raman spectroscopy. Auger electron spectroscopy and atomic force microscopy. Low Fib and high HSA adsorption was observed for all titanium samples except for the radio frequency plasma-treated and water-incubated samples, which adsorbed significantly lower amounts of both. Oxide thickness and carbon contamination showed no influence on protein adsorption or contact activation. Smooth samples with a surface roughness (Rrms) < 1 nm showed some correlation between surface wettability and adsorption of Fib and HSA, whereas rough surfaces (Rrms > 5 nm) did not. To varying degrees, all titanium surfaces indicated activation of the intrinsic pathway of coagulation as determined by their kallikrein formation in plasma.

Protein adsorption studies on model organic surfaces : an ellipsometric and infrared spectroscopic approach

The development of accurate analytical tools to control the interfacial properties of solid substrates is of importance for the design of new biomaterials, as well as for the understanding of biomolecular interactions on surfaces. Considerable research efforts are presently devoted to this area on different levels of molecular complexity, i.e. both in the presence and in the absence of the biomolecules. In this contribution we review briefly applications of infrared reflection-absorption spectroscopy (IRAS) and ellipsometry as tools for analysis of the chemical properties of model surfaces, and their biological response in vitro when in contact with blood plasma or serum, respectively. The strength of the combination of the techniques is demonstrated by determination of protein adsorption patterns on a series of chemically well-defined so-called self-assembled alkanethiolate monolayers (SAMs) of 16-thiohexadecanol (HS-(CH2)16-OH) and n-hexadecanethiol (HS-(CH2)15-CH3) on gold. The protein adsorption patterns after incubations in plasma were determined by the specific binding of antibodies to the surfaces.

Peptide functionalized poly(l-lysine)-g-poly(ethylene glycol) on titanium: resistance to protein adsorption in full heparinized human blood plasma

The graft copolymer poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) and its RGD- and RDG-functionalized derivatives (PLL-g-PEG/PEG-peptide) were assembled from aqueous solutions on titanium (oxide) surfaces. The polymers were characterized by NMR in order to determine quantitatively the grafting ratio, g (Lys monomer units/PEG side chains), and the fraction of the PEG side chains carrying the terminal peptide group. The titanium surfaces modified with the polymeric monomolecular adlayers were exposed to full heparinized blood plasma. The adsorbed masses were measured by in situ ellipsometry. The different PLL-g-PEG-coated surfaces showed, within the detection limit of the ellipsometric technique, no statistically significant protein adsorption during exposure to plasma for 30 min at 22 degrees C or 37 degrees C, whereas clean, uncoated titanium surfaces adsorbed approximately 350 ng/cm2 of plasma proteins. The high degree of resistance of the PEGylated surface to non-specific adsorption makes peptide-modified PLL-g-PEG a useful candidate for the surface modification of biomedical devices such as implants that are capable of eliciting specific interactions with integrin-type cell receptors even in the presence of full blood plasma. The results refer to short-term blood plasma exposure that cannot be extrapolated a priori to long-term clinical performance.

Interaction between hydrogen peroxide and titanium: a possible role in the biocompatibility of titanium

Hydroxyl radicals formed from hydrogen peroxide during an inflammatory response are potent agents for cellular deterioration. The behaviour of implanted material in terms of its ability to sustain or stop free radical formation may be therefore very important. In vitro studies of titanium which is known to be biocompatible and osseointegrates into human bone were carried out. In our model studies, the production of free radicals from H2O2 at Ti and TiO2 surfaces was measured by spin trapping techniques. Our findings suggest that there is no sustained hydroxyl radical production at a titanium (oxide) surface. We propose that this is due to the quenching of the Fenton reaction through both trapping and oxidation of superoxide radicals in a TiOOH adduct.

Titanium gel made from metallic titanium and hydrogen peroxide

Abstract Various titanium-peroxo intermediates may be formed in the Ti-H2O2 redox process. In a catalytically decomposed Ti-H2O2 system, an oxidizing transparent yellow-green gel with a final pH between 3 and 4 is typically formed within 48 h without addition of chelating agents. The redox potential of the gel is larger than that of the Fe(II)/Fe(III) system. The total Ti concentration in the studied gel was spectro-photometrically determined to be about 45 mM. The oxidizing component was determined to be about 15 mM with the use of iodometric titration. A model for the catalytic H2O2 decomposition is discussed, and an explanation for the existence of an ESR-active radical component in the gel is suggested. The possible relation of the gel formation to the known biocompatibility of titanium is speculated on.

Blood protein adsorption onto chitosan

Chitosan was recently indicated to enhance osteogenesis, improve wound healing but to activate the coagulation and the complement systems. In the present study approximately 10 nm thick chitosan film were prepared on aminopropyltriethoxysilane (APTES) coated silicon. The surfaces were incubated in serum or plasma and subsequently in antibodies towards key complement and contact activation of coagulation proteins. The deposited amounts were compared with those on hydrophilic and hydrophobic silicon, APTES and IgG coated reference samples. Although large amounts of serum deposited to chitosan only a weak transient activation of the complement system and no activation of the intrinsic pathway was observed. Upon acetylation the chitosan layer became a strong activator of the alternative pathway of the complement. After incubation in human plasma anti-fibrinogen deposited onto chitosan but not onto the acetylated chitosan, a finding that may explain previous observations of procoagulant activity by chitosan.