Lars Magnus Bjursten

Improved bone-forming functionality on diameter-controlled TiO2 nanotube surface

The titanium dioxide (TiO(2)) nanotube surface enables significantly accelerated osteoblast adhesion and exhibits strong bonding with bone. We prepared various sizes (30-100 nm diameter) of titanium dioxide (TiO(2)) nanotubes on titanium substrates by anodization and investigated the osteoblast cellular behavior in response to these different nanotube sizes. The unique and striking result of this study is that a change in osteoblast behavior is obtained in a relatively narrow range of nanotube dimensions, with small diameter ( approximately 30 nm) nanotubes promoting the highest degree of osteoblast adhesion, while larger diameter (70-100 nm) nanotubes elicit a lower population of cells with extremely elongated cellular morphology and much higher alkaline phosphatase levels. Increased elongation of nuclei was also observed with larger diameter nanotubes. By controlling the nanotopography, large diameter nanotubes, in the approximately 100 nm regime, induced extremely elongated cellular shapes, with an aspect ratio of 11:1, which resulted in substantially enhanced up-regulation of alkaline phosphatase activity, suggesting greater bone-forming ability than nanotubes with smaller diameters. Such nanotube structures, already being a strongly osseointegrating implant material, offer encouraging implications for the development and optimization of novel orthopedics-related treatments with precise control toward desired cell and bone growth behavior.

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 dioxide nanotubes enhance bone bonding in vivo

Implant topography is critical to the clinical success of bone-anchored implants, yet little is known how nano-modified implant topography affects osseointegration. We investigate the in vivo bone bonding of two titanium implant surfaces: titanium dioxide (TiO(2)) nanotubes and TiO(2) gritblasted surfaces. In previous in vitro studies, the topography of the TiO(2) nanotubes improved osteoblast proliferation and adhesion compared with gritblasted titanium surfaces. After four weeks of implantation in rabbit tibias, pull-out testing indicated that TiO(2) nanotubes significantly improved bone bonding strength by as much as nine-fold compared with TiO(2) gritblasted surfaces. Histological analysis confirmed greater bone-implant contact area, new bone formation, and calcium and phosphorus levels on the nanotube surfaces. It is anticipated that further studies will contribute to a better understanding of the effect of implant nanotopography on in vivo bone formation and bonding strength.

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.

Clinical, immunological and bacteriological evaluation of adverse reactions to skin-penetrating titanium implants in the head and neck region.

Between 1977 und October 1989, 445 patients have been treated with bone‐anchored skin‐penetrating titanium implants for anchorage of facial prostheses or bone‐conducting hearing aids, at the Ear. Nose and Throat Department at Sahlgrens Hospital in Gothenburg. The majority of patients had no adverse skin read ions, while a few patients were responsible for the majority of the adverse reactions. The aim of our study was to analyse differences between these groups. We started a clinical study on 9 patients with a clinical history of adverse skin reactions around the titanium implants and 9 patients without adverse skin reactions were used us controls. None of the patients had delayed hypersensitivity to titanium., Microbiological analyses showed that when there was clinical irritation, Staphylococccus aureus could be isolated.

Clinical aspects of osseointegration in joint replacement. A histological study of titanium implants

In an experimental clinical study, 25 implants of pure titanium were inserted into the proximal tibia of 11 volunteer patients, four with rheumatoid arthritis and seven with osteoarthritis. The implants were removed from five weeks to 24 months later and detailed histological analysis was performed. The implants generally healed with direct bone-metal contact, showing so-called osseointegration. Only one of the 21 implants which had been in place for over five months did not show osseointegration, probably because of inadequate primary contact with bone. The presence of rheumatoid disease did not prevent osseointegration, but accompanying osteoporosis seemed to be a risk factor.

Tissue response to commercial silicone and polyurethane elastomers after different sterilization procedures

Two different commercial polymeric materials, a silicone and a polyurethane (PUR), were studied with regard to correlations between the chemical and physical compositions of the polymer surfaces and the biological response on implantation. Test specimens of the materials were manufactured according to standard procedures. The specimens were implanted in rats for 10 and 90 days. Before implantation the polymers were sterilized in three different ways, namely, beta irradiation, ethylene oxide sterilization and steam sterilization. The polymers were characterized before and after the implantation with respect to the chemical composition and the morphology of the surfaces. After implantation the biological response was evaluated by counting numbers of macrophages, giant cells, fibroblasts and other cells present at the surfaces. The thickness of the fibrous capsule surrounding the test specimens was measured at the thickest and thinnest parts. PUR surfaces showed signs of degradation already after sterilization and after 10 to 90 days of implantation, pits and cracks appeared, especially in the ethylene oxide sterilized samples. However, differences in the biological responses were small and independent of the sterilization method. After 10 days of implantation the capsule thickness and the amounts of cell material adhering at the surfaces were different, and it appears that the silicone rubber induces more tissue response than PUR. The differences in the early tissue response evened out after 90 days implantation time and a steady state situation evolved, which was similar for the silicone and the polyurethane.

Rat sciatic nerve regeneration through a micromachined silicon chip.

The capacity of regenerating nerve fibres to grow through a perforated silicon chip was tested using the silicone chamber model for nerve regeneration. The chips were fabricated as circular membranes, 4 mm in diameter, thickness 60 microns, with a perforated area, 2 mm in diameter, in the centre. Three types of chips were fabricated utilizing anisotropic etching. The chips were glued with silicone adhesive between two halves of silicone rubber tubing (total length 8 mm, inner diameter 1.8 mm, outer diameter 3.0 mm) which was used to bridge a 4 mm gap between the proximal and distal nerve stumps of a transected rat sciatic nerve. The capacity of regenerating nerve fibres to grow through the holes of the chip was analysed by light and scanning electron microscopy after 4 or 16 weeks of regeneration. Furthermore, the muscle contractility force of the gastrocnemius muscle was measured after 16 weeks of regeneration and compared as a percentage of the contralateral uninjured side. Nerves generated through chips with hole diameters of 10 or 50 microns were morphological and functional failures. The nerve structures distal to chips with hole diameters of 100 microns contained many myelinated nerve fibres in a minifascicular pattern after both 4 and 16 weeks of regeneration. The muscle contractility force was 56% of that of contralateral control muscles.

Method for ultrastructural studies of the intact tissue-metal interface

Samples were prepared for ultrastructural studies of the intact interface between metallic implants and tissue by transmission electron microscopy. The method is based on plastic embedding of implant and tissue and subsequent removal of the bulk metal by electrochemical dissolution (electropolishing), to facilitate preparation of ultrathin sections for transmission electron microscopy. Surface sensitive spectroscopy (Auger electron microscopy and X-ray photoemission spectroscopy) and transmission electron microscopy EDX results show that the method produces samples with an intact interface, containing the implant surface oxide and the adjacent tissue. Examples of application of the method on titanium, zirconium and aluminium implants in soft tissue are given.

Detection of blood group type glycosphingolipid antigens on thin-layer plates using polyclonal antisera

The conditions for binding of antibodies to glycosphingolipids separated on a thin-layer plate have been optimized for polyclonal antisera. The method has a broad detection range with low background staining. Examples are shown for the detection of blood group A and B active glycosphingolipids.