It have been reported that the wear volume of vitamin E-containing UHMWPE tested with a knee joint simulator was approximately 30% lower than that of virgin UHMWPE at 5 million cycles. However, the wear resistance mechanism of vitamin E-containing UHMWPE has not yet been clarified. The present study examines the effects of the addition of vitamin E on the frictional properties of ultra-high molecular weight polyethylene (UHMWPE) under several different load and serum conditions.

Friction tests were carried out using a computer-controlled pin-on-disk friction test apparatus. The UHMWPE pin was mounted vertically at the tip of the leaf spring and linear reciprocating sliding motion for 2,000 cycles with an amplitude of 1 mm and a frequency of 1 Hz, was applied under 3 MPa or 30 MPa loading against Co-28Cr-6Mo alloy disk. The lubricant bath was filled with 5 ml of ultrapure water, fresh serum, post-friction (PF) serum or diluted-PF (DPF) which were kept at a temperature of 37°C. The friction force between the UHMWPE pin and the Co-28Cr-6Mo alloy disk was calculated from the displacement of the leaf spring during the sliding motion.

Vitamin E-containing UHMWPE showed a significantly higher friction force than that of virgin UHMWPE in fresh serum lubricant at 30 MPa loading, while there were little differences in either ultrapure water or PF serum or DPF serum. And vitamin E-containing UHMWPE tends to exhibit a lower dynamic friction force within the first few hundred cycles in the case of all serum lubricants at 30 MPa loading. These results suggest that some interaction between the UHMWPE surface and the native conformation proteins was specifically affected by the addition of vitamin E and that some weeping of vitamin E might occur at early stage of sliding. Our results also suggest the importance of the conformational changes of serum proteins for the wear testing.

Ex vivo gene transfer of osteogenic factors into multipotential stem cells offers potentially important therapeutic implications in a variety of musculoskeletal diseases. One possible approach is the development of a cellular vehicle, namely bone morphogenetic protein (BMP)-producing bone marrow cells, created using adenoviral gene transfer. These transduced cells provide local delivery of BMP for bone formation. The aims of this study were to study the feasibility of gene transfer to human bone osteoprogenitor cells, using adenoviral vectors. Specifically, the aims were to study the efficacy of transduction with an adenoviral vector expressing BMP-2 and then to determine the ability of the transduced cells to produce active BMP-2 and to generate bone ex vivo.

Primary human bone marrow osteoprogenitor cells were expanded in culture and infected with AxCALacZ, a replication-deficient adenoviral vector carrying the E. coli lacZ gene, with a range of multiplicity of infection (MOI) of 6.25 to100. Transduced cells showed positive staining for β-galactosidase using X-Gal with an efficiency close to 100%. Uninfected cells showed no β-galactosidase activity. Efficiency was independent from MOI, however cells infected at the lower MOIs expressed lower levels of β-galactosidase. Following confirmation that primary bone marrow cells could be infected by adenoviral constructs, additional osteoprogenitors were infected with AxCAOBMP-2, a vector carrying the human BMP-2 gene, at a multiplicity of infection of 10–20. In order to determine BMP-2 activity, conditioned media from bone marrow cells expressing BMP-2 was added to promyoblast C₂C₁₂ cells. The promyoblast C₂C₁₂ cells are exquisitely sensitive to BMP-2 with induction of alkaline phosphatase activity (ED₅₀ 20 nM) in a dose-dependant manner. Alkaline phosphatase activity was induced following culture with conditioned media from BMP-2 expressing cells, in a dose dependant manner, confirming successful secretion of active BMP-2. Immunohistochemical staining for alka- line phosphatase in C₂C₁₂ cells also confirmed the bio-chemical observations. Media from uninfected control human bone marrow cells failed to produce a similar effect. The concentration of BMP-2 in the media was estimated to be 5–10 nM/10⁷ cells.

To examine whether adenoviral transfection affected the osteoblast phenotype and their ability to mineralise in vitro, adenovirally-transduced bone marrow cells expressing BMP-2 were seeded onto poly(-lactic acid co÷glycolic acid) (75:25) porous scaffolds (provided by K. Shakesheff and S. Howdle; Nottingham University) and cultured for up to 6 weeks. Expression of alkaline phosphatase activity, type I collagen formation, as well as the synthesis of osteoblast stimulating factor-1 confirmed bone cell differentiation and maintenance of the osteoblast phenotype in extended culture for up to 6 weeks.

These results indicate the ability to deliver active BMP-2 using human bone marrow osteoprogenitor cells following adenoviral infection. The maintenance of osteoblast phenotype in extended culture and generation of mineralised 3-D scaffolds containing such constructs offers a realistic approach to tissue engineer bone for orthopaedic applications.