2007   Volume No 13– pages 11-25

Title: In-vitro interactions of human chondrocytes and mesenchymal stem cells, and of mouse macrophages with phospholipid-covered metallic implant materials

Author: R Willumeit, M Schossig, H Clemens, F Feyerabend

Address: GKSS Research Center, Max-Planck-Str.1, 21502 Geesthacht, Germany

E-mail: regine.willumeit@gkss.de

Key Words: Phospholipid-coatings, porous Ti-6Al-4V, polished Ti-6Al-7Nb, chondrocytes, mesenchymal stem cells, macrophages, in vitro tests.

Publication date: March 2nd 2007

Abstract: Phospholipid-coatings on metallic implant surfaces were evaluated in terms of adhesion, proliferation and matrix production of skeletal cells, and of macrophage stimulation. The working hypothesis is that mimicking a model biomembrane by phospholipids on surfaces to which cells adhere, the surface recognition by surrounding cells is altered. In this study, 1) mirror-like polished Ti-6Al-7Nb and 2) porous Ti-6Al-4V specimens were covered with the phospholipids POPE (palmitoyl-oleoyl phosphatidyl-ethanolamine) and POPC (palmitoyl-oleoyl phosphatidyl-choline), and the interactions of a) human articular chondrocytes (HAC), b) human mesenchymal stem cells (HMSC), and c) mouse macrophages (RAW 264.7) were tested in vitro.

On POPE-covered polished surfaces adherence of HAC (42% of seeded cells after 2 hrs) and metabolic activity (MTT after 3 days) were reduced, while on porous surfaces 99% HAC adhered, and metabolic activity was significantly increased, compared to respective native surfaces. On both POPE-covered surfaces the chondrocyte phenotype was present. After 3 weeks of chondrogenic differentiation, cartilage matrix production (measuring chondroitin sulphate per HAC number) was significantly increased by about 30% on both POPE-covered metallic surfaces. On both POPC-covered surfaces nearly no adhering and surviving HAC were found.

HMSC grown on POPE-covered porous substrates showed osteogenic differentiation by improved osteopontin and collagen I expression in RT-PCR, and osteocalcin fluorescence and bone nodule formation was only detectable on POPE-covered porous surfaces. In contrast to POPC and other phospholipids used as positive controls, POPE did not stimulate the NO production in mouse macrophage cultures. We therefore conclude that a phospholipid coating by POPE shows potential as surface modification for metallic implant materials.

Article download: Pages 11-25 (PDF file)