Periprosthetic osteolysis depends on the biological activity of wear particles, but there is little known about the distribution of polyethylene wear particles (PE) in the surrounding joint tissue. The purpose of this study was to examine the localisation of wear particles of six different PEs, including four crosslinked polyethylenes (XPE), as well as their biological activity in the murine knee.

Crosslinked polyethylene (XPE) was developed to reduce wear in hip and knee arthroplasty. Periprosthetic osteolysis depends on many factors including biological activity of wear particles. This study examines the relative inflammatory effect of different crosslinked polyethylenes compared to ultra-high-molecular-weight-polyethylene (UHMWPE) particles in vivo.

Materials and Methods: Wear particles of 3 XPE- (1 sequential irradiated/annealed; 2 remelted inserts) and 1 UHMWPE-insert were isolated from a knee joint simulator (20nm-nucleopore-filter;acid digestion method;ISO). Particles were analysed by scanning electron microscopy (n=66000). For all groups the particles were smooth, granular, irregular and less fibrillar. More than 85% of the particles were submicron. After removal of endotoxin the particles were suspended in a phosphate buffered saline solution (0.1% vol/vol (particle volume/PBS volume)). Endotoxin levels were controlled using standardised endotoxin detection tests (Lonza) in all samples.

40 female Balb/c mice were randomly assigned to one of five treatment groups (according to the national guidelines of animal protection laws): control (n=8); XPE1 (95 kGy E-beam, remelted; n=8); XPE2 (65 kGy E-beam, remelted; n=8), XPE3 (3x30 kGy Gamma, annealed and sequential irradiated; n=8) and UHMWPE particles (n=8). 50 μl of the particle suspension were injected into the murine left knee under sterile conditions. The leukocyte–endothelial cell interactions and the synovial microcirculation were performed by intra-vital fluorescence microscopy one week after particle injection to assess the inflammatory reaction to the particles (by measuring the rolling fraction of leukocytes, the adherent cells and the functional capillary density (FCD)). Data analysis was performed using a computer-assisted microcirculation analysis system (Cap-Image).

For the statistical analysis the Kruskal-Wallis test was used to determine differences within the groups, followed by an all pairwise multiple comparison procedure with a Bonferoni correction. The level of significance was set at p< 0.05.

Results: The fraction of the rolling leukocytes, adherent cells and FCD increased significantly (p< 0.05) in all bio-materials compared to control group. However, there was no significant difference between the UHMWPE and the XPE particle groups (p> 0.05).

Conclusion: Our data suggest that crosslinked polyethylene wear particles do not lead to a higher inflammatory reaction in vivo compared to UHMWPE particles.

Background: In regenerative medicine the autologous cartilage implantation (ACI) has been used for the repair of cartilage defects. As modification of ACI, the matrix assisted ACI is used nowadays with varying results. There is a general discussion about whether supporting scaffolds should be used or whether a scaffold-free cartilage repair is the method of choice. The major problem of scaffold-free regenerates is how to keep the cells in place after transplantation. Aim of this study was to examine a new scaffold-free diffusion-culture model, which uses a mega-congregate of chondrocytes cultured at an air-medium interface. This scaffold-free high-density diffusion culture could be used to repair cartilage defects.

Material and methods: Human chondrocytes from passage 1–7 were expanded in monolayer and transferred to pellet-culture or diffusion-culture. After one week cultures were stained with toluidine blue and safranin-O and evaluated by immunohistochemical staining for type II collagen. Quantitative real time reverse transcriptase polymerase chain reaction (qRT-PCR) was performed for the mRNAs of cartilage markers.

Results: Positive alcian blue staining was detectable in diffusion-culture for human chondrocytes up to passage 7. Within passages the amount of proteoglycan production in relationship to the number of cells increased. There was a positive signal for Collagen type II in diffusion-cultures up to passage 7.

In qRT-PCR a redifferentiation of human chondrocytes was shown by the transfer into diffusion-culture. Within passage 1 to 3 human chondrocytes which were cultured in monolayer lost the ability to express Collagen Type II but could regain it if they were transferred to diffusion-culture. At diffusion-culture chondrocytes showed the highest expression of Collagen type II at passage 1 when compared to monolayer or to pellet-culture.

Conclusion: It could be shown that the cultivation in a scaffold-free diffusion-culture can lead to redifferentiation of human chondrocytes Chondrocytes in diffusion-cultures tend to form their own matrix and produce Collagen type II at higher amounts than in monolayer or in normal pellet-cultures. Therefore diffusion-culture congregates might be an appropriate tool to be used for a new scaffold-free cartilage regeneration approach.