ARTICLE IN PRESS 09) 111 b5 54433 050 075 0 左 0 左 0 ys 111 654 766 1.3 0 2 5.5 110 1 5.0 品8- 3. 5 0 000000 50 0 0.0 10 15 Days 443 E5 01血图象自 10 15 6 Fibroblasts (a, c e)and osteoblasts(b, d, f) were cultured for 3, 6, 10 and 15 days in the presence of particles (1000 ug/mL)of conditions in 24-well culture ssay(a, b) sted and centrifuged in order to eliminate residual particles. are expressed as mean* SEM, n=9 for culture in the presence of particles and n= 12 for controls: *p<0.05, ++p<0.01, +++p<0.001(compared to control cultures). described previously in the cell culture medium, alumina and anatomical region. Most of them were found near the patella zirconia particles looked agglomerated (paragraph 3.3). The particular in the folds of the synovial membrane. Our histological amount of particles observed was different depending on the evaluation score exhibited that both alumina and zirconia particles Please cite this article in press as: Roualdes 0, et al, In vitro and in vivo evaluation of an, Biomaterials(2009). doi: 10.1016/ j biomaterials 2009. 11.107described previously in the cell culture medium, alumina and zirconia particles looked agglomerated (paragraph 3.3). The amount of particles observed was different depending on the anatomical region. Most of them were found near the patella, in particular in the folds of the synovial membrane. Our histological evaluation score exhibited that both alumina and zirconia particles 3 6 10 15 0.00 0.25 0.50 0.75 1.00 1.25 1.50 + +++ + + Days 3 6 10 15 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 ++++ a b Days 3 6 10 15 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 1.1 1.2 1.3 1.4 1.5 1.6 c Days 3 6 10 15 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 + + d Days 3 6 10 15 0 5 10 15 20 25 30 35 40 45 50 55 ++ +++++ e Days 3 6 10 15 0 1 2 3 4 5 6 7 8 9 10 11 f Days Cell proliferation (optical density) Human type I collagen (µg/mL) Human fibronectin (µg/mL) Cell proliferation (optical density) Human type I collagen (µg/mL) Human fibronectin (µg/mL) Fig. 5. Fibroblasts (a, c, e) and osteoblasts (b, d, f) were cultured for 3, 6, 10 and 15 days in the presence of particles (1000 mg/mL) of alumina ( ), zirconia ( ) or without particles ( ) under standard cell culture conditions in 24-well culture plates. At each time of culture, cell proliferation was assessed using the MTT assay (a, b) and culture media were harvested and centrifuged in order to eliminate residual particles. Supernatant were used for human type I collagen (c, d) and human fibronectin (e, f) synthesis quantification. Values are expressed as mean  SEM, n ¼ 9 for culture in the presence of particles and n ¼ 12 for controls; þp < 0.05, þþp < 0.01, þþþp < 0.001 (compared to control cultures). O. Roualdes et al. / Biomaterials xxx (2009) 1–12 7 ARTICLE IN PRESS Please cite this article in press as: Roualdes O, et al., In vitro and in vivo evaluation of an..., Biomaterials (2009), doi:10.1016/ j.biomaterials.2009.11.107