Abstract

HMS0014 cells were GBR-engineered to proliferate and differentiate into mature osteoblast(Ob)-like cells, which initiated hard tissue matrix deposition in both monolayer and PuraMatrix 3-D cultures. Subsequently, the osteogenesis initiated with attachment/adhesion of HMS0014 cells on either Titanium (Ti) or Ti alloy discs modified with osteoconductive/ osteoinductive surface textures/substrates (e.g., Disc-AO, Disc-HA, Disc-SPI) was histologically assessed. The results obtained were as follows: 1) The HMS0014 cells actively proliferated and differentiated into mature Obs to initiate mineralisation of the ECM since day 1 in both monolayer and 3-D cultures; mineralization was prominently progressed between day 7 and day 14 of cultures. 2) The SEM of 60-minute(min)s specimens demonstrated a loose distribution of proliferating spherical-to-polygonal (10 to 40 microm in diameter, avg.) cells with a bulging cell body sending out many minute filopodia and some lamellipodia to attach with the substrate in the concavities. 3) In the 180-min specimens, the cultured HMS0014 cells actively proliferated and spread into flat, large polygonal cells with prominent lamellipodia and dendritic filopodia (30 microm x 90 microm to 100 microm x 200 microm, approx.) to employ cell-to-substrate and intercellular attachments. 4) On the other hand, the present immunohistochemistry of the attached HMS0014 cells demonstrated the co-expression of F-actin (actin filaments of the cytoskeleton) and CD51 (aV integrin) in both the 60-min and 180-min specimens. We concluded that the present GBR method enhanced HMS0014 cells to initiate an osteogenesis process with a direct bone-to-substratum contact on Ti discs which were subject to different surface modifications.

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