Enhancement of Cell Ingrowth, Proliferation and Early Differentiation in a 3-D SiC Scaffold using Low-Intensity Pulsed Ultrasound.

Concerns over the use of autografts or allografts have necessitated the development of biomaterials for bone regeneration. Various studies have been performed to optimize the cultivation of osteogenic cells using osteoconductive porous scaffolds. The aim of this study was to evaluate the osteogenic efficiency of bone cell ingrowth, proliferation and early differentiation in a silicon carbide (SiC) porous ceramic scaffold promoted with low-intensity pulsed ultrasound. MC3T3-E1 mouse pre-osteoblasts were seeded onto scaffolds and cultured for 4 and 7 days with daily of 20 min ultrasound treatment. The cells were evaluated for cell attachment, morphology, viability, ingrowth depth, volumetric proliferation and early differentiation. After 4 and 7 days of culture and ultrasound exposure, the cell density was higher in the ultrasound-treated group compared to the sham-treated group on SiC scaffolds. The cell ingrowth depths inside the SiC scaffolds were (149.2±27.3) μm at 1 day, (310.1±12.6) μm for the ultrasound-treated group and (248.0±19.7) μm for the sham control at 4 days, (359.6±18.5) μm for the ultrasound-treated group and (280.0±17.7) μm for the sham control at 7 days. They were significantly increased, i.e. 25% (p=0.0029) and 28% (p=0.0008) increase, respectively, with ultrasound radiation force as compared to those in sham control at 4 and 7 days post-seeding. The dsDNA contents were (583.5±19.1) ng/scaffold at 1 day, (2749.9±99.9) ng/scaffold for the ultrasound-treated group and (2514.9±114.7) ng/scaffold for the sham control at 4 days, (3582.3±325.3) ng/scaffold for the ultrasound-treated group and (2825.7±134.3) ng/scaffold for the sham control at 7 days. There was a significant difference in the dsDNA content between the ultrasound- and sham-treated groups at 4 and 7 days. The ultrasound-treated group with the SiC construct showed a 9% (p=0.00029) and 27% (p=0.00017) increase in the average dsDNA content at 4 and 7 days over the sham control group, respectively. ALP activity was significantly increased by the treatment of ultrasound at 4 (p=0.012) and 7 days (p=0.035). These results suggested that ultrasound treatment with low intensity acoustic energy facilitated the cellular ingrowth, enhanced the proliferation and early differentiation of osteoblasts in SiC scaffolds.
PMID: 24935158