Wu Lin, Lin Liangjun, Qin Yi-Xian
1 Department of Prosthodontics, School of Stomatology, China Medical University , Shenyang, People's Republic of China .
Tissue Eng Part A. 2015 Jan;21(1-2):53-61. doi: 10.1089/ten.TEA.2013.0597. Epub 2014 Jul 24.
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 preosteoblasts 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 with 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, and 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, that is, 25% (p=0.0029) and 28% (p=0.0008) increase, respectively, with ultrasound radiation force as compared with those in sham control at 4 and 7 days postseeding. 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, and 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. Alkaline phosphatase 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 and enhanced the proliferation and early differentiation of osteoblasts in SiC scaffolds.
对自体移植物或同种异体移植物使用的担忧促使人们研发用于骨再生的生物材料。已经进行了各种研究以优化使用具有骨传导性的多孔支架培养成骨细胞。本研究的目的是评估在低强度脉冲超声促进下，碳化硅（SiC）多孔陶瓷支架中骨细胞向内生长、增殖和早期分化的成骨效率。将MC3T3-E1小鼠前成骨细胞接种到支架上，并每天进行20分钟的超声处理，培养4天和7天。对细胞进行细胞附着、形态、活力、向内生长深度、体积增殖和早期分化评估。在培养4天和7天并接受超声照射后，与SiC支架上的假处理组相比，超声处理组的细胞密度更高。SiC支架内细胞在1天时的向内生长深度为149.2±27.3μm，在4天时，超声处理组为310.1±12.6μm，假对照组为248.0±19.7μm；在7天时，超声处理组为359.6±18.5μm，假对照组为280.0±17.7μm。与接种后4天和7天的假对照组相比，超声辐射力分别使细胞向内生长深度显著增加，即分别增加25%（p = 0.0029）和28%（p = 0.0008）。双链DNA含量在1天时为583.5±19.1 ng/支架，在4天时，超声处理组为2749.9±99.9 ng/支架，假对照组为2514.9±114.7 ng/支架；在7天时，超声处理组为3582.3±325.3 ng/支架，假对照组为2825.7±134.3 ng/支架。在4天和7天时，超声处理组与假处理组之间的双链DNA含量存在显著差异。与假对照组相比，含有SiC构建体的超声处理组在4天和7天时的平均双链DNA含量分别增加了9%（p = 0.00029）和27%（p = 0.00017）。在4天（p = 0.012）和7天（p = 0.035）时，超声处理显著提高了碱性磷酸酶活性。这些结果表明，低强度声能的超声处理促进了细胞向内生长，并增强了SiC支架中成骨细胞的增殖和早期分化。
