低强度脉冲超声波作用下羟基磷灰石/磷酸三钙三维支架上成骨细胞的生物学行为的研究

Biological behavior of osteoblasts on hydroxyapatite / tricalcium phosphate three-dimensional scaffold treated with low-intensity pulsed ultrasound

目的评价低强度脉冲超声波对羟基磷灰石(HA)/磷酸三钙(TCP)三维多孔支架材料上的MC3T3-E1成骨细胞的生物学行为的影响。方法将鼠MC3T3-E1成骨细胞与HA/TCP支架材料复合培养4 d和7 d。超声组每天接受20 min的低强度脉冲(1 MHz)超声波辐照,对照组为不开功率源的假辐照。通过观察成骨细胞在支架材料上的附着和细胞活力,检测细胞长入支架深度、DNA含量等指标评价低强度脉冲超声波对成骨细胞生物学行为的影响。结果 14 d和7 d时超声组HA/TCP支架材料上成骨细胞生长密度高于对照组。24 d和7 d时超声组HA/TCP支架材料上成骨细胞长入深度较对照组分别增加了34.45%和23.45%,4 d时二者差异无统计学意义(P=0.057),而7 d差异有统计学意义(P=0.032)。34 d和7 d时超声组较对照组DNA含量分别增加了27.54%(P=0.000)和24.39%(P=0.000),差异均有显著统计学意义。结论加载低强度脉冲超声波有利于成骨细胞在HA/TCP支架材料上的生长与增殖。

Objective To evaluate the effect of low-intensity pulsed ultrasound on the biological behavior of osteoblasts on hydroxyapatite（HA） / tricalcium phosphate（TCP） porous ceramic scaffolds. Methods MC3T3-E1 pre-osteoblasts of rats were seeded onto HA / TCP scaffolds and cultured for 4 days and 7 days. The daily treatment of 20 minutes ultrasound（1 MHz） was performed in ultrasound group, whereas control group was treated without power source. The adhesion and viability of cells on the scaffold material were observed, and in-growth depth and ds DNA contents were detected to evaluate its effect on the biological characteristics of osteoblasts. Results ①After 4 days and 7 days of culture and ultrasound exposure, the cell density on HA /TCP scaffolds of ultrasound group was higher than that of control group. ② 4 days and 7 days post-seeding, the cell in-growth depth of ultrasound group was increased by 34.45 % and 23.45 %, respectively, as compared with control group. At day 4, the difference between the two groups was not significant（P = 0.057）, but was significant at day 7（P = 0.032）. ③The DNA content in ultrasound group with HA / TCP scaffolds at day 4 and day 7 was increased by 27.54 %（P = 0.000） and 24.39 %（P = 0.000）,respectively, and the difference was significant at both day 4 and day 7. Conclusion It is demonstrated that ultrasound treatment with low-intensity acoustic energy facilitates the cellular in-growth, and enhances the proliferation of osteoblasts on HA / TCP scaffolds.