CAJ | 학술논문

背景：研究表明聚乳酸/羟基磷灰石复合材料与自然骨结构和性能相似，具有骨传导性和良好的生物相容性。 　　目的：观察聚乳酸/羟基磷灰石纳米复合物与成骨细胞株MC3T3-E1的生物相容性。 　　方法：分别采用普通完全培养基(对照组)与聚乳酸/羟基磷灰石纳米复合物浸提液(实验组)培养第3代成骨细胞株MC3T3-E1，培养3，5，7 d，采用CCK-8法检测细胞的吸光度值；在培养第7，14天检测细胞碱性磷酸酶、骨钙素、Ⅰ型胶原、核心结合因子α1/成骨特异性转录因子表达。将聚乳酸/羟基磷灰石纳米复合物与第3代成骨细胞株MC3T3-E1共培养，培养7，14 d细胞骨架染色及扫描电镜观察细胞在支架材料上的形态。 　　结果与结论：随着时间的延长，成骨细胞株MC3T3-E1的吸光度值明显增加，两组细胞吸光度值比较差异无显著性意义。实验组培养第7天细胞碱性磷酸酶活性高于对照组(P <0.05)，培养第14天细胞碱性磷酸酶、Ⅰ型胶原、核心结合因子α1/成骨特异性转录因子表达高于对照组(P <0.05)。种植7 d后细胞在材料上贴附生长，细胞呈多角形；种植14 d 后细胞较7 d 前数目明显增多，且完全伸展，呈多角形和梭形。表明聚乳酸/羟基磷灰石纳米复合物对成骨细胞有良好的生物相容性，无细胞毒性。
배경：연구표명취유산/간기린회석복합재료여자연골결구화성능상사，구유골전도성화량호적생물상용성。 　　목적：관찰취유산/간기린회석납미복합물여성골세포주MC3T3-E1적생물상용성。 　　방법：분별채용보통완전배양기(대조조)여취유산/간기린회석납미복합물침제액(실험조)배양제3대성골세포주MC3T3-E1，배양3，5，7 d，채용CCK-8법검측세포적흡광도치；재배양제7，14천검측세포감성린산매、골개소、Ⅰ형효원、핵심결합인자α1/성골특이성전록인자표체。장취유산/간기린회석납미복합물여제3대성골세포주MC3T3-E1공배양，배양7，14 d세포골가염색급소묘전경관찰세포재지가재료상적형태。 　　결과여결론：수착시간적연장，성골세포주MC3T3-E1적흡광도치명현증가，량조세포흡광도치비교차이무현저성의의。실험조배양제7천세포감성린산매활성고우대조조(P <0.05)，배양제14천세포감성린산매、Ⅰ형효원、핵심결합인자α1/성골특이성전록인자표체고우대조조(P <0.05)。충식7 d후세포재재료상첩부생장，세포정다각형；충식14 d 후세포교7 d 전수목명현증다，차완전신전，정다각형화사형。표명취유산/간기린회석납미복합물대성골세포유량호적생물상용성，무세포독성。
BACKGROUND:Studies have shown that polylactic acid/hydroxyapatite (PLA/HA) composite materials have structure and properties similar to natural bone, exhibiting bone conductivity and good biocompatibility. OBJECTIVE:To evaluate the biocompatibility of nano-hydroxyapatite/polylactic acid (n-HA/PLA) biomaterials with MC3T3-E1 cells. METHODS:The isolated third generation MC3T3-E1 cells were cultured in complete medium (control group) and n-HA/PLA extract (experimental group) for 3, 5, and 7 days. The material cytotoxicity was determined by cellCounting Kit-8. Expression of alkaline phosphatase, osteocalcin, type I col agen, core binding factorα1/osteoblast-specific transcription factor were tested by Real Time-PCR at days 7 and 14 after culture. MC3T3-E1 cells were inoculated onto the biomaterials of n-HA/PLA. Immunofluorescence and scanning electron microscope were use to observe MC3T3-E1 cellmorphology and attachment at days 7 and 14 after culture. RESULTS AND CONCLUSION:The osteoblast proliferation rates in the experimental and control groups were increased gradual y with time, and there was no difference in the absorbance between the two groups. At day 4, the activity of alkaline phosphatase was higher in the experimental group than the control group (P<0.05);at day 7, the expressions of alkaline phosphatase, type I col agen, core binding factorα1/osteoblast-specific transcription factor were higher in the experimental group than the control group (P<0.05). cells began to grow adherently on the materials at day 7 after co-culture, and present a polygon shape. At day 14, cells on the scaffold increased significantly in number and ful y extended with polygon and fusiform morphology. These results reveal that n-HA/PLA biomaterials have good cellcompatibility and no cytotoxicity.