波长620nm红光促进前软骨干细胞向软骨细胞分化的实验研究

Enhancing chondrogenic differentiation in precartilaginous stem cells with 620 nm red light

目的探讨620nm波长红光在诱导前软骨干细胞（PSCs）向软骨细胞分化中的作用。方法使用细胞免疫磁珠分选大鼠PSCs并体外培养，使用发光二极管（LED）发出的620nm波长红光，分别予以不同能量照射细胞，使用阿利新蓝染色和Ⅱ型胶原免疫组化染色观察软骨特异性细胞外基质（Ⅱ型胶原和蛋白聚糖）表达情况，光镜下观察细胞形态改变。采用逆转录聚合酶链反应（RT—PCR）技术分别检测各组细胞的Sox9和Ⅱ型胶原mRNA表达并进行半定量分析。结果PSCs经过软骨诱导和红光照射14d后，细胞形态由梭形变为多角形和类圆形，阿利新蓝和Ⅱ型胶原免疫组化染色均呈阳性反应，单纯诱导组呈弱阳性而对照组呈阴性；RT—PCR结果显示红光诱导的各组细胞Sox9和Ⅱ型胶原mRNA表达均明显增加，与对照组比较，差异有统计学意义（P〈0．05），0．5J／cm2组、1J／cm2组和2J／cm2组组间差异具有统计学意义（P〈0．05），但4J／cm2组和2J／cm2组组间差异无统计学意义（P〉0．05）。结论低能量620nm红光照射对诱导PSCs向软骨细胞分化有明显的促进作用。

Objective To investigate the effect of 620 nm red light on chondrogenic differentiation in rat precartilaginous stem cells （PSCs）. Methods Rats＇ PSCs were isolated and purified using magnetically activated cell sorting and cultured in vitro. The PSCs were exposed once to 620 nm wavelength red light from a light-emitting diode （LED） with an irradiation energy of 0.5 J/cm2, 1 J/cm2, 2 J/cm2 or 4 J/cm2, Any effect was confirmed by Alcian blue staining, immunohistochemistry and observing histomorphological changes under a light microscope, as well as detection using a reverse transcription polymerase chain reaction （RT-PCR）. Results After being induced for 14 d, the PSCs exhibited polygonal and round shapes. Alcian blue and type II collagen immunohistochemistry staining showed positive results, but the control group had no significant change. RT-PCR showed that the mRNA expression of Sox9 and type II collagen increased significantly compared with the control group. Conclusion Low energy 620 nm red light can enhance chondrogenic differentiation in PSCs significantly.