Department of Morphology, Medical Science College of China Three Gorges University, Yichang, China.
Department of Morphology, Medical Science College of China Three Gorges University, Yichang, China.
Colloids Surf B Biointerfaces. 2019 Dec 1;184:110499. doi: 10.1016/j.colsurfb.2019.110499. Epub 2019 Sep 10.
The invasive spreading of residual osteosarcoma cells becomes a serious threat to human health, urgently needing new bone regenerative biomaterials for orthopedic therapy. Thus, in this work, selenite-substituted hydroxyapatite (SeHA) nanoparticles were prepared for both inhibiting the recurrence of the tumor and accelerating the regenerative repair of bone defect. Physicochemical characterization showed these synthetic nanoparticles were spherical poly-crystals with the shape of snowflakes. Such structure benefited them to inhibit the cellular viability of osteosarcoma cells by about (58.90 ± 14.37)% during 24 h co-culturing. The expression level of cell growth-related genes such as PTEN, MMP-9, Cyclin D1, Cyclin A2, Annexin A2 and CDC2 decreased. Bisulfite Sequence PCR of PTEN gene exhibited about (22.40 ± 5.39)%, (45.91 ± 6.36)% and (25.90 ± 5.36)% promoter methylation in control, HA and SeHA group. Animal experiment also proved the similar effects. Almost no recurrence were observed in SeHA group. Oppositely, the slowly recurrent growth of the remnant tumor appeared in purely surgical group. The overall survival and toxicity analysis showed that, in the usage dose of 0-0.1 g, the SeHA-0.01 exhibited higher inhibitory recurrence and metastasis potentials, lower renal toxicity and better anti-inflammation function. Immunohistochemistry stain showed the reduced expression of PTEN, MMP-9, Ki-67 and Annexin A2, but slightly increased expression of DNMT1 and BMP-2. Compared the methylation status of PTEN gene in each group, it was confirming that SeHA nanoparticles hardly possessed the de-methylation effect, but the pure HA strikingly increased the methylation level of such gene. It seemed the dopant selenite ions possessed de-methylation effect onto PTEN gene. Therefore, from the viewpoint of inhibiting metastatic potentials, the SeHA-0.01 might be a feasible biomaterial to inhibit the relapse of the tumor post-surgery.
硒代羟磷灰石纳米颗粒抑制骨肉瘤细胞侵袭和促进骨缺损修复
残留的骨肉瘤细胞的侵袭性扩散对人类健康构成严重威胁，迫切需要新的骨再生生物材料进行骨科治疗。因此，本工作制备了亚硒酸根取代的羟磷灰石（SeHA）纳米颗粒，既抑制肿瘤的复发，又加速骨缺损的再生修复。物理化学特性表明，这些合成的纳米颗粒为具有雪花形状的球形多晶体。这种结构使它们在 24 小时共培养过程中能够抑制骨肉瘤细胞的细胞活力，抑制率约为（58.90±14.37）%。细胞生长相关基因的表达水平如 PTEN、MMP-9、Cyclin D1、Cyclin A2、Annexin A2 和 CDC2 降低。PTEN 基因的亚硫酸氢盐测序 PCR 显示，对照组、HA 组和 SeHA 组的启动子甲基化率分别约为（22.40±5.39）%、（45.91±6.36）%和（25.90±5.36）%。动物实验也证明了类似的效果。在 SeHA 组中几乎没有观察到复发。相反，在单纯手术组中，残留肿瘤的生长缓慢复发。总体生存和毒性分析表明，在 0-0.1g 的使用剂量下，SeHA-0.01 具有更高的抑制复发和转移潜力、更低的肾毒性和更好的抗炎功能。免疫组织化学染色显示，PTEN、MMP-9、Ki-67 和 Annexin A2 的表达减少，但 DNMT1 和 BMP-2 的表达略有增加。比较各组 PTEN 基因的甲基化状态，证实 SeHA 纳米颗粒几乎没有去甲基化作用，而纯 HA 显著增加了该基因的甲基化水平。似乎掺杂的亚硒酸根离子对 PTEN 基因具有去甲基化作用。因此，从抑制转移潜力的角度来看，SeHA-0.01 可能是一种抑制术后肿瘤复发的可行生物材料。
