不同形貌羟基磷灰石纳米粒子的体内/体外评价。

PURPOSE: Hydroxyapatite-based nanoparticles have found diverse applications in drug delivery, gene carriers, diagnostics, bioimaging and tissue engineering, owing to their ability to easily enter the bloodstream and target specific sites. However, there is limited understanding of the potential adverse effects and molecular mechanisms of these nanoparticles with varying geometries upon their entry into the bloodstream. Here, we used two commercially available hydroxyapatite nanoparticles (HANPs) with different geometries (less than 100 nm in size each) to investigate this issue. METHODS: First, the particle size, Zeta potential, and surface morphology of nano-hydroxyapatite were characterized. Subsequently, the effects of 2~2000 μM nano-hydroxyapatite on the proliferation, migration, cell cycle distribution, and apoptosis levels of umbilical vein endothelial cells were evaluated. Additionally, the impact of nanoparticles of various shapes on the differential expression of genes was investigated using transcriptome sequencing. Additionally, we investigated the in vivo biocompatibility of HANPs through gavage administration of nanohydroxyapatite in mice. RESULTS: Our results demonstrate that while rod-shaped HANPs promote proliferation in Human Umbilical Vein Endothelial Cell (HUVEC) monolayers at 200 μM, sphere-shaped HANPs exhibit significant toxicity to these monolayers at the same concentration, inducing apoptosis/necrosis and S-phase cell cycle arrest through inflammation. Additionally, sphere-shaped HANPs enhance SULT1A3 levels relative to rod-shaped HANPs, facilitating chemical carcinogenesis-DNA adduct signaling pathways in HUVEC monolayers. In vivo experiments have shown that while HANPs can influence the number of blood cells and comprehensive metabolic indicators in blood, they do not exhibit significant toxicity. CONCLUSION: In conclusion, this study has demonstrated that the geometry and surface area of HANPs significantly affect VEC survival status and proliferation. These findings hold significant implications for the optimization of biomaterials in cell engineering applications.

目的：基于羟基磷灰石的纳米粒子因其易于进入血液并靶向特定部位的能力，在药物输送、基因载体、诊断、生物成像和组织工程等领域得到了广泛的应用。然而，对于具有不同几何形状的这些纳米粒子进入血液后的潜在不良影响和分子机制，人们的了解有限。在这里，我们使用了两种具有不同几何形状（每种尺寸均小于 100nm）的市售羟基磷灰石纳米粒子（HANP）来研究这个问题。

方法：首先，对纳米羟基磷灰石的粒径、Zeta 电位和表面形态进行了表征。随后，评估了 2~2000μM 纳米羟基磷灰石对脐静脉内皮细胞增殖、迁移、细胞周期分布和凋亡水平的影响。此外，通过转录组测序研究了不同形状的纳米粒子对基因差异表达的影响。此外，我们通过给小鼠灌胃纳米羟基磷灰石来研究 HANP 的体内生物相容性。

结果：我们的结果表明，在 200μM 时，棒状 HANP 促进人脐静脉内皮细胞（HUVEC）单层增殖，而同样浓度的球状 HANP 对这些单层具有显著毒性，通过炎症诱导凋亡/坏死和 S 期细胞周期停滞。此外，与棒状 HANP 相比，球状 HANP 增加了 SULT1A3 的水平，促进了 HUVEC 单层中的化学致癌-DNA 加合物信号通路。体内实验表明，HANP 虽然可以影响血液中血细胞的数量和综合代谢指标，但不会表现出明显的毒性。

结论：总之，本研究表明 HANP 的几何形状和表面积显著影响 VEC 的存活状态和增殖。这些发现对细胞工程应用中生物材料的优化具有重要意义。