University of Granada, Faculty of Dentistry, Colegio Máximo de Cartuja s/n, Granada 18071, Spain; Medicina Clínica y Salud Pública PhD Programme, Spain.
Biomedical Group (BIO277). Department of Nursing, Faculty of Health Sciences. University of Granada, Spain; Instituto Investigación Biosanitaria, IBS. Granada, Granada, Spain.
Dent Mater. 2023 Jun;39(6):616-623. doi: 10.1016/j.dental.2023.05.004. Epub 2023 May 10.
To evaluate the effect of doxycycline and dexamethasone doped nanoparticles covering titanium surfaces, on osteoblasts proliferation and differentiation.
Doxycycline and dexamethasone doped polymeric nanoparticles were applied on titanium discs (Ti-DoxNPs and Ti-DexNPs). Undoped NPs and uncovered Ti discs were used as control. Human MG-63 osteoblast-like cells were cultured. Osteoblasts proliferation was tested by MTT assay. Alkaline phosphatase activity was analyzed. Differentiation gene expression was assessed by real-time quantitative polymerase chain reaction. Scanning Electron Microscopy was performed to assess osteoblasts morphology. Mean comparisons were conducted by ANOVA and Wilcoxon or Tukey tests (p < 0.05).
No differences in osteoblasts proliferation were found. Osteoblasts grown on Ti-DoxNPs significantly increased alkaline phosphatase activity. Doxycycline and dexamethasone nanoparticles produced an over-expression of the main osteogenic proliferative genes (TGF-β1, TGF-βR1 and TGF-βR2). The expression of Runx-2 was up-regulated. The osteogenic proteins (AP, OSX and OPG) were also overexpressed on osteoblasts cultured on Ti-DoxNPs and Ti-DexNPs. The OPG/RANKL ratio was the highest when DoxNPs were present (75-fold increase with respect to the control group). DexNPs also produced a significantly higher OPG/RANKL ratio with respect to the control (20 times higher). Osteoblasts grown on titanium discs were mainly flat and polygonal in shape, with inter-cellular connections. In contrast, osteoblasts cultured on Ti-DoxNPs or Ti-DexNPs were found to be spindle-shaped and had abundant secretions on their surfaces.
DoxNPs and DexNPs were able to stimulate osteoblasts differentiation when applied on titanium surfaces, being considered potential inducers of osteogenic environment when performing regenerative procedures around titanium dental implants.
评估载有强力霉素和地塞米松的纳米粒子覆盖钛表面对成骨细胞增殖和分化的影响。
将强力霉素和地塞米松掺杂的聚合物纳米粒子应用于钛盘（Ti-DoxNPs 和 Ti-DexNPs）。未掺杂的 NPs 和未覆盖的 Ti 盘被用作对照。培养人 MG-63 成骨样细胞。通过 MTT 测定法测试成骨细胞增殖。分析碱性磷酸酶活性。通过实时定量聚合酶链反应评估分化基因表达。通过扫描电子显微镜评估成骨细胞形态。通过方差分析和 Wilcoxon 或 Tukey 检验进行均值比较（p < 0.05）。
成骨细胞增殖无差异。在 Ti-DoxNPs 上生长的成骨细胞碱性磷酸酶活性显著增加。强力霉素和地塞米松纳米颗粒产生了主要成骨增殖基因（TGF-β1、TGF-βR1 和 TGF-βR2）的过度表达。Runx-2 的表达上调。在 Ti-DoxNPs 和 Ti-DexNPs 上培养的成骨细胞中，骨形成蛋白（AP、OSX 和 OPG）也过度表达。当存在 DoxNPs 时，OPG/RANKL 比值最高（与对照组相比增加了 75 倍）。与对照组相比，DexNPs 也产生了明显更高的 OPG/RANKL 比值（高 20 倍）。在钛盘上生长的成骨细胞主要呈扁平多角形，细胞间有连接。相比之下，在 Ti-DoxNPs 或 Ti-DexNPs 上培养的成骨细胞呈梭形，表面有丰富的分泌物。
当应用于钛表面时，DoxNPs 和 DexNPs 能够刺激成骨细胞分化，在进行钛牙种植体周围再生手术时被认为是成骨环境的潜在诱导剂。
