Int J Oral Maxillofac Implants. 2019 May/June;34(3):673–680. doi: 10.11607/jomi.6929. Epub 2019 Feb 26.
To evaluate the effects of different titanium particle concentrations on viability of human calvarial osteoblasts and human gingival fibroblasts.
Primary human calvarial osteoblasts (HCO, 3H Biomedical) and human gingival fibroblasts (HGF-1, ATCC) were cultivated and allowed to adhere for 24 hours. Titanium powder concentrations (0.01 to 1.0 mg/mL) were added, and samples were analyzed at three time points (24 hours, 7 days, 21 days). Cell viability was analyzed using living cell count, proliferation (MTT) assay, and a live/dead staining. Cytotoxic effects were evaluated using lactated dehydrogenase assay. Qualitative analysis of cell viability was performed. In addition, scanning electron microscopy (SEM) analysis was performed. Release of interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-±) was estimated with Human IL-6 / Human TNF-± ELISA.
Titanium concentrations of 0.1 mg/mL and 1.0 mg/mL showed medium- and long-term effects on cell growth and proliferation rates. Cytotoxic effects by release of lactate dehydrogenase were observable during the first 24 hours. Human gingival fibroblast cells showed a release factor between 2.6 to 3.4. Titanium powder seemed to be more cytotoxic to human gingival fibroblast cells than to human calvarial osteoblast cells. For human calvarial osteoblasts, only the highest concentration showed cytotoxic effects with a release factor of 2.7. Human calvarial osteoblasts secreted IL-6 only during the first 24 hours and only in the highest titanium concentration, whereas human gingival fibroblasts secreted IL-6 during the entire period. The lowest titanium concentration showed stronger secretion of IL-6 compared to control. Incorporation of smaller and single titanium particles by cells was identified under SEM analysis.
Cell viability is negatively correlated with titanium concentration. Further, titanium debris might lead to an inflammatory biologic response of dental peri-implant tissue. Also, cells interact with the debris, eg, with incorporation of particles.
评估不同钛颗粒浓度对人颅骨成骨细胞和人牙龈成纤维细胞活力的影响。
原代人颅骨成骨细胞（HCO，3H Biomedical）和人牙龈成纤维细胞（HGF-1，ATCC）培养并贴壁 24 小时。加入钛粉浓度（0.01 至 1.0 mg/mL），并在三个时间点（24 小时、7 天、21 天）分析样本。使用活细胞计数、增殖（MTT）测定和活/死染色分析细胞活力。通过乳酸脱氢酶测定评估细胞毒性作用。进行细胞活力的定性分析。此外，还进行了扫描电子显微镜（SEM）分析。通过人 IL-6 /人 TNF-± ELISA 估计白细胞介素 6（IL-6）和肿瘤坏死因子α（TNF-α）的释放。
0.1 mg/mL 和 1.0 mg/mL 的钛浓度对细胞生长和增殖率具有中效和长效作用。在最初的 24 小时内，通过释放乳酸脱氢酶可观察到细胞毒性作用。人牙龈成纤维细胞的释放因子在 2.6 到 3.4 之间。与人类颅骨成骨细胞相比，钛粉似乎对人牙龈成纤维细胞的细胞毒性更大。对于人颅骨成骨细胞，只有最高浓度在释放因子为 2.7 时表现出细胞毒性作用。人颅骨成骨细胞仅在最初的 24 小时内且仅在最高钛浓度下分泌 IL-6，而人牙龈成纤维细胞在整个期间均分泌 IL-6。与对照相比，最低钛浓度表现出更强的 IL-6 分泌。在 SEM 分析中发现细胞内较小和单个钛颗粒的掺入。
细胞活力与钛浓度呈负相关。此外，钛屑可能导致种植体周围组织的炎症生物学反应。此外，细胞与碎片相互作用，例如颗粒的掺入。
