不同等离子体处理氧化锆对人牙龈成纤维细胞黏附能力的影响

目的: 比较以不同种类气体为工作气体的大气压放电冷等离子体处理氧化锆对氧化锆表面人牙龈成纤维细胞生物学行为的影响。方法: 将20个氧化锆试件随机分为4组:对照组(无处理),氦气大气压介质阻挡放电等离子体处理90 s组(He组),氩气大气压介质阻挡放电等离子体处理90 s组(Ar组),95%氩气+5%氧气大气压介质阻挡放电等离子体处理90 s组(Ar+O₂组),每组5个。分组处理后观察氧化锆表面形貌,测定表面粗糙度及润湿性,分析表面元素组成,培养人牙龈成纤维细胞3 h测定细胞在氧化锆表面的初期黏附能力,免疫荧光染色观察细胞黏附初期细胞形态。结果: 等离子体处理后氧化锆表面形貌及粗糙度无明显改变,接触角均显著降低(P<0.05), 其中Ar组及Ar+O₂组接触角与He组具有统计学意义(P<0.05)。X射线光电子能谱分析结果显示,等离子体处理后氧化锆表面C/O比分别下降至0.45(He组)、0.36(Ar组)和0.33(Ar+ O₂组)。经Ar及Ar+O₂混合气体等离子体处理的氧化锆表面人牙龈成纤维细胞黏附初期数量提高,差异具有统计学意义(P<0.05)。相对于对照组,不同等离子体处理后的氧化锆表面成纤维细胞黏附及增殖情况更佳,细胞在黏附初期形态铺展更佳,伸出伪足更多。结论: 不同种类大气压放电冷等离子体处理氧化锆可在不改变材料表面形貌及粗糙度的情况下通过在材料表面引入含氧官能团提高材料表面润湿性;以Ar+O₂混合气体为工作气体的大气压放电冷等离子体可显著提高人牙龈成纤维细胞在氧化锆表面的早期黏附能力。

Effect of different plasma treated zirconia on the adhensive behaviour of human gingival fibroblasts

Objective: To evaluate the effect of different cold atmospheric plasma (CAP) treatment on the surface chemical and physical properties of zirconia and adhensive behaviour of human gingival fibroblasts (HGFs) cultured on zirconia disks.Methods: The zirconia disks were divided into four groups and treated using helium, argon and mixture of argon and oxygen cold atmospheric plasma for 90 s or left untreated. The surface morphology, wettability and chemical elements were analyzed right after treatment. Human gingival fibroblasts were grown from biopsies obtained from a periodontally healthy human subject during periodontal surgery. HGFs were seeded on zirconia disk, and cells density was measured at the time point of 3 hours. Indirect immunofluorescence (IIF) was performed for morphometric examination at the time point of 3 hours.Results: The crystallographic structure of zirconia was analyzed pre-viously and the results suggested that it fitted the properties of zirconium yttrium oxide. After helium, argon and mixture of argon and oxygen cold atmospheric plasma treatment, the surface morphology and roughness of zirconia disks remained the same. The contact angle of zirconia decreased significantly(P<0.05)after CAP treatment: from 68.38° to 17.90°. After different CAP plasmas treatment, the atomic percentage of carbon on the outermost surface of the three groups decreased, as did the surface C/O ratio. The surface C/O ratio of zirconia decreased from 1.07 to 0.33. Fibroblasts density increased on CAP treated disks, especially the ones treated by mixture of argon and oxygen CAP(P<0.05). Cells of the three CAP plasma treatment groups spread better and had more protrusions, as well as larger surficial areas.Conclusion: Based on the results of this study after being treated by different kinds of CAP plasmas for 90 s, the surface wettability increased and the elements changed significantly with no changes in the tomography and roughness of the materials. The CAP treatment enhances the adhensive behavior of fibroblasts on zirconia by increasing the oxygen functional groups and promoting the cell density. Wettability of zirconia, an important physicochemical property, has a vital influence on the cell behaviors.