Yang Y M, Wang S, Li J J, Ning Y J, Li W, Chen J L
Teaching and Research Section of Dental Materials, Stomatologic Hospital and College, Anhui Medical University & Key Laboratory of Oral Diseases Research of Anhui Province, Hefei 230032, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2020 Feb 9;55(2):104-110. doi: 10.3760/cma.j.issn.1002-0098.2020.02.006.
To investigate the antibacterial properties and the osteoblast-compatibility of chlorhexidine (CHX)-modified porous titanium. Smooth pure titanium specimen with diameter of 10.0 mm and thickness of 1.5 mm treated with alkali heat method were set as control group. Those with covalent conjugation of aminosilane were set as silane group, and those with CHX grafted by glutaraldehyde were set as CHX group. Scanning electron microscope (SEM) was used to observe the surface morphology and element compositions were detected by X-ray photoelectron spectroscopy. Hydrophilicity was analyzed by surface water contact angle test (6), while surface amino/imine groups quantification were performed through acid orangeⅡ(5) and the CHX was quantified by optical densitometric method (5). Live/dead bacterial staining, the morphology of adherent bacteria by SEM, plate counting method and inhibition zone method were executed to evaluate the antibacterial property of the samples. Osteoblast compatibility was evaluated by methyl thiazolyl tetrazolium. Cell-bacterial co-culture was conducted to evaluated the cell viability on the samples under the circumstance with bacteria. After CHX grafting, pores on the titanium surface were decreased, while the atom ratio of C, N, Cl increased and the water contact angle decreased to 37.5°±4.0°. The density of CHX on the surface was (5.07±0.39) μg/cm(2). The results of live/dead bacterial staining and the morphology of adherent bacteria showed that only little dead bacterial (bacterial wall rupture) adherent on the surface of CHX group, which proved that the modified surface could inhibit bacteria adhesion and even destroyed bacteria; the plate counting displayed sporadic colonies and a transparent inhibition zone could be observed, which demonstrated that CHX group could suppress bacteria multiplication from surrounding environment. When incubating for 1 and 3 days, the cell viability of CHX group showed no significant difference from that of control group (>0.05) ; when incubating for 5 days, the value of cell viability of CHX group was 0.547±0.087, and this was significantly lower than that of the control group (0.751±0.056) (<0.05), demonstrating a slight inhibition of cell proliferation by CHX. The results of bacteria-cell co-culture for 3 days showed that a mass of bacteria adhered on the surface of the control group while considerable cells adhered on the surface of CHX group and exhibited a good shape. Porous titanium surface grafted by CHX showed an excellent antibacterial properties and allowed cell adhesion in bacterial circumstance, providing immediate implantation options for patients with bad oral health.
研究洗必泰(CHX)修饰的多孔钛的抗菌性能和成骨细胞相容性。将经碱热处理的直径为10.0 mm、厚度为1.5 mm的光滑纯钛试样作为对照组。将氨基硅烷共价偶联的试样作为硅烷组,将通过戊二醛接枝CHX的试样作为CHX组。使用扫描电子显微镜(SEM)观察表面形貌,并用X射线光电子能谱检测元素组成。通过表面水接触角测试分析亲水性,通过酸性橙Ⅱ进行表面氨基/亚胺基团定量,通过光密度法对CHX进行定量。进行活菌/死菌染色、通过SEM观察粘附细菌的形态、平板计数法和抑菌圈法来评估样品的抗菌性能。通过甲基噻唑基四氮唑评估成骨细胞相容性。进行细胞-细菌共培养以评估在有细菌情况下样品上的细胞活力。CHX接枝后,钛表面的孔隙减少,而C、N、Cl的原子比增加,水接触角降至37.5°±4.0°。表面CHX的密度为(5.07±0.39)μg/cm²。活菌/死菌染色结果和粘附细菌的形态表明,CHX组表面仅有少量死菌(细胞壁破裂)粘附,这证明修饰后的表面可抑制细菌粘附甚至破坏细菌;平板计数显示有散在菌落,可观察到透明抑菌圈,这表明CHX组可抑制周围环境中的细菌繁殖。培养1天和3天时,CHX组的细胞活力与对照组无显著差异(>0.05);培养5天时,CHX组的细胞活力值为0.547±0.087,显著低于对照组(0.751±0.056)(<0.05),表明CHX对细胞增殖有轻微抑制作用。细菌-细胞共培养3天的结果表明,对照组表面粘附大量细菌,而CHX组表面粘附大量细胞且形态良好。CHX接枝的多孔钛表面显示出优异的抗菌性能,并允许在有细菌的情况下细胞粘附,为口腔健康不良的患者提供了即时植入选择。