School of Medicine, Department of Biomedical and Surgical Sciences, Odontostomatological University Centre: Chair Prof. Stefano Cianetti, University of Perugia, S. Andrea Delle Fratte, 06156, Perugia, Italy.
Department of Experimental Medicine, Section of Biosciences and Medical Embriology, University of Perugia, S. Andrea Delle Fratte, 06156, Perugia, Italy.
Odontology. 2021 Apr;109(2):524-539. doi: 10.1007/s10266-020-00569-x. Epub 2020 Nov 19.
To analyze the effects of four universal adhesives (Optibond Solo Plus-OB, Universal Bond-UB, Prime&Bond Active-PBA, FuturaBond M + -FB) on human gingival fibroblasts in terms of cytotoxicity, morphology and function. After in vitro exposure for up to 48 h, fibroblast viability was determined by the MTT assay determined, morphology by phase-contrast microscopy and migration by the scratch wound assay. Expression levels of IL1β, IL6, IL8, IL10, TNFα and VEGF genes were assessed by RT-PCR and their protein production by Western blot analysis. Apoptosis and cell cycle were analyzed by flow cytometry. OB and UB induced early morphological changes on fibroblasts (3 h) with extended cell death at 24 h/48 h. Gene expression of collagen type I and fibronectin increased fivefold compared with controls, elastin disappeared and elastase increased threefold, indicating gingival tissue tended to become fibrotic. Only UB and OB increased gene expression of inflammatory markers: IL1β at 3 and 48 h (up to about three times), IL6 and IL8 at 3 h (up to almost four times) which corresponded to the increase of the activated form NF-kB. All adhesives showed an effect on the functionality of fibroblasts with cytotoxic effect time and concentration dependent. Among all the OB and UB adhesives, they showed the greatest cell damage. The in-depth analysis of the effects of universal adhesives and possible functional effects represents an important information for the clinician towards choosing the most suitable adhesive system.
为分析四种通用型黏合剂(Optibond Solo Plus-OB、Universal Bond-UB、Prime&Bond Active-PBA、FuturaBond M + -FB)对人牙龈成纤维细胞的细胞毒性、形态和功能的影响,本研究将细胞与黏合剂在体外共孵育 48 h。采用 MTT 法检测细胞活力,相差显微镜观察细胞形态,划痕实验检测细胞迁移。采用 RT-PCR 检测细胞因子(IL1β、IL6、IL8、IL10、TNFα 和 VEGF)mRNA 表达水平,Western blot 检测蛋白表达水平。采用流式细胞术检测细胞凋亡和细胞周期。OB 和 UB 在 3 h 时引起成纤维细胞早期形态改变,24 h/48 h 时细胞死亡延长。与对照组相比,I 型胶原和纤维连接蛋白的基因表达增加了五倍,弹性蛋白消失,弹性蛋白酶增加了三倍,表明牙龈组织趋于纤维化。只有 UB 和 OB 增加了炎症标志物的基因表达:IL1β 在 3 h 和 48 h(增加约三倍)、IL6 和 IL8 在 3 h(增加近四倍),这与 NF-kB 激活形式的增加相对应。所有黏合剂均对成纤维细胞的功能产生影响,具有时间和浓度依赖性的细胞毒性。在所有 OB 和 UB 黏合剂中,它们表现出最大的细胞损伤。深入分析通用型黏合剂的作用及其可能的功能影响,为临床医生选择最合适的黏合系统提供了重要信息。