School and Hospital of Stomatology, Tianjin Medical University, 12 Observatory Road, Tianjin 300070, PR China.
General Hospital, Tianjin Medical University, 154 An Shan Road, Tianjin 300052, PR China.
Mater Sci Eng C Mater Biol Appl. 2018 Jan 1;82:141-154. doi: 10.1016/j.msec.2017.08.062. Epub 2017 Aug 18.
Titanium (Ti) implants have been commonly used in oral medicine. However, despite their widespread clinical application, these implants are susceptible to failure induced by microbial infection due to bacterial biofilm formation. Immobilization of chimeric peptides with antibacterial properties on the Ti surface may be a promising antimicrobial approach to inhibit biofilm formation. Here, chimeric peptides were designed by connecting three sequences (hBD-3-1/2/3) derived from human β-defensin-3 (hBD-3) with Ti-binding peptide-l (TBP-l: RKLPDAGPMHTW) via a triple glycine (G) linker to modify Ti surfaces. Using X-ray photoelectron spectroscopy (XPS), the properties of individual domains of the chimeric peptides were evaluated for their binding activity toward the Ti surface. The antimicrobial and anti-biofilm efficacy of the peptides against initial settlers, Streptococcus oralis (S. oralis), Streptococcus gordonii (S. gordonii) and Streptococcus sanguinis (S. sanguinis), was evaluated with confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Transmission electron microscopy (TEM) and real-time quantitative PCR (qRT-PCR) were used to study cell membrane changes and the underlying antimicrobial mechanism. Compared with the other two peptides, TBP-1-GGG-hBD3-3 presented stronger antibacterial activity and remained stable in saliva and serum. Therefore, it was chosen as the best candidate to modify Ti surfaces in this study. This peptide inhibited the growth of initial streptococci and biofilm formation on Ti surfaces with no cytotoxicity to MC3T3-E1 cells. Disruption of the integrity of bacterial membranes and decreased expression of adhesion protein genes from S. gordonii revealed aspects of the antibacterial mechanism of TBP-1-GGG-hBD3-3. We conclude that engineered chimeric peptides with antimicrobial activity provide a potential solution for inhibiting biofilm formation on Ti surfaces to reduce or prevent the occurrence of peri-implant diseases.
钛（Ti）植入物已广泛用于口腔医学。然而，尽管这些植入物在临床上得到了广泛的应用，但由于细菌生物膜的形成，它们容易受到微生物感染引起的失效。将具有抗菌特性的嵌合肽固定在 Ti 表面上可能是一种抑制生物膜形成的有前途的抗菌方法。在这里，通过三甘氨酸（G）接头将三个源自人β防御素-3（hBD-3）的序列（hBD-3-1/2/3）与 Ti 结合肽-l（TBP-l：RKLPDAGPMHTW）连接起来设计嵌合肽，以修饰 Ti 表面。使用 X 射线光电子能谱（XPS）评估嵌合肽各个结构域与 Ti 表面结合活性的特性。使用共聚焦激光扫描显微镜（CLSM）和扫描电子显微镜（SEM）评估肽对初始定居者（口腔链球菌（S. oralis），戈登链球菌（S. gordonii）和血链球菌（S. sanguinis））的抗菌和抗生物膜功效。透射电子显微镜（TEM）和实时定量 PCR（qRT-PCR）用于研究细胞膜变化和潜在的抗菌机制。与其他两种肽相比，TBP-1-GGG-hBD3-3 表现出更强的抗菌活性，并且在唾液和血清中保持稳定。因此，它被选为本研究中修饰 Ti 表面的最佳候选物。该肽抑制初始链球菌的生长和 Ti 表面生物膜的形成，对 MC3T3-E1 细胞无细胞毒性。细菌膜完整性的破坏和来自 S. gordonii 的粘附蛋白基因表达的降低揭示了 TBP-1-GGG-hBD3-3 的抗菌机制的一些方面。我们得出结论，具有抗菌活性的工程化嵌合肽为抑制 Ti 表面生物膜的形成提供了一种潜在的解决方案，以减少或预防种植体周围疾病的发生。
