化学合成肽修饰钛种植体表面的抗菌效果研究

Study on the antibacterial effect of chemically synthesized peptides on the surface of titanium implants

目的探讨化学合成肽修饰钛种植体表面的抗菌效果。方法利用源于人类β防御素3(hBD3)的一个片段(hBD3-1)与肽TBP-1化学结合,形成合成肽TBP-1-hBD3-1。Peptide Calculator软件分析合成肽的基本性质;标准微量肉汤稀释法检测TBP-1-hBD3-1对牙龈卟啉单胞菌(P.gingivalis)的最小抑菌浓度(MIC)、最小杀菌浓度(MBC),微量滴定法检测生物膜对照组和生物膜实验组光密度(OD_(600))值差异;共聚焦激光扫描显微镜(CLSM)检测TBP-1-hBD3-1修饰在钛种植体表面的黏附、活性情况。结果TBP-1-hBD3-1具有较好的溶解性及亲水性,并有疏水性基团,带有一定的正电荷数(4+)。TBP-1-hBD3-1对P.gingivalis的MIC值和MBC值分别为400 mg/L和1000 mg/L。生物膜实验组较生物膜对照组OD_(600)值降低(P<0.05)。CLSM结果表明,被AMC标记成蓝色的TBP-1-hBD3-1能够牢固地黏附于钛样本表面。结论合成肽TBP-1-hBD3-1能够修饰在钛样本表面并发挥抗菌和抑制细菌生物膜形成的作用。

Abstact:Objective To explore the bacteriological effects of chemically synthesized peptides on the surface of titanium implants.Methods A fragment(hBD3-1)derived from humanβdefensin 3(hBD3)was chemically combined with peptide TBP-1 to form synthetic peptide TBP-1-hBD3-1.Peptide Calculator software was used to analyze the basic properties of synthetic peptides.The standard micro broth dilution method was used to detect the minimum inhibitory concentration(MIC)and the minimum bactericidal concentration(MBC)of TBP-1-hBD3-1.The microtitration method was used to detect the difference of optical density(OD_(600))between biofilm control group and biofilm experimental group.Confocal laser scanning microscope(CLSM)experiments was used to further test the bacterial adhesion and bacterial activity on the surface of titanium implants.Results TBP-1-hBD3-1 showed good solubility,hydrophilicity and hydrophobicity,and had a certain number of positive charges(+4).The MIC and MBC values of TBP-1-hBD3-1 to P.gingivalis were 400 mg/L and 1000 mg/L,respectively.The OD_(600) value was significantly lower in the biofilm experimental group than that of the biofilm control group(P<0.05).The results of CLSM showed that the TBP-1-hBD3-1 labeled blue by AMC could be modified on the surface of titanium plate and exerted antibacterial effect.Conclusion The synthetic peptide TBP-1-hBD3-1 can be modified on the surface of titanium samples and play an antibacterial and biofilm inhibition effects.