Kalia Paridhi, Jain Ankita, Radha Krishnan Ranjith, Demuth Donald R, Steinbach-Rankins Jill M
Department of Oral Immunology and Infectious Diseases, University of Louisville School of Dentistry.
Department of Microbiology and Immunology, University of Louisville School of Medicine.
Int J Nanomedicine. 2017 Jun 22;12:4553-4562. doi: 10.2147/IJN.S139178. eCollection 2017.
The interaction of g with commensal streptococci promotes colonization of the oral cavity. We previously showed that a synthetic peptide (BAR) derived from potently inhibited the formation of biofilms (IC =1.3 µM) and reduced virulence in a mouse model of periodontitis. Thus, BAR represents a novel therapeutic to control periodontitis by limiting colonization of the oral cavity. Here, we sought to develop drug-delivery vehicles for potential use in the oral cavity that comprise BAR-modified poly(lactic-co-glycolic)acid (PLGA) nanoparticles (NPs).
PLGA-NPs were initially modified with palmitylated avidin and subsequently conjugated with biotinylated BAR. The extent of BAR modification was quantified using a fluorescent-labeled peptide. Inhibition of adherence to by BAR-modified NPs was compared with free peptide using a two-species biofilm model.
BAR-modified NPs exhibited an average size of 99±29 nm and a more positive surface charge than unmodified NPs (zeta potentials of -7 mV and -25 mV, respectively). Binding saturation occurred when 37 nmol BAR/mg of avidin-NPs was used, which resulted in a payload of 7.42 nmol BAR/mg NPs. BAR-modified NPs bound to in a dose-dependent manner and more potently inhibited adherence and biofilm formation relative to an equimolar amount of free peptide (IC of 0.2 µM versus 1.3 µM). BAR-modified NPs also disrupted the preformed biofilms more effectively than free peptide. Finally, we demonstrate that BAR-modified NPs promoted multivalent association with , providing an explanation for the increased effectiveness of NPs.
These results indicate that BAR-modified NPs deliver a higher local dose of peptide and may represent a more effective therapeutic approach to limit colonization of the oral cavity compared to treatment with formulations of free peptide.
G与共生链球菌的相互作用促进口腔定植。我们之前表明，一种源自的合成肽（BAR）能有效抑制生物膜形成（IC =1.3 μM），并降低牙周炎小鼠模型中的毒力。因此，BAR代表一种通过限制口腔定植来控制牙周炎的新型疗法。在此，我们试图开发可用于口腔的药物递送载体，其包含BAR修饰的聚乳酸-乙醇酸共聚物（PLGA）纳米颗粒（NP）。
PLGA-NP最初用棕榈酰化抗生物素蛋白修饰，随后与生物素化的BAR偶联。使用荧光标记肽对BAR修饰程度进行定量。使用双物种生物膜模型将BAR修饰的NP对黏附的抑制作用与游离肽进行比较。
BAR修饰的NP平均尺寸为99±29 nm，表面电荷比未修饰的NP更正（ζ电位分别为-7 mV和-25 mV）。当使用37 nmol BAR/mg抗生物素蛋白-NP时发生结合饱和，这导致NP的有效载荷为7.42 nmol BAR/mg。BAR修饰的NP以剂量依赖性方式与结合，相对于等摩尔量的游离肽更有效地抑制黏附和生物膜形成（IC为0.2 μM对1.3 μM）。BAR修饰的NP也比游离肽更有效地破坏预先形成的生物膜。最后，我们证明BAR修饰的NP促进了与的多价结合，为NP有效性的提高提供了解释。
这些结果表明，与游离肽制剂治疗相比，BAR修饰的NP可递送更高局部剂量的肽，可能代表一种更有效的限制口腔定植的治疗方法。
