Leiva-Sabadini Camila, Berríos Pablo, Saavedra Paula, Carrasco-Rojas Javiera, González-Aramundiz José Vicente, Vera Mario, Tarifeño-Saldivia Estefanía, Schuh Christina M A P, Aguayo Sebastian
Institute for Biological and Medical Engineering, Pontificia Universidad Católica de Chile Santiago Chile
Centro de Medicina Regenerativa, Facultad de Medicina, Clínica Alemana-Universidad del Desarrollo Santiago Chile
Nanoscale Adv. 2025 Jul 24. doi: 10.1039/d5na00248f.
is the major microbial etiological agent of dental caries and can adhere to surfaces such as type-I collagen, which is present in dentin and periodontal tissues. Recent studies have characterized planktonic bacterial extracellular vesicles (bEVs) at the nanoscale range and demonstrated environmental-induced changes due to sugar presence or pH alterations. However, to date, no studies have explored whether surface-derived changes can modulate bEV production in the context of oral biofilm formation in the elderly. Therefore, this work aimed to determine the role of biofilm formation and collagen glycation on the nanoscale morphology and proteomic composition of bEVs. For this, bEVs from biofilms on native and glycated collagen surfaces were isolated, characterized, and compared to bEVs from planktonic cells. Nanoparticle tracking analysis (NTA), atomic force microscopy (AFM), and electron microscopy confirmed bEV production and showed that bEVs from biofilms are smaller in size and less abundant than those from planktonic cells. Furthermore, proteome analysis revealed that biofilm formation on native and glycated collagen led to the enrichment of several key virulence proteins. Also, a shift towards proteins involved in metabolic processes was found in bEVs following biofilm formation on collagen surfaces, whereas glucan metabolism proteins were overexpressed in vesicles from the planktonic state. These results demonstrate that biofilm formation, as well as the glycation of collagen associated with aging and hyperglycaemia, can modulate bEV characteristics and cargo and could play a central role in virulence and the development of diseases such as dental caries and periodontal disease.
是龋齿的主要微生物病原体，可粘附于牙本质和牙周组织中存在的I型胶原蛋白等表面。最近的研究对纳米尺度范围内的浮游细菌细胞外囊泡（bEVs）进行了表征，并证明了由于糖的存在或pH值变化导致的环境诱导变化。然而，迄今为止，尚无研究探讨在老年人口腔生物膜形成的背景下，表面衍生的变化是否能调节bEV的产生。因此，这项工作旨在确定生物膜形成和胶原糖基化对bEVs纳米尺度形态和蛋白质组组成的作用。为此，从天然和糖基化胶原表面的生物膜中分离出bEVs，进行表征，并与浮游细胞来源的bEVs进行比较。纳米颗粒跟踪分析（NTA）、原子力显微镜（AFM）和电子显微镜证实了bEV的产生，并表明生物膜来源的bEVs尺寸更小，数量比浮游细胞来源的更少。此外，蛋白质组分析表明，在天然和糖基化胶原上形成生物膜会导致几种关键毒力蛋白的富集。同样，在胶原表面形成生物膜后，bEVs中发现了向参与代谢过程的蛋白质的转变，而葡聚糖代谢蛋白在浮游状态的囊泡中过度表达。这些结果表明，生物膜的形成以及与衰老和高血糖相关的胶原糖基化可以调节bEV的特征和货物，并可能在龋齿和牙周病等疾病的毒力和发展中发挥核心作用。
