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Nanostructured titanium surfaces exhibit recalcitrance towards Staphylococcus epidermidis biofilm formation.

ABSTRACT: Titanium-based implants are ubiquitous in the healthcare industries and often suffer from bacterial attachment which results in infections. An innovative method of reducing bacterial growth is to employ nanostructures on implant materials that cause contact-dependent cell death by mechanical rupture of bacterial cell membranes. To achieve this, we synthesized nanostructures with different architectures on titanium surfaces using hydrothermal treatment processes and then examined the growth of Staphylococcus epidermidis on these surfaces. The structure obtained after a two-hour hydrothermal treatment (referred to as spear-type) showed the least bacterial attachment at short times but over a period of 6 days tended to support the formation of thick biofilms. By contrast, the structure obtained after a three-hour hydrothermal treatment (referred to as pocket-type) was found to delay biofilm formation up to 6 days and killed 47% of the initially attached bacteria by penetrating or compressing the bacteria in between the network of intertwined nano-spears. The results point to the efficacy of pocket-type nanostructure in increasing the killing rate of individual bacteria and potentially delaying longer-term biofilm formation.

SUBMITTER: Cao Y

PROVIDER: S-EPMC5773551 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature

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Nanostructured titanium surfaces exhibit recalcitrance towards Staphylococcus epidermidis biofilm formation.

Cao Yunyi Y Su Bo B Chinnaraj Subash S Jana Saikat S Bowen Leon L Charlton Sam S Duan Pengfei P Jakubovics Nicholas S NS Chen Jinju J

Scientific reports 20180118 1

Titanium-based implants are ubiquitous in the healthcare industries and often suffer from bacterial attachment which results in infections. An innovative method of reducing bacterial growth is to employ nanostructures on implant materials that cause contact-dependent cell death by mechanical rupture of bacterial cell membranes. To achieve this, we synthesized nanostructures with different architectures on titanium surfaces using hydrothermal treatment processes and then examined the growth of St ...[more]

PMID: 29348582

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Project description:The role of material surface properties in the direct interaction with bacteria and the indirect route via host defense cells is not fully understood. Recently, it was suggested that nanostructured implant surfaces possess antimicrobial properties. In the current study, the adhesion and biofilm formation of Staphylococcus epidermidis and human monocyte adhesion and activation were studied separately and in coculture in different in vitro models using smooth gold and well-defined nanostructured gold surfaces. Two polystyrene surfaces were used as controls in the monocyte experiments. Fluorescent viability staining demonstrated a reduction in the viability of S. epidermidis close to the nanostructured gold surface, whereas the smooth gold correlated with more live biofilm. The results were supported by scanning electron microscopy observations, showing higher biofilm tower formations and more mature biofilms on smooth gold compared with nanostructured gold. Unstimulated monocytes on the different substrates demonstrated low activation, reduced gene expression of pro- and anti-inflammatory cytokines, and low cytokine secretion. In contrast, stimulation with opsonized zymosan or opsonized live S. epidermidis for 1 hour significantly increased the production of reactive oxygen species, the gene expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, and IL-10, as well as the secretion of TNF-α, demonstrating the ability of the cells to elicit a response and actively phagocytose prey. In addition, cells cultured on the smooth gold and the nanostructured gold displayed a different adhesion pattern and a more rapid oxidative burst than those cultured on polystyrene upon stimulation. We conclude that S. epidermidis decreased its viability initially when adhering to nanostructured surfaces compared with smooth gold surfaces, especially in the bacterial cell layers closest to the surface. In contrast, material surface properties neither strongly promoted nor attenuated the activity of monocytes when exposed to zymosan particles or S. epidermidis.

Project description:ObjectivesThe present study aimed to evaluate the potential of the salivary pellicle (SP) formed on titanium (Ti) surfaces to modulate the formation of a biofilm composed of Streptococcus gordonii, Actinomyces naeslundii, Fusobacterium nucleatum, and Porphyromonas gingivalis.Materials and methodsTi substrates were incubated for 2 h with a pool of saliva samples obtained from 10 systemically and periodontally healthy subjects. Enamel substrates were included as a biological reference. Scanning electron microscopy (SEM) and Raman spectroscopy analysis were used to analyze the formation of the salivary pellicle. After the SP formation, the surfaces were incubated for 12 h with a mix of Streptococcus gordonii, Actinomyces naeslundii, Fusobacterium nucleatum, and Porphyromonas gingivalis. The number of bacterial cells attached to each surface was determined by the XTT assay while bacterial viability was analyzed by fluorescence microscopy using the LIVE/DEAD® BacLightTM kit.ResultsThe SEM and Raman spectroscopy analysis confirmed the presence of a salivary pellicle formed on the tested surfaces. Regarding the biofilm formation, the presence of the SP decreases the number of the bacterial cells detected in the test surfaces, compared with the uncover substrates. Even more, the SP-covered substrates showed similar bacterial counts in both Ti and enamel surfaces, meaning that the physicochemical differences of the substrates were less determinant than the presence of the SP. While on the SP-uncover substrates, differences in the bacterial adhesion patterns were directly related to the physicochemical nature of the substrates.ConclusionsThe salivary pellicle was the main modulator in the development of the biofilm consisting of representative oral bacteria on the Ti substrates.Clinical relevanceThe results of this study provide valuable information on the modulatory effect of the salivary pellicle on biofilm formation; such information allows us to understand better the events involved in the formation of oral biofilms on Ti dental implants.

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Nanostructured titanium surfaces exhibit recalcitrance towards Staphylococcus epidermidis biofilm formation.

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Nanostructured titanium surfaces exhibit recalcitrance towards Staphylococcus epidermidis biofilm formation.

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