Mode of killing determines the necrotrophic response of oral bacteria

Zayed, Naiera; Figueiredo, Joana; Holm, Wannes Van; Boon, Nico; Bernaerts, Kristel; Teughels, Wim

doi:10.1080/20002297.2023.2184930

Cited by 7 publications

(7 citation statements)

References 39 publications

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“…Overnight, liquid cultures were prepared in brain heart infusion (BHI) broth (Difco) for S. sobrinus , and BHI supplemented with 0.04% L-Cysteine HCl (BHIC) for F. nucleatum , under the same specific incubation conditions. The overnight cultures were then adjusted to concentrations of 10 8 events/mL each using flow cytometry (FACSVerse cytometer, BD Biosciences, Erembodegem, Belgium) with a 488 nm blue and a 640 nm red laser [ 29 ]. These cultures were used to grow biofilms on the tested surfaces in Brain Hearth Infusion 2 (BHI-2) broth containing BHI supplemented with 2.5 g/L mucin, 1.0 g/L yeast extract, 0.1 g/L cysteine, 2.0 g/L sodium bicarbonate and 0.25% ( v / v ) glutamic acid.…”

Section: Methodsmentioning

confidence: 99%

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pH-Triggered Controlled Release of Chlorhexidine Using Chitosan-Coated Titanium Silica Composite for Dental Infection Prevention

Srivastava,

Kamarudin,

Aktan

et al. 2024

Pharmaceutics

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Peri-implantitis is a growing pathological concern for dental implants which aggravates the occurrence of revision surgeries. This increases the burden on both hospitals and the patients themselves. Research is now focused on the development of materials and accompanying implants designed to resist biofilm formation. To enhance this endeavor, a smart method of biofilm inhibition coupled with limiting toxicity to the host cells is crucial. Therefore, this research aims to establish a proof-of-concept for the pH-triggered release of chlorhexidine (CHX), an antiseptic commonly used in mouth rinses, from a titanium (Ti) substrate to inhibit biofilm formation on its surface. To this end, a macroporous Ti matrix is filled with mesoporous silica (together referred to as Ti/SiO2), which acts as a diffusion barrier for CHX from the CHX feed side to the release side. To limit release to acidic conditions, the release side of Ti/SiO2 is coated with crosslinked chitosan (CS), a pH-responsive and antimicrobial natural polymer. Scanning electron microscopy coupled with energy dispersive X-ray spectroscopy (SEM/EDX) and Fourier transform infrared (FTIR) spectroscopy confirmed successful CS film formation and crosslinking on the Ti/SiO2 disks. The presence of the CS coating reduced CHX release by 33% as compared to non-coated Ti/SiO2 disks, thus reducing the antiseptic exposure to the environment in normal conditions. Simultaneous differential scanning calorimetry and thermogravimetric analyzer (SDT) results highlighted the thermal stability of the crosslinked CS films. Quartz crystal microbalance with dissipation monitoring (QCM-D) indicated a clear pH response for crosslinked CS coatings in an acidic medium. This pH response also influenced CHX release through a Ti/SiO2/CS disk where the CHX release was higher than the average trend in the neutral medium. Finally, the antimicrobial study revealed a significant reduction in biofilm formation for the CS-coated samples compared to the control sample using viability quantitative polymerase chain reaction (v-qPCR) measurements, which were also corroborated using SEM imaging. Overall, this study investigates the smart triggered release of pharmaceutical agents aimed at inhibiting biofilm formation, with potential applicability to implant-like structures.

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Section: Methodsmentioning

confidence: 99%

“…The biofilms were allowed to grow for 24 h under microaerophilic conditions (6% O 2 , 7% CO 2 , 7% H 2 and 80% N 2 ) while shaking at 100 rpm. After that, the biofilms were collected, and bacterial DNA extractions and v-qPCR were performed, according to Zayed et al [ 29 ].…”

Section: Methodsmentioning

confidence: 99%

pH-Triggered Controlled Release of Chlorhexidine Using Chitosan-Coated Titanium Silica Composite for Dental Infection Prevention

Srivastava,

Kamarudin,

Aktan

et al. 2024

Pharmaceutics

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“…Four periopathogens, 2 cariogenic species, 3 anaerobic oral commensals, and 5 oral commensal streptococci (Table 1) were maintained using blood agar with hemin (5.0 mg/mL), menadione (1.0 mg/mL), and 5% sterile horse blood. Brain Heart Infusion (BHI) broth supplemented with 0.04% L-cysteine HCl (BHIC) or BHI alone were used to prepare broth cultures (Zayed et al 2023).…”

Section: Bacterial Strains Media and Culture Conditionsmentioning

confidence: 99%

Oral Biofilm Cryotherapy as a Novel Ecological Modulation Approach

et al. 2023

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Oral cryotherapy is used in dentistry as a safe, simple, and low-cost treatment for a variety of oral lesions. It is well known for its ability to aid in the healing process. However, its effect on oral biofilms is unknown. As a result, the purpose of this study was to assess the effects of cryotherapy on in vitro oral biofilms. In vitro multispecies oral biofilms were grown on the surface of hydroxyapatite discs in symbiotic or dysbiotic states. CryoPen X+ was used to treat the biofilms, whereas untreated biofilms served as control. One set of biofilms was collected for study immediately after cryotherapy, whereas another group was reincubated for 24 h to permit biofilm recovery. Changes in biofilm structure were analyzed with a confocal laser scanning microscope (CLSM) and a scanning electron microscope (SEM), while biofilm ecology and community compositional changes were analyzed with viability DNA extraction and quantitative polymerase chain reaction (v-qPCR) analysis. One cryo-cycle immediately reduced biofilm load by 0.2 to 0.4 log10 Geq/mL, which increased with additional treatment cycles. Although the bacterial load of the treated biofilms recovered to the same level as the control biofilms within 24 h, the CLSM detected structural alterations. Compositional alterations were also detected by SEM, corroborating the v-qPCR findings that showed ≈≤10% incidence of pathogenic species compared to nontreated biofilms that encompassed ≈45% and 13% pathogenic species in dysbiotic and symbiotic biofilms, respectively. Spray cryotherapy showed promising results in a novel conceptual approach to the control of oral biofilms. Acting selectively by targeting oral pathobionts and retaining commensals, spray cryotherapy could modify the ecology of in vitro oral biofilms to become more symbiotic and prevent the evolution of dysbiosis without the use of antiseptics/antimicrobials.

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“…Aliquots were stored at room temperature and at 4°C. Cell concentrations and viability were evaluated before and after lyophilization by flow cytometry (FCM) using a FACSVerse cytometer with a blue 488 nm laser and a red 640 nm laser (BD Biosciences, New Jersey, United States) as described previously (Zayed et al, 2023). Briefly, samples before and after lyophilization were diluted with PBS to 10 5 -10 6 cells/mL and stained with SYBR™ Green I (1× final; Invitrogen, Thermo Fisher Scientific, Massachusetts, United States) and propidium iodide (4 μM final Thermo Fisher Scientific, Massachusetts, United States) and incubated for 20 min at 37°C.…”

Section: Creating Uniformly Lyophilized Probiotics Of Three Commonly ...mentioning

confidence: 99%

Probiotics for oral health: do they deliver what they promise?

Van Holm,

Lauwens,

De Wever

et al. 2023

Front. Microbiol.

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Probiotics have demonstrated oral health benefits by influencing the microbiome and the host. Although promising, their current use is potentially constrained by several restrictions. One such limiting factor lies in the prevailing preparation of a probiotic product. To commercialize the probiotic, a shelf stable product is achieved by temporarily inactivating the live probiotic through drying or freeze drying. Even though a lyophilized probiotic can be kept dormant for an extended period of time, their viability can be severely compromised, making their designation as probiotics questionable. Additionally, does the application of an inactive probiotic directly into the oral cavity make sense? While the dormancy may allow for survival on its way towards the gut, does it affect their capacity for oral colonisation? To evaluate this, 21 probiotic product for oral health were analysed for the number of viable (probiotic), culturable (CFU) and dead (postbiotic) cells, to verify whether the commercial products indeed contain what they proclaim. After isolating and uniformly lyophilizing three common probiotic species in a simple yet effective lyoprotective medium, the adhesion to saliva covered hydroxyapatite discs of lyophilized probiotics was compared to fresh or reactivated lyophilized probiotics. Unfortunately, many of the examined products failed to contain the claimed amounts of viable cells, but also the strains used were inadequately characterized and lacked clinical evidence for that unknown strain, questioning their label of a ‘probiotic’. Additionally, lyophilized probiotics demonstrated low adhesive capacity compared to their counterparts, prompting the question of why fresh or reactivated probiotics are not currently used.

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Mode of killing determines the necrotrophic response of oral bacteria

Cited by 7 publications

References 39 publications

pH-Triggered Controlled Release of Chlorhexidine Using Chitosan-Coated Titanium Silica Composite for Dental Infection Prevention

pH-Triggered Controlled Release of Chlorhexidine Using Chitosan-Coated Titanium Silica Composite for Dental Infection Prevention

Oral Biofilm Cryotherapy as a Novel Ecological Modulation Approach

Probiotics for oral health: do they deliver what they promise?

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