Characterization of an ester-based core-multishell (CMS) nanocarrier for the topical application at the oral mucosa

Objectives

Topical drug administration is commonly applied to control oral inflammation. However, it requires sufficient drug adherence and a high degree of bioavailability. Here, we tested the hypothesis whether an ester-based core-multishell (CMS) nanocarrier is a suitable nontoxic drug-delivery system that penetrates efficiently to oral mucosal tissues, and thereby, increase the bioavailability of topically applied drugs.

Material and methods

To evaluate adhesion and penetration, the fluorescence-labeled CMS 10-E-15-350 nanocarrier was applied to ex vivo porcine masticatory and lining mucosa in a Franz cell diffusion assay and to an in vitro 3D model. In gingival epithelial cells, potential cytotoxicity and proliferative effects of the nanocarrier were determined by MTT and sulphorhodamine B assays, respectively. Transepithelial electrical resistance (TEER) was measured in presence and absence of CMS 10-E-15-350 using an Endohm-12 chamber and a volt-ohm-meter. Cellular nanocarrier uptake was analyzed by laser scanning microscopy. Inflammatory responses were determined by monitoring pro-inflammatory cytokines using real-time PCR and ELISA.

Results

CMS nanocarrier adhered to mucosal tissues within 5 min in an in vitro model and in ex vivo porcine tissues. The CMS nanocarrier exhibited no cytotoxic effects and induced no inflammatory responses. Furthermore, the physical barrier expressed by the TEER remained unaffected by the nanocarrier.

Conclusions

CMS 10-E-15-350 adhered to the oral mucosa and adhesion increased over time which is a prerequisite for an efficient drug release. Since TEER is unaffected, CMS nanocarrier may enter the oral mucosa transcellularly.

Clinical relevance

Nanocarrier technology is a novel and innovative approach for efficient topical drug delivery at the oral mucosa.

     

Cold plasma: a novel approach to treat infected dentin—a combined ex vivo and in vitro study

Objectives

The aim of this study was to evaluate the antimicrobial effect of cold plasma (CP) on infected dentin surfaces in vitro and ex vivo.

Materials and methods

To examine the effect of cold plasma on root surfaces, 24 root surfaces were infected with Streptococcus mitis. Specimens were randomly divided into three groups: Within the control group (C), root surfaces were rinsed with NaCl; root surfaces in the second group were additionally scaled and root planed (SRP), and in the third group, root surfaces were rinsed, scaled, root planed, and in addition, CP was applied (SRP + CP). To examine the effect of CP on root caries lesions (RCLs), 16 freshly extracted teeth with symmetrical carious lesions were equally divided into two groups. In the control group, carious lesions were treated with chlorhexidine (CHX), whereas CHX was applied in conjunction with CP in the test group (CHX + CP). For microbiological analysis, dentin samples were serially diluted and CFU counts were estimated after 24 h of incubation.

Results

Compared to C, mean CFU values for SRP and SRP + CP were significantly lower (p < 0.05). In addition, mean CFUs for SRP + CP were reduced to 0 and, therefore, significantly lower than SRP (2.98 log CFU/mL) alone (p = 0.000, Mann-Whitney U). Regarding RCLs, significantly lower mean CFU values were observed for CHX + CP when compared to CHX (4.45 vs. 2.67 log CFU/mL, p = 0.002, Mann-Whitney U test).

Conclusions

For disinfection of exposed root surfaces, the adjunctive application of CP is promising. In addition, the combined application of CHX + CP has the potential to disinfect root dentin surfaces.

Clinical relevance

It was shown that the combination of cold plasma with CHX is the best available option for the disinfection of root surfaces.

     

Porphyromonas gingivalis Outer Membrane Vesicles Induce Selective Tumor Necrosis Factor Tolerance in a Toll-Like Receptor 4- and mTOR-Dependent Manner

Porphyromonas gingivalis is an important member of the anaerobic oral flora. Its presence fosters growth of periodontal biofilm and development of periodontitis. In this study, we demonstrated that lipophilic outer membrane vesicles (OMV) shed from P. gingivalis promote monocyte unresponsiveness to live P. gingivalis but retain reactivity to stimulation with bacterial DNA isolated from P. gingivalis or AIM2 ligand poly(dA·dT). OMV-mediated tolerance of P. gingivalis is characterized by selective abrogation of tumor necrosis factor (TNF). Neutralization of interleukin-10 (IL-10) during OMV challenge partially restores monocyte responsiveness to P. gingivalis; full reactivity to P. gingivalis can be restored by inhibition of mTOR signaling, which we previously identified as the major signaling pathway promoting Toll-like receptor 2 and Toll-like receptor 4 (TLR2/4)-mediated tolerance in monocytes. However, despite previous reports emphasizing a central role of TLR2 in innate immune recognition of P. gingivalis, our current findings highlight a selective role of TLR4 in the promotion of OMV-mediated TNF tolerance: only blockade of TLR4—and not of TLR2—restores responsiveness to P. gingivalis. Of further note, OMV-mediated tolerance is preserved in the presence of cytochalasin B and chloroquine, indicating that triggering of surface TLR4 is sufficient for this effect. Taking the results together, we propose that P. gingivalis OMV contribute to local immune evasion of P. gingivalis by hampering the host response.