Claudio Cabral-Romero

Zerovalent bismuth nanoparticles inhibit Streptococcus mutans growth and formation of biofilm

Background and methods Despite continuous efforts, the increasing prevalence of resistance among pathogenic bacteria to common antibiotics has become one of the most significant concerns in modern medicine. Nanostructured materials are used in many fields, including biological sciences and medicine. While some bismuth derivatives has been used in medicine to treat vomiting, nausea, diarrhea, and stomach pain, the biocidal activity of zerovalent bismuth nanoparticles has not yet been studied. The objective of this investigation was to analyze the antimicrobial activity of bismuth nanoparticles against oral bacteria and their antibiofilm capabilities. Results Our results showed that stable colloidal bismuth nanoparticles had 69% antimicrobial activity against Streptococcus mutans growth and achieved complete inhibition of biofilm formation. These results are similar to those obtained with chlorhexidine, the most commonly used oral antiseptic agent. The minimal inhibitory concentration of bismuth nanoparticles that interfered with S. mutans growth was 0.5 mM. Conclusion These results suggest that zerovalent bismuth nanoparticles could be an interesting antimicrobial agent to be incorporated into an oral antiseptic preparation.

Bismuth oxide aqueous colloidal nanoparticles inhibit Candida albicans growth and biofilm formation

Multiresistance among microorganisms to common antimicrobials has become one of the most significant concerns in modern medicine. Nanomaterials are a new alternative to successfully treat the multiresistant microorganisms. Nanostructured materials are used in many fields, including biological sciences and medicine. Recently, it was demonstrated that the bactericidal activity of zero-valent bismuth colloidal nanoparticles inhibited the growth of Streptococcus mutans; however the antimycotic potential of bismuth nanostructured derivatives has not yet been studied. The main objective of this investigation was to analyze the fungicidal activity of bismuth oxide nanoparticles against Candida albicans, and their antibiofilm capabilities. Our results showed that aqueous colloidal bismuth oxide nanoparticles displayed antimicrobial activity against C. albicans growth (reducing colony size by 85%) and a complete inhibition of biofilm formation. These results are better than those obtained with chlorhexidine, nystatin, and terbinafine, the most effective oral antiseptic and commercial antifungal agents. In this work, we also compared the antimycotic activities of bulk bismuth oxide and bismuth nitrate, the precursor metallic salt. These results suggest that bismuth oxide colloidal nanoparticles could be a very interesting candidate as a fungicidal agent to be incorporated into an oral antiseptic. Additionally, we determined the minimum inhibitory concentration for the synthesized aqueous colloidal Bi2O3 nanoparticles.

Association of rotavirus viroplasms with microtubules through NSP2 and NSP5

Rotavirus replication and virus assembly take place in electrodense spherical structures known as viroplasms whose main components are the viral proteins NSP2 and NSP5. The viroplasms are produced since early times after infection and seem to grow by stepwise addition of viral proteins and by fusion, however, the mechanism of viropIasms formation is unknown. In this study we found that the viroplasms surface colocalized with microtubules, and seem to be caged by a microtubule network. Moreover inhibition of microtubule assembly with nocodazole interfered with viroplasms growth in rotavirus infected cells. We searched for a physical link between viroplasms and microtubules by co-immunoprecipitation assays, and we found that the proteins NSP2 and NSP5 were co-immunoprecipitated with anti-tubulin in rotavirus infected cells and also when they were transiently co-expressed or individually expressed. These results indicate that a functional microtubule network is needed for viroplasm growth presumably due to the association of viroplasms with microtubules via NSP2 and NSP5.

Antibacterial and antibiofilm activities of the photothermal therapy using gold nanorods against seven different bacterial strains

The objective of this work was to determine the bactericidal and antibiofilm activities of gold nanorods (AuNRs) using plasmonic photothermal therapy (PPTT) against oral microorganisms. AuNRs were synthesized by the seed and growth solution method and the gold nanoclusters were characterized with a size of 33.2 nm ± 2.23 length and 7.33 nm ± 1.60 width. The efficacy of PPTT related to its temperature was done reaching 67°C. Minimum inhibitory concentration (MIC) and minimum bactericide concentration (MBC) of AuNRs and AuNRs PPTT were determined against Enterococcus faecalis, Staphylococcus aureus, Streptococcus mutans, Streptococcus sobrinus, Streptococcus oralis, Streptococcus salivarius, and Escherichia coli growth. The antibiofilm activity of AuNRs was explored by fluorescence microscopy. After experimental analyses, AuNRs PPTT shows better results in MICs and MBCs, when it was compared with AuNRs alone. The laser employed to activate the AuNRs had no antibacterial effect against oral microbes. The MICs and MBCs values were higher for S. aureus and E. coli and lower against S. oralis. Surprisingly, the AuNRs alone presented a high antibiofilm activity, inhibiting the biofilm formation of S. mutans. Altogether, these results strongly suggest that AuNRs could be an interesting option to control oral biofilms.

Effect of lipophilic bismuth nanoparticles on erythrocytes

Lipophilic bismuth dimercaptopropanol nanoparticles (BisBAL NPs) have a very important antimicrobial activity; however their effect on human cells or tissues has not been completely studied. Undesirable effects of bismuth include anemia which could result from suicidal erythrocyte death or eryptosis. The objective of this research was to determine the effect of bismuth dimercaptopropanol nanoparticles on blood cells. The nanoparticles are composed of 53nm crystallites on average and have a spherical structure, agglomerating into clusters of small nanoparticles. Based on cell viability assays and optical microscopy, cytotoxicity on erythrocytes was observed after growing with 500 and 1000 µM of BisBAL NPs for 24 h. AM Calcein was retained inside erythrocytes when they were exposed to 100 µM (or lower concentrations) of BisBAL NPs for 24 h, suggesting the absence of damage in plasmatic membrane. Genotoxic assays revealed no damage to genomic DNA of blood cells after 24 h of exposition to BisBAL NPs. Finally, 100-1000 µM of bismuth nanoparticles promotes apoptosis between blood cells after 24 h of incubation. Hence BisBAL NPs at concentrations lower than 100 µM do not cause damage on blood cells; they could potentially be used by humans without affecting erythrocytes and leukocytes.

Antimicrobial and antibiofilm activities of MTA supplemented with bismuth lipophilic nanoparticles

The objective of this work was to determine the antimicrobial and antibiofilm properties of mineral trioxide aggregate (MTA) supplemented with bismuth lipophilic nanoparticles (BisBAL NPs). The antimicrobial activity of the composite MTA-BisBAL NPs was determined by the disk diffusion assay, while antibiofilm activity was analyzed by fluorescence microscopy. The cytotoxicity of MTA-BisBAL NPs was determined on human gingival fibroblasts by optical microscopy and crystal violet staining. MTA-BisBAL NPs inhibited the growth of Enterococcus faecalis, Escherichia coli, and Candida albicans and also detached the biofilm of fluorescent E. faecalis after 24 h of treatment. The addition of BisBAL nanoparticles did not significantly modify the physical properties of MTA, and cytotoxicity was not observed when MTA-BisBAL NPs was added on human gingival fibroblasts. Altogether these results suggest that BisBAL nanoparticles provide antimicrobial and antibiofilm activities to MTA while it retained their biophysical properties without cause side effects on human gingival fibroblasts.

Green Synthesis of Silver Nanoparticles and Their Bactericidal and Antimycotic Activities against Oral Microbes

Nanotechnology is a new discipline with huge applications including medicine and pharmacology industries. Although several methods and reducing agents have been employed to synthesize silver nanoparticles, reactive chemicals promote toxicity and nondesired effects on the human and biological systems. The objective of this work was to synthesize silver nanoparticles from Glycyrrhiza glabra and Amphipterygium adstringens extracts and determine their bactericidal and antimycotic activities against Enterococcus faecalis and Candida albicans growth, respectively. 1 and 10 mM silver nitrate were mixed with an extract of Glycyrrhiza glabra and Amphipterygium adstringens. Green silver nanoparticles (AgNPs) were characterized by TEM, Vis-NIR, FTIR, fluorescence, DLS, TGA, and X-ray diffraction (XRD) analysis. Bactericidal and antimycotic activities of AgNPs were determined by Kirby and Bauer method and cell viability MTT assays. AgNPs showed a spherical shape and average size of 9 nm if prepared with Glycyrrhiza glabra extract and 3 nm if prepared with Amphipterygium adstringens extract. AgNPs inhibited the bacterial and fungal growth as was expected, without a significant cytotoxic effect on human epithelial cells. Altogether, these results strongly suggest that AgNPs could be an interesting option to control oral biofilms.

Biocompatibility and Surface Characteristics of Resin-Modified Glass Ionomer Cements with Ammonium Quaternary Compounds or Silver Nanoparticles: An In Vitro Study

Glass ionomer cements are materials with diverse clinical applications. Its use is indicated in patients with special needs, pediatric patients, and the elderly; accordingly, it is important to know its properties. The aim of the present study was to determine the cytotoxicity, surface roughness, microhardness, and surface characteristics of GC Fuji ORTHO LC and GC Fuji PLUS resin-modified glass ionomer cements (RMGICs) with 1 and 2% of benzalkonium chloride, cetylpyridinium chloride, hexadecyltrimethylammonium bromide, or silver nanoparticles. All the experimental groups increase or decrease statistically significantly the VHN ( ) compared with the control group, except for GC Fuji PLUS added with hexadecyltrimethylammonium bromide 1 wt%. In the same way, all groups show a statistically significant ( ) increase or decrease in Ra compared with the control group except for GC Fuji ORTHO added with benzalkonium chloride 2 wt%, GC Fuji PLUS added with benzalkonium chloride 2 wt%, and GC Fuji PLUS added with cetylpyridinium chloride 2 wt%. The SEM micrographs show similar surface images between the control and experimental groups. When a dental material is modified, it is important to reevaluate its biological and mechanical characteristics. In the present study, all the additions modified the cytotoxicity and surface characteristics of RMGICs, by increasing or decreasing these properties.

Hydroxyapatite decreases cytotoxicity of a glass ionomer cement by calcium fluoride uptake in vitro

Background: Glass ionomer cements (GICs) are widely used in dentistry because of their remineralizing and cariostatic potential induced by fluoride. In vitro studies have reported cell toxicity triggered by GICs; however, the influence of hydroxyapatite (HAp) must be considered. The aim of this study was to evaluate the effect of HAp in decreasing the cytotoxicity of the GIC 3M Vitrebond in vitro. Methods: Samples of 3M Vitrebond (powder, liquid and light-cured) were incubated in Dulbecco’s modified Eagle’s medium–Ham’s F12 (DMEM-F12) for 24 hours at 37°C. Subsequently, the light-cured medium was treated with 100 mg/mL of HAp overnight. Toxicity of conditioned media diluted 1:2, 1:4, 1:8 and 1:20 was analyzed on human gingival fibroblasts (HGFs) using light microscopy and the fluorometric microculture cytotoxicity assay. The amounts of calcium fluoride (CaF2) were determined by the alizarin red S method. Results: The exposure of HGFs to light-cured induced cell death and morphological changes such as chromatin condensation, pyknotic nuclei and cytoplasmic modifications. Exposure to light-cured treated with HAp, significantly increased cell viability leading to mostly spindle-shaped cells (p<0.001). The concentration of CaF2 released by the light-cured was 200 ppm, although, in the light-cured/HAp conditioned medium, this quantity decreased to 88 ppm (p<0.01). Conclusions: These data suggest that HAp plays a protective role, decreasing the cytotoxic effect of 3M Vitrebond induced by CaF2.

In vitro evaluation of the antitumor effect of bismuth lipophilic nanoparticles (BisBAL NPs) on breast cancer cells

Aim The objective of this study was to evaluate the antitumor activity of lipophilic bismuth nanoparticles (BisBAL NPs) on breast cancer cells. Materials and methods The effect of varying concentrations of BisBAL NPs was evaluated on human MCF-7 breast cancer cells and on MCF-10A fibrocystic mammary epitheliocytes as noncancer control cells. Cell viability was evaluated with the MTT assay, plasma membrane integrity was analyzed with the calcein AM assay, genotoxicity with the comet assay, and apoptosis with the Annexin V/7-AAD assay. Results BisBAL NPs were spherical in shape (average diameter, 28 nm) and agglomerated into dense electronic clusters. BisBAL NP induced a dose-dependent growth inhibition. Most importantly, growth inhibition was higher for MCF-7 cells than for MCF-10A cells. At 1 µM BisBAL NP, MCF-7 growth inhibition was 51%, while it was 11% for MCF-10A; at 25 µM BisBAL NP, the growth inhibition was 81% for MCF-7 and 24% for MCF-10A. With respect to mechanisms of action, a 24-hour exposure of 10 and 100 µM BisBAL NP caused loss of cell membrane integrity and fragmentation of tumor cell DNA. BisBAL NPs at 10 µM were genotoxic to and caused apoptosis of breast cancer cells. Conclusion BisBAL NP-induced growth inhibition is dose dependent, and breast cancer cells are more vulnerable than noncancer breast cells. The mechanism of action of BisBAL NPs may include loss of plasma membrane integrity and a genotoxic effect on the genomic DNA of breast cancer cells.