Comparison of the use of d-enantiomeric and l-enantiomeric antimicrobial peptides incorporated in a calcium-chelating irrigant against Enterococcus faecalis root canal wall biofilms.

OBJECTIVES: To compare the anti-biofilm efficacy of two antimicrobial peptides (AMPs), 1018 and DJK-5, in disrupting canal wall biofilms in the isthmus, canal and dentinal tubules of single-rooted maxillary premolars. METHODS: Enterococcus faecalis single-species biofilms were formed in-situ in the root canal system of the premolars (n = 91). Confocal laser scanning microscopy, bacterial sampling, colony-forming unit counting, XTT assay, lactate dehydrogenase assay and phenol-sulphuric acid method were used to identify the anti-biofilm efficacy of both AMPs and their influence on bacterial metabolic activity. RESULTS: Both AMPs disrupted in-situ E. faecalis biofilms and altered their metabolic activity. At 20 µg/mL, the d-enantiomeric AMP DJK-5 killed 55.5 %, 57.3 % and 55.8 % of biofilm bacteria in the isthmus, canal and dentinal tubules, respectively, in 1 min. In contrast, the l-enantiomeric AMP 1018 only eradicated 25.6 %, 25.5 % and 27.5 % of biofilm bacteria in the isthmus, canal and dentinal tubules, respectively, within the same time. Anti-biofilm efficacy of the root canal irrigants tested were in the order: 6 % NaOCl > 20 µg/mL DJK-5 > 10 µg/mL DJK-5 > 20 µg/mL 1018 > 10 µg/mL 1018 > 0.9 % NaCl. CONCLUSIONS: The present results are confirmatory of previous studies, in that d-enantiomeric AMPs exhibit more potent antibacterial properties than l-enantiomeric AMPs against E. faecalis biofilms within the canal space. Nevertheless, the potency of both AMPs are concentration-dependent. Incorporation of these agents into EDTA, a non-antibacterial calcium-chelating irrigant for removal of the inorganic component of the canal space debris, does not reduce the efficacy of either AMP. CLINICAL SIGNIFICANCE: The present study provides the proof of concept that incorporation of an antimicrobial peptide into a calcium-chelating root canal irrigant enhances the disinfection of intratubular single-species biofilms during smear layer and smear plug removal.

Hollow mesoporous zirconia delivery system for biomineralization precursors.

Strategies based on the combination of nanocarrier delivery systems and scaffolds provide bone tissue engineering scaffolds with multifunctional capability. Zirconia, a biocompatible ceramic commonly used in orthopedic and dental implants, was used to synthesize hollow mesoporous nanocapsules for loading, storage and sustained release of a novel polyamine-stabilized liquid precursor phase of amorphous calcium phosphate (PAH-ACP) for collagen biomineralization and bone marrow stromal cells osteoinduction. Hollow mesoporous zirconia (hmZrO2) nanocapsules loaded with biomimetic precursors exhibited pH-sensitive release capability and good biocompatibility. The PAH-ACP released from loaded hmZrO2 still retained the ability to infiltrate and mineralize collagen fibrils as well as exhibited osteoinductivity. A collagen scaffold blended with PAH-ACP@hmZrO2 supplement and stem cells may be a promising tool for bone tissue engineering. STATEMENT OF SIGNIFICANCE: The advent of nanotechnology has catalyzed the development of bone tissue engineering strategies based on the combination of nanocarrier delivery systems and scaffolds, which provide distinct advantages, including the possibilities of sustained release and protection of the bioactive agents, site-specific pharmacological effects and reduction of side effects. Herein, hollow mesoporous zirconia (hmZrO2) nanocapsules with pH-sensitive capacity were synthesized for loading, storage and sustained release of a novel polyamine-stabilized liquid precursor phase of ACP (PAH-ACP). The loaded nanocapsules show good biocompatibility and demonstrate bioactivities for collagen biomineralization and bone marrow stromal cells osteoinduction. Our results may offer a promising tool for designing bone tissue engineering "cocktail therapy" involving seeding scaffolds with biomineralization precursors loaded hmZrO2 supplement and stem cells.

Biodegradable mesoporous delivery system for biomineralization precursors.

Scaffold supplements such as nanoparticles, components of the extracellular matrix, or growth factors have been incorporated in conventional scaffold materials to produce smart scaffolds for tissue engineering of damaged hard tissues. Due to increasing concerns on the clinical side effects of using large doses of recombinant bone-morphogenetic protein-2 in bone surgery, it is desirable to develop an alternative nanoscale scaffold supplement that is not only osteoinductive, but is also multifunctional in that it can perform other significant bone regenerative roles apart from stimulation of osteogenic differentiation. Because both amorphous calcium phosphate (ACP) and silica are osteoinductive, a biodegradable, nonfunctionalized, expanded-pore mesoporous silica nanoparticle carrier was developed for loading, storage, and sustained release of a novel, biosilicification-inspired, polyamine-stabilized liquid precursor phase of ACP for collagen biomineralization and for release of orthosilicic acid, both of which are conducive to bone growth. Positively charged poly(allylamine)-stabilized ACP (PAH-ACP) could be effectively loaded and released from nonfunctionalized expanded-pore mesoporous silica nanoparticles (pMSN). The PAH-ACP released from loaded pMSN still retained its ability to infiltrate and mineralize collagen fibrils. Complete degradation of pMSN occurred following unloading of their PAH-ACP cargo. Because PAH-ACP loaded pMSN possesses relatively low cytotoxicity to human bone marrow-derived mesenchymal stem cells, these nanoparticles may be blended with any osteoconductive scaffold with macro- and microporosities as a versatile scaffold supplement to enhance bone regeneration.

Collagen intrafibrillar mineralization as a result of the balance between osmotic equilibrium and electroneutrality.

Mineralization of fibrillar collagen with biomimetic process-directing agents has enabled scientists to gain insight into the potential mechanisms involved in intrafibrillar mineralization. Here, by using polycation- and polyanion-directed intrafibrillar mineralization, we challenge the popular paradigm that electrostatic attraction is solely responsible for polyelectrolyte-directed intrafibrillar mineralization. As there is no difference when a polycationic or a polyanionic electrolyte is used to direct collagen mineralization, we argue that additional types of long-range non-electrostatic interaction are responsible for intrafibrillar mineralization. Molecular dynamics simulations of collagen structures in the presence of extrafibrillar polyelectrolytes show that the outward movement of ions and intrafibrillar water through the collagen surface occurs irrespective of the charges of polyelectrolytes, resulting in the experimentally verifiable contraction of the collagen structures. The need to balance electroneutrality and osmotic equilibrium simultaneously to establish Gibbs-Donnan equilibrium in a polyelectrolyte-directed mineralization system establishes a new model for collagen intrafibrillar mineralization that supplements existing collagen mineralization mechanisms.

Effects of different sonic activation protocols on debridement efficacy in teeth with single-rooted canals.

OBJECTIVES: The effects of different EndoActivator® (EA) sonic activation protocols on root canal debridement efficacy were examined. METHODS: Root canals in 48 single-rooted teeth were instrumented, irrigated initially with NaOCl and divided into 6 groups (N=8) based on the application time of QMix (antimicrobial calcium-chelating irrigant), and the time and sequence of EA irrigant activation - Positive Control: 90s QMix; Negative Control: 90s saline; Group 1A: 15s QMix+15s QMix with EA-activation; Group 1B: 30s QMix+30s of QMix with EA-activation; Group 2A: 15s QMix with EA-activation+15s QMix; Group 2B: 30s QMix with EA-activation+30s QMix. Split roots were examined with scanning electron microscopy for assignment of smear and debris scores in locations along the coronal, middle and apical thirds of the canals. The overall cleanliness of pooled canal locations in the Positive Control and the 4 experimental groups were compared with chi-square tests. RESULTS: Significant differences were detected among the 5 groups (P<0.001). Post hoc pairwise comparisons indicated that the overall canal cleanliness was in the order (from best to worst): 1B=2B>2A>1A>Positive Control. Completely clean canals could not be achieved due to the absence of continuous irrigant flow for EA to clear intraradicular debris. CONCLUSIONS: Irrespective of the sonic activation sequence, irrigant activation for 30s during a 60-s period of QMix application appears to maximise the smear layer and debris removal potential of the EndoActivator® system. CLINICAL SIGNIFICANCE: Sonic activation of root canal irrigants produces cleaner root canals and reduces the time required for final delivery of a canal wall smear later-removing irrigant when compared to the use of needle irrigation alone.