The Effectiveness of Different Irrigation Techniques on Debris and Smear Layer Removal in Primary Mandibular Second Molars: An In Vitro Study.

AIMS: The aim of the present in vitro study was to compare the effectiveness of passive ultrasonic irrigation (PUI), sonic irrigation, and mechanic dynamic activation on the removal of debris and smear layer from primary mandibular second molars during pulpectomy. MATERIALS AND METHODS: Mesial roots of 48 primary mandibular second molars were prepared with an R-motion 21 mm file (30/0.04) (FKG Dentaire SA, La Chaux-de-Fonds, Switzerland), irrigated with 1% sodium hypochlorite (NaOCl) and 17% ethylenediaminetetraacetic acid (EDTA), and divided into four groups (n = 24 canals) according to the final irrigation activation technique: control group without activation, PUI with Ultra-X (Eighteeth, Changzhou, China), mechanical activation with XP-endo Finisher (FKG), and sonic irrigation with EQ-S (Meta Biomed, Chungcheongbuk-do, Korea). The roots were split longitudinally and analyzed using scanning electron microscopy (SEM). The presence of debris and smear layer was assessed using a 5-grade scoring scale with 200× and 1000× magnification, respectively. The Kruskal-Wallis and Friedman tests were used for data analysis. RESULTS: The activation of the irrigant significantly improved debris and smear layer removal (p < 0.001). There was no significant difference between Ultra-X, XP-endo Finisher, and EQ-S (p > 0.05). No activation technique was able to completely eliminate debris and smear layer from the root canals of primary mandibular second molars. CONCLUSIONS: During pediatric pulpectomy, the irrigation protocol must include activation of the irrigation solutions using either ultrasonic, sonic, or mechanical activation techniques to enhance the removal of debris and smear layer for a better prognosis. CLINICAL SIGNIFICANCE: During root canal treatment on primary teeth, the clinician must incorporate an activation technique in the irrigation protocol to enhance the removal of debris and smear layer and increase the success of the treatment.

Development of alginate and alginate sulfate/polycaprolactone nanoparticles for growth factor delivery in wound healing therapy.

Connective tissue growth factor (CTGF) holds great promise for enhancing the wound healing process; however, its clinical application is hindered by its low stability and the challenge of maintaining its effective concentration at the wound site. Herein, we developed novel double-emulsion alginate (Alg) and heparin-mimetic alginate sulfate (AlgSulf)/polycaprolactone (PCL) nanoparticles (NPs) for controlled CTGF delivery to promote accelerated wound healing. The NPs' physicochemical properties, cytocompatibility, and wound healing activity were assessed on immortalized human keratinocytes (HaCaT), primary human dermal fibroblasts (HDF), and a murine cutaneous wound model. The synthesized NPs had a minimum hydrodynamic size of 200.25 nm. Treatment of HaCaT and HDF cells with Alg and AlgSulf2.0/PCL NPs did not show any toxicity when used at concentrations <50 µg/mL for up to 72 h. Moreover, the NPs' size was not affected by elevated temperatures, acidic pH, or the presence of a protein-rich medium. The NPs have slow lysozyme-mediated degradation implying that they have an extended tissue retention time. Furthermore, we found that treatment of HaCaT and HDF cells with CTGF-loaded Alg and AlgSulf2.0/PCL NPs, respectively, induced rapid cell migration (76.12% and 79.49%, P<0.05). Finally, in vivo studies showed that CTGF-loaded Alg and AlgSulf2.0/PCL NPs result in the fastest and highest wound closure at the early and late stages of wound healing, respectively (36.49%, P<0.001 on day 1; 90.45%, P<0.05 on day 10), outperforming free CTGF. Double-emulsion NPs based on Alg or AlgSulf represent a viable strategy for delivering heparin-binding GF and other therapeutics, potentially aiding various disease treatments.

Effect of Continuous Chelation Irrigation Using DualRinse HEDP+3% NaOCl with or without High-power Sonic Activation on Debris and Smear Layer Removal.

OBJECTIVE: This study aimed to assess the effect of sodium hypochlorite (NaOCl) combined with a novel chelating agent DualRinse HEDP (Medcem GmbH, Weinfelden, Switzerland), a product consisting of 0.9 g of 1-hydroxyethylidene-1,1-diphosphonic acid (HEDP) powder, with or without high-power sonic activation on debris and smear layer removal. METHODS: Seventy-five mandibular premolars were divided into 5 groups (n=15) and treated with different irrigation protocols: group 1 (D3N), DualRinse HEDP+3% NaOCl without activation; group 2 (D3NA), DualRinse HEDP+3% NaOCl with activation (EDDY, VDW, Munich, Germany) during the final irrigation; group 3 (3NE), 3% NaOCl+17% Ethylenediaminetetracetic acid (EDTA)+3% NaOCl without activation; group 4 (3NEA), 3% NaOCl+17% EDTA+3% NaOCl with activation during the final irrigation; group 5 (NC), negative control group, 0.9% saline. Samples were analysed by scanning electron microscopy (SEM) to evaluate residual debris and smear layer at 3 levels of the root canal: coronal, middle, and apical. Statistical analysis was performed with a level of significance set at p<0.05. The normality distribution of scores within each group was assessed using Kolmogorov-Smirnov and Shapiro-Wilk tests. A Kruskal-Wallis test followed by multiple comparison tests was used to compare scores among the 5 groups on the apical, middle, and coronal levels of the root canal. A Friedman test followed by multiple comparison tests was used to compare scores within the apical, middle, and coronal levels for each treatment group. RESULTS: Debris score was significantly the lowest for D3NA, followed by D3N, 3NEA and 3NE at all root levels (p<0.05). The smear layer score was significantly the lowest for D3NA, followed by D3N, 3NEA and 3NE only at the apical level, while no significant difference was found in the middle and coronal levels between the groups (p<0.05). DualRinse HEDP resulted in less debris and smear layer compared to the classic approach of NaOCl without activation. Implementing sonic activation further improved debris and smear layer removal. CONCLUSION: DualRinse HEDP+3% NaOCl improved debris removal at all levels and smear layer elimination at the apical level of the root canal. These results were further enhanced when adding high-power sonic activation. (EEJ-2022-09-116).

mRNA vaccines: Past, present, future.

mRNA vaccines have emerged as promising alternative platforms to conventional vaccines. Their ease of production, low cost, safety profile and high potency render them ideal candidates for prevention and treatment of infectious diseases, especially in the midst of pandemics. The challenges that face in vitro transcribed RNA were partially amended by addition of tethered adjuvants or co-delivery of naked mRNA with an adjuvant-tethered RNA. However, it wasn't until recently that the progress made in nanotechnology helped enhance mRNA stability and delivery by entrapment in novel delivery systems of which, lipid nanoparticles. The continuous advancement in the fields of nanotechnology and tissue engineering provided novel carriers for mRNA vaccines such as polymeric nanoparticles and scaffolds. Various studies have shown the advantages of adopting mRNA vaccines for viral diseases and cancer in animal and human studies. Self-amplifying mRNA is considered today the next generation of mRNA vaccines and current studies reveal promising outcomes. This review provides a comprehensive overview of mRNA vaccines used in past and present studies, and discusses future directions and challenges in advancing this vaccine platform to widespread clinical use.

Polymeric nanoparticles in the diagnosis and treatment of myocardial infarction: Challenges and future prospects.

Myocardial infarction (MI) is the leading cause of morbidity and mortality worldwide. Despite extensive efforts to provide early diagnosis and adequate treatment regimens, detection of MI still faces major limitations and pathological MI complications continue to threaten the recovery of survivors. Polymeric nanoparticles (NPs) represent novel noninvasive drug delivery systems for the diagnosis and treatment of MI and subsequent prevention of fatal heart failure. In this review, we cover the recent advances in polymeric NP-based diagnostic and therapeutic approaches for MI and their application as multifunctional theranostic tools. We also discuss the in vivo behavior and toxicity profile of polymeric NPs, their application in noninvasive imaging, passive, and active drug delivery, and use in cardiac regenerative therapy. We conclude with the challenges faced with polymeric nanosystems and suggest future efforts needed for clinical translation.

Er,Cr:YSGG Laser Surface Modification Effect on Dentin Bonding to Zirconia: An In Vitro Study.

Objective: This study investigated the effect of dentin surface treatment with the erbium, chromium-doped yttrium, scandium, gallium, and garnet (Er,Cr:YSGG) laser on the bond strength of zirconia to dentin. Background data: Although it is well-known that resin cement (RC) provides adequate bond strength of zirconia restorations to the tooth structure, many clinicians were not convinced in bonding reliability to zirconia materials. So, they preferred cementing their crowns with glass-ionomer cement (GIC). Pretreating the dentin surface is recommended to improve the adhesion of crowns cemented with GIC or RC. Recently, the Er,Cr:YSGG laser has been widely used for a cavity preparation, conditioning the tooth structure. However, there is not enough research on the bond strength of zirconia crowns cemented on a Er,Cr:YSGG laser-treated dentin. Methods: Forty-eight molars were cut horizontally at the crest of curvature. The flat dentin surface of 24 molars was left untreated, whereas the dentin surface of the other half was treated using an Er,Cr:YSGG laser 2.78 µm (4.5 W, 8.18 J/cm2, 90 mJ, 60 µs, 50 Hz, 60% air and 80% water). In addition, 48 plates of zirconia were prepared to be cemented with GIC or RC on the dentin surface. So, the molars were divided into four groups as follows: GIC+untreated dentin; GIC+treated dentin; RC+untreated dentin; and RC+treated dentin. All the specimens were subjected to shear bond strength test. The mode of failure was determined and additional samples were prepared to evaluate the cement-dentin interface using a scanning electron microscope. Data were analyzed with two-way analysis of variance accompanied by univariate analyses. Results: The bond strength of zirconia cemented with GIC or bonded with RC significantly increased on a Er,Cr:YSGG laser-treated dentin surface (p < 0.05). Conclusions: Dentin preconditioning with an Er,Cr:YSGG laser significantly improved the bond strength of zirconia plates to dentin as compared with untreated dentin.

Sulfated alginate/polycaprolactone double-emulsion nanoparticles for enhanced delivery of heparin-binding growth factors in wound healing applications.

Diabetic foot ulcers (DFUs) that are not effectively treated could lead to partial or complete lower limb amputations. The lack of connective tissue growth factor (CTGF) and insulin-like growth factor (IGF-I) in DFUs results in limited matrix deposition and poor tissue repair. To enhance growth factor (GF) availability in DFUs, heparin (HN)-mimetic alginate sulfate/polycaprolactone (AlgSulf/PCL) double emulsion nanoparticles (NPs) with high affinity and sustained release of CTGF and IGF-I were synthesized. The NPs size, encapsulation efficiency (EE), cytotoxicity, cellular uptake and wound healing capacity in immortalized primary human adult epidermal cells (HaCaT) were assessed. The sonication time and amplitude used for NPs synthesis enabled the production of particles with a minimum of 236 ± 25 nm diameter. Treatment of HaCaT cells with up to 50 µg mL-1 of NPs showed no cytotoxic effects after 72 h. The highest bovine serum albumin EE (94.6 %, P = 0.028) and lowest burst release were attained with AlgSulf/PCL. Moreover, cells treated with AlgSulf/CTGF (250 ng mL-1) exhibited the most rapid wound closure compared to controls while maintaining fibronectin synthesis. Double-emulsion NPs based on HN-mimetic AlgSulf represent a novel approach which can significantly enhance diabetic wound healing and can be expanded for applications requiring the delivery of other HN-binding GFs.

Plasma Kallikrein as a Modulator of Liver Injury/Remodeling.

The occurrence and persistence of hepatic injury which arises from cell death and inflammation result in liver disease. The processes that lead to liver injury progression and resolution are still not fully delineated. The plasma kallikrein-kinin system (PKKS) has been shown to play diverse functions in coagulation, tissue injury, and inflammation, but its role in liver injury has not been defined yet. In this study, we have characterized the role of the PKKS at various stages of liver injury in mice, as well as the direct effects of plasma kallikrein on human hepatocellular carcinoma cell line (HepG2). Histological, immunohistochemical, and gene expression analyses were utilized to assess cell injury on inflammatory and fibrotic factors. Acute liver injury triggered by carbon tetrachloride (CCl4) injection resulted in significant upregulation of the plasma kallikrein gene (Klkb1) and was highly associated with the high mobility group box 1 gene, the marker of cell death (r = 0.75, p < 0.0005, n = 7). In addition, increased protein expression of plasma kallikrein was observed as clusters around necrotic areas. Plasma kallikrein treatment significantly increased the proliferation of CCl4-induced HepG2 cells and induced a significant increase in the gene expression of the thrombin receptor (protease activated receptor-1), interleukin 1 beta, and lectin-galactose binding soluble 3 (galectin-3) (p < 0.05, n = 4). Temporal variations in the stages of liver fibrosis were associated with an increase in the mRNA levels of bradykinin receptors: beta 1 and 2 genes (p < 0.05; n = 3-10). In conclusion, these findings indicate that plasma kallikrein may play diverse roles in liver injury, inflammation, and fibrosis, and suggest that plasma kallikrein may be a target for intervention in the states of liver injury.

Modulation of Neuro-Inflammatory Signals in Microglia by Plasma Prekallikrein and Neuronal Cell Debris.

Microglia, the resident phagocytes of the central nervous system and one of the key modulators of the innate immune system, have been shown to play a major role in brain insults. Upon activation in response to neuroinflammation, microglia promote the release of inflammatory mediators as well as promote phagocytosis. Plasma prekallikrein (PKall) has been recently implicated as a mediator of neuroinflammation; nevertheless, its role in mediating microglial activation has not been investigated yet. In the current study, we evaluate the mechanisms through which PKall contributes to microglial activation and release of inflammatory cytokines assessing PKall-related receptors and their dynamics. Murine N9-microglial cells were exposed to PKall (2.5 ng/ml), lipopolysaccharide (100 ng/ml), bradykinin (BK, 0.1 µM), and neuronal cell debris (16.5 µg protein/ml). Gene expression of bradykinin 2 receptor (B2KR), protease-activated receptor 2 (PAR-2), along with cytokines and fibrotic mediators were studied. Bioinformatic analysis was conducted to correlate altered protein changes with microglial activation. To assess receptor dynamics, HOE-140 (1 µM) and GB-83 (2 µM) were used to antagonize the B2KR and PAR-2 receptors, respectively. Also, the role of autophagy in modulating microglial response was evaluated. Data from our work indicate that PKall, LPS, BK, and neuronal cell debris resulted in the activation of microglia and enhanced expression/secretion of inflammatory mediators. Elevated increase in inflammatory mediators was attenuated in the presence of HOE-140 and GB-83, implicating the engagement of these receptors in the activation process coupled with an increase in the expression of B2KR and PAR-2. Finally, the inhibition of autophagy significantly enhanced the release of the cytokine IL-6 which were validated via bioinformatics analysis demonstrating the role of PKall in systematic and brain inflammatory processes. Taken together, we demonstrated that PKall can modulate microglial activation via the engagement of PAR-2 and B2KR where PKall acts as a neuromodulator of inflammatory processes.

The histone deacetylase inhibitor Suberoylanilide Hydroxamic Acid (SAHA) as a therapeutic agent in rhabdomyosarcoma.

Rhabdomyosarcoma (RMS) is an aggressive childhood sarcoma with two distinct subtypes, embryonal (ERMS) and alveolar (ARMS) histologies. More effective treatment is needed to improve outcomes, beyond conventional cytotoxic chemotherapy. The pan-histone deacetylase inhibitor, Suberoylanilide Hydroxamic Acid (SAHA), has shown promising efficacy in limited preclinical studies. We used a panel of human ERMS and ARMS cell lines and xenografts to evaluate the effects of SAHA as a therapeutic agent in both RMS subtypes. SAHA decreased cell viability by inhibiting S-phase progression in all cell lines tested, and induced apoptosis in all but one cell line. Molecularly, SAHA-treated cells showed activation of a DNA damage response, induction of the cell cycle inhibitors p21Cip1 and p27Kip1 and downregulation of Cyclin D1. In a subset of RMS cell lines, SAHA promoted features of cellular senescence and myogenic differentiation. Interestingly, SAHA treatment profoundly decreased protein levels of the driver fusion oncoprotein PAX3-FOXO1 in ARMS cells at a post-translational level. In vivo, SAHA-treated xenografts showed increased histone acetylation and induction of a DNA damage response, along with variable upregulation of p21Cip1 and p27Kip1. However, while the ARMS Rh41 xenograft tumor growth was significantly inhibited, there was no significant inhibition of the ERMS tumor xenograft RD. Thus, our work shows that, while SAHA is effective against ERMS and ARMS tumor cells in vitro, it has divergent in vivo effects . Together with the observed effects on the PAX3-FOXO1 fusion protein, these data suggest SAHA as a possible therapeutic agent for clinical testing in patients with fusion protein-positive RMS.