Physico-chemical and biological characterization of synthetic and eggshell derived nanohydroxyapatite/carboxymethyl chitosan composites for pulp-dentin tissue engineering.

Hybrid nanohydroxyapatite/carboxymethyl chitosan (nHAp-CMC) scaffolds have garnered significant attention in the field of regenerative engineering. The current study comparatively analyzed the physicochemical and biological properties of synthetic nanohydroxyapatite (SnHA)- and eggshell-sourced nanohydroxyapatite (EnHA)- based CMC biocomposites for pulp-dentin regeneration. EnHA and CMC were synthesized through a chemical process, whereas SnHA was commercially obtained. Composite scaffolds of SnHA-CMC and EnHA-CMC (1:5 w/w) were prepared using freeze-drying method. All biomaterials were characterized by FTIR, micro-Raman, XRD, HRSEM-EDX, and TEM analyses, and their in vitro bioactivity was assessed by immersing them in simulated body fluid for 21 days. The biological properties of the composite scaffolds were evaluated by assessing cytocompatibility using MTT assay and biomineralization potential by analyzing the odontogenic gene expressions (ALP, DSPP, DMP-1 and VEGF) in human dental pulp stem cells (DPSCs) using RT-qPCR method. Characterization studies revealed that EnHA displayed higher crystallinity and superior surface morphology compared to SnHA. The composite scaffolds showed a highly interconnected porous microstructure with pore sizes ranging between 60 and 220 µm, ideal for cell seeding. All tested materials, SnHA, EnHA, and their respective composites, displayed high cytocompatibility, increased ALP activity and degree of mineralization with significant upregulation of odontogenic-related genes on DPSCs (p < 0.05). Nevertheless, the odontogenic differentiation potential of EnHA-CMC on DPSCs was significantly higher when compared to SnHA-CMC. The findings from this study highlight the potential of EnHA-CMC as a promising candidate for pulp-dentin engineering.

Comparative evaluation of effect of nisin-incorporated ethylenediamine tetraacetic acid and MTAD on endodontic biofilm eradication, smear layer removal, and depth of sealer penetration.

OBJECTIVES: To comparatively evaluate the nisin-incorporated ethylenediamine tetraacetic acid (N-EDTA) and MTAD on cytotoxicity, endodontic biofilm eradication potential, smear layer removal ability, and sealer penetration depth. MATERIALS AND METHODS: N-EDTA was prepared and characterized using high-performance liquid chromatography (HPLC). Minimum inhibitory, minimum bactericidal, and minimum biofilm inhibitory concentration (MBC, MIC, and MBIC) were determined on Enterococcus faecalis (E. faecalis) strain. The cytocompatibility of N-EDTA and MTAD was evaluated using 3,(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-based colorimetric assay. Dentin specimens (n = 88 for antibacterial analysis, n = 170 for sealer penetration depth) were prepared and subjected to the classical irrigating strategy and obturation, respectively. The scanning electron microscopic evaluation (SEM) was done for the evaluation of biofilm disruption and smear layer removal. Confocal laser scanning microscopy (CLSM) evaluation was done for determining percentage of bacterial viability and sealer penetration depth. Statistical analysis of one-way ANOVA and Tukey's HSD post hoc tests for bacterial viability and Kruskal-Wallis test and Mann-Whitney test for smear layer removal and depth of penetration were done with the significance level set at p < 0.05. RESULTS: MTAD and N-EDTA showed cytocompatibility without any statistical differences from each other. For N-EDTA, the MIC and MBC values were 12.5 µg/ml (1:8), and MBIC values were 36 µg/ml. Biofilm disruption and killed bacterial percentage of N-EDTA was statistically higher than MTAD, whereas both the materials showed similar efficacy in the removal of the smear layer and sealer penetration depth. CONCLUSION: N-EDTA had negligible cytotoxicity with similar smear layer removal ability, sealer penetration, and better antibiofilm potential than MTAD. CLINICAL RELEVANCE: N-EDTA can serve as a viable alternative endodontic irrigant.

Evaluation of Degree of Conversion, Resin-Dentin Bond Strength, and Durability of Polydopamine Incorporated Total Etch Adhesive System.

Objectives: This study aimed to evaluate the degree of conversion (DC%), shear bond strength (SBS), bond durability, and the resin-dentin interface of polydopamine (PDA) incorporated total-etch adhesive system. Materials and Methods: Five percentage by weight (wt%) concentration of PDA incorporated adhesive was prepared and its polymerization was evaluated using Fourier-transform infrared spectroscopy. The results were compared with the DC% of conventional total-etch adhesive (Adper Single Bond 2) using independent t-test. Occlusal surfaces of 80 freshly extracted human premolars were sectioned to expose the dentin, which was acid-etched. The samples were divided into two groups (n=40) of total-etch adhesive and total-etch adhesive+PDA. Composite resin blocks were built up over the bonded surface and cured. Each group was subdivided into two subgroups (n=20) of immediate and post-aging evaluation. Samples were immersed in 10% sodium hypochlorite for five hours, and then, the SBS was evaluated using a universal testing machine at a crosshead speed of 1 mm/minute. The resin-dentin interface was evaluated using confocal laser scanning microscopy. Data were analyzed using two-way analysis of variance and post-hoc Bonferroni test. Results: DC% was not affected by the addition of 5% PDA to the adhesive (group 2). The SBS of group 2A was significantly higher than that of group 1A. There was no significant reduction of SBS in group 2B. The hybrid layer was less degraded after aging in group 2B compared to group 2A. Conclusion: PDA incorporated adhesive increased the immediate bond strength and durability without changing the DC%.

Evaluation of matrix metalloproteinase and cysteine cathepsin activity in dentin hybrid layer by gelatin zymography.

AIM: The aim of this study was to comparatively assess the gelatinolytic activity of matrix metalloproteinases(MMPs) and Cysteine Cathepsins (CCs) in the adhesive interface using etch and rinse adhesive at different time intervals using zymographic technique. METHODOLOGY: Twenty freshly extracted non-carious human third molars were used in this study. Occlusal surfaces were ground flat and 1mm thick horizontal dentin slabs were obtained from each tooth using a diamond disc. The dentin surface was polished with 600-grit silicon-carbide paper. Five out of 20 samples were directly pulverized. In the remaining fifteen samples, the dentin was etched and adhesive was applied and light cured according to the manufacturer's instructions. A 1mm thick flowable composite was build up and light cured. Bonded specimens were cut vertically into 3 to 4 dentin slabs by means of diamond disc to expose the adhesive/dentin interfaces. These were then ground down to 500 µm thick resin-dentin interface using a hard tissue microtome. These sections were then pulverised into powder. Following this, every five samples were subjected to zymographic analysis after 1 day, 7 days and 21 days. RESULTS: Zymograms showed clear, thicker bands on all three isoforms in the etched samples compared to control samples at 1st and 7th day intervals and became inactive at 21st day for all three isoforms. MMP 9 activity was relatively higher when compared to CCs and MMP 2. CONCLUSION: Etch and rinse adhesive activated MMPs and CCs within the hybrid layer that remained active till 7th day and no gelatinolytic activity was found on 21st day and MMPs are more active compared to CCs and MMP-2.

An in-vivo comparative evaluation of two herbal extracts Emblica officinalis and Terminalia Chebula with chlorhexidine as an anticaries agent: A preliminary study.

AIM: To compare the effect of 20% aqueous extract of Terminalia chebula and Emblica officinallis with chlorhexidine as an anticaries mouthwash - an in vivo study. METHODOLOGY: 20% aqueous extract of Terminalia chebula and Emblica officinalis were prepared. Forty five high caries risk patients were divided into group 1,2,3 [ n=15]and asked to rinse with 20% aqueous extract of Terminalia chebula and Emblica officinalis, 0.2% Chlorhexidine for 40 seconds respectively. Salivary samples were collected for pH, buffering capacity before and after rinsing at repeated intervals of 10, 30,60 and 90 minutes and except for microbial analysis for which it was collected before and 90 minutes after rinsing. The pH and the buffering capacity analysis was done using chairside kit and percentage of reduction of Streptococcus mutans and Lactobacillus were tabulated and statistically analyzed. RESULTS: For salivary pH, Group 2 showed the highest peak value followed by group 1. For salivary buffering capacity, there was no significant difference between any of the groups. For microbial count, highest percentage of reduction was seen in group 2 followed by group 1 and 3. However group 3 was more efficient at 90 minutes followed by group 1 and group 2. CONCLUSION: The aqueous extract of Embilica officinalis extract and Terminalia chebula was more effective anticaries mouthwash but with less time of action than chlorhexidine.

Comparative evaluation of propolis and triantibiotic mixture as an intracanal medicament against Enterococcus faecalis.

INTRODUCTION: The aim of this study was to evaluate and compare the antimicrobial activity of calcium hydroxide, triantibiotic mixture (TAM), and an ethanol extract of propolis as intracanal medicaments on Enterococcus faecalis-infected root canals. METHODS: One hundred twenty extracted intact human permanent incisors were decoronated, and chemomechanical preparation of the root canal was performed. After sterilization of the samples, they were inoculated with pure culture of E. faecalis and incubated. After incubation, colony-forming units were recorded before medication. Then, samples were divided randomly into five groups (n = 24). Each group was then exposed to various intracanal medicaments, namely calcium hydroxide (group 1), TAM (group 2), propolis (group 3), ethanol (group 4), and saline as the control group (group V). The antibacterial effectiveness of the different intracanal medicament was recorded by determining the percentage reduction in colony counts (%RCC) at the end of days 1, 2, and 7. The data were statistically analyzed using one-way analysis of variance and Tukey Honestly Significant Difference (HSD) post hoc test. RESULTS: The %RCC was highest for propolis showing 100% reduction on day 2 followed by TAM showing 82.5%, 92.2%, and 98.4% of reduction on days 1, 2, and 3, respectively. Calcium hydroxide showed a gradual increase in antibacterial activity with a maximum of 59.4% on day 7. CONCLUSIONS: Propolis was more effective than TAM against E. faecalis at a 2-day time period, and both were equally effective at 7 days.