MATERIALS AND METHODS: In silico Molecular Dynamics (MD) was used to determine the interactions of K21 inside the pocket of the targeted protein (crystal structure of fibroblast collagenase-1 complexed to a diphenyl-ether sulphone based hydroxamic acid; PDB ID: 966C; Crystal structure of MMP-2 active site mutant in complex with APP-derived decapeptide inhibitor. MD simulations were accomplished with the Desmond package in Schrödinger Drug Discovery Suite. Blood samples (~ 0.5 mL) collected into K2EDTA were immediately transferred for further processing using the Litron MicroFlow® PLUS micronucleus analysis kit for mouse blood according to the manufacturer's instructions. Bacterial Reverse Mutation Test of K21 Molecule was performed to evaluate K21 and any possible metabolites for their potential to induce point mutations in amino acid-requiring strains of Escherichia coli (E. coli) (WP2 uvrA (tryptophan-deficient)).
RESULTS: Molecular Simulation depicted that K21 has a specific pocket binding on various MMPs and SrtA surfaces producing a classical clouting effect. K21 did not induce micronuclei, which are the result of chromosomal damage or damage to the mitotic apparatus, in the peripheral blood reticulocytes of male and female CD-1 mice when administered by oral gavage up to the maximum recommended dose of 2000 mg/kg. The test item, K21, was not mutagenic to Salmonella typhimurium (S. typhimurium) strains TA98, TA100, TA1535 and TA1537 and E. coli strain WP2 uvrA in the absence and presence of metabolic activation when tested up to the limit of cytotoxicity or solubility under the conditions of the test.
CONCLUSION: K21 could serve as a potent protease inhibitor maintaining the physical and biochemical properties of dental structures.
METHODS: Experimental orthodontic adhesive system Transbond™ XT were modified with 1% Sr2+, 0.5% SrF2, 1% strontium, 0.5% Sr2+, 1% F-, 0.5% F-, and no additions were control. Mixing of formulation was monitored using Fourier transform infrared spectroscopy. Small-molecule drug-discovery suite was used to gain insights into Sr2+, F-, and SrF2 binding. Shear bond testing was performed after 6-months of ageing. Enamel blocks were cut, and STEM pictures were recorded. Specimens were indented to evaluate elastic modulus. Raman microscope was used to collect Raman spectra and inspected using a scanning electron microscope. Crystal structural analysis was performed using X-ray diffraction. Effect of material on cellular proliferation was determined. Confocal was performed to evaluate the effect of formulation on biofilms.
RESULTS: FTIR of modified adhesives depicted peak changes within range due to various functional groups existing within samples. TEM represented structurally optimized hexagonal unit-cell of hydroxyapatite. Mean shear bond strength is recorded highest for Transbond XT with 1% SrF2. Dead bacterial percentage appeared higher in 0.5% SrF2 and 1% F- specimens. Crystal lengths showed an increase in 0.5% and 1% SrF2 specimens. Phase contrast within TEM images showed a union of 0.5% SrF2 crystal with enamel crystal with higher elastic modulus and highly mineralized crystalline hydroxyapatite. Intensity of ν1 PO43- and ν1 CO32- along with carbonate - / ν1PO43- ratio displayed good association with strontium fluoride. The formulation showed acceptable cell biocompatibility (p
PURPOSE/OBJECTIVE: To analyze function of new K21 molecule in the invasive process of oral squamous cell carcinoma (OSCC) cell line.
MATERIALS & METHODS: The Fusobacterium (ATCC 23726) streaks were made, and pellets were resuspended in Cal27 (ATCC CRL-2095) OSCC cell line spheroid cell microplate. Cells were seeded and Lysotracker staining performed for CathepsinK red channel. Cell and morphology were evaluated using Transmission Electron microscopy. Thiobarbituric acid assay was performed. OSCC was analyzed for Mic60. Raman spectra were collected from the cancer cell line. L929 dermal fibroblast cells were used for Scratch Assay. ELISA muti arrays were used for cytokines and matrix molecules. Internalization ability of fibroblast cells were also analyzed. Structure of K21 as a surfactant molecule with best docked poses were presented.
RESULTS: Decrease in lysosomal staining was observed after 15 and 30 min of 0.1% treatment. Tumor clusters were associated with cell membrane destruction in K21 primed cells. There was functional silencing of Mic60 via K21, especially with 1% concentration with reduced cell migration and invasiveness. Raman intensity differences were seen at 700 cm-1, 1200 cm-1 and 1600 cm-1 regions. EVs were detected within presence of fibroblast cells amongst K21 groups. Wound area and wound closure showed the progress of wound healing.
CONCLUSION: Over expression of CatK can be reduced by a newly developed targeted K21 based drug delivery system leading to reduced migration and adhesion of oral squamous cell carcinoma cells. The K21 drug formulation can have great potential for cancer therapies due to targeting and cytotoxicity effects.
METHODS: Fourier Transform Infrared Spectroscopy (FTIR) was performed after complete hydrolysis of K21 solution. Human teeth were inoculated with biofilms for 7-days followed by treatment with various irrigants. The irrigant groups were Sodium hypochlorite [NaOCl (6%)], Chlorhexidine [CHX (2%)], K21 (0.5%), K21 (1%) and Saline. Scanning electron microscopy (SEM) was performed for biofilm and resin-dentin penetration. Transmission Electron Microscopy (TEM) of biofilms was done to evaluate application of K21. For in vivo evaluation, Albino wistar rats were injected subcutaneously and sections were stained with haematoxylin/eosin. Macrophage, M1/M2 expression were evaluated along with molecular simulation. Raman measurements were done on dried biofilms.
RESULTS: FTIR K21 specimens demonstrated presence of ethanol/silanol groups. Raman band at 1359 cm-1 resemble to -CH2- wagging displaying 29Si atoms in Nuclear Magnetic Resonance (NMR). 0.5%K21 showed cells exhibiting folded membranes. SEM showed staggering amount of resin tags with 0.5% K21 group. TEM showed membrane disruption in K21-groups. K21 groups were initially irritant, which subsided completely afterwards showing increased CD68. K21 and MMP/collagen complex was thermodynamically favourable.
CONCLUSION: K21 root canal irrigant was able to penetrate bacterial wall and can serve as a potential irrigant for therapeutic benefits. Expression of M2 polarized subsets showed K21 can serve in resolving inflammation and potentiate tissue repair.
MATERIALS AND METHODS: REVEAL dental fluorescence loupes and headlight system were used. Occlusal enamel was removed, and mid-coronal dentine was exposed. Carious artificial lesion was created. Streptococcus mutans, Actinomyces naeslundii, and Streptococcus sanguis were used. The assessment was performed using two diagnostic methods: naked eye and Design for Vision Glasses with inter examiner blinding using two calibrated examiners. After 7 days, Raman measurements were made on dentin disc specimens with 785 nm wavelength. The bacterial counts in colony-forming units (CFU) were used to examine the growth kinetics of biofilms. The collagen fibril structure within the discs was performed using Transmission Electron Microscope. Scanning Electron Microscope was used to image samples at various magnifications. FISH was performed with specimens fixed in 4% paraformaldehyde in phosphate-buffered saline. Reproducibility was measured by Cohen kappa scores, values of which range from 0 for less than chance agreement to 1 for almost perfect agreement (p
METHODS: Root canals were instrumented and randomly divided into the following groups: irrigation with saline, 6% NaOCl (sodium hypochlorite), 6% NaOCl+2% CHX (Chlorhexidine), 2% CHX, 0.5% k21/E (k21 - quaternary ammonium silane) and 1% k21/E. E. faecalis were grown (3-days) (1×107CFU mL-1), treated, and further cultured for 11-days. Specimens were subjected to SEM, confocal and Raman analysis and macrophage vesicles characterized along with effect of lipopolysaccharide treatment. 3T3 mouse-fibroblasts were cultured for alizarin-red with Sortase-A active sites and Schrödinger docking was performed. TEM analysis of root dentin substrate with matrix metalloproteinases profilometry was also included. A cytotoxic test analysis for cell viability was measured by absorbance of human dental pulp cells after exposure to different irrigant solutions for 24h. The test percentages have been highlighted in Table 1.
RESULTS: Among experimental groups, irrigation with 0.5% k21/E showed phase separation revealing significant bacterial reduction and lower phenylalanine 1003cm-1 and Amide III 1245cm-1 intensities. Damage was observed on bacterial cell membrane after use of k21/E. No difference in exosomes distribution between control and 0.5%k21/E was observed with less TNFα (*p<0.05) and preferential binding of SrtA. TEM images demonstrated integrated collagen fibers in control and 0.5%k21/E specimens and inner bacterial membrane damage after k21/E treatment. The k21 groups appeared to be biocompatible to the dental pulpal cells grown for 24h.
SIGNIFICANCE: Current investigations highlight potential advantages of 0.5% k21/E as irrigation solution for root canal disinfection.
METHODS: A 24-item validated questionnaire including closed and open questions on the teaching of posterior composites was emailed to faculty members in all 13 Dental Schools in Malaysia. Responses were compiled on Excel and analysed.
RESULTS: All 13 dental schools responded to the survey yielding a 100 % response. All schools indicated the use of posterior composites for 2- and 3-surface cavities in premolars and molars. The didactic teaching time devoted to composites was greater than for amalgam (38 h vs 29 h). Clinically, most posterior restorations placed by students were composites (average 74.1 %, range 10 %-100 %); the remaining 25.9 % were amalgams (range, 0 %-50 %). Slot-type cavities were the preparation techniques most commonly taught (n = 11,84.6 %). The use of rubber dam for moisture control was mandatory in most schools (n = 11, 84.6 %). History of adverse reaction to composites was found to be the most common contraindication to composite placement. The phase down of teaching and use of amalgam in Malaysia is expected to occur within the next six years.
CONCLUSION: The trend to increase the teaching of posterior composites reported for other countries is confirmed by the findings from Malaysian dental schools. Notwithstanding this trend, the use of amalgam is still taught, and future studies are required to investigate the implications of the phase down of amalgam in favour of posterior composites.
CLINICAL SIGNIFICANCE: Notwithstanding the increase in the teaching of posterior composites there is a pressing need to update and refine clinical guidelines for the teaching of posterior composites globally.
METHODOLOGY: Dentin blocks were sterilized and E. faecalis and C. albicans microbial colonies were counted for colony-forming-units against 2%k21, 2%CHX and Ca(OH)2 medicaments. Biofilm colonies after 7 days on dentin were analysed using confocal laser scanning microscopy with live/dead bacterial viability staining. TEM was done to study dentin collagen matrix. Dentin discs from 3rd day and 7th day well plate was used for Raman spectra and observed under fluorescent-microscope. Docking studies were carried out on MMP-2 S1 binding-domain with k21.
RESULTS: There was reduction of E. faecalis/C. albicans when k21, chlorhexidine and calcium hydroxide were used with highest percentage in 2%k21 treated specimens. 2%k21 showed dense and regular collagen network with intact cross-banding and decreased Raman intensity for 2%k21 on 3rd day. NaOCl + k21 showed least adherence, whereas saline groups showed highest adherence of E. faecalis and C. albicans to root-canal dentin. Alizarin red staining of hDPSCs revealed calcium deposition in all groups with significant difference seen amongst 2%k21 groups. MMP-2 ligand binding was seen accurately indicating possible target sites for k21 intervention.
CONCLUSION: 2%k21 can be considered as alternative intracanal medicament.