A multi-platform analysis of human gingival crevicular fluid reveals ferroptosis as a relevant regulated cell death mechanism during the clinical progression of periodontitis

Ferroptosis is implicated in the pathogenesis of numerous chronic-inflammatory diseases,yet its association with progressive periodontitis remains unexplored.To investigate the involvement and significance of ferroptosis in periodontitis progression,we assessed sixteen periodontitis-diagnosed patients.Disease progression was clinically monitored over twelve weeks via weekly clinical evaluations and gingival crevicular fluid(GCF)collection was performed for further analyses.Clinical metrics,proteomic data,in silico methods,and bioinformatics tools were combined to identify protein profiles linked to periodontitis progression and to explore their potential connection with ferroptosis.Subsequent western blot analyses validated key findings.Finally,a single-cell RNA sequencing(scRNA-seq)dataset(GSE164241)for gingival tissues was analyzed to elucidate cellular dynamics during periodontitis progression.Periodontitis progression was identified as occurring at a faster rate than traditionally thought.GCF samples from progressing and non-progressing periodontal sites showed quantitative and qualitatively distinct proteomic profiles.In addition,specific biological processes and molecular functions during progressive periodontitis were revealed and a set of hub proteins,including SNCA,CA1,HBB,SLC4A1,and ANK1 was strongly associated with the clinical progression status of periodontitis.Moreover,we found specific proteins-drivers or suppressors-associated with ferroptosis(SNCA,FTH1,HSPB1,CD44,and GCLC),revealing the co-occurrence of this specific type of regulated cell death during the clinical progression of periodontitis.Additionally,the integration of quantitative proteomic data with scRNA-seq analysis suggested the susceptibility of fibroblasts to ferroptosis.Our analyses reveal proteins and processes linked to ferroptosis for the first time in periodontal patients,which offer new insights into the molecular mechanisms of progressive periodontal disease.These findings may lead to novel diagnostic and therapeutic strategies.

Present status and future directions:Apexification

The success rate of apexification is primarily determined by multiple factors,including the material used,the size of the open apex compared to the length of the root,and the technique used in each case.The main objective of this review was to provide an update on the present management of open apex to identify factors and circumstances that may influence the success of apexification using different materials and techniques.Future research on apexification should focus on how to treat open apices with wide periapical lesions without surgery.Previously,the predictability of these parameters with non-surgical procedures was uncertain,but now,with the use of a dental operating microscope,it has become more predictable.Another reason could be that extra visits are no longer required due to major advances in the armamentarium and materials used for apexification.

Long-term follow-ups of revascularized immature necrotic teeth:three case reports

Revascularization of immature necrotic teeth is a reliable treatment alternative to conventional apexogenesis or apexification. In case 1, a 12-year-old boy had his necrotic, immature mandibular left second premolar treated with a revascularization technique. At a 24-month follow-up, periapical radiolucency had disappeared and thickening of the root wall was observed. In cases 2 and 3, a lO-year-old boy had his necrotic, immature, bilateral mandibular second premolars treated with the same modality. At 48-month （in case 2） and 42-month （in case 3） follow-ups, loss of periapical radiolucencies and increases in the root wall thickness were also observed.

Long-term observation of the mineral trioxide aggregate extrusion into the periapical lesion:a case series

One-step apexification using mineral trioxide aggregate(MTA) has been reported as an alternative treatment modality with more benefits than the use of long-term calcium hydroxide for teeth with open apex.However,orthograde placement of MTA is a challenging procedure in terms of length control.This case series describes the sequence of events following apical extrusion of MTA into the periapical area during a one-step apexification procedure for maxillary central incisor with an infected immature apex.Detailed long-term observation revealed complete resolution of the periapical radiolucent lesion around the extruded MTA.These cases revealed that direct contact with MTA had no negative effects on healing of the periapical tissues.However,intentional MTA overfilling into the periapical lesion is not to be recommended.

Antimicrobial effect of alexidine and chlorhexidine against Enterococcus faecalis infection

A previous study demonstrated that alexidine has greater affinity for the major virulence factors of bacteria than chlorhexidine.The aim of this study was to compare the antimicrobial activity of 1%alexidine with that of 2%chlorhexidine using Enterococcus faecalis-infected dentin blocks.Sixty bovine dentin blocks were prepared and randomly divided into six groups of 10 each.E.faecalis was inoculated on 60 dentin blocks using the Luppens apparatus for 24 h and then the dentin blocks were soaked in 2%chlorhexidine or 1%alexidine solutions for 5 and 10 min,respectively.Sterile saline was used as a control.The antimicrobial efficacy was assessed by counting the number of bacteria adhering to the dentin surface and observing the degradation of bacterial shape or membrane rupture under a scanning electron microscope.Significantly fewer bacteria were observed in the 2%chlorhexidine-or 1%alexidine-soaked groups than in the control group(P<0.05).However,there was no significant difference in the number of bacteria adhering to the dentinal surface between the two experimental groups or between the two soaking time groups(P>0.05).Ruptured or antiseptic-attached bacteria were more frequently observed in the 10-min-soaked chlorhexidine and alexidine groups than in the 5-min-soaked chlorhexidine and alexidine groups.In conclusion,10-min soaking with 1%alexidine or 2%chlorhexidine can be effective against E.faecalis infection.

Recent advances in imaging technologies in dentistry

Dentistry has witnessed tremendous advances in all its branches over the past three decades. With these advances, the need for more precise diagnostic tools,specially imaging methods, have become mandatory.From the simple intra-oral periapical X-rays, advanced imaging techniques like computed tomography, cone beam computed tomography, magnetic resonance imaging and ultrasound have also found place in modern dentistry. Changing from analogue to digital radiography has not only made the process simpler and faster but also made image storage, manipulation(brightness/contrast, image cropping, etc.) and retrieval easier. The three-dimensional imaging has made the complex cranio-facial structures more accessible for examination and early and accurate diagnosis of deep seated lesions. This paper is to review current advances in imaging technology and their uses in different disciplines of dentistry.

Inhibition of matrix metalloproteinases expression in human dental pulp cells by all-trans retinoic acid

All-trans retinoic acid（ATRA） inhibits matrix metalloproteinase（MMP）-2 and MMP-9 in synovial fibroblasts, skin fibroblasts,bronchoalveolar lavage cells and cancer cells, but activates MMP-9 in neuroblast and leukemia cells. Very little is known regarding whether ATRA can activate or inhibit MMPs in human dental pulp cells（HDPCs）. The purpose of this study was to determine the effects of ATRA on the production and secretion of MMP-2 and-9 in HDPCs. The productions and messenger RNA（mRNA） expressions of MMP-2 and-9 were accessed by gelatin zymography and real-time polymerase chain reaction（PCR）, respectively. ATRA was found to decrease MMP-2 level in a dose-dependent manner. Significant reduction in MMP-2 mRNA expression was also observed in HDPCs treated with 25 mmol？L21ATRA. However, HDPCs treated with ATRA had no effect on the pattern of MMP-9 produced or secreted in either cell extracts or conditioned medium fractions. Taken together, ATRA had an inhibitory effect on MMP-2 expression in HDPCs,which suggests that ATRA could be a candidate as a medicament which could control the inflammation of pulp tissue in vital pulp therapy and regenerative endodontics.

Role of the CXCR4-SDF1-HMGB1 pathway in the directional migration of cells and regeneration of affected organs

In recent years,several studies have reported positive outcomes of cell-based therapies despite insufficient engraftment of transplanted cells.These findings have created a huge interest in the regenerative potential of paracrine factors released from transplanted stem or progenitor cells.Interestingly,this notion has also led scientists to question the role of proteins in the secretome produced by cells,tissues or organisms under certain conditions or at a particular time of regenerative therapy.Further studies have revealed that the secretomes derived from different cell types contain paracrine factors that could help to prevent apoptosis and induce proliferation of cells residing within the tissues of affected organs.This could also facilitate the migration of immune,progenitor and stem cells within the body to the site of inflammation.Of these different paracrine factors present within the secretome,researchers have given proper consideration to stromal cell-derived factor-1(SDF1)that plays a vital role in tissue-specific migration of the cells needed for regeneration.Recently researchers recognized that SDF1 could facilitate site-specific migration of cells by regulating SDF1-CXCR4 and/or HMGB1-SDF1-CXCR4 pathways which is vital for tissue regeneration.Hence in this study,we have attempted to describe the role of different types of cells within the body in facilitating regeneration while emphasizing the HMGB1-SDF1-CXCR4 pathway that orchestrates the migration of cells to the site where regeneration is needed.

Maxillary first molar with an O-shaped root morphology:report of a case

This case report is to present a maxillary first molar with one O-shaped root, which is an extended C-shaped canal system. Patient with chronic apical periodontitis in maxillary left first molar underwent replantation because of difficulty in negotiating all canals. Periapical radiographs and cone-beam computed tomography （CBCT） were taken. All roots were connected and fused to one root, and all canals seemed to be connected to form an O-shape. The apical 3 mm of the root were resected and retrograde filled with resin-modified glass ionomer. Intentional replantation as an alternative treatment could be considered in a maxillary first molar having an unusual O-shaped root.

Antibacterial activity of nanostructured Ti-45S5 bioglass-Ag composite against Streptococcus mutans and Staphylococcus aureus

Mechanical alloying and annealing at 1150 °C for 2 h under an argon atmosphere were used to prepare Ti-45S5 bioglass nanocomposites. Ti-45S5 bioglass material was chemically modified by silver. The antibacterial activity of Ti-10% 45S5 bioglass nanocomposite containing silver against Streptococcus mutans and Staphylococcus aureus was studied. Nanocomposites were characterized by X-ray diffraction, scanning electron microscopy equipped with an electron energy dispersive spectrometer and transmission electron microscopy to evaluate phase composition, crystal structure and grain size. In vitro bacterial adhesion study indicated a significantly reduced number of Streptococcus mutans and Staphylococcus aureus on the bulk nanostructured Ti-45S5 bioglass-Ag plate surface in comparison to that on microcrystalline Ti plate surface. Nanostructured Ti-based biomaterials can be considered to be the future generation of dental implants.

Microscopic removal of type Ⅲ dens invaginatus and preparation of apical barrier with mineral trioxide aggregate in a maxillary lateral incisor:A case report and review of literature

BACKGROUND Invaginated teeth pose greater challenges in clinical management because of their complex configuration.With advancements in equipment and materials,such as the dental operation microscope,cone-beam computed tomography and mineral trioxide aggregate,the preservation rate of type Ⅲ dens invaginatus could be greatly increased.CASE SUMMARY This case report presented a 31-year-old woman with complaints of spontaneous swelling and pain in the right maxillary lateral tooth.With the aid of cone-beam computed tomography,type Ⅲ dens invaginatus with apical periodontitis was diagnosed and confirmed.Three-visit endodontic treatment was performed.In the first visit,the invagination was carefully removed under the dental operation microscope,and chemomechanical preparation was done.In the second visit,mineral trioxide aggregate apical barrier surgery was performed in this tooth.In the third visit,the canal was finally obturated with thermoplastic gutta-percha to recover the crown morphology.A 26-mo follow-up revealed a satisfied outcome both in the radiographic and oral examinations.CONCLUSION In this case,removal of the entire abnormal structure provided great convenience for the follow-up treatment.When confronted with the same clinical case in the future,we can take a similar approach to address it.

Antibacterial activity of calcium hydroxide combined with chitosan solutions and the outcomes on the bond strength of RealSeal sealer to radicular dentin

The purpose of this study was to investigate the antibacterial activity of calcium hydroxide [Ca（OH）2] combined with chitosan solutions against Enterococcus faecalis-infected root canal dentin and the effect of this new intraca- nal medicament on the bond strength of RealSeal sealer to radicular dentin. An experimental intracanal medicament was prepared by mixing different concentrations of chitosan solution （25%, 50%, and 100%, W/V） to Ca（OH）2 powder. Antibacterial activity was evaluated and the total numbers of colony forming units were determined. Bonding ability of RealSeal sealer to radicular dentin was evaluated using push-out bond strength test. Data were analyzed using one-way analysis of variance （ANOVA） and Tukey＇s multiple comparison tests. We found that Ca（OH）2 combined with different concentrations of chitosan solutions showed better antibacterial activity than Ca（OH）2 mixed with saline, without significantly affecting the bond strength of RealSeal sealer to radicular dentin （P 〉 0.05）. The findings suggest that Ca（OH）2 combined with chitosan is a promising intracanal medicament and may be effective in endodontic therapy.

Comparison of the rheological properties of four root canal sealers

The flowability of a root canal sealer is clinically important because it improves the penetration of the sealer into the complex root canal system. The purpose of this study was to compare the flowabilities of four root canal sealers, measured using the simple press method （ISO 6876）, and their viscosities, measured using a strain-controlled rheometer. A newly developed, calcium phosphate-based root canal sealer （Capseal） and three commercial root canal sealers （AH Plus, Sealapex and Pulp Canal Sealer EWT） were used in this study. The flowabilities of the four root canal sealers were measured using the simple press method （n= 5） and their viscosities were measured using a strain-controlled rheometer （n=5）. The correlation between these two values was statistically analysed using Spearman＇s correlation test. The flow diameters and the viscosities of the root canal sealers were strongly negatively correlated （p= -0.8618）. The viscosity of Pulp Canal Sealer EWT was the lowest and increased in the following order： AH Plus〈Sealapex〈Capseal （P〈0.05）. All of the tested root canal sealers showed characteristic time- and temperature-dependent changes in their rheological properties. The viscosities measured using the strain-controlled rheometer were more precise than the flowabilities measured using the simple press method, suggesting that the rheometer can accurately measure the rheological properties of root canal sealers.

Fracture resistance of posterior teeth restored with modern restorative materials

We studied the fracture resistance of maxillary premolars restored with recent restorative materials. Fifty maxillary premolars were divided into five groups： Group 1 were unprepared teeth; Group 2 were teeth prepared without restoration; Group 3 were teeth restored with tetric ceram HB; Group 4 were teeth restored with InTen S; and Group 5 were teeth restored with Admira. The samples were tested using a universal testing machine. Peak loads at fracture were recorded. The teeth restored with Admira had the highest fracture resistance followed by those restored with InTen-S and tetric ceram HB. Prepared, unrestored teeth were the weakest group. There was a significant difference between the fracture resistance of intact teeth and the prepared, unrestored teeth. There was also a significant difference among the tested restorative materials. Teeth restored with Admira showed no significant difference when compared with the unprepared teeth. It was concluded that the teeth restored with Admira exhibited the highest fracture resistance.

Immediate and delayed micro-tensile bond strength of different luting resin cements to different regional dentin

We sought to evaluate immediate and delayed micro-tensile bond strength of Panavia F2.0 and Multilink Sprint resin cement to superficial, deep and cervical dentin. Thirty-six freshly extracted non-carious human molars were sectioned in the mesiodistal direction to expose three different dentin regions including superficial dentin （1 mm below the dentine-enamel junction）, deep dentin （1 mm above the highest pulp horn） and cervical dentin （0.5 mm above the cemento-enamel junction and 0.5 mm below the dentine-enamel junction）. Resin cements were applied on dentin surfaces and composite blocks were luted under constant seating pressure. Each group was divided into three subgroups according to time intervals. Specimens were sectioned to obtain sticks of 1 mm2 in diameter and subjected to microtensile bond strength testing at a cross head speed of 1 mrn/min. Both resin cements showed higher micro-tensile bond strength to superficial dentin than that to deep or cervical dentin （P 〈 0.001）. Micro-ten- sile bond strengths of Panavia F2.0 were higher than those of Multilink Sprint at different dentin regions （P 〈 0.001）. Immediate ＂micro-tensile bond strengths were higher than those of delayed micro-tensile bond strengths for both resin cements （P 〈 0.001）. It was concluded that resin cements with different chemical formulations and applications yield significantly different micro-tensile bond strengths to different dentin regions.

Non-surgical management of dens invaginatus type IIIB in maxillary lateral incisor with three root canals and 6-year follow-up:A case report and review of literature

BACKGROUND The presence of dens invaginatus(DI)complicates treatment of any tooth,from diagnosis to access cavity and biomechanical preparation and obturation.Reports of successful non-surgical management of DI type IIIB in maxillary lateral incisor are rare.Here,we report such a case,with three root canals and a long follow-up.CASE SUMMARY A 13-year-old female patient presented with mild pain in the maxillary right lateral incisor(#7)for 10-15 d.On examination,the tooth was slightly rotated,with slight tenderness on percussion and grade I mobility but with no caries,pockets or restorations and non-vital pulp(via vitality tests).Radiographic examination revealed unusual configuration of the tooth’s root canals,with an enamel-lined invagination extending to the apex,suggesting the possibility of DI Oehler’s type IIIB and a periapical radiolucency.Widening the access cavity lingually revealed one distinct buccal orifice and two distinct palatal orifices;under higher magnification of a dental operating microscope(DOM),the mesio palatal and disto-palatal orifices were observed as connected by a C-shaped groove.The root canals were prepared with hand K-files following a step-back technique,and obturated using a combination technique of lateral condensation and vertical compaction.At the 6-year follow-up,the patient was asymptomatic,and the periapical radiography displayed significant healing around the apical end of the root.CONCLUSION Proper knowledge of unusual root canal anatomy is required in treating DI.Conventional methods of root canal treatment can successfully resolve such complex cases,facilitated by DOM and cone-beam computed tomography.

3D cellular visualization of intact mouse tooth using optical clearing without decalcification

Dental pulp is composed of nerves,blood vessels,and various types of cells and surrounded by a thick and hard enamel-dentin matrix.Due to its importance in the maintenance of tooth vitality,there have been intensive efforts to analyze the complex cellularlevel organization of the dental pulp in teeth.Although conventional histologic analysis has provided microscopic images of the dental pulp,3-dimensional (3D) cellular-level visualization of the whole dental pulp in an intact tooth has remained a technically challenging task.This is mainly due to the inevitable disruption and loss of microscopic structural features during the process of mechanical sectioning required for the preparation of the tooth sample for histological observation.To accomplish 3D microscopic observation of thick intact tissue,various optical clearing techniques have been developed mostly for soft tissue,and their application for hard tissues such as bone and teeth has only recently started to be investigated.In this work,we established a simple and rapid optical clearing technique for intact mouse teeth without the time-consuming process of decalcification.We achieved 3D cellular-level visualization of the microvasculature and various immune cell distributions in the whole dental pulp of mouse teeth under normal and pathologic conditions.This technique could be used to enable diverse research methods on tooth development and regeneration by providing 3D visualization of various pulpal cells in intact mouse teeth.

Comparison of Apical Leakage in Root Canal Obturation Using Bioceramic and Polydimethylsiloxane Sealer (<i>In Vitro</i>)

Background: One of the ideal properties of a root canal sealer is to have a good sealing ability, especially at the apical third of the root. Objective: To evaluate the comparison of the apical leakage between obturation using bioceramic sealer (BS) and polydimethylsiloxane sealer (PS). Materials and Methods: Thirty-six mandibular premolars were equally divided into two groups and were obturated with single cone technique. The sealers used for Group I and Group II were BS and PS respectively. After obturation, the samples were incubated (37&deg;C, 24 h), sealed with two coats of nail polish except for 2 mm from the apex, submerged in the Indian ink for 7 days, decalcified, dehydrated, and made transparent according to Robertson technique. Dye penetration was evaluated under stereomicroscope. Samples without dye penetration were given score 0, dye penetration ≤ 0.5 mm were given score 1, 0.51 - 1 mm were given score 2, and >1 mm were given score 3. Result: The largest proportion distribution in BS group was at the score 1 (55.6%), whereas in PS group was at the score 2 (44.4%). Conclusion: Bioceramic sealer showed similar apical leakage to polydimethylsiloxane sealer. Clinical Relevance: Based on this study, bioceramic sealer can be recommended to be used as sealer with low level of apical leakage as well as polydimethylsiloxane sealer.

The Ability of Lysate-PRF Induces Proliferation of Fibroblast Cells in Endodontic Regenerative Therapy

Lysate-platelet rich fibrin (Lysate-PRF) is a scaffold that contains growth factor. Aim: To analyze the ability of two types of Lysate-PRF in inducing the proliferation of fibroblast cells. Material & Methods: Microplate with 24 wells each filled 200 μl suspension DMEM + 10% FBS and 10 × 104 fibroblasts. Then FBS is replaced with a concentration of 1%, for serum starvation process. Each 3 wells were exposed with 50%, 25%, 12.5% Lysate-PRF and 50%, 25%, 12.5% Lysate A-PRF respectively. Three other wells are exposed to 10% FBS. Microplate was incubated for 24 hour at 37°C with 5% CO2. The growth of fibroblast was calculated by automatic cell counter. Results: The highest mean value of the Lysate-PRF group was at 12.5% (312.833) while the Lysate A-PRF group at 25% (303.500). If all the groups compared did not show any significant differences. Conclusions: Lysate-PRF and Lysate A-PRF have the same ability as 10% FBS in inducing fibroblast cell proliferation or the same as physiological condition.

Influence of glass-based dental ceramic type and thickness with identical shade on the light transmittance and the degree of conversion of resin cement

The purpose of this study was to assess the influence of the types and thicknesses of glass ceramic plates on light transmittance and compare the degrees of conversion(DC) of resin cement under the ceramic materials. Three ceramic plates with thicknesses of 0.5, 1.0, 2.0, and 4.0 mm were fabricated from each of five commercial ceramic blocks in shade A2: high-translucency and low-translucency IPS Empress CAD(EmpHT and EmpLT); high-translucency and low-translucency IPS e.max CAD(EmxHT and EmxLT); and Vita Mark Ⅱ(Vita). The translucency parameter was obtained using a colorimeter. The light transmittance rate was measured using a photodetector attached to an optical power meter. The DC of a resin cement(Variolink N) underneath the ceramic plates was examined by Fourier transform infrared spectroscopy. The translucency parameter, light transmittance rate, and DC showed significant differences by ceramic type and thickness(P < 0.05). The EmpHT specimens showed the highest light transmission and DCs, and the EmxLT showed the least light transmission and the lowest DCs. The high-translucency Empress showed significantly higher DCs than the low-translucency types(P < 0.05), but there was no significant difference in e.max(P >0.05). Both type and thickness of the glass ceramics significantly influenced the light transmittance and DC of the light-cured resin cement beneath the ceramic of the same shade.