Radiotherapy in the head and neck region influences the chemical and mechanical properties of intraradicular dentin.

OBJECTIVES: To investigate the chemical and mechanical properties of intraradicular dentin submitted to radiotherapy. MATERIALS AND METHODS: Sixteen mandibular incisors were divided into two groups (n = 8): non-irradiated and irradiated. The irradiated teeth were obtained from head and neck radiotherapy patients, with a total dose ranging from 70.2 to 72 Gy divided into 1.8 Gy daily. After sample preparation, intraradicular dentin slices of each root third were evaluated by Raman spectroscopy, energy dispersive spectroscopy and Knoop microhardness test. Data were analyzed by Two-way ANOVA and Tukey's test (α = 0.05). RESULTS: In Raman spectroscopy, carbonate and amide III showed a significant difference for irradiation and third (carbonate p = 0.021 and p < 0.001; amide III p < 0.001 and p = 0.001, respectively). For amide I, there was a significant difference for third (p < 0.001). For carbonate/mineral ratio, there was a significant difference for irradiation (p = 0.0016) and third (p < 0.001), with the irradiated middle third showing the lowest values. For amide I/amide III ratio, there was a significant difference for irradiation (p = 0.005) in the cervical third. In energy dispersive spectroscopy, carbon (p = 0.004; p = 0.020), phosphorus (p < 0.001; p = 0.009) and calcium (p = 0.008; p = 0.007) showed differences for irradiation and third, with the irradiated groups presenting lower values in cervical and middle thirds. For calcium/phosphorus ratio, there was a significant difference for irradiation (p < 0.001) in cervical and middle thirds. Regarding microhardness, there was a significant difference for irradiation (p < 0.001), with all irradiated groups showing lower microhardness values. CONCLUSIONS: The radiotherapy altered the chemical and mechanical properties of intraradicular dentin, mainly in the cervical and middle root thirds.

Fracture resistance and bonding performance after antioxidants pre-treatment in non-vital and bleached teeth.

This study aimed to evaluate the effect of antioxidant solutions on fracture strength and bonding performance in non-vital and bleached (38% hydrogen peroxide) teeth. One hundred and eighty dentin specimens were obtained, 60 for each test: fracture strength, hybrid layer thickness, and bond strength. The groups (n=10) were randomly composed according to post-bleaching protocol: REST - restoration, without bleaching; BL - bleaching + restoration; SA - bleaching, 10% sodium ascorbate solution, and restoration; AT - bleaching, 10% α-tocopherol solution, and restoration; CRAN - bleaching, 5% cranberry solution, and restoration; CAP - bleaching, 0.0025% capsaicin solution, and restoration. Data were analyzed with ANOVA, Kruskal-Wallis, Dunn, and Qui-Square tests (α=0.05). The highest fracture strength values were observed in REST (1508.96 ±148.15 N), without significant difference for the bleached groups (p>0.05), regardless of the antioxidant use. The hybrid layer thickness in the group that was not subjected to bleaching (REST) was significantly higher than in any other group. The bond strength in the bleached and antioxidants-treated groups (SA, AT, CRAN, CAP) has no differences with the bleached group without antioxidants (BL). Adhesive failures were predominant in the groups that did not receive the antioxidant application. In conclusion, the evaluated antioxidants did not show an effect on the fracture strength, hybrid layer thickness, or bond strength of dentin bleached after endodontic treatment. The application of 10% sodium ascorbate, 10% alpha-tocopherol, 5% cranberry, or 0.0025% capsaicin solutions is not an effective step and should not be considered for the restorative protocols after non-vital bleaching.

Minimally invasive management of vital teeth requiring root canal therapy.

The present study aimed to investigate the possible use of a non-instrumentation technique including blue light irradiation for root canal cleaning. Extracted human single rooted teeth were selected. Nine different groups included distilled water, NaOCl, intra-canal heated NaOCl, and NaOCl + EDTA irrigation after either instrumentation or non-instrumentation, and a laser application group following non-instrumentation technique. The chemical assessment of the root canal dentine was evaluated using energy dispersive spectroscopy (EDS) and Fourier transform infrared (FT-IR) spectroscopy. Surface microstructural analyses were performed by using scanning electron microscopy (SEM). The antimicrobial efficacy of different preparation techniques was evaluated using microbial tests. Light application didn't change the calcium/phosphorus, carbonate/phosphate and amide I/phosphate ratios of the root canal dentin. The root canal dentin preserved its original chemistry and microstructure after light application. The instrumentation decreased the carbonate/phosphate and amide I/phosphate ratios of the root canal dentin regardless of the irrigation solution or technique (p < 0.05). The application of light could not provide antibacterial efficacy to match the NaOCl irrigation. The NaOCl irrigation both in the non-instrumentation and instrumentation groups significantly reduced the number of bacteria (p < 0.05). The use of minimally invasive root canal preparation techniques where the root canal is not instrumented and is disinfected by light followed by obturation with a hydraulic cement sealer reduced the microbial load and preserved the dentin thus may be an attractive treatment option for management of vital teeth needing root canal therapy.

Caries affected disinfection using Phycocyanin activated by PDT, Holy Basil, and Ti-sapphire laser on adhesive bond strength, microleakage, and bond failure.

AIM: To investigate the effect of different cavity disinfectants, Phycocyanin (PC), Ocimum Sanctum (OS), and Ti Sapphire Laser, on the bond integrity and microleakage of resin restorations. MATERIAL AND METHOD: 60 human mandibular molars were extracted and prepared based on ICDAS scores of 4 and 5. To obtain the CAD surface, a visual examination was supported by tactile sensation and a dye for caries detection. Samples were randomly allocated into 4 groups based on cavity disinfectants applied (n = 15). Group 1: Specimens disinfected with CHX, Group 2: Specimens disinfected with Ti sapphire laser, Group 3: Specimens disinfected with Phycocyanin activated by Photodynamic therapy, and Group 4: Specimens disinfected with OS. Following the disinfection of the CAD surfaces, composite bulk-fill restorative material was bonded to each specimen and all samples were subjected to thermocycling. Ten samples from each group underwent SBS testing performed on a universal testing machine. Five samples were subjected to a microleakage analysis. RESULT: The maximum microleakage scores were displayed by Group 3: PC (0.521 nm) treated specimens. Whereas, and minimum microleakage was exhibited by Group 4: OS (0.471 nm). Group 4: OS (23.06±0.21 MPa) treated group displayed the maximum bond scores of resin adhesive to the CAD surface. However, Group 3: PC (21.67±0.24 MPa) treated specimens exhibited the lowest bond scores. Failure mode analysis revealed that among all the investigated groups the predominant type of failure was cohesive failure i.e., Group 1 (80%), Group 2 (80%), Group 3 (70%), and Group 4 (90%). CONCLUSION: Ocimum Sanctum, Phycocyanin activated by Photodynamic therapy, and Ti-sapphire laser for disinfection of caries-affected dentin have shown promise in terms of improved bond strength and reduced microleakage.

Adhesive bond of Bioactive and bulk fill resin to disinfected caries affected dentin (lactobacillus specie) with laser and photodynamic therapy.

AIMS: To evaluate the antimicrobial effectiveness of cavity disinfectants chlorhexidine gluconate (CHX), Er, Cr, YSGG laser (ECL), and curcumin photosensitizer (CP) against Lactobacillus and shear bond strength (SBS) of Bioactive (BA) and bulk fill composite (BFC) restorative material bonded to carious affected dentin (CAD). MATERIALS AND METHODS: Sixty human mandibular molars scored 4 and 5 on the International Caries Detection and Assessment System (ICDAS) were included. After inoculating the specimens with lactobacillus specie all the samples were arbitrarily divided into three groups based on the disinfection regime used (n=20). Groups 1 and 2: CAD disinfection using ECL, Groups 3 and 4: CAD disinfection using CP, and Groups 5 and 6: CAD disinfection using CHX. After cavity sterilization, the survival rate was estimated and each group was further divided into two sub-groups based on the restorative material used. Groups 1, 3, and 5 (n = 10) were restored using BFC restorative material, and groups 2, 4, and 6 (n = 10) were restored using a conventional bulk-fill resin material. A universal testing machine (UTM) was used to determine the SBS and debonded surfaces were examined under a stereomicroscope to determine the modes of failure. Kruskal-Walis, ANOVA, and Post Hoc Tukey were applied to investigate the survival rate and bond strength values. RESULTS: ECL group displayed the highest survival rate (0.73±0.13) of Lactobacillus. The least survival rate (0.17±0.09) was demonstrated by CP activated by PDT. Group 1 (ECL+ BA) treated specimens exhibited the maximum value of SBS (18.31 ± 0.22 MPa). However, group 3 (CP + BA) revealed the minimum values of bond strength (14.05 ± 1.02 MPa). The intergroup comparison revealed that group 1, group 2 (ECL+BFC) (18.11 ± 0.14 MPa), group 5 (CHX+ BA) (18.14 ± 0.36 MPa), and group 6 (CHX+BFC) (18.18 ± 0.35 MPa) displayed comparable outcomes of bond integrity (p>0.05). CONCLUSION: Caries-affected dentin disinfected with Er, Cr: YSGG, and chlorhexidine improve bond scores of bioactive and conventional bulk-fill restorative material.

Impact of radiotherapy on the morphological and compositional structure of intra-radicular dentin.

Considering the side effects in the oral cavity and dental structures of radiotherapy (RDT) for head and neck cancer, this study aimed to evaluate the effects of RDT on the root dentin concerning the obliteration of dentinal tubules, the inorganic composition of intra-radicular dentin, and the integrity of collagen fibers. Thirty human canines were selected from a biobank and randomly divided into two groups (n=15). The samples were sectioned buccolingually, and a hemisection was used for structural analysis by scanning electron microscopy (SEM) and energy-dispersive X-ray spectrometer (EDS). Low-vacuum SEM images were obtained at 2000-x magnification to observe the obliteration of the dentinal tubules. Moreover, compositional evaluation was performed using EDS. After RDT, the SEM and EDS analyses were repeated using the same methodology. RDT was applied fractionally at 2 Gy per day, 5 days per week, for 7 weeks, resulting in a total dose of 70 Gy. The collagen integrity of the irradiated and non-irradiated samples was analyzed using Masson's trichrome and picrosirius red staining polarization microscopy. Samples subjected to RDT exhibited dentinal tubule obliteration (p < 0.001); low integrity of type I and III collagen fibers (p < 0.05); compositional reduction of calcium (p = 0.012), phosphorus (p = 0.001), and magnesium (p < 0.001); an increased Ca/P ratio (p < 0.001). RDT affects the structure of dentinal tubules, the inorganic composition of intra-radicular dentin, and the collagen fiber integrity in the root dentin, which may interfere with the effectiveness and durability of dental procedures.

Effect of dentin biomodification using natural collagen cross-linkers on the durability of the resin-dentin bond and demineralized dentin stiffness.

OBJECTIVE: The purpose of this study was to evaluate the effect of using natural cross-linkers as sumac and curcumin on the durability of the resin-dentin bond and stiffness of demineralized dentin matrix. METHODS: Thirty sound molars were divided into 5 groups: Control (CO), Grape Seed extract (GSE), Cacao seed extract (CSE), Sumac extract (SE) and Curcumin extract (CE). The teeth had their coronal dentin exposed, etched, and pre-treated for 1 min with the extracts. Teeth were then bonded using Single-Bond II adhesive and 4 mm composite was built up on dentin surface. Teeth were sectioned into 1 × 1 × 8mm beams and their micro-tensile bond strength (µTBS) was tested after 24 h and 6 months of water storage. For stiffness testing, 15 teeth were sectioned to obtain dentin beams (1 × 1 × 6.5 mm), the beams were demineralized in 10% phosphoric acid then rinsed and divided into 5 groups. Beams were then immersed in their respective extract solution for 1 min after which they were subjected to a 3- point loading test using a universal testing machine to calculate their modulus of elasticity. RESULTS: After 24 h, no significant difference in µTBS was shown between all groups. After 6 Months, GSE, CE, and SE showed significantly higher µTBS compared to CO (p ≥ 0.05). For the modulus of elasticity; only GSE showed a significantly higher modulus compared to other groups. CLINICAL RELEVANCE: The application of grape seed extract, curcumin and sumac extract as dentin pre-treatments appear to be a promising approach to enhance the durability of the resin-dentin bond in a clinically relevant application time.

Collagen-depletion strategies in dentin as alternatives to the hybrid layer concept and their effect on bond strength: a systematic review.

Strategies aiming to improve the longevity of resin-dentin adhesive interface developed so far have only been able to retard the problem. Different approaches are thus needed. The objective of this review was to determine whether the use of collagen-depletion strategies after acid-etching procedures may improve the bond strength of resin-based materials to dentin. A systematic review was planned following 2021 PRISMA statement guidelines, with a search strategy performed in five electronic databases: PubMed/Medline, Scopus, EMBASE, SciELO and IADR Abstract Archive (last search: 17/01/2022). Inclusion criteria encompassed studies which evaluated a collagen-depletion strategy in acid-etched human dentin and tensile/shear bond strength tests. Risk of bias assessment was carried out by two reviewers, working independently on an adapted five-domain risk of bias (RoB) checklist for laboratory studies. Results were synthesized qualitatively, as a meta-analysis was not possible due to limited number of studies and their RoB. A total of eight studies were eligible for inclusion in the systematic review after inclusion/exclusion criteria application. Out of these, two evaluated the effect of using NaOCl followed by an antioxidant, and the remaining six evaluated different enzymatic treatments (bromelain, chondroitinase ABC, papain, and trypsin). None of the studies reported a decrease of bond strength when a collagen-depletion strategy was used, in comparison to traditional hybrid layers (control). All enzymatic treatment studies which respected the inclusion criteria improved the bond strength to dentin. Some specific collagen-depletion strategies seem to play a favorable role in improving immediate bond strengths to dentin. Further research with sound methodology is required to consolidate these findings, since limitations in RoB and a low number of studies were found. The assessment of further proteolytic agents and long-term outcomes is also required.

Functional role of inorganic trace elements in dentin apatite tissue-part III: Se, F, Ag, and B.

Dentin hydroxyapatite possesses a unique versatile structure which allows it to undergo ionic substitutions. Trace elements play pivotal roles within the oral cavity, especially in dentin apatite tissue. Therefore, it is critical to explore the role of these elements in dentin apatite structure. The roles of other inorganic elements in dentin apatite were discussed in part I (Mg, Sr, Zn, and Fe) and part II (Cu, Mn, Si, and Li) of these series. In the last part of the review series, the role of selenium, fluorine, silver, and boron in the regulation of dentin apatite structure and function was discussed. We evaluated how these elements affect the overall size, morphology, and crystallinity of dentin apatite crystals. Moreover, we investigated the importance of these elements in regulating the solubility of dentin apatite. An electronic search was performed on the role of these trace elements in dentin apatite from January 2010 to January 2022. The concentration of selenium in teeth has been explored only recently, particularly its incorporation into dentin apatite. Silver nanomaterials inhibit the growth of cariogenic microorganisms as well as arrest the degradation of collagen. Fluorine was found to have important roles in dentin remineralization and dentinal tubule occlusion, making it widely used for hydroxyapatite doping. Boron is critical for mineralized tissues like bone, dentin, and enamel, but its exact role in dentin apatite is unknown. Therefore, understanding the impact of these elements on dentin apatite is potentially transformative, as it may help to fill a significant knowledge gap in teeth mechanics.

Noncarious cervical lesion pretreated using antimicrobial photodynamic therapy and diode laser in reducing dentin hypersensitivity bonded to different restorative material: Valuation of bond values and invitro dye leakage.

AIM: The present study aimed to appraise the effect of Diode laser (DL), riboflavin, and curcumin on the bond strength of adhesive restorative material and the effects of pretreatment on microleakage scores. MATERIAL AND METHODS: Ninety caries-free human permanent premolars were obtained and disinfected. All specimens were mounted till CEJ. To provide standardization, a 2 × 2 mm flat non-carious cervical lesion (NCCL) dentin surface was exposed. This was followed by dentin surface roughening/cervical groove preparation with a round bur. After tooth surface preparation, ninety specimens were divided arbitrarily into three groups. Group 1: NCCL exposed to DL; group 2: NCCL pretreated with riboflavin photosensitizer (RFP) and group 3 pretreated with curcumin photosensitizer (CP). All the three groups were further divided into two subgroups A1, A2, B1, B2, C1, and C2 based on the restorative material used (n = 15). Subgroup A1, B1, and C1 were restored with composite resin (CR). Subgroup A2, B2, and C2 were restored with RMGIC. Ten samples from each subgroup were evaluated for shear bond strength (SBS) testing under a universal testing machine (UTM). Five from each subgroup were evaluated for microleakage assessment. Bond strength and microleakage analysis were performed using analysis of variance (ANOVA) with a Post-hoc test (Tukey's post hoc test) at a significance level (p = 0.05). RESULTS: The group A1, DL-CR shows the highest SBS (16.41± 0.4 MPa). However, CP-RMGIC groups demonstrated the lowest bond integrity. RFP-CR and the DL-CR group displayed comparable outcomes of SBS (p>0.05). The highest microleakage was observed with CP-RMGIC. The least microleakage was displayed by DL-CR. CONCLUSION: Noncarious cervical lesions pretreated with diode laser and riboflavin photosensitizer showed better shear bond strength and reduced microleakage when bonded to composite resin than non-carious cervical lesions pretreated with curcumin.

Long-term effect of curcuminoid treatment on resin-to-dentin bond strength.

Endogenous dentin proteases contribute to the degradation of collagen fibrils in the hybrid layer. Recently, inhibition of host-derived proteases by curcuminoids has shown promising results. The aim of this study was to evaluate the effect of curcuminoid treatment on the microtensile bond strength (µTBS) after 24 h or 12 months of storage. Fifty-four extracted sound human molars were flattened to mid-coronal dentin and divided into nine groups. After phosphoric acid-etching for 15 s, the dentin was experimentally treated for 60 s using 100 µM or 200 µM of curcumin, diflourobenzocurcumin, or demethoxycurcumin dissolved in 1% and 2% dimethyl sulfoxide (DMSO)/water solutions. Untreated and DMSO-treated groups served as controls. After bonding agent application, each tooth was restored with dental composite. The molars were sectioned into 0.9 × 0.9 × 6 mm beams. The µTBS testing was performed after 24 h and 12 months of storage in artificial saliva. Data were analyzed using regression analyses. Failure patterns were evaluated using scanning electron microscopy. Dentin treatment with curcuminoids did not adversely affect 24-h µTBS compared to controls. After 12 months, the µTBS of curcuminoid groups was statistically significantly higher than the controls. This study indicates the feasibility of using curcuminoids as protease inhibitors.

Sequential Application of Calcium Phosphate and ε-Polylysine Show Antibacterial and Dentin Tubule Occluding Effects In Vitro.

In this study, ε-polylysine and calcium phosphate precipitation (CPP) methods were employed to induce antibacterial effects and dentin tubule occlusion. Antibacterial effects of ε-polylysine were evaluated with broth dilution assay against P. gingivalis. CPP solution from MCPM, DCPD, and TTCP was prepared. Four concentrations of ε-polylysine(ε-PL) solutions (0.125%, 0.25%, 0.5%, 1%) were prepared. Dentin discs were prepared from recently extracted human third molars. Dentin discs were incubated with P. gingivalis (ATCC 33277) bacterial suspension (ca. 105 bacteria) containing Brain Heart Infusion medium supplemented with 0.1 g/mL Vitamin K, 0.5 mg/mL hemin, 0.4 g/mL L-cysteine in anaerobic jars (37 °C) for 7 days to allow for biofilm formation. P. g-infected dentin specimens were randomly divided into four groups: CPP + 0.125% ε-PL, CPP + 0.25% ε-PL, CPP + 0.5% ε-PL, CPP + 1% ε-PL. On each dentin specimen, CPP solution was applied followed by polylysine solution with microbrush and immersed in artificial saliva. Precipitate formation, antibacterial effects, and occlusion of dentinal tubules were characterized in vitro over up to 72 h using scanning electron microscopy. ε-PL showed 34.97% to 61.19% growth inhibition levels against P. gingivalis (P. g) after 24 h of incubation. On P. g-infected dentin specimens, DCPD + 0.25% ε-PL, and DCPD + 0.5% ε-PL groups showed complete bacterial inhibition and 78.6% and 98.1% dentin tubule occlusion, respectively (p < 0.001). The longitudinal analysis on fractured dentin samples in DCPD and TTCP groups revealed deeply penetrated hydroxyapatite-like crystal formations in dentinal tubules after 72 h of incubation in artificial saliva.

Studies of Peculiar Mg-Containing and Oscillating Bioapatites in Sheep and Horse Teeth.

New types of biological apatites have been discovered in molar sheep and horse teeth and are divided in two types. In the first and more general type, the release of Mg ions is parallel to the changes in composition of apatite leading to a final stoichiometric ratio of Ca to P ions, going from dentin depth towards the boundary of enamel with air. Inside dentin, another apatite sub-types were discovered with alternating layers of Mg-rich and C-rich apatites. The approximate formal stoichiometric relationships for these peculiar types of bioapatites are suggested. We identified two kinds of ion-exchanges responsible for formation of peculiar apatites. Various combinations of main and minor elements lead to new versions of biological apatites.

Influence of Dentin Priming with Tannin-Rich Plant Extracts on the Longevity of Bonded Composite Restorations.

OBJECTIVE: This in vitro study evaluated the influence of bioactive plant extracts as dentin biomodifying agents to improve the longevity of bonded restorations. For that, plant extracts were applied to the dentin surface prior to the adhesive system. MATERIALS AND METHODS: Bovine incisors were ground flat to obtain 2 mm thick slices in which conical preparations were made (N = 10). Tannin-containing plant extracts were applied to dentin before the application of the restorative system, as follows: control group (untreated, CTL), chlorhexidine 0.12% (CHX), mastruz (Dysphania ambrosioides, MTZ), cat's claw (Uncaria tomentosa, CTC), guarana (Paullinia cupana, GUA), galla chinensis (Rhus chinensis, GCH), and tannic acid (extracted from Acacia decurrens, TNA). The push-out bond strength test was conducted (0.5 mm/min). Dentin biomodification was assessed by the modulus of elasticity and mass change in bovine tooth sections (0.5 × 1.7 × 7.0 mm). The dentin staining after extract treatments of dentin slices was compared. The dentin surface wettability was also evaluated by means of the contact angles of the adhesive system with the dentin surface and compared with the untreated control group. Data were subjected to ANOVA/Tukey's test (α = 0.05). RESULTS: The bond strength of the restoratives to dentin was not significantly improved by the plant extracts, irrespective of the evaluation time (p > 0.05). Except for TNA, the elastic modulus of demineralized dentin significantly reduced after treatment with the plant extracts (p < 0.05). The dentin staining correlated with the tannin content of the extracts. The contact angle was significantly reduced when treated with CTC, GCH, and TNA. CONCLUSIONS: The tannin-containing extracts had a questionable effect on the longevity of bonded restorations. The dentin modulus was negatively affected by the extract treatments. Although some of the extracts changed the contact angle, which seems to improve the adhesive monomer permeation, the tannin-rich plant extract application prior to adhesive application was proven to be clinically unfeasible due to dentin staining.

The Stability of Dentin Surface Biobarrier Consisting of Mesoporous Delivery System on Dentinal Tubule Occlusion and Streptococcus Mutans Biofilm Inhibition.

BACKGROUND: The dentin exposure always leads to dentin hypersensitivity and/or caries. Given the dentin's tubular structure and low mineralization degree, reestablishing an effective biobarrier to stably protect dentin remains significantly challenging. This study reports a versatile dentin surface biobarrier consisting of a mesoporous silica-based epigallocatechin-3-gallate (EGCG)/nanohydroxyapatite delivery system and evaluates its stability on the dentinal tubule occlusion and the Streptococcus mutans (S. mutans) biofilm inhibition. MATERIALS AND METHODS: The mesoporous delivery system was fabricated and characterized. Sensitive dentin discs were prepared and randomly allocated to three groups: 1, control group; 2, casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) group; and 3, the mesoporous delivery system group. The dentin permeability, dentinal tubule occlusion, acid and abrasion resistance, and S. mutans biofilm inhibition were determined for 1 week and 1 month. The in vitro release profiles of EGCG, Ca, and P were also monitored. RESULTS: The mesoporous delivery system held the ability to sustainably release EGCG, Ca, and P and could persistently occlude dentinal tubules with acid and abrasion resistance, reduce the dentin permeability, and inhibit the S. mutans biofilm formation for up to 1 month compared with the two other groups. The system provided prolonged stability to combat oral adverse challenges and served as an effective surface biobarrier to protect the exposed dentin. CONCLUSION: The establishment of the dentin surface biobarrier consisting of a mesoporous delivery system indicates a promising strategy for the prevention and the management of dentin hypersensitivity and caries after enamel loss.

Shear Bond Strength and Film Thickness of a Naturally Antimicrobial Modified Dental Luting Cement.

Although several natural plants and mixtures have been known and used over the centuries for their antibacterial activity, few have been thoroughly explored in the field of dentistry. Thus, the aim of this study was to enhance the antimicrobial activity of a conventional glass ionomer cement (GIC) with natural plant extracts. The effect of this alteration on the bond strength and film thickness of glass ionomer cement was evaluated and related to an 0.5% chlorohexidine modified GIC. Olive leaves (Olea europaea), Fig tree (Ficus carica), and the leaves and roots of Miswak (Salvadora persica) were used to prepare an alcoholic extract mixture. The prepared extract mixture after the evaporation of the solvent was used to modify a freeze-dried glass ionomer cement at three different extracts: water mass ratios 1:2, 1:1, and 2:1. An 0.5% chlorhexidine diacetate powder was added to a conventional GIC for the preparation of a positive control group (CHX-GIC) for comparison. The bond strength to dentine was assessed using a material-testing machine at a cross head speed of 0.5 mm/min. Failure mode was analyzed using a stereomicroscope at 12× magnification. The cement film thickness was evaluated in accordance with ISO standard 9917-1. The minimum number of samples in each group was n = 10. Statistical analysis was performed using a Kruskal-Wallis test followed by Dunn's post hoc test for pairwise comparison. There was a statistically insignificant difference between the median shear bond strength (p = 0.046) of the control group (M = 3.4 MPa), and each of the CHX-GIC (M = 1.7 MPa), and the three plant modified groups of 1:2, 1:1, 2:1 (M = 5.1, 3.2, and 4.3 MPa, respectively). The CHX-GIC group showed statistically significant lower median values compared to the three plant-modified groups. Mixed and cohesive failure modes were predominant among all the tested groups. All the tested groups (p < 0.001) met the ISO standard of having less than 25 µm film thickness, with the 2:1 group (M = 24 µm) being statistically the highest among all the other groups. The plant extracts did not alter either the shear bond strength or the film thickness of the GIC and thus might represent a promising additive to GICs.

Molecular clocks in ancient proteins: Do they reflect the age at death even after millennia?

Age at death estimation in cases of human skeletal finds is an important task in forensic medicine as well as in anthropology. In forensic medicine, methods based on "molecular clocks" in dental tissues and bone play an increasing role. The question, whether these methods are applicable also in cases with post-depositional intervals far beyond the forensically relevant period, was investigated for two "protein clocks", the accumulation of D-aspartic acid (D-Asp) and the accumulation of pentosidine (Pen) in dentine. Eight teeth of skeletons from different burial sites in Austria and with post-depositional intervals between c. 1216 and c. 8775 years were analysed. The results of age at death estimation based on D-Asp and Pen in dentine were compared to that derived from a classical morphological examination. Age at death estimation based on D-Asp resulted consistently in false high values. This finding can be explained by a post-mortem accumulation of D-Asp that may be enhanced by protein degradation. In contrast, the Pen-based age estimates fitted well with the morphological age diagnoses. The described effect of post-mortem protein degradation is negligible in forensically relevant time horizons, but not for post-depositional intervals of thousands of years. That means that the "D-Asp clock" loses its functionality with increasing post-depositional intervals, whereas Pen seems to be very stable. The "Pen-clock" may have the potential to become an interesting supplement to the existing repertoire of methods even in cases with extremely long post-depositional intervals. Further investigations have to test this hypothesis.

Desperately seeking stress: A pilot study of cortisol in archaeological tooth structures.

OBJECTIVES: Cortisol is a glucocorticoid hormone produced through activation of the hypothalamic pituitary adrenal axis. It is known as the "stress hormone" for its primary role in the body's stress response and has been the focus of much modern clinical research. Within archaeology, only a few studies have analyzed cortisol in human remains and these have been restricted to hair (Webb et al., 2010; Webb, White, van Uum, & Longstaffe, 2015a; Webb, White, van Uum, & Longstaffe, 2015b). This study examines the utility of dentine and enamel, which survive well archaeologically, as possible reservoirs for detectable levels of cortisol. MATERIALS AND METHODS: Then, 69 teeth from 65 individuals from five Roman and Post-Roman sites in France were tested via competitive enzyme-linked immunosorbent assay (ELISA) to assess and quantify the cortisol concentrations present within tooth dentine and enamel. RESULTS: In both tooth dentine and enamel, detectable concentrations of cortisol were identified in multiple teeth. However, concentrations were low and not all teeth yielded results that were measurable through cortisol ELISA. Differences in cortisol values between dentine and enamel could suggest different uptake mechanisms or timing. DISCUSSION: These results suggest that cortisol is incorporated within tooth structures and merits further investigation in both modern and archaeological contexts. Analysis of the results through liquid chromatographic-mass spectrometry would verify current results and might yield values that could be better integrated with published cortisol studies. Future studies of cortisol in tooth structures would greatly expand the research potential of cortisol in the past and could have implications for studies of human stress across deep time.

Growing up in Ancient Sardinia: Infant-toddler dietary changes revealed by the novel use of hydrogen isotopes (δ2H).

Detailed information about the lives and deaths of children in antiquity is often in short supply. Childhood dietary histories are, however, recorded and maintained in the teeth of both juveniles and adults. Primary tooth dentinal collagen does not turn over, preserving a sequential record of dietary changes. The use of nitrogen (δ15N) and carbon (δ13C) isotope values of incrementally sampled dentin are used in the study of breastfeeding practices but evidence for the addition of weaning foods, both in terms of mode and, particularly, duration, has remained analytically inaccessible to date. Here, we demonstrate how the novel use hydrogen isotope (δ2H) values of sequentially micro-sampled dentin collagen, measured from individuals excavated from a Punic cemetery, in Sardinia, Italy, can serve as a proxy for weaning food type and duration in ancient childhood diet. The weaning rate and age, based on the decline in δ15N and δ13C values of permanent first molars and the concomitant increase in δ2H, appears to be broadly similar among six individuals. Hydrogen isotopes vary systematically from a low value soon after birth, rising through early childhood. The early post-birth values can be explained by the influence of 2H-depleted lipids from mother's breastmilk and the later δ2H rise is consistent with, among other things, a substantial portion of boiled foodstuffs, such as the higher δ2H values observed in porridge. Overall δ2H in dentin shows great promise to elucidate infant and childhood feeding practices, and especially the introduction of supplementary foods during the weaning process.

Chemical mapping of teeth in 2D and 3D: X-ray fluorescence reveals hidden details in dentine surrounding fillings.

X-rays are frequently used for characterizing both tooth tissues and dental materials. Whereas radiographs and tomography utilize absorption contrast for retrieving details, chemical mapping is usually achieved by energy dispersive X-ray (EDX) analysis that is stimulated under vacuum in electron microscopes. However, the relatively dense mineralized composition of teeth, and the frequent inclusion of a large range of elements in filling materials raise the possibility that other X-ray based techniques such as X-ray fluorescence (XRF) spectroscopy may strongly contribute to investigations of a large variety of dental structures. By exploiting the fluorescence excited by micron sized X-rays (µXRF) it is possible to map minute quantities of a large range of elements (from aluminum to uranium), where spectra containing signals from multiple different elements can be resolved non-destructively and concomitantly. The high penetration depth of X-rays makes XRF highly effective at detecting variable compositions with information emerging from tooth tissues situated well beneath the sample surface. The method supports minimal sample preparation and, different from electron microscopy, it facilitates investigation of hydrated dental materials. Direct comparison of µXRF and confocal µXRF (CµXRF) with SEM-EDX reveals micro zones of chemical heterogeneity in the complex 3D architecture of root canal fillings. These methods reproducibly clarify the mutual arrangement of biomaterials in both fresh fillings as well as in repeatedly treated old teeth of unknown history. The results showcase the complementarity of X-ray and electron based elemental mapping for dental materials research. STATEMENT OF SIGNIFICANCE: Chemical characterization of mineralized tissues such as tooth dentine is often performed using energy dispersive X-ray spectroscopy (EDS/EDX) analysis by scanning electron microscopy (SEM). The widespread use of electron microscopes and simplified detector designs have made this form of chemical and structural analysis extremely popular. However, excitation by electrons is limited to the upper microns of the tissue, and these may not well represent the chemical composition of the bulk. Especially when heavier elements are of interest and when dental filling materials exhibit diffusion into the tooth, little is known about the spatial distribution. Here we show how complementary X-ray fluorescence data originating by electron and X-ray excitation can help visualize the distribution and impregnation of heavy elements through teeth, e.g. for root canal treatment.