Maria Giovanna Gandolfi

Apatite-forming ability (bioactivity) of ProRoot MTA

AIM Apatite-forming ability, considered as an index of bioactivity (bond-to-bone ability), was tested on ProRoot MTA cement after immersion in phosphate-containing solution (DPBS). METHODOLOGY Disk samples were prepared and immersed in DPBS for 10 min, 5 h, 1 and 7 days. The cement surface was studied by attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, by micro-Raman spectroscopy and by environmental scanning electron microscope with energy dispersive X-ray (ESEM-EDX) analyses. The pH of the storage solution was also investigated. RESULTS Spectroscopic analyses revealed calcium phosphate bands after 5-h immersion in DPBS. After 1 day, an even coating composed of apatite spherulites (0.1-0.8 micron diameter) was observed by ESEM/EDX. After 7 days, its thickness had increased. Apatite nucleation had already occurred after 5-h immersion. At this time, the presence of portlandite (i.e. Ca(OH)(2) , calcium hydroxide) on the cement surface was also observed; at longer times, this component was released into the medium, which underwent a remarkable pH increase. CONCLUSIONS The study confirms the ability of ProRoot MTA to form a superficial layer of apatite within hours. The excellent bioactivity of ProRoot MTA might provide a significant clinical advantage over the traditional cements used for root-end or root-perforation repair.

Environmental Scanning Electron Microscopy Connected with Energy Dispersive X-ray Analysis and Raman Techniques to Study ProRoot Mineral Trioxide Aggregate and Calcium Silicate Cements in Wet Conditions and in Real Time

INTRODUCTION ProRoot mineral trioxide aggregate (MTA) and calcium silicate cements are able to set in a moist environment. The aim of the study was to examine the surface structure and composition of a cement paste under wet conditions and in real time during setting by environmental scanning electron microscopy connected with energy dispersive x-ray analysis (ESEM-EDX) and micro-Raman techniques. METHODS White ProRoot MTA and experimental white tetrasilicate cement (wTC) and wTC containing bismuth oxide (wTC-Bi) were studied. Cement disks were analyzed 10 minutes after powder-liquid mixing (freshly prepared samples) and after immersion in Dulbecco phosphate-buffered saline at 37 degrees C for 24 hours (24-hour-aged samples). RESULTS Freshly prepared wet cements at ESEM-EDX exposed an irregular surface (displaying calcium, silicon, aluminum, chlorine reflexes, and bismuth traces in MTA and wTC-Bi) with needle-like and cubic-hexagonal shaped crystals. Aggregates of spheroidal Ca-P-rich crystals (spherulites) appeared on the surface of 24-hour-aged samples. The starting unhydrated powders displayed the typical Raman bands of Portland cement components: alite, belite, and calcium sulfate (only as anhydrite in MTA and as both anhydrite and gypsum in wTC and wTC-Bi). MTA powder showed higher amount of calcium carbonate and lower quantities of anhydrite and higher crystallinity of the silicate component, leading to a slower hydration reaction. Products/markers of hydration reactions were present on fresh samples; ettringite formed on the surface of all the cements; calcium hydroxide (portlandite) was detected only on the surface of wTC, but no conclusion can be drawn on wTC-Bi and MTA because of the interference of bismuth oxide. Calcium phosphate and calcite/aragonite bands were detected on all 24-hour-aged cements; portlandite was no longer detected on wTC. CONCLUSIONS ESEM and micro-Raman are powerful and suitable techniques to investigate endodontic calcium silicate hydrated cements in real time and in their humid state without inducing artifacts by sample preparation. The formation of apatite spherulites on calcium silicate cements might have clinical relevance.

Evaluation of the radiopacity of calcium silicate cements containing different radiopacifiers

AIM To identify the suitable ratio of alternative radiopacifiers to impart the necessary radiopacity to calcium silicate cements (CSC) and assess the purity of the radiopacifying agents. METHODOLOGY Alternative radiopacifying materials for incorporation into CSC included barium sulphate, titanium oxide, zinc oxide, gold powder and silver/tin alloy. The chemical composition of the alternative radipacifying materials and bismuth oxide, which is used in mineral trioxide aggregate (MTA), was determined using energy dispersive X-ray analysis. In addition, using an aluminium step-wedge and densitometer, the radiopacity of each material was evaluated as recommended by international standards. The optical density was compared with the relevant thickness of aluminium (Al). A commercial MTA and CSC were used as controls. Statistical analysis comparing the radiodensity of the different cements to MTA was performed using anova with P = 0.05 and post hoc Tukey test. RESULTS All percentage replacements of bismuth oxide, gold and silver-tin alloy powder, and the 25% and 30% replacements with barium sulphate and zinc oxide had radiopacities greater than 3 mm thickness of aluminium (Al) recommended by ISO 6876 (2002). The 25% replacement of cement with gold powder and 20% replacement of cement with silver/tin alloy powder exhibited radiopacity values of 8.04 mm Al and 7.52 mm Al, respectively, similar to MTA (P > 0.05). The cement replaced with 20% bismuth oxide showed a radiopacity of 6.83 mm Al, lower than MTA (P = 0.003). CONCLUSIONS Silver/tin alloy and gold powder imparted the necessary radiopacity to a calcium silicate-based cement. Barium sulphate was also a suitable radiopacifier together with a lower concentration of silver/tin alloy and gold powder that achieved the radiodensity recommended by ISO 6876. Further research is required to investigate the broader properties of the calcium silicate-based cement with the different radiopacifiers.

Development of the foremost light-curable calcium-silicate MTA cement as root-end in oral surgery. Chemical–physical properties, bioactivity and biological behavior

AIM An innovative light-curable calcium-silicate cement containing a HEMA-TEGDMA-based resin (lc-MTA) was designed to obtain a bioactive fast setting root-end filling and root repair material. METHODS lc-MTA was tested for setting time, solubility, water absorption, calcium release, alkalinizing activity (pH of soaking water), bioactivity (apatite-forming ability) and cell growth-proliferation. The apatite-forming ability was investigated by micro-Raman, ATR-FTIR and ESEM/EDX after immersion at 37°C for 1-28 days in DPBS or DMEM+FBS. The marginal adaptation of cement in root-end cavities of extracted teeth was assessed by ESEM/EDX, and the viability of Saos-2 cell on cements was evaluated. RESULTS lc-MTA demonstrated a rapid setting time (2min), low solubility, high calcium release (150-200ppm) and alkalinizing power (pH 10-12). lc-MTA proved the formation of bone-like apatite spherulites just after 1 day. Apatite precipitates completely filled the interface porosities and created a perfect marginal adaptation. lc-MTA allowed Saos-2 cell viability and growth and no compromising toxicity was exerted. SIGNIFICANCE HEMA-TEGDMA creates a polymeric network able to stabilize the outer surface of the cement and a hydrophilic matrix permeable enough to allow water absorption. SiO(-)/Si-OH groups from the mineral particles induce heterogeneous nucleation of apatite by sorption of calcium and phosphate ions. Oxygen-containing groups from poly-HEMA-TEGDMA provide additional apatite nucleating sites through the formation of calcium chelates. The strong novelty was that the combination of a hydraulic calcium-silicate powder and a poly-HEMA-TEGDMA hydrophilic resin creates the conditions (calcium release and functional groups able to chelate Ca ions) for a bioactive fast setting light-curable material for clinical applications in dental and maxillofacial surgery. The first and unique/exclusive light-curable calcium-silicate MTA cement for endodontics and root-end application was created, with a potential strong impact on surgical procedures.

Chemical-physical properties of TheraCal, a novel light-curable MTA-like material for pulp capping.

AIM To evaluate the chemical-physical properties of TheraCal, a new light-curable pulp-capping material composed of resin and calcium silicate (Portland cement), compared with reference pulp-capping materials (ProRoot MTA and Dycal). METHODOLOGY Calcium (Ca) and hydroxyl (OH) ion release over 28 days, solubility and water uptake (weight percentage variation, Δ%) at 24 h, cure depth and radiopacity of TheraCal, ProRoot MTA and Dycal were evaluated. Statistical analysis (P < 0.05) of release of ion was carried out by two-way repeated measures anova with Tukey, whilst one-way anova with Tukey test was used for the other tests. RESULTS TheraCal released significantly more calcium than ProRoot MTA and Dycal throughout the test period. TheraCal was able to alkalinize the surrounding fluid initially to pH 10-11 (3 h-3 days) and subsequently to pH 8-8.5 (7-14 days). TheraCal had a cure depth of 1.7 mm. The solubility of TheraCal (Δ-1.58%) was low and significantly less than that of Dycal (Δ-4.58%) and ProRoot MTA (Δ-18.34%). The amount of water absorbed by TheraCal (Δ +10.42%) was significantly higher than Dycal (Δ +4.87%) and significantly lower than ProRoot MTA (Δ +13.96%). CONCLUSIONS TheraCal displayed higher calcium-releasing ability and lower solubility than either ProRoot MTA or Dycal. The capability of TheraCal to be cured to a depth of 1.7 mm may avoid the risk of untimely dissolution. These properties offer major advantages in direct pulp-capping treatments.

Setting time and expansion in different soaking media of experimental accelerated calcium-silicate cements and ProRoot MTA.

OBJECTIVES The setting time and the expansion in deionized water, phosphate-buffered saline (PBS), 20% fetal bovine serum (FBS)/80% PBS or hexadecane oil of experimental accelerated calcium-silicate cements and ProRoot MTA were evaluated. STUDY DESIGN Different compounds such as sodium fluoride, strontium chloride, hydroxyapatite, and tricalcium phosphate were separately added to a basic experimental calcium-silicate cement to test their effect on setting and expansion. The initial and final setting times were determined using appropriate Gilmore needles. A linear variable differential transformer (LVDT) device was used to test the restricted hygroscopic linear expansion over 180 minutes of cements immersed in different solutions. Results were statistically compared using a 2-way ANOVA test (cement type versus solution type). RESULTS All experimental cements showed initial setting times between 28 and 45 minutes and final setting times between 52 and 80 minutes. MTA showed a final setting time of 170 minutes. Final setting time of all experimental cements was faster than MTA. All cements showed slight (0.04%-0.77%) expansion in water, PBS, or FBS/PBS. Only fluoride-containing cement showed a significant expansion in water (6.68%) and in PBS (6.72%). The PBS/FBS contamination significantly reduced the expansion of fluoride-containing cement (2.98%) and MTA (0.07%). In contrast, cements showed a slight shrinkage when immersed in hexadecane, especially fluoride-containing cement. CONCLUSIONS The study demonstrated that: (1) the setting time of calcium-silicate cements may be effectively reduced; (2) the expansion is a water dependent mechanism owing to water uptake, because no expansion occurred in cements immersed in oil; (3) a correlation between setting time and expansion in water and PBS exists; (4) fluorine-containing cement showed a significant expansion in water and in PBS; (5) the immersion in FBS/PBS strongly reduced the expansion of MTA and fluoride-doped cement suggesting that fluid contamination (ie, blood) during surgical procedures may greatly affect the expansion of some calcium-silicate cements.

Calcium silicate coating derived from Portland cement as treatment for hypersensitive dentine

OBJECTIVES To evaluate the in vitro effectiveness on dentine permeability and dentine morphology of a calcium silicate treatment based on Portland cement (DSC). METHODS The experimental treatment consisted of a calcium silicate paste based on Portland cement that was applied on dentine surface for 3 min. A professional re-mineralizing treatment (GC Tooth Mousse), two in office desensitizing agents (D/Sense Crystal, and By Sealant) and a commercial toothpaste Dentosan S were studied as comparison materials. All materials were applied accordingly with manufacturer directions on wet dentine. The quantitative changes in the hydraulic conductance i.e., permeability through tubules of dentine discs samples produced by treatment were quantified in vitro using a hydrostatic device working at 6.9 kPa. SEM/EDX analyses of dentine were carried out to obtain qualitative information on dentine morphology and surface deposits and to study their relationship with the hydraulic conductance. After treatment, each dentine sample was immersed in artificial saliva and permeability re-evaluated. Finally, each sample was exposed to 0.02 M citric acid solution and the final permeability was assessed. RESULTS The experimental treatment and both oxalate-based products (D/Sense Crystal and By Sealant) significantly decreased dentine permeability and created crystals and precipitates on the dentine surface that reduced the diameter of dentinal tubules. Artificial saliva immersion and citric acid challenge increased dentine permeability and partially modified the treated surfaces. Dentosan S and GC Tooth Mousse treatments partially reduced dentine permeability and created small amount of precipitates that were removed by saliva immersion and citric acid exposure. EDX revealed the presence of calcium-rich layer after DSC experimental treatment. CONCLUSIONS The application of the experimental calcium silicate paste and oxalate-based treatments was determined to be effective on dentine permeability reduction and tubules occlusion. The clinical use as desensitizing agent of materials derived from Portland cement as desensitizing agent should be considered for dentine hypersensitivity treatment.

Innovative silicate‐based cements for endodontics: A study of osteoblast‐like cell response

Silicate-based filling materials were designed to obtain new endodontic sealers and root-end filling materials with adequate workability and consistency. Four different formulations (TC, TC 1%, TCf 1%, and TCf) were prepared incorporating calcium chloride as accelerant agent. A plasticizing compound (phyllosilicate) was added to TC 1% and TCf 1%. TC and TC 1% were prepared with water, whereas TCf and TCf 1% were mixed with a latex polymer as fluidizing agent. The aim of this study was to assess the in vitro biological compatibility of designed materials. White-MTA and AH Plus were tested as reference materials. Human osteoblast-like Saos-2 cells were challenged in short-term cultures (72 h) with solid materials and with material extracts in culture medium, and cell viability and number, cellular adhesion, and morphology were assessed. The new cements exerted no acute toxicity in the assay systems. Saos-2 like cells adhered and proliferated on solid samples of the experimental cements and MTA whilst AH Plus did not allowed cell growth. The extracts from the latex-containing cements showed some toxicity. By SEM analysis, osteoblast-like cells appeared adherent and spread on the new materials, and showed the maintenance of polygonal osteoblastic phenotype. Similar morphology was observed for cells on MTA, whereas only few cells were noted on the AH Plus surface. In conclusion, the new materials proved non toxic and supported the growth of bone-like cells, and resulted suitable to be used as endodontic sealers and root-end filling materials.

MTA and F-doped MTA cements used as sealers with warm gutta-percha. Long-term study of sealing ability

AIM To evaluate the long-term sealing ability (up to 6 months) of two experimental calcium silicate MTA cements used as root canal sealers in association with warm gutta-percha. METHODOLOGY Calcium silicate (MTA) and calcium-fluoro-silicate powders were prepared. Sodium fluoride was included in FMTA (Fluoride-doped Mineral Trioxide Aggregate) as an expansive and retardant agent. Single-rooted teeth were instrumented with NiTi rotary instruments, filled with warm gutta-percha in association with one of the experimental sealers or with AH Plus as a control (n = 20 for each sealer) and stored at 37 °C. Sealing was assessed at 24, 48 h, 1, 2 weeks and 1, 3, 6 months by a fluid filtration method. Scanning electron microscopy with energy dispersive analysis (SEM/EDX) was used to study the dentine/sealer interface of roots stored for 6 months and the surface of cement disks stored for 24 h. RESULTS All sealers revealed a statistically significant reduction (P < 0.05) in fluid filtration after the first 2 weeks. No statistically significant differences were observed between FMTA and AH Plus at all analysis times. At short times (24, 48-h), no statistically significant differences were found between the experimental cements and AH Plus. At long-term evaluations (1, 3, 6 months), FMTA and AH Plus sealed significantly better (P < 0.05) than MTA. FMTA was associated with lower fluid filtration rates, and the seal was stable from 48 h to 6 months, thus proving the most effective material. Scanning electron microscopy with energy dispersive analysis of root sections filled with calcium silicate sealers revealed the formation of a blend layer of gutta-percha and cement consequent to the warm gutta-percha condensation technique. Scanning electron microscopy with energy dispersive analysis of 24-h-stored disks identified a Ca-rich coating on the outer surface consisting of globular particles (calcium hydroxide and calcium carbonate), and a deeper internal Ca- and Si-rich region consisting of needle-like ettringite crystals and round formations of calcium silicate hydrate gel. CONCLUSION Fluoride-doped MTA demonstrated stable sealing during a period of up to 6 months and significantly better than conventional calcium silicate MTA cements and comparable to AH Plus. The study supports the suitability of calcium silicate MTA cements as sealers in association with warm gutta-percha for root filling.

Biointeractivity‐related versus chemi/physisorption‐related apatite precursor‐forming ability of current root end filling materials

Commercial root end filling materials, namely two zinc oxide eugenol-based cements [intermediate restorative material (IRM), Superseal], a glass ionomer cement (Vitrebond) and three calcium-silicate mineral trioxide aggregate (MTA)-based cements (ProRoot MTA, MTA Angelus, and Tech Biosealer root end), were examined for their ability to: (a) release calcium (Ca(2+) ) and hydroxyl (OH(-) ) ions (biointeractivity) and (b) form apatite (Ap) and/or calcium phosphate (CaP) precursors. Materials were immersed in Hanks balanced salt solution (HBSS) for 1-28 days. Ca(2+) and OH(-) release were measured by ion selective probes, surface analysis was performed by environmental scanning electron microscopy/energy dispersive X-ray analysis, micro-Raman, and Fourier transform infrared spectroscopy. IRM and Superseal released small quantities of Ca(2+) and no OH(-) ions. Uneven sparse nonapatitic Ca-poor amorphous CaP (ACP) deposits were observed after 24 h soaking. Vitrebond did not release either Ca(2+) or OH(-) ions, but uneven nonapatitic Ca-poor CaP deposits were detected after 7 days soaking. ProRoot MTA, MTA Angelus, and Tech Biosealer root end released significant amounts of Ca(2+) and OH(-) ions throughout the experiment. After 1 day soaking, nanospherulites of CaP deposits formed by amorphous calcium/magnesium phosphate (ACP) Ap precursors were detected. A more mature ACP phase was present on ProRoot MTA and on Tech Biosealer root end at all times. In conclusion, zinc oxide and glass ionomer cements had little or no ability to release mineralizing ions: they simply act as substrates for the possible chemical bonding/adsorption of environmental ions and precipitation of nonapatitic Ca-poor ACP deposits. On the contrary, calcium-silicate cements showed a high calcium release and basifying effect and generally a pronounced formation of more mature ACP apatitic precursors correlated with their higher ion-releasing ability.