Paediatric endodontics Part 2: Pulp regeneration: current approaches.

Dental pulp regeneration: An overview of the current approaches. Regenerative Endodontic Procedures (REPs) are biologically based procedures aimed at restoring the damaged structures and physiological functions of the pulp-dentine complex. Clinically, two strategies have been proposed so far to induce REP: cell transplantation and cell homing. REPs success relies primarily on the clinical and biological conditions of the tooth; therefore, cell homing strategies will not be consistently successful in every condition. Root canal treatment remains the standard of care for mature teeth with necrotic pulps and closed apex.

Wear Properties of Different Additive Restorative Materials Used for Onlay/Overlay Posterior Restorations.

The purpose of this laboratory study was to compare the two-body wear resistance of different restorative materials commonly used for the indirect restoration of posterior teeth. The tested materials, based on ceramic (Imagine Press X, IPS e.max CAD, Milled Celtra Duo, Glaze-Fired Celtra Duo, Vita Mark II) and composite (Enamel Plus HRi, Enamel Plus HRi Bio-Function, Filtek Supreme XTE, Lava Ultimate), were compared with the wear properties of a type III gold alloy (Aurocast 8). Flat samples were prepared with a 6-mm thickness (n=10). Composite samples were tested after a heat polymerization cycle. All samples were exposed to a two-body wear test in a dual axis chewing simulator performing over 120,000 loading cycles. The opposing abrader cusps were fabricated from yttria-stabilized tetragonal zirconia polycrystal. The vertical substance loss (mm) and the volume loss (mm3) were recorded, as was the wear of the antagonist cusp (mm). Mean values were analyzed by one-way analysis of variance. Significant differences among materials were detected. The heat-cured resin-based composite material Enamel Plus Bio-Function and the type III gold alloy demonstrated similar mean values for wear depth and volumetric loss.

Near-infrared LEDS provide persistent and increasing protection against E. faecalis.

In a previous study, we have shown that non-coherent light-emitting diodes (LEDs) are able to significantly decrease Enterococcus faecalis viability, in vitro. In particular, the concurrent treatment for 5 min with sodium hypochlorite (NaOCl) 1.0% and LED permitted to obtain the total inactivation of the culture. The primary outcome of this study was to verify if the photoinactivation was only temporary or stable over 1 week. The secondary outcome was to verify if using a minor concentration of NaOCl together with LED irradiation, it was possible to reach the total inactivation of the culture. Bacterial suspensions were subjected to different protocols. NaOCl samples were treated with 0.5% sodium hypochlorite for 5 min; LED 5 were subjected to 880-nm LED irradiation for 5 min and LED+NaOCl were contemporary subjected to LED irradiation and 0.5% sodium hypochlorite for 5 min. Then the colony forming units per milliliter (CFU/mL) were analyzed and recorded at 5 min, 24 h, 48 h and 1 week. The data were submitted to ANOVA and LSD post hoc tests at a level of significance of 0.05. Live dead assay and fluorescent microscope observations were also performed for all group at all timing points. The results showed that the 3 protocol treatments were able to decrease significantly the bacterial count respect controls, however, after 5 min, 24 h and 48 h the most effective reduction was measured for NaOCl and NaOCl+LED 5. In the first 48 h LED 5 alone reduced significantly the bacterial count respect controls, but it was higher respect the other tested groups. On the contrary, after 1 week we found an interesting result: samples treated with LED were characterized by a decrement of bacterial count, independently from the addition of sodium hypochlorite. Samples treated with NaOCl alone were characterized by a mild increment of bacterial count, although results were still significant respect C+. The secondary outcome was not confirmed: a concentration of NaOCl 0.5% together with LED irradiation did not permit to reach the total inactivation of the culture.

Wear Evaluation of Prosthetic Materials Opposing Themselves.

The purpose of the present in vitro study was to compare the two-body wear resistance of a type 3 gold alloy (Aurocast8), two lithium disilicate glass ceramics (IPS e.max CAD and IPS e.max Press), a heat-pressed feldspathic porcelain (Cerabien ZR Press), an yttria-stabilized tetragonal zirconia polycrystal ceramic (Katana Zirconia ML), and three heat-cured composite resins (Ceram.X Universal, Enamel Plus Function, and Enamel Plus HRi) opposing antagonistic cusps made out of the same restorative materials. Ten 6-mm-thick samples and 10 cusp-shaped abraders were manufactured with each test material (n=10) according to standard laboratory procedures. All sample/antagonist pairs made out of the same material were subjected to a two-body wear test in a dual-axis chewing simulator for up to 120,000 loading cycles. The total vertical wear (mm) and the total volumetric loss (mm3) for each sample/antagonist pair were calculated. Data were statistically analyzed using one-way analysis of variance tests. The total vertical wear for the gold alloy was not significantly different compared to Ceram.X Universal, Enamel Plus Function, IPS e.max CAD, and Cerabien ZR Press. Significantly increased wear values were observed for Enamel Plus HRi and IPS e.max Press. The lowest values for total vertical wear and volumetric loss were recorded on the monolithic zirconia.

Conservative approach for the management of congenital bilateral agenesis of permanent mandibular incisors: case report and literature review.

BACKGROUND: Agenesis of mandibular incisors can lead to compromised dental and facial aesthetics and therefore requires appropriate treatment. When retained primary incisors are associated to this condition, a medium-term restoration may be provided before any definitive treatment to provide aesthetic relief to the young patient. CASE REPORT: The aim of this paper is to discuss the aesthetic management of a case with bilateral congenital agenesis of permanent incisors and retained primary incisors, which were successfully treated with direct composite restorations. CONCLUSION: This appears a viable technique that restores function and aesthetics with a very conservative approach.

Influence of a repeated preheating procedure on mechanical properties of three resin composites.

The aim of this study was to assess the flexural strength, flexural elastic modulus and Vickers microhardness of three resin composites prepared at room temperature or cured after one or repeated preheating cycles to a temperature of 39°C. Three resin composites were evaluated: Enamel Plus HFO (Micerium), Opallis (FGM), and Ceram X Duo (Dentsply DeTrey). For each trial, one group of specimens of each material was fabricated under ambient laboratory conditions, whereas in the other groups, the composites were cured after 1, 10, 20, 30, or 40 preheating cycles to a temperature of 39°C in a preheating device. Ten rectangular prismatic specimens (25 × 2 × 2 mm) were prepared for each group (N=180; n=10) and subjected to a three-point bending test for flexural strength and flexural modulus evaluation. Vickers microhardness was assessed on 10 cylindrical specimens from each group (N=180; n=10). Statistical analysis showed that, regardless of the material, the number of heating cycles was not a significant factor and was unable to influence the three mechanical properties tested. However, a significant main effect of the employed material on the marginal means of the three dependent variables was detected.

Wear properties of a novel resin composite compared to human enamel and other restorative materials.

The purpose of this in vitro study was to compare the two-body wear resistance of human enamel, a pressable glass-ceramic (Imagine PressX), a type 3 gold alloy (Aurocast8), three resins composites currently available on the market (Enamel plus HRi, Filtek Supreme XTE, Ceram.X duo), and one recently introduced resin composite (Enamel plus HRi-Function). Resin composites were tested after simple light curing and after a further heat polymerization cycle. Ten cylindrical specimens (7 mm in diameter) were manufactured with each dental material according to standard laboratory procedures. Ten flat enamel specimens were obtained from freshly extracted human molars and included in the control group. All samples were subjected to a two-body wear test in a dual-axis chewing simulator over up to 120,000 loading cycles, against yttria stabilized tetragonal zirconia polycrystal cusps. Wear resistance was analyzed by measuring the vertical substance loss (mm) and the volume loss (mm(3)). Antagonist wear (mm) was also recorded. Data were statistically analyzed using one-way analysis of variance (ANOVA) (wear depth and volume loss) and Kruskal-Wallis one-way ANOVA on ranks (antagonist wear). Heat-cured HRi function and Aurocast8 showed similar mean values for wear depth and volumetric loss, and their results did not statistically differ in comparison with the human enamel.

Pro-inflammatory cytokine release and cell growth inhibition in primary human oral cells after exposure to endodontic sealer.

AIM: To assay the toxicity of the single-methacrylate-based sealer urethane dimethacrylate (UDMA) (EndoRez) in terms of cell growth and pro-inflammatory cytokines release, in expanded ex vivo human dental pulp stem cells (hDPSCs), human periodontal ligament stem cells (hPDLSCs), human gingival fibroblasts (hGFs) and human osteoblasts (hOSTs). METHODOLOGY: Dental pulp and periodontal ligament stem cells, osteoblasts and fibroblasts were derived from five young donors. After in vitro isolation, hDPSCs, hPDLSCs, hGFs and hOSTs were seeded to resin-based sealers for 24, 48, 72 h up to 1 week. The morphological features and the cell growth and the release of pro-inflammatory interleukin (IL)6, IL8, IL12 and tumour necrosis factor (TNF) α were analysed. Differences in cell growth and in interleukin secretion were analysed for statistical significance with two-way anova tests for multiple comparisons. RESULTS: Exposure to endodontic sealer based on UDMA resulted in a 50% decrease in survival oral cells at 24 h of incubation. No evident morphological changes were present in cell cultures examined. After 48 h, 72 h and 1-week culture time, a progressive cell growth was evident. A significant up-regulation of IL6, IL8, IL12 and TNFα cytokines in cells in contact with the dental sealer compared to the control was observed. CONCLUSION: In vitro, EndoRez interacted with primary human hDPSCs, hPDLSCs, hGFs and hOSTs causing damage to biological system evidenced through cell growth inhibition and up-regulation of IL6, IL8, IL12 and TNFα proinflammatory mediators.

Copper-zinc superoxide dismutase activity in dental pulp after dental preparation.

The superoxide dismutases (SODs) are the major enzymatic defence mechanism against toxic reactive oxygen species generated during normal oxidative metabolism and during the respiratory burst associated with inflammation. To further clarify the potential role of copper-zinc (Cu/Zn)-SOD during inflammation of pulp tissue in humans, the aim was to determine whether significant changes in Cu/Zn-SOD activity occur in healthy dental pulp after dental preparation. The condition of the pulp was assessed using clinical and radiographic evaluation. Thirty systemically healthy patients were the source of the pulp tissue, which was collected by longitudinally grooving and splitting teeth that were matched between the control dental pulp and the prepared tooth (test) dental pulp. Cu/Zn-SOD activity was determined through spectrophotometric methods, with Mann-Whitney tests used to assess the significance of the differences between the groups. The Cu/Zn-SOD activity was 168.2+/-46.4 mU.mg−1 total protein (range: 96-212 mU.mg−1) in the control group, and 328.2+/-84.2 mU.mg−1 total protein (range: 280-420 mU.mg−1) in the test group. The difference between the groups was statistically significant, at P <0.001. These results demonstrate a potential role for Cu/Zn-SOD during dental pulp inflammation in humans after dental preparation.

Endodontic-orthodontic relationships: expression of no synthase in human dental pulp during orthodontic tooth movement.

Inflamed human pulp tissue presents an increase in the level of nitric oxide synthase (NOS). The aim of this study is to verify the presence of NOS in human pulp of teeth that are subject to orthodontic force. 20 healthy subjects, wearers of fixed braces on the upper arch, were selected. An open coil-spring in NiTi was applied on the upper premolar test tooth (TT); the controlateral control tooth (CCT) was subjected to orthodontic treatment but not to the further force of the open coil-spring; the antagonist control tooth (ACT) did not undergo any orthodontic treatment. Pulps were taken from test, contralateral control and antagonist control teeth immediately after the extractions which were done at 15 and 30 days from the start of application of the orthodontic force. The pulp tissue was analyzed through immunohistochemical and molecular biology examinations. The results showed tooth pulps subject to orthodontic treatment were very inflamed in the first 15 days with high levels of iNOS and low levels of eNOS; after 30 days a decrease of the inflammation and an increase of the pulp vascularization were observed together with a reduction of iNOS and an increase of eNOS respectively.

Conservative restorations of endodontically compromised anterior teeth in paediatric patients: physical and mechanical considerations.

AIM: Objective of this in vitro study was to evaluate the influence of fiber post placement on fracture resistance of pulpless anterior teeth restored with standardised Class III and Class IV resin composite fillings. MATERIALS AND METHODS: One hundred and five human maxillary central incisors were selected and randomly divided into 7 (n=15) experimental groups (endodontic therapy/ endodontic therapy and one Class III resin composite filling/ endodontic therapy and one Class IV resin composite filling/ endodontic therapy and two Class III resin composite fillings/ endodontic therapy, fiber post and one Class III resin composite filling/ endodontic therapy, fiber post and one Class IV resin composite filling/ endodontic therapy, fiber post and two Class III resin composite fillings). Specimens underwent fracture strength test. Means (N) were calculated and data were analysed using 1-way ANOVA and Tukey multiple comparisons tests (p=0.05). RESULTS: Concerning teeth with two Class III, fiber post placement significantly increased fracture strength values from 603.59 to 864.24 N. Specimens restored with one Class III (795.21 N without post, 936.68 N with post) showed higher fracture strength values if compared with specimens with two Class III, with significant differences just concerning specimens without a fiber post. Fracture strength was not significantly influenced by fiber post placement in Class IV groups (720.71 N without post, 799.69 N with post). CONCLUSION: Data suggest that fiber post placement may significantly improve anterior teeth fracture strength when at least two Class III composite fillings are associated to the endodontic treatment. No significant effect of fiber post placement could be recorded when just one Class III or one Class IV composite filling were present.

Morphological analysis and interleukin release in human gingival fibroblasts seeded on different denture base acrylic resins.

The development of different types of materials with application in practice dentistry is an area of intense growth and research due to its importance in oral health. Among the diverse materials currently used in restoration or in dentures, the acrylic based resins have been widely employed. The release of toxic components and the changes on their physical and mechanical properties actually represent a goal of intensive research. In vivo analysis showed that the surface roughness of the acrylic resin represents a factor that could stimulate bacteria colonization and soft tissue inflammation. For this purpose, in this work, we have analyzed the cell response to acrylic based resins Ivoclar, Tokuso and Coldpack in basal conditions, unpolished, and after the polished procedure performed to reduce the surface roughness. Our in vitro results using human gingival fibroblasts (HGFs) showed a decrease of cell growth, evaluated by MTT assay starting at 24 h of incubation, in samples seeded on resins in basal conditions and after the polished procedure. This cell growth reduction was associated to evident morphological changes in unpolished materials. After 24 h of culture in presence of polished and unpolished resins a spontaneous release was present of pro-inflammatory cytokines such as interleukin-6 (IL-6) and -8 (IL-8), which was higher in unpolished resins, indicating that the polished procedure, minimizing the cytotoxicity process, could contribute to reduce the gingival inflammation processes.

Overexpression of interleukin-6 and -8, cell growth inhibition and morphological changes in 2-hydroxyethyl methacrylate-treated human dental pulp mesenchymal stem cells.

AIM: To evaluate morphological features, cell growth and interleukin-6 (IL-6) and interleukin-8 (IL-8) secretion in expanded ex vivo human dental pulp mesenchymal stem cells (DP-MSCs) after exposure to 2-hydroxyethyl methacrylate (HEMA). METHODOLOGY: Dental pulp mesenchymal stem cells were derived from the dental pulps of 10 young donors. After in vitro isolation, DP-MSCs were treated with 3 and 5 mmol L(-1) HEMA, and after 24, 48 and 72 h of incubation, their morphological features, cell growth, IL-6 and IL-8 secretion were analysed. Differences in the cell growth and in the interleukin secretion were analysed for statistical significance with two-way anova tests and the Holm-Sidak method for multiple comparisons. RESULTS: Dental pulp mesenchymal stem cells revealed a decrease in cell growth with both treatments (P < 0.05), more evident at 5 mmol L(-1) . Microscopic analysis displayed extensive cytotoxic effects in treated cells, which lost their fibroblastoid features and became retracted, even roundish, with a large number of granules. An up-regulation of IL-6 and IL-8 in treated cells cytokines was evident (P < 0.05). CONCLUSIONS: 2-Hydroxyethyl methacrylate exhibited cytotoxicity, inhibited cell growth and induced morphological changes in cultured DP-MSCs. Moreover, in treated samples, an up-regulation of soluble mediators of inflammation such as IL-6 and IL-8 cytokines was found. The direct application of HEMA potentially induces an inflammation process that could be the starting point for toxic response and cell damage in DP-MSCs.

Histomorphologic alterations of human enamel after repeated applications of a bleaching agent.

The objective of the study was to analyse the histomorphology of enamel after repeated applications of a professional bleaching regimen. Enamel specimens were obtained from buccal surfaces of 20 extracted human incisors. Two specimens were obtained for each tooth. Half of each tooth was maintained in distilled water and served as control; the other part was treated with a 38 percent hydrogen peroxide professional bleaching agent. The treated specimens were divided in four groups: in group 1, the professional bleaching protocol suggested by the manufacturer was applied once; in group 2, the same protocol was repeated twice; in group 3, three times; in group 4, four times. Between bleaching applications and before SEM evaluation, enamel specimens were maintained in artificial saliva for 1 week. Enamel specimens for each group were submitted to a qualitative scanning electron microscopy (SEM) analysis (200X, 500X, 1000X, 3000X), comparing the treated specimens with the related control enamel. Results of the SEM analysis showed no relevant alteration on the enamel surfaces, when the bleaching protocol was applied once or twice. However, significant changes of enamel surface morphology were SEM observed in groups 3 and 4, suggesting a predominance of depressions when bleaching procedure was repeated three or four times. From the results of this in vitro study, it is possible to state that bleaching procedures should not be carried out indiscriminately. SEM analysis showed important alterations of the prismatic structure of the enamel when the bleaching protocol was applied three and four times.

Bond strength evaluation of three self-adhesive luting systems used for cementing composite and porcelain.

Self-adhesive resin cements were recently introduced with the purpose of simplifying the cementation technique, as they combine the use of adhesive and cement in a single application, eliminating the need for pretreatment of the tooth. In the present study a microtensile bond strength test (µ-TBS) was used to compare three self-adhesives, an etch-and-rinse and a self-etch luting system, in the cementation of resin-based composite (RBC) and ceramic disks to dentin. Freshly extracted molars were transversally sectioned to expose flat, deep dentin surfaces. Cylindrical specimens (5 mm in diameter and 10 mm in height), consisting of RBC disks and leucite-based glass ceramic disks, were produced. The RBC disks were sandblasted with 50-µm Al2O3. The ceramic disks were conditioned with 9.5% hydrofluoric acid gel and silane application. All of the disks were then bonded to dentin surfaces employing five different luting agents: iCEM Self Adhesive (Heraeus Kulzer), MaxCem (Kerr Corporation), RelyX UniCem (3M ESPE), EnaCem HF (Micerium), and Panavia F2.0 (Kuraray-Dental). The products were applied according to the manufacturers' instructions. The specimens were sectioned perpendicular to the adhesive interface to produce multiple beams measuring approximately 1 mm2 in cross section. For each experimental group 12 beams were tested. The preterm failures were also taken into account. All of the specimen preparations were performed by the same operator. The beams were tested under tension at a crosshead speed of 0.5 mm/min until failure. Mean µ-TBS values were calculated for each group. Data were analyzed by a two-way analysis of variance, and multiple comparisons were performed using a Tukey test (α=0.05). The UniCem group showed the lowest number of preterm failures among the tested self-adhesive systems. When premature debondings were included in the mean value calculation, bond strength values for the UniCem group were statistically equal to or even higher than those achieved with the other self-adhesives, although these values were still statistically worse than those obtained using traditional multi-step luting agents.

The cytotoxic effects of resin-based sealers on dental pulp stem cells.

AIM: To evaluate the effect of four current resin-based adhesives on expanded ex vivo human dental pulp mesenchymal stem cells (DP-MSCs). METHODOLOGY: Dental pulp mesenchymal stem cells were derived from dental pulps of ten donors. After in vitro isolation, dental pulp stem cells were analysed using flow cytometry. The immunophenotype of DP-MSCs disclosed the homogeneous expression of the mesenchymal-related antigens CD29, CD44, CD73, CD90, CD105, CD166. DP-MSCs were exposed to four different commercially available bonding systems (CMF Bond, Prime&Bond NT, Clearfil S(3) Bond, XP Bond), and after 24, 48 and 72 h of incubation the morphological features and the cell growth were analysed. Moreover, the cell viability was evaluated at the same times by MTT assay. Data were statistically analysed using a two-way anova and Holm-Sidak method (alpha set at 0.05). RESULTS: Significant differences were observed between the four groups when comparing DP-MSCs appearance. DP-MSCs survived and proliferated without inhibition in the presence of CMF Bond adhesive. On the contrary, microscopic evaluation of the other three groups revealed extensive cytotoxic effects from the dentine bonding agents. The MTT assay revealed no statistically significant differences in cell viability after 72 h between the control group and CMF Bond group. All the other experimental groups had statistically lower optical density values. CONCLUSIONS: CMF Bond adhesive allowed human dental pulp stem cells to survive and proliferate. All of the other dentine bonding agents had extensive cytotoxic effects.

Bond strengths of three types of fibre-reinforced post systems in various regions of root canals.

AIM: To evaluate the bond strengths of three adhesive/resin cement/fibre post systems to coronal, middle and apical thirds of post space dentine. METHODOLOGY: Three types of glass-fibre post systems were selected for the study. Saremco Post Non-stop Fibre (Saremco), FRC Postec Plus (Ivoclar Vivadent) and Anatomical Post (Dentalica) were luted in prepared root canals using adhesive system and resin cement provided by the respective manufacturer. The luting agent was placed into the root canal using a specific syringe with needle. A push-out test was performed on sections from the apical, middle and coronal parts of each specimen to measure bond strength. All fractured specimens were observed using a stereomicroscope to identify the modes of failure. RESULTS: Bond strength values were significantly affected both by the post-adhesive-cement system used and by root region (P < 0.05). Additionally, there was no significant interaction between the post system and region of canal factors (P > 0.05). The highest bond strength values were found in the coronal third for all experimental groups. The middle and apical thirds exhibited similar bond strengths with no statistically significant differences amongst them. Saremco and Precision post systems had the highest retentive strengths, whilst the Ivoclar post system had the lowest values. Microscopic analysis of the specimens revealed a prevalence of post-cement and mixed failures. CONCLUSIONS: Type of post-adhesive-cement system and root position had a significant effect on fibre post retention. The coronal region of the canals was characterized by significantly higher bond strengths.

Pulp-dissolving ability of several endodontic irrigants: a spectrophotometric evaluation.

The aim of this study is to test the dissolving action of different concentrations of NaOCl (0.5%, 2.5% and 5%) and EDTA 17% on bovine pulp tissue, testing solutions at three different times (1, 5, 10 minutes), 100 mg.of dental pulp placed into a becker with 20 ml of solution was evaluated using a spectrophotometer. A solution of NaOCl 5% showed the highest solubility value. Samples treated with 2.5% NaOCl showed solubility values of 0.98 in one minute, 29.1 in five minutes and 57 in ten minutes. Groups treated with NaOCl 0.5% showed small dissolving modifications. 17% EDTA showed the lowest values of solubility. 5% NaOCl solution showed highest pulp dissolving ability, even a 2.5% solution showed high dissolving ability but in longer times.

NOS evaluations in human dental pulp-capping with MTA and calcium-hydroxide.

The aim of this study is to compare mineral trioxide aggregate (MTA) with calcium hydroxide when used as pulp-capping material in human teeth. 40 teeth were divided into groups based on clinical diagnosis: healthy and hyperaemic. The teeth were pulp capped with MTA and calcium hydroxide. We localized the eNOS and iNOS by immunohistochemistry, tested their mRNA expression by RT-PCR and protein levels by western blots. The evaluation of the samples was based on the cell inflammatory response and on the pulp tissue organization. In particular, evaluation of eNOS and iNOS differences between the various groups and the cellular evolution after the first 7 days from the treatment, and at a distance of 28 days. Our results suggest that there are differences in localization and expression between eNOS and iNOS in dental pulp. Our study has helped us to better understand the effects that calcium hydroxide and MTA have on pulp tissue.

The performance of human periodontal ligament mesenchymal stem cells on xenogenic biomaterials.

Mesenchymal stem cells from periodontal ligament (PDL-MSCs) hold great promise for bone regeneration. Most studies regarding the osteogenic differentiation of stem cells from periodontal tissue suggest that PDL cells may have many osteoblast-like properties, including the ability to form calcified nodules in vitro. This study investigated the morphological and histochemistry aspects of human PDL-MSCs, induced for osteogenic differentiation and seeded on a xenogenic porcine bone substitute in vitro, at different times of incubation. This biomaterial seems physically identical to human bone, and it has been reported to be osteoconductive. Our results indicated that the cells had a high affinity for the three-dimensional biomaterials; in fact, cellular proliferation and colonization was evident, and after 21 days the adherent cells started to detach themselves from the substrate, and at 30 days of incubation in differentiation medium, the cells completely lost the adhesion to the Petri's disk, englobing all bioparticles. In conclusion, the in vitro behaviour of PDL-MSCs and their relationship with three-dimensional scaffold biomaterials encourage in vivo investigations for their use in dental tissue regeneration.