Oxidative stress (OS) is a common feature of many inflammatory diseases, oral pathologies, and aging processes. The impact of OS on periodontal ligament cells (PDLCs) in relation to oral pathologies, including periodontal diseases, has been investigated in different studies. However, its impact on orthodontic tooth movement (OTM) remains poorly understood. This study used an in vitro model with human PDLCs previously exposed to H2O2 to investigate the effects of OS under a static compressive force which simulated the conditions of OTM. Human PDLCs were treated with varying concentrations of H2O2 to identify sub-lethal doses that affected viability minimally. To mimic compromised conditions resembling OTM under OS, the cells were pretreated with the selected H2O2 concentrations for 24 h. Using an in vitro loading model, a static compressive force (2 g/cm2) was applied for an additional 24 h. The cell viability, proliferation, and cytotoxicity were evaluated using live/dead and resazurin assays. Apoptosis induction was assessed based on caspase-3/7 activity. The gene expression related to bone remodeling (RUNX2, TNFRSF11B/OPG, BGLAP), inflammation (IL6, CXCL8/IL8, PTGS2/COX2), apoptosis (CASP3, CASP8), and autophagy (MAP1LC3A/LC3, BECN1) was analyzed using RT-qPCR. This study suggests an altering effect of previous OS exposure on static-compression-related mechanosensing. Further research is needed to fully elucidate these mechanisms.

ABSTRACTObjectiveTo investigate the association between interleukin‐8 (IL‐8) levels in gingival crevicular fluid (GCF) and total oral fluid (TOF) and the responsiveness to steps 1 and 2 of periodontal therapy.Materials and MethodsOne‐hundred and fifty‐nine patients affected by periodontitis received steps 1 and 2 of periodontal therapy. At baseline, TOF and GCF samples were collected and analysed for IL‐8 (Il‐8TOF/IL‐8GCF) using flow cytometry. Therapy outcomes were relative proportions of residual periodontal pockets (PPD%), pocket closure (PC) rates and pocket probing depth (PPD) reductions; these were associated with IL‐8TOF/IL‐8GCF.ResultsHigh IL‐8TOF was significantly associated with higher residual PPD% (p = 0.044) and lower PPD reduction compared to low IL‐8TOF (high 0.79 ± 1.20 mm vs. low 1.20 ± 1.20 mm, p < 0.001) in non‐smokers, while in smokers high IL‐8GCF was related to lower PPD reduction (high 0.62 ± 1.22 mm vs. low 0.84 ± 1.12 mm, p = 0.009). Furthermore, high baseline IL‐8TOF was significantly associated with poorer PC rates compared to medium and low concentrations in both non‐smokers (high 41% vs. medium 55% vs. low 58%, p < 0.001) and smokers (high 34% vs. medium 44% vs. low 46%, p < 0.001).ConclusionHigh IL‐8 concentrations at baseline had a significant impact on residual PPD%, PC rates and PPD reduction. The findings suggest that, especially in non‐smokers, baseline IL‐8 levels collected from the TOF could serve as a component in the estimation of responsiveness to steps 1 and 2 of periodontal therapy.

AIM:To analyse whether some selected inflammatory biomarkers collected from venous blood and gingival crevicular fluid (GCF) were associated with the outcome of non-surgical periodontal therapy. MATERIALS AND METHODS:Two-hundred and nine patients affected by periodontitis were enrolled in the study, who had undergone steps I and II therapy as well as a non-surgical re-instrumentation (NSRI) of periodontal pockets after 6 months. Serum (SE), plasma (PL) and GCF samples were quantitatively analysed for the following inflammatory biomarkers: active matrix metalloproteinase-8 (aMMP-8), prostaglandin E2 (PGE2) and surfactant protein D (SP-D). Therapy outcomes were evaluated using a 'treat-to-target' endpoint (T2T) at the patient level, defined as ≤ 4 sites with pocket depth ≥ 5 mm. RESULTS:Patients presented with 23 ± 6 teeth (mean ± SD) at baseline. After steps I and II therapy, 41.6% of the patients reached T2T and after NSRI 47.4%. Univariate analysis identified a potential association between high levels of PL-SP-D and more favourable treatment outcomes. Multivariate binary logistic regression adjusted for sex, mean baseline probing depth, diabetes and current smoking status confirmed an independent relationship between baseline PL-SP-D and the T2T after steps I and II therapy (aOR 0.432, p = 0.011), implying that a higher level PL-SP-D at baseline is associated with a > 50% reduced risk of failing T2T. However, no such association was found for PL-SP-D and NSRI. CONCLUSION:Higher baseline PL-SP-D levels might be associated with more favourable treatment outcomes after steps I and II therapy. This may be due to its role in the regulation of neutrophil function. However, further investigation is required to confirm this hypothesis. If proven, PL-SP-D could play a role as a biomarker for identifying individuals who respond differentially to primary therapy.

This study aims to analyse the association between the baseline microbial load of selected periodontopathogenic bacteria collected from gingival crevicular fluid (GCF) and the primary outcome of steps I and II therapy. 222 patients with stage III periodontitis were included into this retrospective analysis that received steps 1 and 2 periodontal therapy without adjunctive systemic antibiotics. Baseline GCF samples were quantitatively analysed using ELISA-based kits for levels of periodontopathogens (Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa), Prevotella intermedia (Pi), Fusobacterium nucleatum (Fn), Treponema denticola (Td), and Tannerella forsythia (Tf)) and associated with the primary therapy outcome using a “treat-to-target” therapy endpoint (TE) defined as ≤ 4 sites with PD ≥ 5 mm six months after therapy. 38.2

While HA is present naturally in periodontal tissues, its molecular weight can vary widely in vivo. The objective of this study was to directly compare the biological reactions of periodontal ligament cells to four distinct molecular weights of hyaluronic acid (HA). Immortalized human periodontal ligament cells (PDL-hTERT) were cultured for 21 days in culture medium alone (control) or enriched with osteogenic supplements (OS group). Other 4 experimental groups were cultured in OS medium with the addition of HA with different molecular weights (HMW, MMW, LMW, and ULMW). The cell morphology was examined daily. WST1 assays were performed to evaluate metabolic activity. Von Kossa staining and calcium deposition assay were used to analyze osteogenic differentiation and mineralization. Cell morphology remained unaltered in all groups. Cells stimulated with OS alone or with the addition of hyaluronan showed all the typical microscopic appearance of osteogenic differentiation. Metabolic activity increased in all groups over time. Hyaluronan stimulated greater metabolic activity than the control group, with LMW HA and MMW HA showing the most significant increase. All groups showed mineral deposits and calcium deposition after 21 days of stimulation. Our results suggest that hyaluronan can promote metabolic activity and mineralization of PDL-hTERT cells, with LMW HA being the most effective. These results shed light on how the various molecular weight fractions of HA promote tissue regeneration and repair, as well as help to identify an optimal molecular weight range for this application in periodontal tissues.

Objectives: Amelogenins are clinically used in periodontal regeneration as main components of root surface modifying agents, even without specifically preventing the premature colonization of the healing tissue defect by means of a physical barrier membrane. The objective of this study was to investigate the effects of human amelogenin on the proliferation, migration, and morphology of Immortalized Human Oral Keratinocytes (iHOKs).Methods: Immortalized Human Oral Keratinocytes were expanded in Keratinocyte Growth Medium-2 (KGM-2). Full-length recombinant amelogenin protein was diluted in KGM-2 in five concentrations (10 ng/ml, 100 ng/ml, 1.000 ng/ml, 5.000 ng/ml and 10.000 ng/ml). iHOKs were cultured in medium supplemented with the amelogenin dilutions. Samples without amelogenin served as control. Cell metabolism and cell proliferation together with cell migration were evaluated at day 7, 14, 21.Results: At day 7, iHOKs treated with 10,000 ng/ml showed a significant decrease in keratinocytes ' proliferation. The metabolic activity at this timepoint was significantly lower for concentrations > 1000 ng/ml. At days 14 and 21, both the addition of 5000 ng/ml and even more 10,000 ng/ml amelogenin reduced significantly the proliferation of keratinocytes. The effects on the metabolic activity for these timepoints were visible already with 100 ng/ml. Treatment of iHOKs with amelogenin of > 1000 ng/ml led to inhibitory effects on cell migration already after 24 h.Conclusions: The full-length recombinant amelogenin has a significant biological impact on iHOKs. The increasing dose dependent inhibitory effects of amelogenin shown on iHOKs might explain the disruption of the apical migration of the junctional epithelium during regenerative healing.Clinical significance: Amelogenin, presents time- and dose-dependent inhibitory effects on the growth of keratinocytes, which might explain the biological rationale behind its application in periodontal regeneration.

Objectives: Various dental resin materials are available for the fabrication of temporary restorations using modern additive printing methods. Albeit these materials are placed for several months in intimate contact with dental hard and soft tissues, including the gingival crevice, there exists only insufficient evidence on the biocompatibility of these materials. This in vitro study aimed to delineate the biocompatibility of 3D printable materials on periodontal ligament cells (PDL-hTERTs). Methods: Samples of four dental resin materials for additive fabrication of temporary restorations using 3D printing (MFH, Nextdent; GC Temp, GC; Freeprint temp, Detax; 3Delta temp, Deltamed), one material for subtractive fabrication (Grandio disc, Voco) and one conventional temporary material (Luxatemp, DMG) were prepared with a standardized size according to the manufacturer's instructions. Human PDL-hTERTs were exposed to resin specimens or eluates of the material for 1, 2, 3, 6 and 9 days. For determination of cell viability, XTT assays were performed. In addition, the expression of the proinflammatory cytokines interleukin 6 and 8 (IL6 and 8) was assessed in the supernatants with ELISA. Cell viability and the expression of IL-6 and 8 in presence of the resin material or their eluates was compared with untreated controls. Immunofluorescence staining for IL6 and IL-8, as well as scanning electron microscopy of the discs after culturing, were performed. Differences between groups were analyzed with Student ' s t-test for unpaired samples. Results: Compared to untreated control samples, the exposure against the resin specimen induced strong reduction of cell viability in case of the conventional material Luxatemp (p < 0.001) and the additive material 3Delta temp (p < 0.001) irrespective of the observation period. On the contrary, the presence of eluates of the various materials induced only minor changes in cell viability. Considering IL-6 (day 2: p = 0.001; day 6 and 9: p < 0.001) and IL-8 (day 1: p = 0.001; day 2, 3, 6, 9: p < 0.001) their expression was strongly reduced in presence of the eluate of Luxatemp. Except for IL-6 at day 1 and 6 also the material 3Delta temp caused significant reduction of both proinflammatory mediators at any time point. Significance: The conventional material Luxatemp and the additive material 3Delta temp appear to severely affect cell viability when in direct contact with PDL-hTERTs. The other tested materials of this new category of additive materials and the subtractive material Grandio seem to induce only minor changes in direct contact with these cells. Therefore, they could serve as a viable alternative in the fabrication of temporary restorations.

Objectives Evaluation of cumulative survival and complication rate of monolithic lithium disilicate inlays and partial crowns performed by supervised undergraduate students up to 8.3 years of clinical service. Materials and methods In this retrospective clinical study 143 lithium disilicate posterior restorations (IPS e.max Press) were examined according to the FDI criteria. A standardised questionnaire was used to determine patient satisfaction. The aesthetic outcome was evaluated by dentists and dental technicians using intraoral photographs. Data were descriptively analysed. Cumulative survival and success rates were calculated using Kaplan–Meier estimation. Results The cumulative survival rate of lithium disilicate restorations was 97.5% after a mean service time of 5.9 years and 95.0% after 8.3 years. The cumulative success rate decreased from 94.4% after 5.9 years to 30.7% after 8.3 years. Repairs were required for 7 restorations (4.9%), and 5 (3.5%) were classified as failures. The results of the questionnaire indicate a high level of patient satisfaction. The subjective aesthetics were assessed more critically by dental technicians compared to dentists. Conclusion Lithium disilicate posterior restorations survived successfully up to 8.3 years when carried out by undergraduate students. Clinical relevance Pressed lithium disilicate glass ceramic inlays and partial crowns are reliable treatment options in posterior teeth.

Objective: In this retrospective clinical study, we investigated the long-term survival and success of indirect gold restorations, placed by undergraduate students. Material and Methods: The study included 72 gold restorations in 45 patients, placed in a five-year period between 2009 and 2013. Two experienced dentists performed the evaluation following the FDI criteria after 5 to 10 years. Kaplan-Meier test was used for longevity analysis of success and survival. Data were tested for normality with Shapiro-Wilk test. Cox regression analysis, with significance level appointed at p <= .05, was performed using success and survival of restorations as dependent variables. Annual failure rates (AFRs) were calculated. Results: Overall, 72 indirect gold restorations prepared in posterior teeth were assessed, with an observation time from 63 months to 117 months. The mean service time was 88.1 months and the median 87 months. No failures, but 2 repairs, were observed during the first five years. From 5 to 10 years 1 replacement and 1 more repair occurred. The survival and success rates of gold restorations were calculated at 9 years with 98.6% and 91%, respectively. Restorations placed on premolars had higher survival and success rates compared to molars. Three cases were categorized as repairs: in two cases, a root canal treatment was needed and, in one other, an occlusal perforation of the restoration was observed. Conclusions: Posterior indirect gold restorations placed by dental undergraduate students showed a highly predictable clinical performance in terms of success and survival. These results are comparable with survival rates reported in literature for restorations cemented in general dental practices. Clinical significance: Casted gold restorations placed from dental students in an undergraduate setting show high survival rates, similar to professional dentists.

Objectives: Self-adhesive dual cured resin cements provide easier clinical application than conventional resin cements but release higher amounts of unreacted monomers, potentially affecting their biocompatibility. This study aimed to compare the cytotoxic effects of self-adhesive dual cured resin cements with two conventional resin cements. Methods: Samples of four resin cements, two self-adhesive dual cured cements (group A: RelyX Unicem, group B: SmartCem), and two conventional resin cements (group C: Panavia 2.0, group D: Variolink Esthetic DC) were prepared with a similar dimension under standardized polymerization conditions and stored in water. For each material 18 samples were used and cell cultures of human mesenchymal stem cells (hMSCs) or periodontal ligament cells (PDL-hTERT) were added under appropriate conditions. One experimental group (group E) was left untreated as control. A cell viability WST test, was performed in each experimental group at day 1, 7, 14 and 21. Moreover, microscopic examination of cells was performed using cell viability staining. Results: Viability of both cell types as determined by WST test was significantly impaired at all time periods by the four different cement materials compared to the untreated control. Comparison between the four materials revealed different inhibition of the viability of both, PDL-hTERT and hMSC cells (group C > group B > group A > group D; p < 0.0001). Significance: All resin-based cements caused significant impairment of cell viability, reflecting considerable cytotoxicity. Variolink caused significantly smaller changes of viability than the other tested materials. (c) 2021 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.

OBJECTIVE This study investigated the effect of eluates of conventional and 3D-printed resin materials for manufacturing temporary dental restorations on gingival keratinocytes. METHODS Three-dimensional (3D)-printed resin materials: 3Delta temp (Deltamed), NextDent MFH (Nextdent), Freeprint temp (Detax), GC temp (GC), were compared to Grandio disc (Voco) and Luxatemp (DMG). Human gingival keratinocytes (IHGKs) were exposed to eluates of the materials and XTT assays were performed at 24 h, 48 h, 72 h, or 144 h. For quantification of the proinflammatory response, the protein amount of IL-6 and 8 was determined in the supernatants using ELISA. One-way ANOVA with post hoc analysis was used to compare differences in cell viability and IL-6 and IL-8 levels between groups. RESULTS At 24 h, and more remarkably at 48 h, a significant decrease in cell viability occurred for the 3D-printed materials compared to the untreated IHGKs, but also compared to Grandio disc and Luxatemp. Except for the expression of IL-8 in presence of the eluate of Grandio disc at 24 and 48 h, all tested materials caused attenuation of IL-6 and 8 from IHGKs for any observation period. CONCLUSIONS The materials for additive manufacturing affect cell proliferation differently than the subtractive manufactured material Grandio disc and the conventional material Luxatemp. CLINICAL SIGNIFICANCE In comparison to conventional and subtractive manufactured restorations, 3D printed temporary restorations might induce more negative effects on the gingival and probably also on pulpal health since viability and the proinflammatory response of oral keratinocytes are more intensively affected by these materials.

Objectives Neutrophil granulocytes have been proposed to play a major role in the mediation of periodontitis-associated tissue destruction. Their recruitment and activation are regulated by the chemokine CXCL8. This study aimed to delineate the dependency of CXCL8 expression in gingival crevicular fluid (GCF) and saliva on periodontal status, bacterial infection, and smoking, in patients with periodontitis. Methods The study cohort comprised 279 subjects with untreated periodontitis. Probing pocket depth (PPD), gingival recession, bleeding on probing (BOP), plaque index, and bone loss were evaluated. CXCL8 was determined in saliva and GCF using flow cytometry. Results Considering the entire study sample, CXCL8 levels were correlated with the mean PPD (rho = 0.131; p = 0.029), severity of periodontitis (rho = 0.121; p = 0.043), BOP (rho = 0.204; p = 0.001), and smoking (rho = -0.219; p < 0.0001) in GCF; and, in whole saliva, with mean PPD (rho = 0.154; p = 0.010) severity of periodontitis (rho = 0.140; p = 0.020), gender (rho = 0.178; p = 0.003), and smoking (rho = -0.156; p = 0.010). Subgroup analysis among non-smokers revealed significantly higher amounts of CXCL8 in GCF (p = 0.012) and saliva (p = 0.026) comparing subjects with mean PPD <= 3mm and >3mm. Conclusion The current study revealed a strong dependency of CXCL8 expression in GCF on the severity and activity of periodontal disease. Smoking causes a significant reduction in CXCL8 expression in saliva and GCF.

To determine the ultrastructural changes of titanium surfaces of dental implants induced by the tip of periodontal probes. A total of 40 samples of smooth and rough surfaces of titanium implants were randomly assigned for the treatment with metal or plastic periodontal probes under application angles of 20° and 60°. Titanium surfaces have been evaluated with CLSM prior and following to experimental probing determining various standardized 2D and 3D roughness parameters. The average profile and surface roughness (Ra and Sa) showed no significant difference between treated and untreated samples on smooth and rough surface areas irrespective of the probe material. On smooth surfaces several amplitude roughness parameters were increased with metal probes but reached significance only for Rp (p = 0.007). Rough surface parts showed a slight but not significant reduction of roughness following to the contact with metal probes. The surface roughness remained almost unchanged on smooth and rough implant surfaces using plastic probes. The surface roughness on implant surfaces was not dependent on the application angle irrespective of the probe material. Probing of titanium implants with metal probes and even less with plastic probes causes only minor changes of the surface roughness. The clinical significance of these changes remains to be elucidated. Using plastic probes for the clinical evaluation of the peri-implant sulcus might avoid ultrastructural changes to titanium implant surfaces.

OBJECTIVES:Long-term survival of two highly viscous glass ionomer cements (Fuji IX GP Fast and Equia Fil) over a period of 6 years in vivo.METHODS:A total of 85 two- or three-surface class II restorations, comprising 43 Equia Fil / Equia Coat and 42 Fuji IX GP Fast / Fuji Coat LC, were placed in 34 patients. The restorations were re-evaluated after 6 years using the FDI criteria. The statistical analysis was performed with Fisher's exact test, the Wilcoxon signed-rank test, the Mann-Whitney U test and the Kaplan-Meier method.RESULTS:Forty-four restorations (22 Equia Fil and 22 Fuji IX GP Fast) could be assessed at the 6-year follow-up. During the whole study period, eight failures, four for each material, were observed. The main reasons for failure were material fractures and retention loss, which were partly combined with poor marginal adaptation or poor proximal anatomical form. Two failures may be attributed to insufficient application of the materials, as suspected according to the radiographs. The Kaplan-Meier survival proportion for Equia Fil restorations at 6 years was 86.5% and that for Fuji IX GP Fast at 6 years was 86.8% (log-rank p = 0.907). During the period from 3 to 6 years, only one filling in each group failed.CONCLUSION:Both materials showed acceptable and comparable survival rates after 6 years.CLINICAL SIGNIFICANCE:Highly viscous glass ionomer cement can be an acceptable restoration material for smaller class II cavities.

OBJECTIVES:To analyze retrospectively the clinical performance and survival of lithium disilicate inlays, partial crowns and crowns placed by undergraduate students. METHODS:250 lithium disilicate posterior restorations (IPS™Emax Press) were evaluated. Mean service time was 6.6 years (79 months) with a maximum observation time of 8.5 years (102 months). Clinical performance was assessed using FDI Criteria. Descriptive statistics were used to display the distribution of the criteria. Success and survival of the restorations were calculated with the Kaplan Meier method. A Cox regression analysis was performed to detect influencing parameters for the survival or the success of the restorations. RESULTS:Survival and success rates of lithium disilicate indirect restorations were calculated at 6.6 years with 96.3 % and 93.8 %, respectively. After 8.5 years the survival rate was calculated with 94 % and the success rate with 83.8 %. 10 out of 250 restorations (4%) failed, while the main reasons for failure were fracture of material, debonding/loss of restoration, endodontic complications as well as recurrence of caries. A Cox regression analysis failed to show a correlation between restoration failure and the experience level of the students, an existing root canal treatment, type of tooth, bruxism or the use of rubber dam. CONCLUSIONS:Lithium disilicate indirect posterior restorations demonstrate a high success and survival rate after a mean service of 6.6 years when placed by undergraduate students with different levels of experience. Co-factors influencing the survival or success rate of these restorations could not be identified. CLINICAL SIGNIFICANCE:Lithium disilicate restorations show a good clinical performance up to 8.5 years when prepared by undergraduate students.

Objective: To compare the clinical performance of two glass-ionomer cements in combination with two light-cured coatings in Class II cavities. Method and materials: In total, 43 Equia Fil/Equia Coat and 42 Fuji IX GP Fast/Fuji Coat LC restorations were placed, in 34 patients. Only Class II cavities with two-or three-surface restorations were included. The clinical performance was evaluated at baseline, after 1, 2, and 3 years using the FDI criteria. For the statistical analysis, Fisher exact test, Wilcoxon signed-rank test and Mann-Whitney U test were applied. The success and survival of the restorations were calculated with the Kaplan-Meier method and a log-rank test was used to detect significant differences between the tested materials. Results: At the 3-year recall, no significant differences could be detected between the two glass-ionomer cements for any of the evaluated criteria (Mann-Whitney U test, P> .05). When comparing the baseline with the recall data at 3 years for each material separately, significant changes could be observed in both materials for the criteria "surface luster," "fracture of material and retention," and "approximal anatomical form" (Wilcoxon signed-rank test, P < .05). Additionally, the Equia Fil restorations showed a notable change for the criteria "marginal adaptation" (Wilcoxon signed-rank test, P= .039). At the 3-year follow up, the Equia Fil and the Fuji IX GP Fast restorations exhibited an overall survival rate of 86.21% and 84.0%, respectively. Conclusion: The two glass-ionomer cements tested performed similarly in Class II cavities in adult patients, with a moderate to high failure rate after 3 years.

Oral bacteria and odontogenic oral infections are responsible for a high portion of cases with infective endocarditis. Hence, oral health in patients with congenital heart disease (CHD) gains particular importance. This case-control study compared the oral health status in 112 adults with CHD and 168 healthy control subjects. In addition, the patient group was stratified according to the complexity of the heart defect and the recommendation for antibiotic prophylaxis during invasive dental procedures. Considering caries experience, a significantly lower mean DMFT (decayed missing filled teeth) score (7.91 ± 6.63 vs. 13.6 ± 8.15; p < 0.0001) was found in patients with CHD compared to healthy controls. Healthy controls had a higher average number of decayed teeth (0.33 ± 0.76 vs. 1.76 ± 2.61; p < 0.0001). In female subjects a significant lower relative amount of teeth with apical periodontitis was found among CHD patients (3.4% ± 0.9%) as compared to healthy controls (5.6% ± 1.9%) (p = 0.053). Regarding periodontal health, patients with CHD had lower rate of sulcus bleeding (0.32 ± 0.65 vs. 0.71 ± 0.60; p < 0.0001) and less alveolar bone loss than heart healthy individuals (% root length: multi rooted teeth: 8.97 ± 10.64 vs. 23.22 ± 20.70; p < 0.0001; single rooted teeth: 5.59 ± 6.25 vs. 17.30 ± 17.17; p = 0.003). On the contrary, CHD patients presented with higher amount of plaque in comparison to healthy controls (Quigley & Hein index: 2.22 ± 0.67 vs. 1.25 ± 0.72; p < 0.0001). Based on the current results, it can be concluded that adults with CHD have better oral health than heart healthy individuals.

Regenerative therapies of pathogenic tissue defects are gaining increasing importance in periodontology. Among others, the osteogenic effect of BMP-7 seems to play a major role in the development of teeth and alveolar bone. Human periodontal ligament stem cells (hPDLSC), as well as human mesenchymal stem cells (hMSC), show the ability to differentiate into various types of tissues. Regarding prostaglandin E2, many studies have confirmed that it is involved in the inflammation associated to periodontitis stimulating osteoclasts, which ultimately leads to resorption of tooth supporting bone. Herein, we aimed to investigate how PGE2 influences regenerative processes. The influence of PGE2 and BMP-7 on the osteogenic differentiation of hMSC and hPDLSC was determined in a 3D cell culture model using qRT-PCR, immunocytochemistry and REM. BMP-7 enhanced the expression of osteogenic markers in hMSC and lowered it in hPDLSC-TERT. BMP-7 had a lower osteogenic effect on hPDLSC-hTERT than on hMSC, while PGE2 decreases the osteogenic differentiation in both cell types, thus, inhibiting anabolic processes. Both cell types presented good proliferation and adhesion onto the scaffolds. The well-developed structural morphology and the support of osteogenic differentiation suggest that the scaffolds are potential candidate materials for bone regeneration. The positivity for Cap in hPDLSC and more in hMSC immunostaining samples indicates the initiation of neocementogenesis as part of periodontal regeneration. In conclusion, BMP7, in particular combined with MSC, seems to have a favourable application also in periodontal regeneration. Our results show that inflammation plays an important role in periodontal regeneration. PGE2 is a key mediator, which stimulates bone resorption also via a mechanism involving the inhibition of osteogenic differentiation of MSC as well as PDLSC. Therefore, regenerative approaches should always be conducted in combination with anti-inflammatory measures oriented to control inflammation.

Background: Amelogenin is an extracellular matrix protein well known for its role in the organization and mineralization of enamel. Clinically, it is used for periodontal regeneration and, due to its finding also in predentin and intercellular spaces of dental pulp cells, it has recently been suggested for pulp capping procedures. The aim of this study was to analyse in vitro the effect of the recombinant human full-length amelogenin on the growth and differentiation of human dental pulp stem cells (hDPSCs).Methods: Human DPSCs were treated with a supplement of amelogenin at a concentration of 10 ng/ml, 100 ng/ml and 1000 ng/ml. The groups were compared to the unstimulated control in terms of cell morphology and proliferation, mineralization and gene expression for ALP (alkaline phosphatase), DMP1 (dentin matrix protein-1) and DSPP (dentin sialophosphoprotein).Results: Amelogenin affects hDPSCs differently than PDL (periodontal ligament) cells and other cell lines. The proliferation rate at two weeks is significantly reduced in presence of the highest concentration of amelogenin as compared to the unstimulated control. hDPSCs treated with low concentrations present a downregulation of DMP1 and DSPP, which is significant for DSPP (p = 0.011), but not for DMP1 (p = 0.395).Conclusions: These finding suggest that the role of full-length amelogenin is not restricted to participation in tooth structure. It influences the differentiation of hDPSC according to various concentrations and this might impair the clinical results of pulp capping.