Periodontal grading-estimation of responsiveness to therapy and progression of disease.

OBJECTIVE: To investigate the capability of periodontal grading to estimate the progression of periodontal disease and the responsiveness to therapy. MATERIALS AND METHODS: Eighty-four patients who underwent non-surgical therapy (NST) were included. Direct and indirect evidence of progression were determined according to the current classification. Responsiveness to therapy was examined using mean pocket probing depths reduction (PPDRed), reduction of bleeding on probing (BOPRed), and the rate of pocket closure (%PC) after six months. RESULTS: Statistical analysis revealed no agreement between direct and indirect evidence in grading periodontitis (κ = 0.070). The actual rate of progression as determined by longitudinal data was underestimated in 13% (n = 11), overestimated in 51% (n = 43) and correctly estimated in 30% (n = 36) by indirect evidence. No significant differences in responsiveness to therapy were observed in patients graded according to direct evidence. Using indirect evidence, patients assigned grade C showed more PPDRed but less BOPRed and lower %PC compared to grade B. CONCLUSION: The present data indicate that indirect evidence may lead to inaccuracies compared to direct evidence regarding the estimation of periodontal progression. However, indirect evidence seems to be more suitable in the estimation of responsiveness to therapy than direct evidence, helping to identify cases that are more likely to require additional therapies such as re-instrumentation or periodontal surgery. CLINICAL RELEVANCE: Regarding the estimation of disease progression and responsiveness to periodontal therapy, accuracy and reliability of both direct and indirect evidence are limited when grading periodontitis.

MRI-detected intraosseous bone marrow edema recedes after effective therapy of periodontitis.

OBJECTIVES: T2 STIR MRI sequences can detect preclinical changes associated with periodontal inflammation, i.e. intraosseous edema in the tooth-supporting bone. In this study, we assessed whether MRI can be used for monitoring periodontal disease. MATERIAL AND METHODS: In a prospective cohort study, we examined 35 patients with periodontitis between 10/2018 and 04/2019 by using 3D isotropic T2-weighted short tau inversion recovery (STIR) and Fast Field Echo T1-weighted Black bone sequences. All patients received standardized clinical exams before and three months after non-surgical periodontal therapy. Bone marrow edema extent was quantified in the STIR sequence at 922 sites before and after treatment. Results were compared with standard clinical findings. Non-parametric statistical analysis was performed. RESULTS: Non-surgical periodontal treatment caused significant improvement in mean probing depth (p < 0.001) and frequency of bleeding on probing (p < 0.001). The mean depth of osseous edema per site was reduced from a median [IQR] of 2 [1, 3] mm at baseline to 1 [0, 3] mm, (p < 0.001). Periodontal treatment reduced the frequency of sites with edema from 35 to 24% (p < 0.01). CONCLUSION: The decrease of periodontal bone marrow edema, as observed with T2 STIR MR imaging, is indicative of successful periodontal healing. CLINICAL RELEVANCE STATEMENT: T2 STIR hyperintense bone marrow edema in the periodontal bone decreases after treatment and can therefore be used to evaluate treatment success. Furthermore, MRI reveals new options to depict hidden aspects of periodontitis. KEY POINTS: • T2 STIR hyperintense periodontal intraosseous edema was prospectively investigated in 35 patients with periodontitis before and after treatment and compared to clinical outcomes. • The frequency of affected sites was reduced from 35 to 24% (p < 0.001), and mean edema depth was reduced from a median [IQR] of 2 [1, 3] mm at baseline to 1 [0, 3] mm 3 months after treatment. (p < 0.001). • T2 STIR sequences can be used to monitor the posttreatment course of periodontitis.

Influence of decision-making algorithms on the diagnostic accuracy using the current classification of periodontal diseases-a randomized controlled trial.

OBJECTIVES: To examine the influence of the decision-making algorithms published by Tonetti and Sanz in 2019 on the diagnostic accuracy in two differently experienced groups of dental students using the current classification of periodontal diseases. MATERIALS AND METHODS: Eighty-three students of two different clinical experience levels were randomly allocated to control and study group, receiving the staging and grading matrix, resulting in four subgroups. All diagnosed two patient cases with corresponding periodontal charts, panoramic radiographs, and intraoral photographs. Both presented severe periodontal disease (stage III, grade C) but considerably differed in complexity and phenotype according to the current classification of periodontal diseases. Controls received the staging and grading matrix published within the classification, while study groups were additionally provided with decision-trees published by Tonetti and Sanz. Obtained data was analyzed using chi-square test, Spearman's rank correlation, and logistic regression. RESULTS: Using the algorithms significantly enhanced the diagnostic accuracy in staging (p = 0.001*, OR = 4.425) and grading (p < 0.001**, OR = 30.303) regardless of the clinical experience. In addition, even compared to the more experienced control, less experienced students using algorithms showed significantly higher accuracy in grading (p = 0.020*). No influence on the criteria extent could be observed comparing study groups to controls. CONCLUSION: The decision-making algorithms may enhance diagnostic accuracy in dental students using the current classification of periodontal diseases. CLINICAL RELEVANCE: The investigated decision-making algorithms significantly increased the diagnostic accuracy of differently experienced under graduated dental students and might be beneficial in periodontal education.

The molecular weight of hyaluronic acid influences metabolic activity and osteogenic differentiation of periodontal ligament cells.

OBJECTIVE: 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). MATERIALS AND METHODS: 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. RESULTS: 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. CONCLUSION: Our results suggest that hyaluronan can promote metabolic activity and mineralization of PDL-hTERT cells, with LMW HA being the most effective. CLINICAL RELEVANCE: 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.

Visualization of clinically silent, odontogenic maxillary sinus mucositis originating from periapical inflammation using MRI: a feasibility study.

OBJECTIVES: Maxillary sinus mucositis is frequently associated with odontogenic foci. Periapical inflammation of maxillary molars and premolars cannot be visualized directly using radiation-based imaging. The purpose of this study was to answer the following clinical question: among patients with periapical inflammatory processes in the maxilla, does the use of magnetic resonance imaging (MRI), as compared to conventional periapical (AP) and panoramic radiography (OPT), improve diagnostic accuracy? METHODS: Forty-two subjects with generalized periodontitis were scanned on a 3 T MRI. Sixteen asymptomatic subjects with mucosal swelling of the maxillary sinus were enrolled in the study. Periapical edema was assessed using short tau inversion recovery (STIR) sequence. Apical osteolysis and mucosal swelling were assessed by MRI, AP, and OPT imaging using the periapical index score (PAI). Comparisons between groups were performed with chi-squared tests with Yates' correction. Significance was set at p < 0.05. RESULTS: Periapical lesions of maxillary premolars and molars were identified in 16 subjects, 21 sinuses, and 58 teeth. Bone edema and PAI scores were significantly higher using MRI as compared to OPT and AP (p < 0.05). Using the STIR sequence, a significant association of PAI score > 1 and the presence of mucosal swelling in the maxillary sinus was detected (p = 0.03). CONCLUSION: Periapical inflammation and maxillary mucositis could be visualized using STIR imaging. The use of MRI may help detect early, subtle inflammatory changes in the periapical tissues surrounding maxillary dentition. Early detection could guide diagnostic criteria, as well as treatment and prevention.

Evaluation of 3D MRI for early detection of bone edema associated with apical periodontitis.

OBJECTIVES: To detect and evaluate early signs of apical periodontitis using MRI based on a 3D short-tau-inversion-recovery (STIR) sequence compared to conventional panoramic radiography (OPT) and periapical radiographs in patients with apical periodontitis. MATERIALS AND METHODS: Patients with clinical evidence of periodontal disease were enrolled prospectively and received OPT as well as MRI of the viscerocranium including a 3D-STIR sequence. The MRI sequences were assessed for the occurrence and extent of bone changes associated with apical periodontitis including bone edema, periradicular cysts, and dental granulomas. OPTs and intraoral periapical radiographs, if available, were assessed for corresponding periapical radiolucencies using the periapical index (PAI). RESULTS: In total, 232 teeth of 37 patients (mean age 62±13.9 years, 18 women) were assessed. In 69 cases reactive bone edema was detected on MRI with corresponding radiolucency according to OPT. In 105 cases edema was detected without corresponding radiolucency on OPT. The overall extent of edema measured on MRI was significantly larger compared to the radiolucency on OPT (mean: STIR 2.4±1.4 mm, dental radiograph 1.3±1.2 mm, OPT 0.8±1.1 mm, P=0.01). The overall PAI score was significantly higher on MRI compared to OPT (mean PAI: STIR 1.9±0.7, dental radiograph 1.3±0.5, OPT 1.2±0.7, P=0.02). CONCLUSION: Early detection and assessment of bone changes of apical periodontitis using MRI was feasible while the extent of bone edema measured on MRI exceeded the radiolucencies measured on OPT. CLINICAL RELEVANCE: In clinical routine, dental MRI might be useful for early detection and assessment of apical periodontitis before irreversible bone loss is detected on conventional panoramic and intraoral periapical radiographs.

Influence of DLP and SLA printer technology on the accuracy of surgical guides for implant dentistry in free-end situations.

AIM: The present in vitro study aimed to compare the accuracy of the implant position inserted by surgical guides manufactured by three different 3D printers (one SLA and two DLP) and a subtractive manufacturing technique for a free-end situation. MATERIALS AND METHODS: Surgical guides were manufactured using three different 3D printers, one SLA printer (Form 2; Formlabs) and two DLP printers (D20 II; Rapid Shape, and SolFlex 350; W2P Engineering). Surgical guides manufactured using the conventional CAD/CAM-based subtractive method served as controls. In total, 48 surgical guides were printed and assessed for accuracy. The guides were used for the placement of implants at tooth position 37. The postoperative implant position was digitally scanned for metric comparison with the preoperatively planned implant position. RESULTS: Significant differences were found when the SLA and DLP printers were compared. The SLA printer showed significantly lower accuracy in implant displacement at crest (P = 0.032) and angular displacement (P = 0.049) compared with the two DLP printers. The DLP printers showed comparable values to the control group. The results were within an acceptable clinical range. CONCLUSION: DLP technology seems to be superior to conventional SLA technology in terms of the accuracy of 3D-printed surgical guides when used for placement of implants in free-end situations.

Salivary and gingival CXCL8 correlation with periodontal status, periodontal pathogens, and smoking.

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 (ρ = 0.131; p = 0.029), severity of periodontitis (ρ = 0.121; p = 0.043), BOP (ρ = 0.204; p = 0.001), and smoking (ρ = -0.219; p < 0.0001) in GCF; and, in whole saliva, with mean PPD (ρ = 0.154; p = 0.010) severity of periodontitis (ρ = 0.140; p = 0.020), gender (ρ = 0.178; p = 0.003), and smoking (ρ = -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.

Effects of resin materials dedicated for additive manufacturing of temporary dental restorations on human gingival keratinocytes.

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.

Effect of Different Parameters of In Vitro Static Tensile Strain on Human Periodontal Ligament Cells Simulating the Tension Side of Orthodontic Tooth Movement.

This study aimed to investigate the effects of different magnitudes and durations of static tensile strain on human periodontal ligament cells (hPDLCs), focusing on osteogenesis, mechanosensing and inflammation. Static tensile strain magnitudes of 0%, 3%, 6%, 10%, 15% and 20% were applied to hPDLCs for 1, 2 and 3 days. Cell viability was confirmed via live/dead cell staining. Reference genes were tested by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) and assessed. The expressions of TNFRSF11B, ALPL, RUNX2, BGLAP, SP7, FOS, IL6, PTGS2, TNF, IL1B, IL8, IL10 and PGE2 were analyzed by RT-qPCR and/or enzyme-linked immunosorbent assay (ELISA). ALPL and RUNX2 both peaked after 1 day, reaching their maximum at 3%, whereas BGLAP peaked after 3 days with its maximum at 10%. SP7 peaked after 1 day at 6%, 10% and 15%. FOS peaked after 3 days with its maximum at 3%, 6% and 15%. The expressions of IL6 and PTGS2 both peaked after 1 day, with their minimum at 10%. PGE2 peaked after 1 day (maximum at 20%). The ELISA of IL6 peaked after 3 days, with the minimum at 10%. In summary, the lower magnitudes promoted osteogenesis and caused less inflammation, while the higher magnitudes inhibited osteogenesis and enhanced inflammation. Among all magnitudes, 10% generally caused a lower level of inflammation with a higher level of osteogenesis.

Influence of 3D-printing method, resin material, and sterilization on the accuracy of virtually designed surgical implant guides.

STATEMENT OF PROBLEM: Three-dimensional printing has introduced new manufacturing methods. However, information on the influence of the specific printing technology, material, sterilization, and the comparison between printing and milling on the accuracy of surgical guides is lacking. PURPOSE: The purpose of this in vitro study was to evaluate the influence of the manufacturing method (printing and milling), printing technology stereolithography (SLA) and digital light processing (DLP), material, and sterilization on the accuracy of digitally designed surgical implant guides. MATERIAL AND METHODS: Resin patient replicas with a single edentulous space were used to place 132 implants with digitally designed surgical guides. The accuracy of postoperative implant position was analyzed for the manufacturing method (printing and milling), resin materials, and preoperative autoclaving. To determine 3D accuracy, angular displacement, mean horizontal crestal, apical displacement, and the linear vertical displacement at the apex were calculated separately for each group (n=12). In addition, the surgical guides were qualitatively analyzed by using field emission scanning electron micrograph. RESULTS: The postoperative angular deviation ranged from 0.76 ±0.52 degrees (Rapidshape D20II with NextDent SG) to 2.43 ±0.64 degrees (Form2 with NextDent SG) (P<.001). Linear horizontal displacement at the crest was smallest for Rapidshape D20II with 3Delta Guide (0.27 ±0.08 mm) and highest for Form2 with NextDent SG (0.54 ±0.10 mm) (P<.001). Linear horizontal displacement at the apex ranged from 0.36 ±0.10 mm (SolFlex 350 with V-Print SG) to 0.89 ±0.32 mm (Form2 with NextDent SG) (P<.001). Considering the vertical position displacement was no more than 0.43 ±0.07 mm (Form2 with NextDent SG) short of the apex, none of the implant tips were displaced apically. Preoperative autoclaving differentially impaired the accuracy of surgical guides. CONCLUSIONS: The specific manufacturing technique, the 3D printing device, the resin material, and the application of preoperative sterilization all affected the accuracy of the postoperative implant position. Irrespective of the manufacturing method, all implants were placed within the commonly accepted safety distance.

Magnetic resonance imaging as a diagnostic tool for periodontal disease: A prospective study with correlation to standard clinical findings-Is there added value?

AIM: To evaluate the correlation between standard clinical findings, radiographic (OPT) and magnetic resonance imaging (MRI) as well as to assess whether MRI is capable of providing additional information related to the severity and extent of periodontal disease. METHODS: 42 patients with generalized periodontitis received pre-interventional MRI scans. These were compared to MR images of a periodontal healthy control group (n = 34). The extent of the osseous oedema, detected by MRI, was set in correlation with clinical periodontitis-associated findings. RESULTS: A highly significant correlation between bone oedema and clinical testings such as probing depth (p < 0.0001) and bleeding on probing (p < 0.0001) was revealed. The oedema exceeded the extent of demineralized bone. Patients with a positive BOP test showed a 2.51-fold increase in risk of already having a bone oedema around the respective tooth even if probing depth was ≤3 mm (logistic binary regression analysis, OR 2.51; 95% CI: 1.54-4.11; p < 0.0001). CONCLUSION: MRI findings correlated with standard clinical findings, and MRI was able to depict intraosseous changes before any osseous defect had occurred.

Influence of the tooth position, guided sleeve height, supporting length, manufacturing methods, and resin E-modulus on the in vitro accuracy of surgical implant guides in a free-end situation.

PURPOSE: The study aims to evaluate the effect of tooth position, sleeve height, supporting length, resin E-modulus, and manufacturing method on the accuracy of dental implants placed in vitro fully guided in a lower jaw free-end situation. MATERIAL AND METHODS: Using resin clones of a patient case providing a free-end situation 384 implants has been experimentally placed guided. The accuracy of postoperative implant position analyzed depending on the tooth position (first and second molar), sleeve height (2 and 6mm), supporting length (tooth no. 37-33 and 37 43), resin E-modulus (<2,000, >2,000, and >3,000 MPa), and manufacturing method (milled, printed). To determine the three-dimensional accuracy angular deviation, mean crestal, apical deviation, and the linear vertical deviation at apex were calculated separately for each group (n = 12). RESULTS: The accuracy of implant placement using milled guides was affected only by the tooth position, indicating stronger deviation of implants replacing second molars than first molars. Considering printed guides, the implant position was influenced by tooth position, sleeve height, and supporting length. Linear vertical deviation >1 mm was found for printed materials with the lowest E-modulus at tooth position no. 37. Logistic regression analysis revealed a significant lower odds for linear vertical deviation >1 mm for materials with higher E-modulus (OR: 0.04; 95% CI:0.01-0.97; p = .048) and higher ratio for smaller sleeve height (OR: 2.77; 95% CI:1.20-6.38; p = .017). CONCLUSION: Accuracy of implant placement in a free-end situation lacking distal tooth support is superior for milled as compared to 3D-printed surgical guides. When using 3D-printed surgical guides, smaller sleeve heights and extended tooth support improve the accuracy of implant placement.

Ultrastructural changes of smooth and rough titanium implant surfaces induced by metal and plastic periodontal probes.

OBJECTIVES: To determine the ultrastructural changes of titanium surfaces of dental implants induced by the tip of periodontal probes. MATERIALS AND METHODS: 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. RESULTS: 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. CONCLUSION: 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. CLINICAL RELEVANCE: Using plastic probes for the clinical evaluation of the peri-implant sulcus might avoid ultrastructural changes to titanium implant surfaces.

Temperature changes on the root surface during application of warm vertical compaction using three different obturation units.

This study aimed to determine the temperature increase on the root surface during warm vertical compaction using three different obturation systems. Forty-five human single-rooted premolars were randomly assigned to one of three experimental groups of 15 teeth each for obturation with a System B unit, an Elements Obturation Unit, or a B&L SuperEndo Alpha II unit. All teeth were filled using the continuous wave of condensation technique with a set temperature of 200 °C. The temperature measurements were made with an infrared camera at 3 s (T0) and 1 min (T1) following activation of the heat plugger. T0 ranged between 48.1 and 84 °C depending on the obturation system. The mean value for T0 showed significant differences between various obturation systems (p = 0.001). The temperature increase on the root surface during the application of vertical compaction shows considerable variability depending on the obturation system.

Differences in RUNX2 and P2RX7 gene expression between mono- and coculture of human periodontal ligament cells and human osteoblasts under compressive force application.

OBJECTIVES: Objectives were (a) to establish an in vitro coculture model of human PDL-derived fibroblasts (HPDFs) and human alveolar bone-derived osteoblasts (HABOs), and (b) to measure RUNX2 and P2RX7 gene expression at multiple time points after compressive force (CF) stimulation for different durations. SETTING/SAMPLE POPULATION: Human PDL-derived fibroblasts and HABOs from two individuals isolated from extracted teeth of healthy donors due to orthodontic reasons at LMU's Dental School were used. MATERIALS AND METHODS: Mono- and cocultured cells were subjected to CF by centrifugation (47.4 g/cm2 ) for 1, 2 and 4 hours at 30°C. Total RNA was isolated before and 2, 4, 8, 16 minutes after CF application. Expression of RUNX2 and P2RX7 was determined using quantitative real-time polymerase chain reaction. RESULTS: In monocultured cells, an upregulation of RUNX2 and P2RX7 expression was found after all CF durations: RUNX2 twofold in HPDFs after 4 hours (P = 0.002) and 1.7-fold in HABOs after 1 hour (P = 0.009) or 2 hours (P = 0.002) of CF. P2RX7 was significantly upregulated following 1 hour (P = 0.004) and 2 hours (P = 0.002) of CF application in HPDFS. In HABOs, a transient regulation was observed. In cocultured cells, a significant alleviation in gene expression was found for both loci after CF application. In both cell types, RUNX2 expression was significantly reduced in cocultured cells compared to cells in monoculture. P2RX7 expression showed significant attenuation in HPDFs only. CONCLUSIONS: Cocultivation attenuated the CF induced increase of RUNX2 and P2RX7 gene expression in comparison to monoculture in both cell types. This suggests that intercellular communication may exist between HPDFs and HABOs.

Influence of static forces on the expression of selected parameters of inflammation in periodontal ligament cells and alveolar bone cells in a co-culture in vitro model.

OBJECTIVE: Aim of this study was to investigate the impact of human PDL-derived fibroblasts (HPDF) and human alveolar bone-derived osteoblasts (HABO) co-culture on the expression of cytokines involved in tissue remodeling using an in vitro compressive force (CF) model. MATERIALS AND METHODS: Static compressive force (CF) of 47.4 g/cm2 was applied on mono- and co-cultured HPDFs and HABOs for 1, 2, or 4 h at 30 °C. TNFA, PTGS2, and IL6 gene expressions were determined by quantitative real-time polymerase chain reaction. TNF, PGE2, and IL6 concentrations were measured using enzyme-linked immunosorbent assay. RESULTS: In mono-culture, TNFA, PTGS2, and IL6 gene expressions were upregulated under CF as compared to controls for each time period in both cell types. PGE2 increased at 1 and 2 h in both cell types, and IL6 increased only at 2 and 4 h in HPDFs. Co-culture alleviated the force-induced increase of the expression of TNFA, PTGS2, IL6, PGE2, and IL6 in HPDFs at any time point. In HABOs, co-cultivation decreased the expression of PGE2 after 1 h and 4 h, and that of IL6 after 1 h compared to mono-culture. CONCLUSIONS: CF application on co-cultures of HPDFs and HABOs causes significant changes of TNFA, PTGS2, and IL6 gene expressions and PGE2 and IL6 production in comparison to mono-culture indicating intercellular communication. CLINICAL RELEVANCE: Mechanical stimulation of HPDFs and HABOs in co-culture induces a different gene expression pattern than stimulation of individual cell types alone. Co-culture might therefore be a relevant method to elucidate periodontal regeneration during orthodontic therapy.

A haplotypic variant at the IRGM locus and rs11747270 are related to the susceptibility for chronic periodontitis.

OBJECTIVE AND DESIGN: Immunity-regulated GTPase M (IRGM) plays a critical role in the defense against intracellular bacteria by regulating autophagy formation. This direct genetic association study aimed to determine whether variants at the IRGM genetic locus are associated with chronic periodontitis. MATERIALS AND SUBJECTS: Using PCR and melting curve analysis 390 periodontitis patients and 770 healthy controls have been genotyped regarding six polymorphisms in the IRGM gene (rs13361189, rs10065172, rs4958847, rs1000113, rs11747270, rs931058). RESULTS: Frequency distribution of alleles and genotypes for the six polymorphisms were not significantly different between the periodontitis and the control group. Also following stratification according to gender and smoking no significant linkage was found for any of the IRGM variants with periodontitis. Analysis of a subsample of patients revealed a significant association for rs11747270 with severe periodontitis (p = 0.003). Pairwise linkage analysis revealed one block composed of rs13361189, rs10065172, rs4958847, rs1000113 and 11747270 with strong or even complete linkage disequilibrium (r 2 > 0.9). Four haplotypes showed a frequency of > 1%, among which the haplotype C-T-A-T-G was significantly associated with chronic periodontitis (p = 0.0051; OR 4.66, 95% CI 1.41-15.42). CONCLUSIONS: One rare haplotype of the IRGM locus is significantly associated with chronic periodontitis in a German cohort.

Clinical association between chronic periodontitis and the leukocyte extravasation inhibitors developmental endothelial locus-1 and pentraxin-3.

This clinical study aimed to determine whether periodontal disease is associated with expression of developmental endothelial locus-1 (Del-1) and pentraxin-3 (PTX-3), endogenous inhibitors of leukocyte extravasation in humans. Expression of DEL1, PTX3, interleukin-17A (IL17A), and lymphocyte function-associated antigen-1 (LFA1) was determined, using RT-PCR and melting curve analysis, in biopsies of gingival tissues from 95 patients: 42 with moderate periodontitis; 40 with severe periodontitis; and 13 healthy controls. Relative expression of DEL1 and PTX3 was statistically significantly weaker in patients with periodontitis than in the control subjects. On the contrary, both IL17A and LFA1 showed statistically significant stronger expression in patients with periodontitis than in healthy controls. Correlation analysis, performed using Spearman's test, showed that expression of DEL1 was statistically significantly linked to periodontitis (ρ = -0.103) and to age (ρ = -0.134), but not to the gender of the patient, and that expression of PTX3 was significantly correlated with periodontitis (ρ = -0.354). Expression of neutrophil extravasation inhibitors DEL1 and PTX3 show significant, but weak, association with the clinical manifestation of chronic periodontitis.