Immunohistochemical expression of non-collagenous extracellular matrix molecules involved in tertiary dentinogenesis following direct pulp capping: a systematic review.

BACKGROUND: Extracellular matrix molecules (ECMM) expression during tertiary dentinogenesis provides useful information for regenerative applications and efficacy of pulp capping materials. AIM: To identify and review the expression and roles of non-collagenous ECMM after successful direct pulp capping (DPC), following mechanical pulp exposures, via immunohistochemistry (IHC). The study addressed the question of where will successful DPC impact the IHC expression of these molecules. DATA SOURCES: In vivo animal and human original clinical studies reporting on ECMM in relation to different follow-up periods were screened and evaluated via descriptive analysis. The electronic literature search was carried out in three databases (MEDLINE/PubMed, Web of Science, Scopus), followed by manual screening of relevant journals and cross-referencing, up to December 2018. STUDY ELIGIBILITY CRITERIA, PARTICIPANTS, AND INTERVENTIONS: Randomized and non-randomized controlled trials, conducted in humans and animals, were selected. Histological evidence for tertiary dentine formation was a prerequisite for IHC evaluation. STUDY APPRAISAL AND SYNTHESIS METHODS: The methodological quality of the included articles was independently assessed using the Systematic Review Centre for Laboratory animal Experimentation (SYRCLE) and the Cochrane risk of bias tool (RoB 1), respectively. RESULTS: From a total of 1534 identified studies, 18 were included. Thirteen papers evaluated animal subjects and five studies were carried out on humans. In animals and humans, fibronectin and tenascin expressions were detected in pulp and odontoblast-like cells (OLC); dentine sialoprotein was expressed in both soft and newly-formed mineralized tissue. In animals, bone sialoprotein was early expressed, in association with OLC and predentin; the immunoreactivity for dentine sialophosphoprotein and dentine matrix protein-1 was associated with the OLC and dentine bridge; osteopontin was expressed in OLC, predentine and reparative dentine. A considerable heterogeneity was found in the methodologies of the included studies, as well as interspecies variability of results in terms of time. CONCLUSIONS AND IMPLICATIONS OF KEY FINDINGS: Within the limited scientific evidence, all non-collagenous ECMM expressions during tertiary dentinogenesis are active and related to soft and hard tissues. There is a shortage of human studies, and future research directions should focus more on them. PROSPERO Protocol: CRD42019121304.

Evaluation of a hydrophobic gel adhering to the gingiva in comparison with a standard water-soluble 1% chlorhexidine gel after scaling and root planing in patients with moderate chronic periodontitis. A randomized clinical trial.

OBJECTIVES: To evaluate the clinical, microbiological and enzymatic activity of a hydrophobic chlorhexidine-based gingiva-adhering gel containing herbal ingredients, compared with a commercially available 1% chlorhexidine water-soluble gel, during non-surgical therapy of moderate chronic periodontitis. METHODS: A total of 34 subjects participated in this 6-month blinded randomized parallel controlled trial (ISRCTN35210084). After scaling and root planing (SRP), test group received the gel, by rubbing on the gingiva, once every second day, for 14 days. The control group received the control gel twice daily. Clinical parameters considered were the approximal plaque index, simplified oral hygiene index, modified gingival index, bleeding on probing, probing depth and clinical attachment level (primary outcome), assessed at baseline, 3 and 6 months, together with the frequency of detection of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis (P.g.), Prevotella intermedia, Treponema denticola (T.d.), Tannerella forsythia (T.f.), and activity of neutrophil elastase and myeloperoxidase (secondary outcomes). RESULTS: At 3 and 6 months, all clinical parameters improved significantly, without significant intergroup differences, except OHI-S, which improved at 3 months (P < 0.05). Microbiological data resulted in no significant intergroup differences at baseline and 6 months. At 3 months, significant differences for P.g., T.f. and T.d. were noted. A significant reduction of neutrophil elastase after 3 and 6 months was observed (P < 0.005), without significant intergroup differences. For myeloperoxidase, significant reductions were noted in both groups (P < 0.005 and P < 0.05), but no significant intergroup differences. The tested product seemed to have an increased efficacy, due to longer persistence on the gingiva, with reduced application frequency. CONCLUSIONS: Both products had a relatively similar influence on the clinical, microbiological and enzymatic outcomes at 3 and 6 months after SRP.

The impact of some behavioral aspects on periodontal disease in a group of Romanian students - an epidemiological survey.

AIMS: To investigate the prevalence of periodontitis in a young population representative for the North-western part of Romania (Transylvania) and to identify possible risk indicators of periodontitis. METHODS: The study is a cross-sectional epidemiological survey. The subjects were students randomly sampled from three universities in Cluj-Napoca and high school students from the neighboring city of Bistrita. The sample size of the population was calculated. Overall, 623 subjects aged 16-35 years were evaluated, of which 488 were university students and 135 high school students. A structured questionnaire was administrated to collect information on socio-behavioral status and oral hygiene habits. Periodontal data was collected using a full-mouth methodology by trained examiners. A recent introduced case definition was used to pick up periodontitis cases. RESULTS: The older the subject, the more frequent toothbrushing, dental visits, and use of interproximal hygiene devices, but also addiction to tobacco and alcohol consumption. The prevalence of periodontitis was 0.96% (n=6). Half of these subjects (n=3, 0.48%) were considered to have aggressive periodontitis (AP). Low frequency of toothbrush changing was identified to influence the development of periodontitis. Smoking and lower socioeconomic level did not seem to correlate with periodontal disease in the present study. CONCLUSIONS: In order to better understand the prevalence of periodontal diseases and identify periodontitis cases as well as to evaluate the impact of specific behavioral factors on the disease development in individual and population levels, further extensive screenings are obviously required. Periodontal prevention programs focusing on oral health behavior are mandatory.

Bone tissue response to an oily calcium hydroxide suspension in tibial defects. An experimental pilot study in minipigs.

OBJECTIVES: The present study was conducted to evaluate the bone tissue response after the application of an oily calcium hydroxide suspension (OCHS) into defects created in the tibial bone of minipigs. MATERIALS AND METHODS: Standardized defects (2 ccm) were created into the tibia of 4 Goettinger minipigs. Defects in the test group (n = 4) were filled with OCHS (Osteora, DFS-Diamon, Riedenburg, Germany). Defects in the control group (n = 4) were filled with venous blood. Animals were sacrificed after healing periods of 4 and 8 weeks. Tibias were dissected, soft tissues removed and processed for histological analysis. Digital images (×200) were evaluated using the software CellD (Soft Imaging System, Münster, Germany). The following histomorphometrical landmarks were identified: defect size, mineralized tissue, non-mineralized tissue and residual OCHS. RESULTS: Healing was uneventful in all four animals. In the test group, new bone formation was observed in the vicinity of the defect margins whereas the centre of the defect was dominated by non-mineralized tissue. Mean percentages of mineralized tissue after 4 weeks were 23.01% in the test group vs. 43.45% in the control group. The mean value for residual OCHS was 7,11% at 4 weeks. After 8 weeks mean percentages of mineralized tissue were 28.15% in the test group vs. 44.39% in the control group as well as 7.05% for residual OCHS. CONCLUSION: Within the limits of the present pilot study it can be concluded that OCHS did not have a beneficial effect on new bone formation. To prove an osteoinductive potential of OCHS further studies based on a higher number of samples are needed.

Antibacterial action of Chlorhexidine/thymol containing varnishes in vitro and in vivo.

OBJECTIVES: The antibacterial activity of two different formulations of a chlorhexidine/thymol varnish should be elucidated in vitro and in vivo. METHODS: The agar diffusion assay with Cervitec(®) and CervitecPlus(®) and three reference strains each of streptococci, lactobacilli, actinomyces and periodontal pathogens was performed. In a split-mouth study, 40 volunteers applied the test (CervitecPlus(®), solvent water and ethanol) and control (Cervitec(®), solvent ethyl acetate) varnish at buccal recessions of premolar teeth at baseline as well as after two, four and seven days. Supra- and subgingival plaques were collected 2 weeks before baseline and at the screening appointments. Supragingival plaque was analysed for mutans streptococci and lactobacilli and subgingival samples for Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Porphyromonas intermedia. Friedman/Wilcoxon tests and U-test were used for statistical analysis (P < 0.05). RESULTS: Most reference strains were susceptible with inhibition zones (mm) as follows: Cervitec(®)/CervitecPlus(®) streptococci 27 ± 1.7/21.3 ± 2.5, lactobacilli 26 ± 9.2/23.7 ± 4.9, actinomyces 36.3 ± 6.6/27.3 ± 1.5, periodontal pathogens 18.7 ± 7.6/18 ± 1.7. Both varnishes reduced significantly the counts of mutans streptococci and lactobacilli in the patients. However, no significant differences were found between test and control sides at any time. The total counts of periodontal pathogens were low. A tendency to higher counts of A. actinomycetemcomitans at the control side could be shown; the test side did not harbour significantly higher counts. CONCLUSION: Both varnishes may influence the plaque formation and reduce mutans streptococci in supragingival plaque.

In vitro and in vivo characterization of porcine acellular dermal matrix for gingival augmentation procedures.

BACKGROUND AND OBJECTIVE: Recently, porcine acellular dermal matrix (PADM) has been proposed as a possible alternative to autogenous grafts in periodontal plastic surgery. The aim of the present study was to investigate the in vitro responses of four different oral cell lines cultured on a novel PADM. Furthermore, tissue reaction to PADM was evaluated histologically after subcutaneous implantation in mice. MATERIAL AND METHODS: Human gingival fibroblasts (HGF), human osteoblast-like cells, human umbilical vein endothelial cells and human oral keratinocytes (HOK) were cultured and transferred on to the PADM. A tissue culture polystyrene surface served as the control. The viability of all tested cell lines on PADM was measured by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide colorimetric assay and PrestoBlue(®) reagent. The ToxiLight(®) assay was performed to analyze the effect of PADM on adenylate kinase release. PADM was implanted into nude mice subcutaneously and subjected to histological analysis after 21 d. RESULTS: Using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide colorimetric assays, all tested cell lines cultured on PADM demonstrated a significant increase of viability compared to the control group (each p < 0.001) with the exception of HGF and HOK after 3 d (each p > 0.05). According to the PrestoBlue(®) analysis, all cell lines demonstrated a significant increase of viability compared to the control group at the particular points of measurement after 18 h (HGF p < 0.01; human osteoblast-like cells, human umbilical vein endothelial cells, HOK each p < 0.001). No significant cytotoxic effects of PADM on the tested cell lines could be observed, as assessed by changes in adenylate kinase release. Subcutaneous implantation of PADM into nude mice demonstrated good integration with surrounding tissues and significant revascularization of its collagen structure. CONCLUSION: Overall, the results suggest that PADM is a promising substitute for autogenous soft tissue grafts in periodontal surgery.

Subepithelial connective tissue graft with or without enamel matrix derivative for the treatment of Miller class I and II gingival recessions: a controlled randomized clinical trial.

BACKGROUND: The aim of this study was to evaluate whether the combination of enamel matrix derivative (EMD) with subepithelial connective tissue graft (SCTG) plus coronally advanced flap (CAF) would improve the treatment outcomes of Miller class I and II gingival recessions when compared with the same technique (SCTG plus CAF) alone. METHODS: The study was designed as a randomized, parallel, controlled, double-blinded clinical trial. Forty-two patients were randomly assigned in the test group (SCTG plus EMD) and in the control group (SCTG). Patients had at least one gingival recession ≥ 2 mm. The clinical parameters were evaluated at baseline and at 14 d, 1, 3, 6 and 12 mo follow-up time points. RESULTS: Forty-two patients, 21 in the test group (SCTG plus EMD) and 21 in the control group (SCTG), aged 21-48 years (mean age 31 ± 8.56) were initially included in the study. Both treatments, STCG plus EMD and SCTG, resulted in a significant final mean root coverage (2.91 ± 0.95mm and 2.91 ± 1.29 mm, respectively) (p < 0.001) and in a high mean percentage of root coverage (82.25 ± 22.20% and 89.75 ± 17.33%, respectively) (p < 0.001), 1 year after surgery. The differences in mean root coverage recorded for the two techniques after 1 year, were not statistically significant (p = 0.19). Complete root coverage was achieved in 56.5% of patients treated with SCTG plus EMD and in 70.6% of patients treated with SCTG (p = 0.275), 1 year after treatment. CONCLUSIONS: The present study failed to demonstrate any additional clinical benefits when EMD was added to SCTG plus CAF.

The influence of enamel matrix derivative on the angiogenic activity of primary endothelial cells.

BACKGROUND AND OBJECTIVE: Angiogenesis plays a crucial role in early wound healing and tissue regeneration. Although enamel matrix derivative (EMD) has demonstrated the potential to stimulate periodontal regeneration, the biological effects of EMD on angiogenesis and underlying mechanisms have not been fully elucidated. The aim of the present study was to examine the angiogenic effects of EMD in vitro. MATERIAL AND METHODS: Human umbilical vein endothelial cells (HUVECs) were used to assess the effect of EMD on proliferation, survival, adhesion and migration. The effect of EMD on HUVEC angiogenesis was assessed by a three-dimensional sprouting assay. In order to understand the signalling mechanism of altered cell proliferation of HUVECs caused by EMD, the phosphorylation status of ERK1/2 and of the serine/threonine protein kinase Akt was analysed by western blot using phospho-specific antibodies. RESULTS: The proliferation of HUVECs was stimulated by 50 µg/mL EMD, whereas higher concentrations (≥100 µg/mL) resulted in an increased apoptotic rate. The mitogenic response to EMD was associated with the activation of ERK1/2. Enamel matrix derivative did not affect cell adhesion, but all concentrations of EMD tested (0.1-250 µg/mL) promoted migration of HUVECs. Furthermore, EMD induced capillary-like sprout formation from HUVEC spheroids in a dose-dependent manner. CONCLUSION: Our data indicate that EMD acts as a proangiogenic factor in vitro and, as such, might contribute to periodontal tissue regeneration by stimulation of vessel formation during wound healing.

Investigation of six selected bacterial species in endo-periodontal lesions.

AIM: To investigate and determine possible associations of six tested bacteria belonging to 'orange' and 'green' complexes, in endo-periodontal lesions: Parvimonas micra, Fusobacterium nucleatum, Campylobacter rectus, Eubacterium nodatum, Eikenella corrodens and Capnocytophaga sputigena. METHODOLOGY: Forty-six patients presenting with different types of endo-periodontal lesions were investigated. Clinical examinations, periapical radiographs and microbiological sampling from the canal system (endo) and periodontal pockets (perio) were performed. Qualitative and semiquantitative evaluation of bacteria was performed by polymerase chain reaction (PCR) and DNA-DNA hybridization (micro-IDent plus; Hain Lifescience, Germany). RESULTS: Extremely high bacterial loads in endodontic samples were recorded for P. micra, F. nucleatum and C. sputigena, while periodontal samples were often colonized by the same species, plus C. rectus. Significant association was recorded between F. nucleatum-endo and P. micra-endo (P = 0.03, Fisher's exact test). There was marginal evidence of associations between: (i) C. sputigena-endo and C. sputigena-perio (P = 0.06, Fisher's exact test); (ii) P. micra-endo and P. micra-perio (P = 0.05, Fisher's exact test). Sensitivity to percussion was associated with an increased chance of cases with P. micra-endo (P = 0.03, Pearson chi-square test). CONCLUSION: The findings suggest that F. nucleatum, P. micra and C. sputigena may play a role in the pathogenesis of endo-periodontal lesions.

How accurate replicates the Thermafil System the morphology of the apical endodontic space? An ex vivo study.

AIM: To evaluate the morphology of the root canal in its apical third and the capacity of the Thermafil System to reproduce the entire morphology of the cleaned and shaped root canal. MATERIALS AND METHODS: Thirty-two roots of periodontally compromised teeth were prepared using the ProTaper System to an apical size 30 and filled with the Thermafil obturation technique and sealer. The roots were surgically amputated and prepared for metallographic evaluation by incremental reductions of 0.5 mm each, starting with the apical foramen. Photomicrographs of each section were taken at a magnification of 500x and 100x. The images were analyzed and processed. The position of the apical foramen with respect to the anatomical apex was identified and marked. Additional morphological details as lateral canals and recesses were also recorded. The cross-sectioned area of the canal and gutta-percha, the total perimeter, the shaped perimeter and the filled perimeter were recorded for each sample and the results were expressed as percentages. Multiple images of successive sections were used to create a 3D reconstruction of the apical anatomy of the tooth. The ANOVA test was performed to assess mean differences between evaluations of perimeters/areas at different levels. RESULTS: The anatomical apical foramen was found at the tip of the root in 50% of the evaluated samples. In the remaining samples, the foramen was located between 0.5 and 2.5 mm from the centre of the apex. Lateral canals, which opened in accessory foramens, were recorded in 25% of the evaluated samples. Statistical significant differences (p<0.05) were found between different levels of preparation and obturation. CONCLUSIONS: The complex morphology of the apical third of the root canal is satisfactory microstructurally replicated by the Thermafil System. Moreover, polarized light microscopy and the 3D reconstruction offered a discriminative vision of morphological details as lateral canals, recesses, the gutta-percha and debris.

Computer-assisted photomicrographic evaluation of root canal morphology after removal of the filling material during retreatment.

AIM: The purpose of this study was to evaluate the morphology of the apical third of the root canal and the effectiveness of rotary instrumentation for the removal of lateral condensed gutta-percha during endodontic retreatment. MATERIALS AND METHODS: Thirty roots with canals with regular morphology were prepared to apical size #30 and were filled with gutta-percha/AHPlus using lateral condensation. Digital radiographs of teeth were captured. After two weeks, canals were retreated to size 40 using ProTaper rotary instruments, without solvent. Roots were embedded in resin blocks and reduced incrementally at four different apical levels (at 0.5, 1.0, 1.5 and 2.0 mm from the apical foramen). The sectioned surfaces were observed under a metallographic optical microscope and digital micrographic images were captured and processed. Perimeter covered with root-filling residue was expressed as percentage of total canal perimeter for all specimens. RESULTS: Residue percentage was greater at 2.00 mm from apical level than at other levels. Most residue of filling material in all specimens was observed at 0.5 and at 2.0 mm from apex. The use of ProTaper instruments allowed the removal of gutta-percha and AHPlus sealer in the apical 2 mm in average 50% of the cases. CONCLUSIONS: The resulting morphology of the prepared canals allowed root fillings performed with gutta-percha and AHPlus sealer that were efficiently removed by using rotary instruments. More material residue was found adhering to the canal walls in the apical segments of canals. The removal of this residue was enhanced by apical enlargement beyond the diameter of the canal before retreatment.

[An interdisciplinary approach to complex orthodontic-periodontal clinical situations in adult patients]. / O abordare interdisciplinara a unor situatii clinice complexe ortodontico-parodontale la pacientii adulti.

Between orthodontics and periodontology there are many correlations, especially in the treatment of adults with already-damaged periodontal tissues and malocclusions. Aim of this paper was to emphasize that an interdisciplinary periodontal-orthodontic treatment could be beneficial even in a case that seemed hopelessly. A typical case of an adult patient with extensive bone loss due to periodontal disease, occlusal trauma caused by dental malpositions, spacing due to early loss of teeth in the lateral segments and pathologic tooth migration will be presented. The orthodontic treatment was initiated at 3 months after the stabilization of the periodontal therapy and was done with a fixed appliance. During the orthodontic stage periodontal maintenance visits continued at 2-month intervals. The therapy of"black triangles" occurred due to the loss of interdental gingiva was done by stripping followed by a space-closure procedure. A permanent retainer was applied. Benefits and problems of the comprehensive periodontal-orthodontic therapy and treatment protocols will be presented. In conclusion the interdisciplinary therapy of this patient, with a good compliance, was beneficial, improved esthetics, the periodontal status, the ability of the patient to clean the teeth and also led to best position of the abutments for optimal placement of prosthetic reconstructions.

Prospective clinical study evaluating the long-time adjunctive use of chlorhexidine after one-stage full-mouth SRP.

OBJECTIVES: Scaling and root planing are the causal procedure in the treatment of periodontitis. Many attempts have been made to improve the outcome. The aim of this study was to verify the influence of the extended use of chlorhexidine after one-stage full-mouth (FM) SRP in patients with chronic periodontitis on the clinical outcome after 3 months. METHODS: Eighty-one patients with pockets > or =5 mm were treated by FM. All patients rinsed additionally with 0.2% chlorhexidine (CHX) twice daily over 3 months. Plaque index, bleeding on probing, probing depth (PD) and clinical attachment level (CAL) were recorded at baseline and after 1 and 3 months. RESULTS: In the test group, all variables were significantly improved after 1 and 3 months. Mean reduction of PD and CAL gain was 2.25 +/- 1.08 and 1.67 +/- 1.08 after 1 and 2.99 +/- 1.11 and 2.33 +/- 1.31 after 3 months respectively. CONCLUSIONS: Over 3 months of extended use of CHX mouth rinse after SRP showed slightly but statistically significant better results.