Enamel Matrix Derivative Group Research Articles

Effects of Advanced Platelet Rich Fibrin (A-PRF+), Enamel Matrix Derivative (EMD) and Open Flap Debridement on clinical and wound healing parameters in molar furcation sites: A case series from a RCT study

AimTo study effects of advanced platelet rich Fibrin (A-PRF+) and enamel matrix derivative (EMD) compared to open flap debridement (OFD) alone in molar furcation sites grade II on clinical and wound healing parameters.Materials and methodsA randomized controlled trial was designed. Eligible patients were randomly allocated in one of three treatment groups: A-PRF+, EMD or OFD. The patients and clinical examiners were blinded for the treatment received. A minimally invasive microsurgical approach was performed for the three modalities. Clinical measurements were scored at baseline and 6 months postoperatively. The clinical healing of each furcation was scored via Early Wound Healing Index on day 3, 1 week, 2 weeks and 6 weeks.Results17 patients (A-PRF+ n = 6, EMD n = 5, OFD n = 6) completed the 6 months follow-up. The further completion of the trial had to be cancelled due to the COVID-19 pandemic. In three patients in the A-PRF+ group, the grade II of the treated furcation regressed to grade I; the corresponding number in the EMD and OFD groups were zero and one respectively. Further, 3, 1 and 4 patients in the PRF, EMD and OFD groups respectively, showed a gain of bone level ≥1 mm. The defects in the A-PRF+ group showed delayed early healing compared to EMD and OFD groups.ConclusionThe case series (RCT design) suggest a slight advantage for A-PRF+ over EMD and OFD, regarding the regressing of a furcation II to grade I, however this treatment showed delayed early wound healing compared to EMD or OFD.Clinical Trial Registrationhttps://www.isrctn.com/, identifier ISRCTN13520922.

Enhancing attachment of human gingival fibroblasts to periodontally compromised teeth: A comparative analysis of hyaluronic acid, EDTA, enamel matrix derivatives

BackgroundGingival recession is a characteristic indicator of periodontitis and one of the factors that contributes to increased tooth clinical crown length. Patients with root exposure frequently undergo periodontal surgical procedures in addition to adjunctive therapy to increase root coverage area and soft tissue stability. PurposeThis study aimed to evaluate fibroblast-root surface adhesion and determine whether periodontitis-damaged root surface microstructure can be restored using ethylenediaminetetraacetic acid (EDTA) and an enamel matrix derivative (EMD), individually or in combination. Material and methodsTeeth extracted from patients with periodontal disease were used to create 60 samples, with each group containing six specimens. The test groups were provided root planing or root condition-specific materials (hyaluronic acid [HA], 24% EDTA, EMD, or EDTA/EMD) for varying treatment time periods. In contrast, the control group did not undergo any surface modifications. The samples and fibroblast cells were incubated for 72 h. The number of living cells on the root surface in each group was calculated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (cell viability assessment). ResultsThe control, root planing, and EMD groups showed that the root surfaces treated with EDTA for 4 min had significantly better cell adhesion. Surface EDTA treatment for 2 min significantly promoted cell attachment compared to root planing treatment. The root surfaces modified with EDTA/EMD for 2 and 4 min showed significantly improved cellular migration and adhesion compared to the root surface treated with root planing. ConclusionEDTA and EDTA/EMD substantially affected the root surface, which was related to the length of the treatment process. This effect shifts the surface properties, alters fibroblast interactions with the root surface, and recruits more cells to cover a larger area.

Comparison of early wound healing using modified papilla preservation technique between enamel matrix derivative and recombinant human fibroblast growth factor.

Enamel matrix derivative (EMD) has demonstrated beneficial effects on wound healing following surgery. However, the effects of recombinant human fibroblast growth factor 2 (rhFGF-2) in periodontal regeneration therapy have not been extensively studied. This retrospective study was conducted to compare the wound healing outcomes of the modified papilla preservation technique (mPPT) between EMD and rhFGF-2 therapies. A total of 79 sites were evaluated for early wound healing using the modified early wound healing index (mEHI), which included 6 items: incision, fibrin clotting, step, redness, swelling, and dehiscence. A numeric analog scale, along with postoperative images of the 6 mEHI items, was established and used for the evaluations. The inter-rater reliability of the mEHI was assessed via intraclass correlation coefficients (ICCs). After adjusting for factors influencing the mPPT, the differences in mEHI scores between the EMD and rhFGF-2 groups were statistically analyzed. Additionally, radiographic bone fill (RBF) was evaluated 6 months after surgery. The ICC of the mEHI was 0.575. The mEHI, redness score, and dehiscence scores were significantly higher in the rhFGF-2 group (n=33) than in the EMD group (n=46). Similar results were observed in the subgroup of patients aged 50 years or older, but not in those younger than 50 years. In the subgroup with non-contained bone defects, related results were noted, but not in the subgroup with contained bone defects. However, early wound healing did not correlate with RBF at 6 months after surgery. Within the limitations of this study, the findings suggest that early wound healing following the use of mPPT with rhFGF-2 is somewhat superior to that observed after mPPT with EMD. However, mEHI should be improved for use as a predictive tool for early wound healing and to reflect clinical outcomes after surgery.

Clinical Outcome of Treating Peri-Mucositis and Peri-implantitis with Adjunctive Enamel Matrix Derivative Application: A Systematic Review

<p>Purpose: Enamel matrix derivative (EMD), the active component of Emdogain®, has been demonstrated to affect cytokines related to fibroplasia, angiogenesis, inflammation, and chemotaxis both in vitro and in vivo. However, whether the adjunctive application of EMD has clinical benefits on treating peri-mucositis and peri-implantitis has not been investigated thoroughly. The aim of this systematic review is to investigate the clinical outcomes of treating peri-implant diseases with adjunctive EMD application. Materials and Methods: A systematic search of multiple electronic databases including PubMed, Cochrane library, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Embase, and Scopus was carried out based on the PICOs structure for articles published up to September 2020. The primary outcomes were bleeding on probing (BoP), probing depth (PD), and radiographic bone level (RBL). Data and risk of bias were explored qualitatively. Results: The search identified 942 studies with 7 studies included for qualitative analysis. These studies reported 200 participants with 270 implants diagnosed with peri-mucositis (n=41) and peri-implantitis (n=229). For implants with peri-mucositis, BoP reduced significantly (83 to 25%) at the 3-month follow-up period for those treated with non-surgical mechanical debridement combined with adjunctive EMD (EMD group), while there was limited improvement for those treated by mechanical debridement alone (control group). PD also decreased (4.5 to 3.12 mm) in EMD group whereas an increase to 5.17 mm was demonstrated in the control group. For cases diagnosed with peri-implantitis, the high prevalence rate (83% - 100%) of BoP at baseline was significantly reduced in 2 prospective studies at the 2-10-years follow-up after surgical debridement, surface decontamination with bone graft, and EMD application. However, there was no statistical difference between surgical treatment with or without adjunctive EMD application in the other randomized controlled trials (RCTs) and half of the implants still demonstrated persistent BoP. Moreover, the PD can also be reduced significantly from 8.12 to 3.17 mm for those treated with adjunctive EMD while the RBL reduced from 4.41 to 2.31 mm at 2-10 year follow-up. The overall survival rate was 85-100% at 2-10 years follow-up. Conclusion: This review investigated the efficacy of adjunctive EMD for treating peri-implant diseases and was able to demonstrate its effect in reducing PD, reducing BoP, and increasing survival rate of treated implants from 2 to 10 years. The beneficial impacts of traditional surgical therapy on implants diagnosed with peri-implantitis were investigated; however, in terms of treating peri-implantitis, the definite efficacy of adjunctive usage of EMD should be determined based on more quantitative analysis and research in future.</p> <p> </p>

Effect of Root Surface Biomodification on Multiple Recession Coverage with Modified Coronally Advanced Tunnel Technique and Subepithelial Connective Tissue Graft: A Retrospective Analysis.

To improve treatment efficacy of gingival recessions (GR), chemical preparation of the exposed root surface was advocated. The aim of this study was to compare the additional influence of root biomodifications with 24% ethylenediaminetetraacetic acid (EDTA) alone or with enamel matrix derivative (EMD) on the 12 month outcomes of modified coronally advanced tunnel (MCAT) with subepithelial connective tissue graft in the treatment of multiple GR. Average root coverage (ARC), complete root coverage (CRC), reduction in GR, reduction in recession width (RW), gain in clinical attachment level (CAL), increase in gingival thickness (GT), increase in keratinized tissue width (KTW) and changes in root coverage esthetic score (RES) were evaluated. A total of 60 patients with 215 GR were enrolled. In 70, GR root surfaces were treated with EDTA + EMD, in other 72, with EDTA, while in the remaining 73 saline solution was applied. ARC was 94%, 89%, and 91% in the EDTA + EMD, the EDTA and the saline groups, respectively (p = 0.8871). Gains in clinical attachment level (CAL; 2.1 ± 1.1 mm) and RES values (9.6 ± 0.9) were significantly higher in the EDTA + EMD group, when compared with two other groups. The differences between other preoperative and postoperative parameters showed statistical significance only within but not between groups. MCAT outcomes may benefit from adjunctive use of EDTA + EMD regarding 12 month CAL gain and professionally assessed esthetics using RES following treatment of GR.

Effect of Enamel Matrix Derivative and N-Acetyl Cysteine on Proliferation and Osteogenic Activity of Dental Pulp Stem Cells.

Successful bone regeneration often requires induction by signaling molecules. Enamel matrix derivative (EMD) is said to enhance initial phases of healing. N-acetyl cysteine (NAC) is a molecule assumed to enhance osteogenesis and induce osteoblastic differentiation. This study sought to compare effects of EMD and NAC on proliferation, mineralization, and enzymatic activity of dental pulp mesenchymal stem cells (DPSCs). DPSCs were cultured on mineralized bone allograft (MBA) powder. After 24 hours, EMD in concentrations of 10, 50, and 100 μg/mL and NAC in 5 mM concentration were added. Methyl thiazolyl tetrazolium (MTT) assay was used for cell proliferation assessment at 1, 2, and 3 days. Osteoblastic differentiation of DPSCs was evaluated at 30 days, by alizarin red staining and assessment of alkaline phosphatase (ALP) activity. Both EMD and NAC caused time-dependent reduction of cell proliferation compared with the negative control. Maximum proliferation of DPSCs was observed in the 10 μg/mL EMD group at all time points, whereas NAC caused higher ALP activity and mineralization of DPSCs compared with EMD. In vitro application of NAC, as a signaling molecule, may effectively enhance bone regeneration by the induction of mineralization and enzymatic activity, despite the resultant reduction in cell proliferation rate.

Efficacy of a new-generation platelet-rich fibrin in the treatment of periodontal intrabony defects: a randomized clinical trial

BackgroundThe aim of the study was to clinically evaluate the healing of intrabony defects after treatment with a new generation of platelet-rich fibrin (A-PRF+) respect to enamel matrix derivative (EMD).MethodsThirty (30) intrabony defects of 18 patients (9 males, 9 females) were randomly treated with A-PRF+ (test, n = 15) or EMD (control, n = 15). The following clinical parameters were recorded at baseline and 6 months after surgery: pocket depth (PD), gingival recession (GR) and clinical attachment level (CAL). After debridement the intrabony defects were filled with A-PRF+ in the test group, respectively with EMD in the control group, and fixed with sutures to ensure wound closure and stability.ResultsBoth treatment methods resulted in statistically significant PD reductions, respectively CAL gains six months post-operatively. No statistically significant differences were found between the two groups as the mean CAL gain was 2.33 ± 1.58 mm in the A-PRF+ group, respectively 2.60 ± 1.18 mm in the EMD group (p < 0.001).ConclusionWithin the limits of this study the new-generation platelet-rich fibrin seems to be as clinically effective as EMD during surgical treatment of intrabony defects. Treatment with A-PRF+ or EMD resulted in reliable clinical outcomes. The use of A-PRF+ as a human autologous product can give a positive impact on periodontal healing.Clinical Relevance A-PRF+ may be suitable for the treatment of intrabony periodontal defects.Trial registration number (TRN) NCT04404374 (ClinicalTrials.gov ID).

Enamel matrix derivative (EMD) enhances the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs)

ABSTRACT To investigate the EMD’s capacity in BMSCs osteogenic differentiation. In vivo and in vitro, BMSCs were treated with EMD, scanning electron microscopy, and Alizarin Red staining were used to detect the changes in the osteogenic ability of BMSCs, and the proliferation ability of BMSCs was evaluated by CCK8. In addition, by adding xav939, a typical inhibitor of Wnt/β-catenin signaling pathway, the regulatory function of Wnt/β-catenin signaling was clarified. The results showed that EMD promote cell proliferation and 25 μg/ml EMD had the most significant effect. Cells inducing osteogenesis for 2 and 3 even 4 weeks, the cell staining is deeper in EMD treated group than that of the control (P < 0.05) by alizarin Red staining, suggesting more mineralization of BMSCs. In vivo implanting the titanium plate wrapped with 25 μg/ml EMD treated-BMSC film into nude mice for 8 weeks, more nodules were formed on the surface of the titanium plate than that the control (P < 0.05). HE showed that there is a little blue-violet immature bone-like tissue block. Besides, the expression of RUNX Family Transcription Factor 2 (Runx2), Osterix, Osteocalcin (OCN), collagen I (COLI), alkaline phosphatase (ALP) and β-catenin were inhibited in xav939 group (P < 0.05); Inversely, all were activated in EMD group (P < 0.05). In conclusion, EMD promoted the proliferation and osteogenic differentiation of BMSCs. EMD’s function on BMSCs might be associated with the Wnt/β-catenin signaling pathway.

Clinical, radiographic, and patient-centered outcomes after use of enamel matrix proteins for the treatment of intrabony defects in patients with aggressive periodontitis: A 12-month multicenter clinical trial.

The aim of this study was to evaluate the clinical, radiographic and patient-centered results of enamel matrix derivative (EMD) therapy in intrabony defects in aggressive periodontitis (AgP) patients and compare them with those in chronic periodontitis (CP) patients. Sixty intrabony defects in AgP and CP patients associated with ≥ 6 mm residual probing pocket depth (PPD) were included and randomly assigned to one of three groups: AgP+CS (conservative surgery) (n = 20); AgP+CS/EMD (n = 20); CP+CS/EMD (n = 20). Clinical parameters were measured at baseline and after 6 and 12 months. Defect resolution (DR) and bone filling (BF) were used for radiographic analysis. The quality of life was recorded at baseline and 6 months using OHIP-14 and VAS scale in the early post-therapy period. PPD and relative clinical attachment level (rCAL) improved for all groups during follow-up (P ≤ 0.05), and AgP+CS/EMD presented a higher rCAL gain (2.4 ± 1.0 mm) when compared to AgP control patients (1.6 ± 1.6 mm, P ≤ 0.05) after 12 months. No difference was observed between AgP+CS/EMD and CP+CS/EMD groups (P > 0.05). No radiographic differences were observed among groups at any time point (P > 0.05). All the groups reported a positive impact on OHIP-14 total score, without differences among them. EMD therapy of intrabony defects promotes additional benefits in AgP patients, presenting a similar regeneration rate compared to CP patients, and has proven to be a viable therapy for the treatment of individuals with AgP.

Healing of periodontal suprabony defects following treatment with open flap debridement with or without an enamel matrix derivative: A randomized controlled clinical study.

To compare the healing of suprabony defects following treatment with either open flap debridement (OFD) and application of an enamel matrix derivative (EMD) with OFD alone. Eighty patients with suprabony periodontal defects were randomly assigned to treatment with OFD + EMD (test) or OFD alone (control). The primary outcome variable was the difference in clinical attachment level (CAL) gain. At baseline and after 12 months, full-mouth plaque scores (FMPS), full-mouth bleeding scores (FMBS), probing depths (PD), gingival recessions (GR), and CAL were recorded. Sixty-five patients were available for the 12-month follow-up examination. At 12 months, the mean FMPS was 21.9 ± 3.0% in the OFD + EMD and 21.1 ± 2.4% in the OFD group, respectively (p = 0.30), while mean FMBS measured 20.4 ± 3.4% in the OFD + EMD group and 19.9 ± 2.9% in the OFD group (p = 0.48). Mean CAL gain at sites treated with OFD + EMD was statistically significantly different (p = 0.0001) compared with sites treated with OFD alone (3.4 ± 0.6 mm vs 1.8 ± 0.6 mm). A statistically significant difference (p = 0.0001) was found between mean PD change in the OFD + EMD (3.9 ± 0.6 mm) and OFD alone (3.2 ± 0.6 mm) treated groups and also in terms of mean GR change between treatment with OFD + EMD (0.5 ± 0.7 mm) and OFD alone (1.4 ± 1.0 mm) (p = 0.001). Within their limits, the present results indicate that in suprabony periodontal defects, the application of EMD in conjunction with OFD may additionally improve the clinical outcomes compared with OFD alone. In periodontal suprabony defects, the application of EMD in conjunction with OFD may additionally enhance the clinical outcomes in terms of CAL gain and PD reduction.

Local application of enamel matrix derivative prevents acute systemic inflammation after periodontal regenerative surgery: A randomized controlled clinical trial.

The aim of this study was to compare surgical treatment of periodontal intra-bony defects (IBD) with or without the adjunct of enamel matrix derivative (EMD) in terms of acute-phase responses in healthy patients. Thirty-eight periodontitis-affected subjects, one IDB each, were randomized to minimally invasive periodontal surgery (MIS) with or without EMD. Periodontal parameters were recorded at baseline and 6-months. Blood samples were collected at baseline, 1, 7 and 180days after treatment. At 24hr, the group treated MIS with EMD showed lower values of C-reactive protein (CRP; p<.01) as no inflammatory perturbation was noticed. Conversely, MIS group resulted in an acute inflammatory response at 24hr (p<.05) that regressed to its baseline values at day 7. The EMD group showed a higher number of cases without residual BOP or PPD≥5mm 6months after surgery (p<.05), and post-surgical gingival recession was lower (p<.05). The adjunctive application of EMD during surgical treatment resulted in a minor increase in serum CRP 24-hr after surgery. These findings suggest a possible systemic anti-inflammatory effect of EMD. Within its limitations, this pilot trial confirmed better clinical periodontal outcomes in the EMD group. NCT03590093.

Treatment of dehiscence-type defects with collagen matrix and/or enamel matrix derivative: Histomorphometric study in minipigs.

This study aimed to evaluate, histomorphometrically, the use of collagen matrix (CM) and/or enamel matrix derivative (EMD) for the treatment of dehiscence-type recession defects in minipigs. Eight healthy, male, young BR-1 minipigs, with no periodontal disease were treated. Bilateral dehiscence-type defects were surgically created on the buccal of the mandibular premolars (PI and PII). After 30 days, the defects were randomly assigned to four groups: coronally advanced flap (CAF); CAF + CM; CAF + EMD; and CAF + CM + EMD (split-mouth design). The evaluated parameters (mm): total defect length; new cementum (NC); new bone (NB); gingival margin position; total epithelium length; epithelium on the root; connective tissue adaptation; and soft tissue thickness (STT). The EMD-treated groups showed a superior length of NC [4.13±1.22 (CAF+ EMD); 3.95±1.11 (CAF + CM + EMD); 2.94±0.77 (CAF + CM); 2.72±0.81 (CAF), P=0.02] and NB [3.21±0.68 (CAF + CM + EMD); 3.01±0.56 (CAF + EMD); 2.15±0.47 (CAF + CM); 2.29±0.82 (CAF), P=0.005]. The CAF and CAF + CM groups showed a superior epithelial length when compared to EMD-treated groups after 3 months. A superior STT was observed for CAF + CM + EMD group (1.5±0.33) when compared with the other groups [1.09±0.26 (CAF + EMD); 1.04±0.34 (CAF + CM); and 1.14±0.29 (CAF), P=0.03]. The results of the present study indicate that EMD application, irrespective of the combination with CM, may improve the periodontal regeneration of dehiscence-type defects in this animal model.

Early wound healing outcomes after regenerative periodontal surgery with enamel matrix derivatives or guided tissue regeneration: a systematic review

BackgroundProper wound healing after regenerative surgical procedures is an essential issue for clinical success. Guided tissue regeneration (GTR) and application of enamel matrix derivatives (EMD) are common means to regenerate periodontal tissues. Both methods bear considerable advantages due to their special characteristics, but also go along with certain disadvantages. Today, there is no consensus in the literature whether GTR or EMD show better results regarding early wound healing, which is considered a crucial stage in periodontal regeneration. Therefore, the aim of the present systematic review was to compare the early wound healing after regenerative periodontal surgery with either EMD or GTR treatment.MethodsAn electronic literature search in PubMed was performed to identify randomized clinical trials (RCTs) or clinical trials (CTs) comparing regenerative surgery employing EMD and/or GTR in patients with chronic periodontitis. Among the finally included studies, a qualitative and quantitative data extraction regarding early wound healing parameters was performed. Primary outcome parameters were early wound healing index (EWH), flap dehiscence, membrane exposure, suppuration and abscess formation during the first 6 weeks. As secondary parameters, swelling and allergic reactions were assessed.ResultsSeven studies reporting 220 intrabony periodontal defects in 199 patients were analysed.Flap dehiscence was observed in two studies in 12% of the GTR treated sites and in 10.3% of those treated with EMD. Membrane exposure was evaluated in five studies and was registered in the 28.8% of the defects, while no dehiscence was reported on the EMD group. Swelling was reported only in one study in 8/16 GTR sites and 7/16 EMD sites. Due to considerable heterogeneity of parameters no meta-analysis was possible.ConclusionsDue to considerable heterogeneity of the published studies a clear beneficial effect of the EMD on the early wound healing outcomes after surgical treatment of periodontal intrabony defects cannot be confirmed.Standardized RCT studies are needed in order to allow for proper comparison of early wound healing after both types of surgical approaches.

Enamel matrix derivative stabilizes blood clot and improves clinical healing in deep pockets after flapless periodontal therapy: A Randomized Clinical Trial.

An acute phase response is induced after non-surgical periodontal treatment (SRP). The main aim of this study was to compare acute phase (24-hr) and medium-term (3-months) inflammation and clinical outcomes after SRP with or without application of enamel matrix derivative (EMD) in sites with probing pocket depth (PPD)≥6mm. Thirty-eight periodontitis-affected subjects were randomized to SRP or SRP+EMD. Periodontal parameters were recorded at baseline and 3months. Serum samples were collected at baseline, 1 and 90days after treatment. Both treatments triggered an intense acute inflammation on day 1, which regressed to baseline values at 3months. D-dimer and cystatin C levels did not show sharp increases in SRP+EMD group 24hr after treatment, compared to SRP. Significant difference between groups was observed for D-dimer (p<0.001). EMD application was also associated with better periodontal healing as shown by greater PPD reduction and clinical attachment level gain in sites with PPD≥6mm, and higher number of cases with no residual PPD≥6mm (p<0.05) at 3months. EMD application after non-SRP resulted in lower fibrinolysis, and better periodontal healing of deep pockets. These initial observations warrant further investigations on the potential to modulate both local and systemic outcomes of non-SRP. NCT03544931.

Surgical treatment of peri‐implantitis using enamel matrix derivative, an RCT: 3‐ and 5‐year follow‐up

To assess the clinical and radiographic outcomes 3 and 5years after the surgical treatment of peri-implantitis per se or in combination with an enamel matrix derivative (EMD). At baseline, 29 patients were randomized to surgical treatment with adjunctive EMD or no EMD. One year after the surgical treatment of peri-implantitis, 25 patients remained eligible for survival analyses at the 3- and 5-year follow-up. The primary outcomes were implant loss and bone level (BL) change measured on radiographs, and the secondary outcomes, bleeding on probing, pus and plaque at each implant were analysed in 18 and 14 patients at the 3- and 5-year follow-up, respectively. After exclusion of four patients who discontinued the study, at the 3-year follow-up, 13 (100%) implants survived in the EMD group, and 10 of 12 (83%) in the non-EMD group. At the 5-year follow-up, 11 of 13 (85%) implants in the EMD group and nine of 12 (75%) in the non-EMD group survived. In multivariate modelling, BL changes and EMD treatment were positively associated with implant survival. Similarly, the same trend was seen in univariate analysis. An exploratory analysis suggests that adjunctive EMD is positively associated with implant survival up to 5 years, but larger studies are needed.

Effect of EMD on the orthodontically induced root resorption repair process in rats.

While different levels of root resorption may occur in orthodontic treatment, several preventive approaches have been reported. Nevertheless, little is known about the effect of enamel matrix derivative (EMD) on root repair during orthodontic tooth movement. To evaluate the effect of EMD on root resorption repair following the application of orthodontic force. Aforce of 100 g was exerted for 14days on the left maxillary first molars of twenty 10-week-old Sprague-Dawley rats divided into the EMD and control groups (n= 10 per group). In the EMD group, repeatedly injection of Emdogain® was administered after the appliance was removed, while phosphate-buffered saline was administered in the control group. In vivo microcomputed tomography (CT), haematoxylin and eosin (H&E) staining, and immunohistochemistry were then used to evaluate the effect of EMD on the process of root repair. In the EMD group, the observed decrease in root resorption crater volume and increase in both the bone volume fraction and trabecular thickness were significantly greater than those in the control group. H&E staining showed that the periodontal fibres were relatively regular in arrangement and that the surface of the cementum was smooth in the EMD group. Immunohistochemical analysis showed higher bone morphogenetic protein 2 (BMP-2) and bone sialoprotein (BSP) expression levels in the EMD group than in the control group. In addition, the osteoprotegerin (OPG) and receptor activator of NF-κB ligand (RANKL) expression levels were similar in both groups. EMD enhanced the repair process following orthodontically induced root resorption in rats.

Clinical usability of aspartate aminotransferase to evaluate the prognosis of periodontal regeneration therapies: prospective, longitudinal study.

To evaluate the degree of periodontal tissue destruction, aspartate aminotransferase (AST) levels in the gingival crevicular fluid (GCF) are utilized as a predictor of periodontal therapy. We have previously shown that the usefulness of AST activities [periodontal tissue monitor (PTM) values] using a PTM-kit to evaluate the effects of initial periodontal therapy and periodontal regeneration therapy by enamel matrix derivative (EMD). This prospective, longitudinal study was conducted using 38 healthy and 80 periodontitis sites with probing depth (PD) of 5-10mm for guided tissue regeneration (GTR) and EMD from 36 patients. GCF samples were used to evaluate PTM values at base line (BL) and after 6months of surgeries (re-evaluation: RE), and periodontal examinations were performed concurrently. PTM values at BL were statistically improved at RE, accompanied by the improvement of periodontal parameters in both groups. PTM values and PD, and the clinical attachment level (CAL) showed high correlations. PD, CAL and bleeding on probing (BOP) were highly correlated with PTM values in both groups, whereas only PD showed a significant correlation with PTM values at RE in the GTR group. Change in the amounts of PD, CAL and BOP between BL and RE in both groups showed no correlation with PTM values. In the negative PTM value sites at BL in EMD group, the mean PD was significantly reduced at RE compared with positive PTM sites at BL. PTM values are able to be utilized as the biochemical predictor of prognosis after periodontal regeneration therapy.

Effect of the combined use of enamel matrix derivative and atelocollagen sponge scaffold on osteoblastic differentiation of mouse induced pluripotent stem cells in vitro.

Induced pluripotent stem cells (iPSCs) are a candidate cell source in periodontal regenerative therapy. Enamel matrix derivative (EMD) has been shown to regenerate periodontal tissues, and atelocollagen sponge (ACS) is considered a suitable scaffold or carrier for growth factors. This study aimed to investigate the effect of combined use of EMD and an ACS scaffold on cell behaviors and differentiation of mouse iPSCs (miPSCs) in vitro. Following embryonic body formation from miPSCs, dissociated cells (miPS-EB-derived cells) were seeded onto ACS with or without EMD, and cultured in osteoblast differentiation medium. Scanning electron microscopy and histological analyses were used to assess cell morphology and infiltration within the ACS. Cell viability (metabolism) was determined using an MTS assay, and expression of mRNA of osteoblastic differentiation markers was assessed by quantitative RT -PCR. Alkaline phosphatase (ALP) staining intensity and activity were evaluated. Mineralization was assessed by von Kossa staining, and calcium content was quantitated using the methylxylenol blue method. By 24hours after seeding, miPS-EB-derived cells in both the EMD and control groups had attached to and infiltrated the ACS scaffold. Scanning electron microscopy images revealed that by day 14, many cytoplasmic protrusions and extracellular deposits, suggestive of calcified matrix, were present in the EMD group. There was a time-dependent increase in cell viability up to day 3, but no difference between groups was observed at any time point. The levels expressed of ALP and osterix genes were significantly higher in the EMD group than in the control group. Expression of runt-related transcription factor 2 was increased in the EMD group compared with the control group on day 7. EMD upregulated the expression of bone sialoprotein and osteopontin on day 14, whereas expression of osteocalcin was lower at all time points. The staining intensity and activity of ALP were higher in the EMD group than in the control group. Mineralization levels and calcium contents were significantly higher in the EMD group throughout the observation period. These data suggest that combining ACS with EMD increases levels of osteoblastic differentiation and mineralization in miPS-EB-derived cells, compared with ACS used alone.

Biofunctionalization of porcine-derived collagen matrix using enamel matrix derivative and platelet-rich fibrin: influence on mature endothelial cell characteristics in vitro.

The present study evaluated the effect of an enamel matrix derivative (EMD) and platelet-rich fibrin (PRF)-modified porcine-derived collagen matrix (PDCM) on human umbilical vein endothelial cells (HUVEC) in vitro. PDCM (mucoderm®) was prepared to 6mm (±0.1mm) diameter discs. PDCM samples were incubated with either EMD, PRF, or control solutions for 100min at 4°C before the experiments. Cell-inducing properties of test materials on HUVEC cells were tested with cell proliferation assays (MTT, PrestoBlue®), a cytotoxicity assay (ToxiLight®), a Boyden chamber migration assay, and a cell attachment assay. Scanning electron microscopy (SEM) imaging was performed to determine the surface and the architecture of the modified matrices. Cell proliferation was elevated in the EMD and PRF groups compared with control (p each ≤0.046). PRF modification increased HUVEC migration ability by 8-fold compared with both control and EMD groups (p each <0.001). Both treatments significantly promoted the cell attachment of HUVEC to PDCM, as assessed by direct cell counts on the matrices (p each <0.001). HUVEC cell characteristics were overall improved by EMD- and PRF- modified PDCM. Adsorbed bioactive molecules to the PDCM surface may have contributed to a more preferable environment to surrounding cells. The results may give evidence that PDCM modification with EMD or PRF, respectively, might be a useful approach to improve clinical outcomes, to prevent inflammatory reactions and wound-healing disturbances, and to expand the clinical application area of PDCM.

Treatment of Non-Contained Infrabony Defects With Enamel Matrix Derivative Alone or in Combination With Biphasic Calcium Phosphate Bone Graft: A 12-Month Randomized Controlled Clinical Trial.

Use of enamel matrix derivative (EMD) when dealing with non-contained defects may be limited because EMD does not maintain a space itself. Use of combined therapy has been proposed, using a bone graft in combination with EMD to avoid collapse of the flap into the bony defect during healing time. Therefore, the aim of this study is to evaluate the clinical and radiologic healing response of non-contained infrabony defects after treatment with a combination of EMD and biphasic calcium phosphate (BC) or EMD alone. Fifty-two patients with at least one infrabony defect ≥3 mm in depth with a probing depth (PD) ≥6 mm were randomly treated with EMD/BC or EMD alone. Clinical and radiographic parameters were evaluated at baseline, 6, and 12 months after surgery. To standardize the procedure, an acrylic stent and millimeter radiographic grid were used. The primary outcome was the change in clinical attachment level (CAL). Analysis of the data demonstrated a statistically significant difference from baseline within each group (P <0.05), with a difference in clinical and radiographic parameters at 6 and 12 months. After 1 year, mean PD reductions of 3.14 ± 1.95 mm (39.6%) in the EMD/BC group and 3.30 ± 1.89 mm (48.7%) in the EMD group were achieved. A mean CAL gain of 2.38 ± 2.17 mm (24.9%) in the EMD/BC group and 2.65 ± 2.18 mm (36.2%) in the EMD group were obtained. Reduction in the infrabony component was 2.71 ± 1.79 mm (57.9%) in the test group and 2.60 ± 2.03 mm (28.5%) in the control group. There were no statistically significant differences between treatment groups. It was concluded that treatment of non-contained infrabony defects with EMD, with or without BC, resulted in statistically significantly better results after 12 months compared with baseline measurements. In contrast, the combined approach did not result in a statistically significant improvement.