Melatonin-Medicated Neural JNK3 Up-Regulation Promotes Ameloblastic Mineralization

Introduction: Melatonin, an endogenous neurohormone, modulates the biological circadian rhythms of vertebrates. It functions have been reported in previous stomatological studies as anti-inflammation, antioxidant, osseointegration of dental implants and stimulation to dental pulp stem cells differentiation, but its role in ameloblastic differentiation and mineralization has been rarely studied.Objective: To reveal the effects of melatonin on the mineralization of ameloblast lineage cells (ALCs), and to identify the change in gene expression and the potential mechanism based on ribonucleic acid sequencing (RNA-seq) analysis.Method: ALCs were induced in melatonin-conditioned medium. After 7-days culture, Western blot, real-time PCR, alkaline phosphatase (ALP) activity test, RNA-seq were accordingly used to detect the change in molecular level. After 1-month odontogenic induction in melatonin medium, Alizarin Red-S (ARS) staining showed the changes of mineral nodules. Differentially expressed genes (DEGs), enrichment of functions and signaling pathways analysis based on Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) database were performed. The JNK3 antagonist (JNK3 inhibitor IX, SR3576) and β-arrestin1 (Arrb1) overexpression were applied to confirm the fluctuation of melatonin-medicated JNK3 and Arrb1 expression.Results: In this study, we found out melatonin contributed to the ameloblastic mineralization, from which we can observed the elevated expression of enamel matrix protein, and increased ALP activity and mineralized nodules formation. RNA-seq analysis showed the up-regulation of neural JNK3 and down-regulation of Arrb1 in ALCs. Meanwhile, phosphorylated JNK3 deficiency (phosphorylated JNK3 inhibitor---SR3576 added to culture medium) led to mineralization delay, and Arrb1 overexpression proved Arrb1 takes bridge between melatonin receptors (MTNR) and JNK3 in MAPK signaling pathway.

EXPERIENCE OF USING ENAMEL MATRIX PROTEIN IN COMBINATION WITH A CORONARY ADVANCED FLAP AND A CONNECTIVE TISSUE GRAFT TO ELIMINATE GUM RECESSION

The article presents the experience of complex managing a patient with gum recession in the area of the maxillary premolars. Periodontal treatment of gum recession consisted of initial therapy, including supra- and subgingival scaling, root planing, pharmacotherapeutic support (broad-spectrum antiseptics, drugs that stimulate regeneration), surgical combined method based on the use of enamel matrix protein to stimulate regenerative processes. The clinical material Emdogaine is a combination of an enamel matrix derivative and propylene glycol alginate, which performs the function of a carrier. Biora company has been producing it for more than 20 years (Emdogain, BIORA): now the rights to this material belong to Straumann). This preparation was developed to stimulate regeneration of periodontal tissues. Amelogenin is a specific enamel protein capable of stimulating regeneration of lost periodontal tissues by influencing cell differentiation. Dental cement is an osteoid connective tissue that covers the roots of teeth and serves to attach periodontal fibers. Application of an enamel matrix derivative to the sanitized root surface leads to regeneration of non-cellular cement and to the formation of periodontal tissues, which was observed in our study. Transplantation of free connective tissue grafts to eliminate gum recessions makes it possible to effectively eliminate gum recession in most clinical cases (complete root coverage, an increase in the level of clinical attachment from the keratinized attached gum area). In our clinical case, free grafts were obtained in the donor area of the maxillary tuberosity. Transplantation of free connective tissue grafts provides an aesthetic result and is currently considered a standard technique for eliminating gum recession. According to the clinical study, a combined use of connective tissue grafts and the enamel matrix derivative, an additional use of enamel matrix derivative enabled to obtain a satisfactory result and to improve the appearance of the dentition. Such combined surgeries are indicated in an aesthetically significant area. The described materials, methods and stages of patient's follow up can serve as the basis for the protocol of managing patients with gum recession.

Hopeless tooth? Prognosis and comprehensive treatment. A case report

A hopeless tooth from a periodontal point of view, with severe bone resorption, mobility and abnormal tooth migration, is often extracted. In advanced cases, function and esthetics are impaired, and an interdisciplinary treatment is requested. Retaining or not these teeth is based on clinician judgment. A growing body of evidence claims that prognosis has great potential to be improved in a motivated patient with good oral hygiene and regular maintenance. This case report aims to present a periodontal regenerative technique combining enamel matrix protein derivatives and a particulated xenograft to treat intraosseous defects caused by periodontitis. The patient healed uneventfully, and no complications were recorded after the surgical procedure. To correct abnormal tooth migration and improve function and esthetics, orthodontic treatment was instituted. Tooth prognosis improved from hopeless to questionable. This approach extended the life span of a compromised tooth, improving periodontal support and decreasing tooth mobility. This could be an alternative to extraction and implant.

Regenerative Potential of Enamel Matrix Protein Derivative and Acellular Dermal Matrix for Gingival Recession: A Systematic Review and Meta-Analysis

Objective: The purpose of this study was to assess the clinical effectiveness of using a combination of enamel matrix protein derivative and acellular dermal matrix in comparison to acellular dermal matrix alone for treating gingival recessions. Methods: The Cochrane Library (Wiley), PubMed by Medline (NLM), Medline (EBSCO), and Embase (Ovid) databases were searched for entries up to April 2020. Only clinical trials were included. Primary outcomes were root coverage (%), changes in keratinized tissue width and recession (mm). Meta-analysis was conducted for root coverage, changes in keratinized tissue width, recession, clinical attachment level and probing depth. Results: Four studies were selected for the analysis. In primary outcomes, root coverage, change in keratinized tissue width and recession analysis showed a mean difference of 4.99% (p = 0.11), 0.20 mm (p = 0.14) and 0.13 mm (p = 0.23) respectively between the two groups. Secondary outcomes analysis also exhibited a statistically insignificant difference between the test and control group with mean difference of 0.11 mm (p = 0.32) in clinical attachment level gain and -0.03 mm (p = 0.29) in probing depth reduction analysis. Conclusions: Within the limits of this study, enamel matrix protein derivative combined with acellular dermal matrix used for treating gingival recession defects resulted in no beneficial effect clinically than acellular dermal matrix only.

Comparative evaluation of fluoride varnishes, self-assembling peptide-based remineralization agent, and enamel matrix protein derivative on artificial enamel remineralization in vitro

Background One of the most unfavorable side effects of fixed orthodontic treatment is white spot lesions (WSLs). Although the most important approach is prevention of WSLs, it is also essential to evaluate the efficacy of the remineralization agents. However, there is no concurrence in the literature with respect to the remineralization process of these agents. The objective of the present study was to evaluate the effects of different fluoride varnishes, enamel matrix protein, and self-assembling peptide derivatives with varying chemical compositions on remineralization of artificially created WSLs in vitro using quantitative light-induced fluorescence (QLF). Methods Artificial WSLs were created on bovine enamel samples using acidic buffer solution (pH 5, 10 days). Specimens were randomly allocated to six groups (n = 10/group): (1) Emdogain (Straumann, Basel, Switzerland), (2) Curodont Repair (Credentis AG, Switzerland), (3) Duraphat (Colgate-Palmolive, New York, NY), (4) Clinpro XT (3 M ESPE, Pymble, New South Wales, Australia), (5) Enamel Pro Varnish (Premier Dental Products, PA, USA), and (6) control (untreated). The agents were applied to the WSLs according to the manufacturers’ instructions. Fluorescence loss (ΔF), lesion area (area), and impact (ΔQ) values of enamel surfaces were quantified by QLF-D BiluminatorTM (Inspektor-Pro, Amsterdam, The Netherlands) at baseline and after 7, 14, and 21 days of application of the respective materials. Results ΔF value presented a significantly decreasing trend throughout the 21 days for all groups except the Duraphat and Enamel Pro varnishes. The changes between 14th and 21st days of the Clinpro XT varnish application were significantly higher than Emdogain, Curodont, and Enamel Pro. The Curodont group showed higher lesion area changes between the first and second week in comparison to the Emdogain, Clinpro XT, and Enamel Pro groups, whereas Clinpro XT assured the highest reduction from the second to the third week of the observation period. Conclusions The fluorescence loss was significantly reduced with enamel matrix protein, self-assembling peptide, and light-curable fluoride varnishes in the analysis for 21 days. Curodont and Clinpro XT were effective in diminishing the fluorescence loss and lesion area compared to the Duraphat, Enamel Pro fluoride varnishes, and Emdogain in different time points.

Amelogenin phosphorylation regulates tooth enamel formation by stabilizing a transient amorphous mineral precursor

Dental enamel comprises interwoven arrays of extremely long and narrow crystals of carbonated hydroxyapatite called enamel rods. Amelogenin (AMELX) is the predominant extracellular enamel matrix protein and plays an essential role in enamel formation (amelogenesis). Previously, we have demonstrated that full-length AMELX forms higher-order supramolecular assemblies that regulate ordered mineralization in vitro, as observed in enamel rods. Phosphorylation of the sole AMELX phosphorylation site (Ser-16) in vitro greatly enhances its capacity to stabilize amorphous calcium phosphate (ACP), the first mineral phase formed in developing enamel, and prevents apatitic crystal formation. To test our hypothesis that AMELX phosphorylation is critical for amelogenesis, we generated and characterized a hemizygous knockin (KI) mouse model with a phosphorylation-defective Ser-16 to Ala-16 substitution in AMELX. Using EM analysis, we demonstrate that in the absence of phosphorylated AMELX, KI enamel lacks enamel rods, the hallmark component of mammalian enamel, and, unlike WT enamel, appears to be composed of less organized arrays of shorter crystals oriented normal to the dentinoenamel junction. KI enamel also exhibited hypoplasia and numerous surface defects, whereas heterozygous enamel displayed highly variable mosaic structures with both KI and WT features. Importantly, ACP-to-apatitic crystal transformation occurred significantly faster in KI enamel. Secretory KI ameloblasts also lacked Tomes' processes, consistent with the absence of enamel rods, and underwent progressive cell pathology throughout enamel development. In conclusion, AMELX phosphorylation plays critical mechanistic roles in regulating ACP-phase transformation and enamel crystal growth, and in maintaining ameloblast integrity and function during amelogenesis.