Effect of Conditioned Medium from Preameloblasts on Regenerative Cellular Differentiation of the Immature Teeth with Necrotic Pulp and Apical Periodontitis

• Published in: Journal of endodontics Vol. 40; no. 9; pp. 1355 - 1361
• Main Authors: Yoo, Yeon-Jee, DDS, PhD, Lee, WooCheol, DDS, PhD, Cho, Young-Ah, DDS, PhD, Park, Joo-Cheol, DDS, PhD, Shon, Won-Jun, DDS, PhD, Baek, Seung-Ho, DDS, PhD
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.09.2014
• Subjects: Ameloblasts - physiology; Animals; Anti-Bacterial Agents - therapeutic use; Apexification - methods; Cell Culture Techniques; Cell Differentiation - physiology; Cellular differentiation; Cellular Microenvironment - physiology; conditioned medium; Core Binding Factor Alpha 1 Subunit - analysis; Culture Media, Conditioned; Dental Cementum - pathology; Dental Cementum - physiopathology; Dental Pulp - pathology; Dental Pulp - physiopathology; Dental Pulp Necrosis - pathology; Dental Pulp Necrosis - physiopathology; Dental Pulp Necrosis - therapy; Dentin - pathology; Dentin - physiopathology; Dentistry; Dogs; Endocrinology & Metabolism; Mesenchymal Stromal Cells - physiology; Mice; Mice, Inbred C57BL; Osteoblasts - physiology; osterix; Periapical Periodontitis - pathology; Periapical Periodontitis - physiopathology; Periapical Periodontitis - therapy; preameloblast; Random Allocation; Regeneration - physiology; revascularization; Root Canal Preparation - methods; Runx2; Time Factors; Tissue Scaffolds; Tooth Apex - pathology; Tooth Apex - physiopathology; Transcription Factors - analysis; Zinc Fingers; conditioned medium; revascularization; Cellular differentiation; preameloblast; osterix; Runx2
• ISSN: 0099-2399; 1878-3554
• DOI: 10.1016/j.joen.2014.02.009

Abstract Introduction The purpose of this study was to investigate the effect of conditioned medium (CM) from murine preameloblasts on the cellular differentiation of mesenchymal stem cells (MSCs) in immature teeth with necrotic pulp and apical periodontitis. Methods Pulp necrosis and apical periodontitis were induced in 30 immature permanent double-rooted premolars of 3 beagles and were randomly assigned to the following treatment groups: group CM ( n  = 10), revascularization treatment was performed using CM from preameloblasts of C57BL/6 mice apical bud cells; group CR ( n  = 10), conventional revascularization treatment was performed; positive control group ( n  = 5), left infected; and negative control group ( n  = 5), untreated. The dogs were followed up for 12 weeks and assessed for treatment outcomes with radiographic and histologic analyses. The effect of the CM on sequential Runx2 and osterix messenger RNA gene expression during the differentiation of MG63 osteoblastlike cells was analyzed with real-time polymerase chain reaction. Results The overall treatment outcomes were not significantly different between the 2 treatment groups. However, the teeth in the CM group showed significantly more mature apices and a higher degree of hard tissue formation with projections intercalating into the pre-existing root dentin ( P  < .05). In CM-treated teeth, regenerated pulplike tissue was more frequently observed ( P  < .05). During differentiation, the CM induced early peak expression of Runx2 followed by sustained osterix overexpression. Conclusions CM from preameloblasts rendered a favorable effect in providing a physiologic microenvironment for the differentiation of MSCs after revascularization treatment.