Effect of Emdogain enamel matrix derivative and BMP-2 on the gene expression and mineralized nodule formation of alveolar bone proper- derived stem/progenitor cells Karim M. Fawzy El-Sayed a, b, * , Christof Dörfer a , Hendrick Ungefroren c , Neemat Kassem d , Jörg Wiltfang e , Sebastian Paris f a Clinic for Conservative Dentistry and Periodontology (Head: Prof. Dr. C. Dörfer), School of Dental Medicine, Christian Albrechts-University of Kiel, Arnold-Heller-Str. 3, Haus 26, 24105 Kiel, Germany b Oral Medicine and Periodontology Department, Faculty of Oral and Dental Medicine, Cairo University, Egypt c Clinic for Applied Cellular Therapy, Christian Albrechts-University of Kiel, Kiel, Germany d Department of Clinical Pathology, Faculty of Medicine, Cairo University, Egypt e Department of Oral and Maxillofacial Surgery, Christian Albrechts-University of Kiel, Kiel, Germany f Department of Operative and Preventive Dentistry, Charité - Universitätsmedizin Berlin, Berlin, Germany article info Article history: Paper received 26 April 2013 Accepted 31 July 2013 Keywords: Stem cells Tissue engineering Alveolar bone proper EMD BMP-2 abstract The objective of this study was to evaluate the effect of Emdogain (Enamel Matrix Derivative, EMD) and Bone Morphogenetic Protein-2 (BMP-2), either solely or in combination, on the gene expression and mineralized nodule formation of alveolar bone proper-derived stem/progenitor cells. Stem/progenitor cells were isolated from human alveolar bone proper, magnetically sorted using STRO-1 antibodies, characterized flowcytometrically for their surface markers’ expression, and examined for colony formation and multilineage differentiation potential. Subsequently, cells were treated over three weeks with 100 mg/ml Emdogain (EMD-Group), or 100 ng/ml BMP-2 (BMP-Group), or a combi- nation of 100 ng/ml BMP-2 and 100 mg/ml Emdogain (BMP/EMD-Group). Unstimulated stem/progenitor cells (MACS þ -Group) and osteoblasts (OB-Group) served as controls. Osteogenic gene expression was analyzed using RTq-PCR after 1, 2 and 3 weeks (N ¼ 3/group). Mineralized nodule formation was eval- uated by Alizarin-Red staining. BMP and EMD up-regulated the osteogenic gene expression. The BMP Group showed significantly higher expression of Collagen-I, III, and V, Alkaline phosphatase and Osteonectin compared to MACS þ - and OB-Group (p < 0.05; Two-way ANOVA/Bonferroni) with no mineralized nodule formation. Under in-vitro conditions, Emdogain and BMP-2 up-regulate the osteogenic gene expression of stem/ progenitor cells. The combination of BMP-2 and Emdogain showed no additive effect and would not be recommended for a combined clinical stimulation. Ó 2013 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved. 1. Introduction Periodontitis, as a bacterial induced inflammatory disorder, is characterized by the destruction of the periodontal attachment apparatus, resulting in apical migration of epithelial attachment with pocket formation and possible tooth loss (Page et al., 1997). The ideal goal of periodontal therapy is to regenerate these lost tissues to their original form, architecture and function, which due to the complexity of the periodontal environment poses a challenging task (Minabe, 1991; Schroeder, 1992; Garrett, 1996), requiring the harmonization of many events at cellular and molecular levels (Bartold et al., 2000). A range of trials to regenerate the lost tissue have been attempted, however with limited regeneration success and unpredictable clinical outcomes (Needleman et al., 2006; Esposito et al., 2009; Ivanovski, 2009). Recently the principle of tissue engineering, employing stem/progenitor cells in combina- tion with various growth/differentiation factors has emerged as a promising approach for oral tissue regeneration (Behnia et al., 2012; Fawzy El-Sayed et al., 2012a,b; Warnke et al., 2013). Different dental tissues, including the periodontal ligament, the dental pulp, the tooth follicle, the apical papilla, the gingiva and the * Corresponding author. Clinic for Conservative Dentistry and Periodontology, School of Dental Medicine, Christian Albrechts-University of Kiel, Arnold-Heller-Str. 3, Haus 26, 24105 Kiel, Germany. Tel.: þ49 431 597 2814; fax: þ49 431 597 4108. E-mail address: karim.fawzy@gmail.com (K.M. Fawzy El-Sayed). Contents lists available at ScienceDirect Journal of Cranio-Maxillo-Facial Surgery journal homepage: www.jcmfs.com 1010-5182/$ e see front matter Ó 2013 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.jcms.2013.07.028 Journal of Cranio-Maxillo-Facial Surgery xxx (2013) 1e9 Please cite this article in press as: Fawzy El-Sayed KM, et al., Effect of Emdogain enamel matrix derivative and BMP-2 on the gene expression and mineralized nodule formation of alveolar bone proper-derived stem/progenitor cells, Journal of Cranio-Maxillo-Facial Surgery (2013), http:// dx.doi.org/10.1016/j.jcms.2013.07.028