Please cite this article in press as: H. Bakhtiar, M. Pezeshki-Modaress, Z. Kiaipour et al.. Pulp ECM-derived macroporous scaffolds for stimulation of dental-pulp regeneration process. Dent Mater (2019), https://doi.org/10.1016/j.dental.2019.10.011 ARTICLE IN PRESS DENTAL-3446; No. of Pages 12 d e n t a l m a t e r i a l s x x x ( 2 0 1 9 ) xxx–xxx Available online at www.sciencedirect.com ScienceDirect jo ur nal home p ag e: www.intl.elsevierhealth.com/journals/dema Pulp ECM-derived macroporous scaffolds for stimulation of dental-pulp regeneration process Hengameh Bakhtiar a,b,c , Mohammad Pezeshki-Modaress d , Zahra Kiaipour c , Mahdieh Shafiee c , Mohammad Reza Ellini c , Amir Mazidi c , Sarah Rajabi e , Soheila Zamanlui Benisi c , Seyed Naser Ostad f , Kerstin Galler g , Pardis Pakshir b , Amir Azarpazhooh b,h,i , Anil Kishen b,i,∗ a Department of Endodontics, Faculty of Dentistry, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran b Faculty of Dentistry, University of Toronto, Toronto, Ontario, Canada c Stem Cell Research Center, Tissue Engineering and Regenerative Medicine Institute, Tehran Central Branch, Islamic Azad University, Tehran, Iran d Burn Research Center, Iran University of Medical Sciences, Tehran, Iran e Department of Cell Engineering, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran f Department of Toxicology-Pharmacology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran g Department of Conservative Dentistry and Periodontology, University Hospital Regensburg, Regensburg, Germany h Clinical Epidemiology and Health Care Research, Institute of Health Policy, Management and Evaluation, University of Toronto, Toronto, Ontario, Canada i Department of Dentistry, Mount Sinai Hospital, Toronto, Ontario, Canada a r t i c l e i n f o Article history: Received 15 July 2019 Accepted 18 October 2019 Available online xxx Keywords: Tissue engineering Dentin-pulp complex Pulp ECM Crosslinked scaffold a b s t r a c t Objective. Recent studies suggest xenogeneic extracellular matrices as potential regener- ative tools in dental pulp regeneration. This study aimed to fabricate and characterize a novel three-dimensional macroporous pulp-derived scaffold that enables the attachment, penetration, proliferation and differentiation of mesenchymal stem cells. Method. Bovine pulp was decellularized and characterized with histological and DNA con- tent methods. This scaffold was prepared using finely milled lyophilized decellularized pulp extracellular matrix (ECM) digested with pepsin. Three different concentrations of ECM (1.50, 2.25 and 3.00 mg/ml) were freeze-dried and were tested with/without chemical crosslink- ing. The specimens were subjected to physicochemical characterization, cell viability and quantitative real time polymerase chain reaction assessments with human bone marrow mesenchymal stem cells (hBMMSCs). All scaffolds were subcutaneously implanted in rats for two weeks and histological and immunostaining analyses were performed. ∗ Corresponding author at: Faculty of Dentistry, University of Toronto, 124 Edward St, Toronto, ON M5G 1G6, Canada. E-mail address: Anil.Kishen@dentistry.utoronto.ca (A. Kishen). https://doi.org/10.1016/j.dental.2019.10.011 0109-5641/© 2019 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.