Synergistic effect of Wnt modulatory small molecules and an osteoinductive ceramic on C2C12 cell osteogenic differentiation

• Published in: Bone (New York, N.Y.) Vol. 67; pp. 109 - 121
• Main Authors: Chen, Sigeng, Ryan, Daniel A, Dwyer, Mary A, Cashman, John R
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Inc 01.10.2014; Elsevier
• Subjects: Biological and medical sciences; Bone Morphogenetic Protein 2 - metabolism; C2C12; Calcium Phosphates - pharmacology; Cell Differentiation - drug effects; Cell differentiation, maturation, development, hematopoiesis; Cell Line; Cell physiology; Ceramics; Core Binding Factor Alpha 1 Subunit - metabolism; Drug Synergism; Fundamental and applied biological sciences. Psychology; Humans; Mesenchymal stromal cells; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - drug effects; Molecular and cellular biology; Orthopedics; Osteogenesis; Osteogenesis - drug effects; Skeleton and joints; Small molecules; Tricalcium phosphate ceramic; Vertebrates: anatomy and physiology, studies on body, several organs or systems; Vertebrates: osteoarticular system, musculoskeletal system; Wnt; TCP; PP; ALP; hMSC; Small molecules; Tricalcium phosphate ceramic; C2C12; phenyl; tricalcium phosphate; Ph; Mesenchymal stromal cells; alkaline phosphatase; THQ; tetrahydroquinoline; Wnt; Osteogenesis; human mesenchymal stem cell; pyrazolopyrimidine; Phosphates; Morphology; Mesenchymal cell; Stromal cell; Cell differentiation
• ISSN: 8756-3282; 1873-2763
• DOI: 10.1016/j.bone.2014.06.032

Abstract Although osteoinductive ceramics can induce osteoblast differentiation in vitro and bone regeneration in vivo, their effects rely solely on the limited number of endogenous stem cells. More recently, ceramic carriers seeded with culture-expanded stem cells have been reported as implants capable of in vivo bone formation. However, effective and safe signaling agents that promote cell differentiation to the osteogenic lineage are still needed. In the present report, two osteogenic small-molecules THQ-1a and PP-9 were identified by testing a series of compounds for Runx2 and BMP2 expression in C2C12 cells. Compounds THQ-1a and PP-9 modulated Wnt signaling and enhanced the expression of molecular markers of osteoblast differentiation. To probe the utility of these compounds for use with ceramic cell implants, the effect of THQ-1a and PP-9 on C2C12 cell osteogenic differentiation was investigated in the presence of a tricalcium phosphate (TCP) ceramic. The effect of THQ-1a and PP-9 on markers such as Osteocalcin and Collagen I was significantly increased in the presence of TCP ceramic. Additionally, THQ-1a or PP-9 in the presence of TCP ceramic gave a synergistic increase in alkaline phosphatase activity in the differentiation of C2C12 cells. Taken together, the results suggest an approach to directing cell lineage commitment for bone regeneration by the application of small-molecule osteogenic agents to cells in the presence of osteoinductive ceramics.