Rapid Osteogenic Enhancement of Stem Cells in Human Bone Marrow Using a Glycogen-Synthease-Kinase-3-Beta Inhibitor Improves Osteogenic Efficacy In Vitro and In Vivo

Abstract

Non‐union defects of bone are a major problem in orthopedics, especially for patients with a low healing capacity. Fixation devices and osteoconductive materials are used to provide a stable environment for osteogenesis and an osteogenic component such as autologous human bone marrow (hBM) is then used, but robust bone formation is contingent on the healing capacity of the patients. A safe and rapid procedure for improvement of the osteoanabolic properties of hBM is, therefore, sought after in the field of orthopedics, especially if it can be performed within the temporal limitations of the surgical procedure, with minimal manipulation, and at point‐of‐care. One way to achieve this goal is to stimulate canonical Wingless (cWnt) signaling in bone marrow‐resident human mesenchymal stem cells (hMSCs), the presumptive precursors of osteoblasts in bone marrow. Herein, we report that the effects of cWnt stimulation can be achieved by transient (1–2 hours) exposure of osteoprogenitors to the GSK3β‐inhibitor (2′Z,3′E)‐6‐bromoindirubin‐3′‐oxime (BIO) at a concentration of 800 nM. Very‐rapid‐exposure‐to‐BIO (VRE‐BIO) on either hMSCs or whole hBM resulted in the long‐term establishment of an osteogenic phenotype associated with accelerated alkaline phosphatase activity and enhanced transcription of the master regulator of osteogenesis, Runx2. When VRE‐BIO treated hBM was tested in a rat spinal fusion model, VRE‐BIO caused the formation of a denser, stiffer, fusion mass as compared with vehicle treated hBM. Collectively, these data indicate that the VRE‐BIO procedure may represent a rapid, safe, and point‐of‐care strategy for the osteogenic enhancement of autologous hBM for use in clinical orthopedic procedures. stem cells translational medicine 2018;7:342–353

6-Bromoindirubin-3′-oxime Promotes Osteogenic Differentiation of Periodontal Ligament Stem Cells and Facilitates Bone Regeneration in a Mouse Periodontitis Model

“…A previous study demonstrated that BIO can promote the formation of cell mineralized nodules, which is consistent with our results. Moreover, BIO can promote osteogenic differentiation of bone marrow mesenchymal stem cells and dental pulp stem cells by upregulating ALP activity and expression of osteogenic genes. − The potential mechanism of stimulation of osteogenesis by BIO may involve the activation of the Wnt signaling pathway in PDLSCs. Wnt family proteins inhibit apoptosis of mature osteoblasts through the classical and nonclassical pathways, thus extending the lifespan of osteoblasts .…”

Dissociation of nanosilicates induces downstream endochondral differentiation gene expression program

“…To validate the effect of ionic dissolution products on some of the key signaling pathways, we treated hMSCs with Si, Mg, and Li for 14 days in the presence and absence of cWnt inhibitor (10 M cardamonin; Cayman Chemicals) and MAPK inhibitor (5 M PD185342; Abcam). Both cWnt signaling (39,40) and MAPK/ERK signaling (41) are implicated in osteogenesis and are highlighted in our GO analysis. Western blot was performed on day 14 to evaluate the effect on ALP production (fig.…”

6-Bromoindirubin-3′-Oxime Regulates Colony Formation, Apoptosis, and Odonto/Osteogenic Differentiation in Human Dental Pulp Stem Cells

“…BIO is a small molecule that targets the glycogen synthase kinase-3 (GSK-3) [ 26 ]. A previous report demonstrated that BIO enhanced alkaline phosphatase (ALP) activity and osteogenesis-related gene expression in osteoprogenitor cells [ 27 ]. Similarly, BIO accelerated the in vitro chondrogenesis of mouse bone marrow mesenchymal stem cells (BMSCs), demonstrated by the upregulation of cartilage-specific gene expression [ 28 ].…”