Abstract
ABSTRACTBone fractures occur in horses following traumatic and non-traumatic (bone overloading) events. They can be difficult to treat due to the need for the horse to bear weight on all legs during the healing period. Regenerative medicine to improve fracture union and recovery could significantly improve horse welfare. Equine induced pluripotent stem cells (iPSCs) have previously been derived. Here we show that equine iPSCs cultured for 21 days in osteogenic induction media on an OsteoAssay surface upregulate the expression of osteoblast associated genes and proteins, including COL1A1, SPARC, SPP1, IBSP, RUNX2 and BGALP. We also demonstrate that iPSC-osteoblasts are able to produce a mineralised matrix with both calcium and hydroxyapatite deposition. Alkaline phosphatase activity is also significantly increased during osteoblast differentiation. Although the genetic background of the iPSC donor animal affects the level of differentiation observed after 21 days of differentiation, less variation between lines of iPSCs derived from the same horse was observed. The successful, direct, differentiation of equine iPSCs into osteoblasts may provide a source of cells for future regenerative medicine strategies to improve fracture repair in horses undergoing surgery. iPSC-derived osteoblasts will also provide a potential tool to study equine bone development and disease.
Highlights
Fractures occur in horses from a direct trauma, such as a kick, as well as from bone overloading
Equine induced pluripotent stem cells (iPSCs) differentiate into cells with the molecular signature of osteoblasts When cultured in osteogenic media on a surface designed to promote osteogenic differentiation [OsteoAssay (OA)], iPSCs differentiated into cells that expressed COL1A1, SPARC, SPP1, IBSP, RUNX2 (Runt related transcription factor 2) and BGALP (Fig. 1)
Equine iPSCs produce osteoblast-associated proteins following differentiation Undifferentiated iPSCs do not express any detectable osteoblastassociated proteins, whereas following 21 days in osteoblast induction media, COL1A1, SPARC, SPP1, IBSP, RUNX2 and BGALP are all detected in the differentiated cells (Fig. 1)
Summary
Fractures occur in horses from a direct trauma, such as a kick, as well as from bone overloading. They occur in all types of horses (Morgan and Dyson, 2012), with horses that take part in strenuous activities such as racing being at high risk of having a fracture resulting from overloading (Verheyen and Wood, 2004). Received 13 February 2018; Accepted 11 April 2018 genetic (Bulathsinhala et al, 2017; Korvala et al, 2010; Zhao et al, 2016) and environmental (Warden et al, 2006, 2007) factors have been implicated in stress fractures in human athletes and military personnel
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