Acta Vet. Brno 2016, 85: 77-83

Three-dimensional bone tissue substitute based on a human mesenchymal stem cell culture on a nanofiber carrier and inorganic matrix

Martin Krbec1, Lukáš Plíštil2, Eva Matoušková2, Václav Mandys3, Jakub Ježek1, Markéta Sedlinská4, Valér Džupa1

1University Hospital Královské Vinohrady, Department of Orthopaedics and Traumatology, Prague, Czech Republic
2University Hospital Královské Vinohrady, Centre for Applied Bioimplantology, Prague, Czech Republic
3University Hospital Královské Vinohrady, Institute of Pathology, Prague, Czech Republic
4University of Veterinary and Pharmaceutical Sciences Brno, Faculty of Veterinary Medicine, Equine Clinic, Department of Reproduction, Brno, Czech Republic

Received December 4, 2015
Accepted February 10, 2016

The aim was to construct a composite structure for bone tissue substitute on the basis of a degradable composite of an organic nanofiber carrier and an inorganic matrix in 3D, and to achieve subsequent colonisation by differentiated human mesenchymal stem cells (hMSC) towards osteocytes. We developed an active bone tissue substitute using nanofiber technology for a polycaprolactone (PCL) scaffold with the addition of hydroxyapatite and the colonisation of both components with hMSC with the ability of differentiation towards osteocytes. The constructed composition included the components necessary for bone healing (inorganic and cellular) and it also forms a spatially-oriented 3D structure. We used polycaprolactone Mw 70,000 with electrostatic spinning for the formation of nanofibers using a modified NanospiderTM method. For the inorganic component we used orthophosphate-calcium silicate with a crystal size of 1-2 mm which the nanofiber membrane was coated with. Both components were connected together with a tissue adhesive based of fibrin glue. Cultivated hMSC cells at a concentration of 1.2 × 104/cm2 were multiplied in vitro and then cultivated in the expansion medium. HMSC overgrew both the PCL membrane and the Si-CaP crystals. After colonisation with cultivated cells, this composite 3D structure can serve as a three-dimensional bone tissue replacement.


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