Purpose: Poor bone quality is a common challenge to orthopaedic surgeons and frequently leads to complications such as non union and implant failure, particularly the elderly whose capacity for tissue repair is significantly reduced. The current study was designed to determine if bone marrow derived mesenchymal stem cells (MSC) seeded in dense collagen scaffolds and delivered to a surgically-induced femoral defect will expedite bone healing.
Method: Ex Vivo: MSC isolated from four month old donor mice were expanded ex vivo, seeded into hydrated type I collagen, which was subjected to unconfined compression to generate dense collagen scaffolds. The cell-seeded scaffolds were then cultured for up to 21 days. MSC viability was monitored using the AlamarBlue® metabolic assay and differentiation into osteoblasts using alkaline phosphatase (ALP) and von Kossa stain. In Vivo: A 3mm x 1mm window defect was drilled in the femur of elderly recipient C57Bl6 and C3H mice. The C3H mice were assigned to one of two study groups:
LEFT femur drill hole alone; RIGHT femur acellular scaffold.
LEFT femur acellular scaffold; RIGHT femur cell-seeded scaffold.
The quantity and quality of bone regeneration was assessed after 2 and 4 weeks using micro computed tomography (mCT) and histology.
Results: Ex Vivo: The dense collagen scaffold had superior mechanical properties and supported the survival and differentiation of MSC into osteoblasts up to 21 days in culture. Cells in uncompressed gels and those in compressed gels in non-osteogenic medium, had fewer ALP-positive cells at early time point and less mineral deposited at later times compared with those in compressed gels in osteogenic medium. In Vivo: A high incidence of postoperative fracture was seen in C57Bl6 mice compared with age matched C3H mice in the first study group. Furthermore, the empty surgical defect healed more rapidly than that containing the dense collagen scaffold, in which bone volume compared with tissue volume (BV/TV), trabecular number (Tb.N.) and connectivity were lower. In study group two, bone regeneration was evident at 2 weeks post operative and transplantation of MSC-seeded dense collagen scaffolds resulted in higher BV/TV, Tb.N. and trabecular connectivity compared with the acellular dense collagen scaffold.
Conclusion: Bone fragility in elderly C57Bl6 mice led to post operative fracture after generation of a non-critical sized drill hole defect in the proximal femur whereas age-matched C3H mice with higher bone mass sustained no fractures. Dense collagen scaffolds supported the survival and osteoblast differentiation of bone marrow derived MSC in 3D culture. Their superior mechanical properties allowed for transplantation into non-critical sized femoral defects, suggesting the approach shows promise as adjunct therapy for use with bone grafts and implants in patients with poor quality bone.
