Generating 3D tissue constructs with mesenchymal stem cells and a cancellous bone graft for orthopaedic applications

Abstract

Bone matrix (BM) is an acellular crosslinked porcine-derived cancellous bone graft, and therefore may provide advantages over other synthetic and naturally derived materials for use in orthopaedic surgery. Here, we analysed the potential of BM to support the growth and differentiation of primary human multipotent stromal cells/mesenchymal stem cells (MSCs) in order to predict in vivo bone regeneration events. Imaging with laser scanning confocal microscopy and scanning electron microscopy showed that 1 day after static seeding, a dense population of viable MSCs could be achieved on scaffolds suggesting they could be used for in vivo delivery of cells to the implant site. Long-term growth analysis by confocal imaging and histology demonstrated that BM was permissive to the growth and the 3D population of primary MSCs and an enhanced green fluorescent protein expressing osteosarcoma cell line, eGFP.MG63s, over several days in culture. Measurement of alkaline phosphatase (ALP) activities and mRNA expression levels of osteogenic markers (Runx-2, ALP, collagen type I, osteonectin, osteocalcin and osteopontin) indicated that BM supported osteogenesis of MSCs when supplemented with osteogenic stimulants. Upregulation of some of these osteogenic markers on BM, but not on tissue culture plastic, under non-osteogenic conditions suggested that BM also had osteoinductive capacities.

Three‐Dimensional Osteosarcoma Models for Advancing Drug Discovery and Development

“…However, the composition and architecture of these scaffolds do not fully reconstitute the in vivo ECM and thus the native cellular function cannot be strictly recapitulated. [87,88] The ability of MSCs to differentiate into osteoblast-like cells without osteogenic media highlighted the osteoinductive properties of the decellularized bone matrix. [86] Bone grafts have been explored to bone regeneration induced by osteogenic differentiation of MSCs in vitro and in vivo.…”

“…[86] Hence, 3D decellularized matrices emerged as the 3D scaffold that resemble in vivo ECM composition and structure as closely as possible and, despite the difficulties related to decellularization process, they have been applied in several tissues. [87] Furthermore, bone matrix-based particles seeded with MSCs was showed to be an effective support for new bone formation when implanted into a rat calvaria. [87,88] The ability of MSCs to differentiate into osteoblast-like cells without osteogenic media highlighted the osteoinductive properties of the decellularized bone matrix.…”

“…[87,88] The ability of MSCs to differentiate into osteoblast-like cells without osteogenic media highlighted the osteoinductive properties of the decellularized bone matrix. [87] Furthermore, bone matrix-based particles seeded with MSCs was showed to be an effective support for new bone formation when implanted into a rat calvaria. [88] Metastasis development toward bone is dependent of the attachment of cancer cells to proteins presented in bone ECM.…”

Embroidery technology for hard-tissue scaffolds

“…There is an abundant supply of animal bone material, but due to the high antigenicity of the graft and the risk of infection, these materials are denaturised to remove cellular components while maintaining the mineralised structure (Arca et al, 2011;Calori et al, 2011). Xenografts have successfully been used to fill segmental defects in oral and maxillofacial surgery.…”

Stem Cell Research: A New Era for Reconstructive Surgery

“…[18] Arca et al used acellular crosslinked porcine-derived cancellous bone graft with BM-MSCs for in vitro bone tissue engineering and an osteoinductive capacity was found in such material. [19] The choosing of scaffold material is largely determined by the bone defect itself. Fast resorbing materials, like tricalcium phosphates (TCP) can be used in a wound without special requirements of mechanical support, and slower degradation can be achieved by using materials like hydroxyapatite (HA) or through a combination of different materials, such as TCP and HA, or polyglycolic acid (PGA) and polylactic acid (PLA).…”