Human autologous plasma‐derived clot as a biological scaffold for mesenchymal stem cells in treatment of orthopedic healing

Abstract

Recent advances in the isolation, expansion, and characterization of human mesenchymal stem cells (hMSCs) have raised the possibility of using them in cell therapies and tissue engineering for bone reconstruction. hMSCs, isolated from the bone marrow of eight normal adult patients, were minimally expanded ex vivo and pulsed twice toward osteogenic lineage. The cells were then included into autologous plasma-derived clots. Cytofluorimetric analysis, immunocytochemistry (osteopontin), histochemistry (alkaline phosphatase, Alcian blue, Von Kossa, and alizarin red staining), and viable/proliferation tests were performed to study both stem and differentiating cells. Although two short inductions increased osteogenic markers in hMSCs, inside the clot the cells were able to terminally differentiate into osteoblasts. Moreover, we show that the clot is able to sustain cell proliferation under appropriate cell culture conditions. Our results suggested that clot could be useful for hMSC delivery into the site of the lesion to promote bone formation. Moreover, the plasticity of this material allowed good in vitro hMSC spreading and proliferation. The advantages of using this autologous biological material are its biocompatibility and reabsorption; furthermore, using a gel as scaffold, it is possible to mold it to the shape of a bone cavity. ß

Injection of Demineralized Bone Matrix With Bone Marrow Concentrate Improves Healing in Unicameral Bone Cyst

“…This procedure takes approximately 22 minutes. At the end of the procedure, the product included mononuclear progenitor cells, fibrin, and platelet-rich plasma [37]. When mixed with the demineralized bone powder (Musculoskeletal Tissue Bank of the Rizzoli Orthopaedic Institute), the marrow concentrate remains in a semisolid phase, thus maintaining this form also in the cystic cavity after injection.…”

Fibrin matrices: The versatile therapeutic delivery systems

“…Plasma-derived clots have been investigated earlier as potential scaffold for Human Mesenchymal Stem Cells (hMSCs) in treatment of orthopedic healing [106]. Vedakumari and Sastry [107] investigated the suitability of physiologically clotted fibrin, obtained by churning of whole blood, as an inexpensive scaffold for MG-63 human osteosarcoma cells and drug delivery.…”

Good manufacturing practice–grade fibrin gel is useful as a scaffold for human mesenchymal stromal cells and supports in vitro osteogenic differentiation

“…21 The objective of this study was to evaluate the possible use as a biologic scaffold of a standardized human fibrin clot, generated from thrombin and purified fibrinogen good manufacturing practice prepared from plasma pools (supplied by Kedrion SpA, Lucca, Italy) for therapeutic purposes.…”