title: 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.
authors.affiliation: Department of Oncology, Transplant and Advanced Technologies in Medicine, Hematology Division, University of Pisa, Via Roma 67, 56126, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Trombi
authors.forename: Letizia
authors.initials: L
authors.lastname: Mattii
authors.forename: Simone
authors.initials: S
authors.lastname: Pacini
authors.forename: Delfo
authors.initials: D
authors.lastname: D'Alessandro
authors.forename: Barbara
authors.initials: B
authors.lastname: Battolla
authors.forename: Enrico
authors.initials: E
authors.lastname: Orciuolo
authors.forename: Gabriele
authors.initials: G
authors.lastname: Buda
authors.forename: Rita
authors.initials: R
authors.lastname: Fazzi
authors.forename: Sara
authors.initials: S
authors.lastname: Galimberti
authors.forename: Mario
authors.initials: M
authors.lastname: Petrini
mesh_terms: D000328:Adult; D000368:Aged; D001672:Biocompatible Materials / Q000737:chemistry; D001777:Blood Coagulation; D001854:Bone Marrow Cells / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D002470:Cell Survival; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D017102:Fracture Healing; D006622:Hip Prosthesis; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D008875:Middle Aged; D008954:Models, Biological; D010006:Osteoblasts / Q000378:metabolism; D010012:Osteogenesis; D013234:Stem Cells / Q000378:metabolism
chemical_list: D001672:Biocompatible Materials