title: Towards an intraoperative engineering of osteogenic and vasculogenic grafts from the stromal vascular fraction of human adipose tissue.
abstract: Grafts generated by cultivation of progenitor cells from the stromal vascular fraction of human adipose tissue have been proven to have osteogenic and vasculogenic properties in vivo. However, in vitro manufacture of such implants is challenged by complex, impractical and expensive processes, and requires implantation in a separate surgery. This study investigates the feasibility of an intraoperative approach to engineer cell-based bone grafts with tissue harvest, cell isolation, cell seeding onto a scaffold and subsequent implantation within a few hours. Freshly isolated adipose tissue cells from a total of 11 donors, containing variable fractions of mesenchymal and endothelial progenitors, were embedded at different densities in a fibrin hydrogel, which was wrapped around bone substitute materials based on beta-tricalcium phosphate (ChronOS), hydroxyapatite (Engipore), or acellular xenograft (Bio-Oss). The resulting constructs, generated within 3 hours from biopsy harvest, were immediately implanted ectopically in nude mice and analysed after eight weeks. All explants contained blood vessels formed by human endothelial cells, functionally connected to the recipient's vasculature. Human origin cells were also found within osteoid structures, positively immunostained for bone sialoprotein and osteocalcin. However, even with the highest loaded cell densities, no frank bone tissue was detected, independently of the material used. These results provide a proof-of-principle that an intraoperative engineering of autologous cell-based vasculogenic bone substitutes is feasible, but highlight that - in the absence of in vitro commitment--additional cues (e.g., low dose of osteogenic factors or orthotopic environmental conditions) are likely needed to support complete osteoblastic cell differentiation and bone tissue generation.
authors.affiliation: 1Departments of Surgery and of Biomedicine, University Hospital Basel, CH-4031 Basel, Switzerland. imartin@uhbs.ch
authors.forename: A M
authors.initials: AM
authors.lastname: Müller
authors.forename: A
authors.initials: A
authors.lastname: Mehrkens
authors.forename: D J
authors.initials: DJ
authors.lastname: Schäfer
authors.forename: C
authors.initials: C
authors.lastname: Jaquiery
authors.forename: S
authors.initials: S
authors.lastname: Güven
authors.forename: M
authors.initials: M
authors.lastname: Lehmicke
authors.forename: R
authors.initials: R
authors.lastname: Martinetti
authors.forename: I
authors.initials: I
authors.lastname: Farhadi
authors.forename: M
authors.initials: M
authors.lastname: Jakob
authors.forename: A
authors.initials: A
authors.lastname: Scherberich
authors.forename: I
authors.initials: I
authors.lastname: Martin
mesh_terms: D000273:Adipose Tissue / Q000166:cytology / Q000502:physiology / Q000637:transplantation*; D000818:Animals; D018786:Bone Substitutes / Q000494:pharmacology*; D016025:Bone Transplantation / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology / Q000637:transplantation; D006085:Graft Survival / Q000187:drug effects / Q000502:physiology; D006801:Humans; D020100:Hydrogels / Q000737:chemistry / Q000494:pharmacology; D007432:Intraoperative Period; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology; D051379:Mice; D008819:Mice, Nude; D018919:Neovascularization, Physiologic / Q000187:drug effects / Q000502:physiology; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects / Q000502:physiology; D012795:Sialoglycoproteins / Q000378:metabolism; D033581:Stem Cell Transplantation / Q000379:methods*; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology / Q000637:transplantation*; D023822:Tissue Engineering / Q000379:methods*; D014183:Transplantation, Heterologous / Q000379:methods
chemical_list: D018786:Bone Substitutes; D020100:Hydrogels; D012795:Sialoglycoproteins; D015675:Osteocalcin