title: Fabrication, vascularization and osteogenic properties of a novel synthetic biomimetic induced membrane for the treatment of large bone defects.
abstract: The induced membrane has been widely used in the treatment of large bone defects but continues to be limited by a relatively lengthy healing process and a requisite two stage surgical procedure. Here we report the development and characterization of a synthetic biomimetic induced membrane (BIM) consisting of an inner highly pre-vascularized cell sheet and an outer osteogenic layer using cell sheet engineering. The pre-vascularized inner layer was formed by seeding human umbilical vein endothelial cells (HUVECs) on a cell sheet comprised of a layer of undifferentiated human bone marrow-derived mesenchymal stem cells (hMSCs). The outer osteogenic layer was formed by inducing osteogenic differentiation of hMSCs. In vitro results indicated that the undifferentiated hMSC cell sheet facilitated the alignment of HUVECs and significantly promoted the formation of vascular-like networks. Furthermore, seeded HUVECs rearranged the extracellular matrix produced by hMSC sheet. After subcutaneous implantation, the composite constructs showed rapid vascularization and anastomosis with the host vascular system, forming functional blood vessels in vivo. Osteogenic potential of the BIM was evidenced by immunohistochemistry staining of osteocalcin, tartrate-resistant acid phosphatase (TRAP) staining, and alizarin red staining. In summary, the synthetic BIM showed rapid vascularization, significant anastomoses, and osteogenic potential in vivo. This synthetic BIM has the potential for treatment of large bone defects in the absence of infection.
authors.affiliation: Department of Orthopaedic Surgery, Stanford University, 300 Pasteur Drive, Stanford, CA 94305,USA.|School of Stomatology, Lanzhou University, 199 Donggang West Road, Lanzhou, Gansu 730000, China.
authors.forename: Liling
authors.initials: L
authors.lastname: Ren
authors.affiliation: Department of Orthopaedic Surgery, Stanford University, 300 Pasteur Drive, Stanford, CA 94305,USA.
authors.forename: Yunqing
authors.initials: Y
authors.lastname: Kang
authors.affiliation: Department of Orthopaedic Surgery, Stanford University, 300 Pasteur Drive, Stanford, CA 94305,USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Browne
authors.affiliation: Department of Orthopaedic Surgery, Stanford University, 300 Pasteur Drive, Stanford, CA 94305,USA.
authors.forename: Julius
authors.initials: J
authors.lastname: Bishop
authors.affiliation: Department of Orthopaedic Surgery, Stanford University, 300 Pasteur Drive, Stanford, CA 94305,USA.|Department of Materials Science and Engineering, Stanford University, 300 Pasteur Drive, Stanford, CA 94305,USA.
authors.forename: Yunzhi
authors.initials: Y
authors.lastname: Yang
mesh_terms: D032701:Biomimetics; D001842:Bone and Bones / Q000002:abnormalities*; D005455:Fluorescent Antibody Technique; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D010012:Osteogenesis / Q000187:drug effects*; D054457:Tissue Scaffolds
keywords: Cell sheet; HUVEC; Induced membrane; Vascularization; hMSC
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