OBJECTIVETo evaluate the therapeutic effect of bone morphogenetic protein 2 (BMP-2) gene modified tissue engineering bone (GMB) combined with vascularized periosteum in the reconstruction of segmental bone defect.

METHODSAdenovirus carrying BMP-2 gene (Ad-BMP-2) was transfected into the isolated and cultured rabbit bone marrow stromal cells (MSCs). The transfected MSCs were seeded on bovine cancellous bone scaffolds (BCB) to construct gene modified tissue engineering bone (GMB). The bilateral rabbits radial defects (2.5 cm long) were created as animal model. The rabbits were divided into five groups to reconstruct the defects with CMB combined with vascularized periosteum (group A); or GMB combined with vascular bundle implantation (group B); or GMB combined with free periosteum (group C); or GMB only (group D); or BCB scaffolds only (group E). Angiogenesis and osteogenesis were observed by X-ray, histological examination, biomechanical analysis and capillary ink infusion.

RESULTSIn group A, the grafted GMB was revascularized rapidly. The defect was completely reconstructed at 8 weeks. The mechanism included both intramemerbrane and endochondral ossification. In group B, the vascular bundle generated new blood vessels into the grafted GMB, but the osteogenesis process was slow in the central zone, which healed completely at 12 weeks. In group C, the free graft of periosteum took at 4 weeks with angiogenesis. The thin extremal callus was formed at 8 weeks and the repairing process almost finished at 12 weeks. Better osteogenesis was found in group D than in group E, due to the present of BMP2 gene-transfected MSCs. The defects in group D were partial repaired at 12 weeks with remaining central malunion zone. The defects in group E should nonunion at 12 weeks with only fibre tissue.

CONCLUSIONSBMP-2 gene modified tissue engineering bone combined with vascularized periosteum which provides periosteum osteoblasts as well as blood supply, has favorable ability of osteogenesis, osteoinduction and osteoconduction. It is an ideal method for the treatment of segmental bone defect.

Animals ; Bone Marrow Cells ; cytology ; Bone Morphogenetic Protein 2 ; genetics ; Bone Regeneration ; Bone Substitutes ; Bone Transplantation ; methods ; Bone and Bones ; pathology ; Cattle ; Mesenchymal Stromal Cells ; cytology ; Periosteum ; blood supply ; transplantation ; Rabbits ; Surgical Flaps ; blood supply ; Tissue Engineering ; methods ; Tissue Scaffolds ; Transfection

Repair of skeletal defects with vascularized bone grafts has many advantages over non-vascularized free grafts, but the availability of these grafts is extremely limited. This study was designed to determine whether new vascularized bone could be engineered by transplantation of osteoblasts around existing vascular pedicles using biodegradable, synthetic polymer as a cell delivery vehicle. Cells were isolated from the periosteum of fetal bovine humerus, and then seeded onto non-woven multifilament, polyglycolic acid polymer. The polymers provided three dimensional support during in vitro culture. The cell-polymer constructs were maintained in vitro for two weeks and then implanted around the right femoral vessels of twelve athymic nude rats. The polymer templates without the cells were implanted around left femoral vessels of each mouse as a control. Twelve rats were sacrificed at the following intervals: three rats at six, and nine rats at nine weeks. New bone formation was evident in 10 out of the 12 periosteal-derived cell seeded implants. At six weeks, the tissue was primarily composed of what appeared both grossly and histologically to be cartilage enveloping small islands of osteoid. The degree of osteoid and bone formation progressed with time, as blood vessels invaded the tissue. This tissue ultimately underwent morphogenesis to become an organized trabeculated bone with a vascular pedicle. We believe that this technique may prove to be useful in the reconstruction of bony defect.
Animals ; Animals, Newborn ; Bone and Bones/*blood supply/cytology ; Cattle ; Cells, Cultured ; Implants, Experimental ; Osteoblasts/*cytology ; Osteogenesis ; *Polyglycolic Acid ; Time Factors ; Tissue Engineering/*methods