Tissue Engineered Bone Repair of Calvarial Defects Using Cultured Periosteal Cells

Abstract

Periosteum has been demonstrated to have cell populations, including chondroprogenitor and osteoprogenitor cells, that can form both cartilage and bone under appropriate conditions. In the present study, periosteum was harvested, expanded in cell culture, and used to repair critical size calvarial defects in a rabbit model. Periosteum was isolated from New Zealand White rabbits, grown in cell culture, labeled with the thymidine analog bromodeoxyuridine for later localization, and seeded into resorbable polyglycolic acid scaffold matrices. Thirty adult New Zealand White rabbits were divided into groups, and a single 15-mm diameter full-thickness calvarial defect was made in each animal. In group I, defects were repaired using resorbable polyglycolic acid implants seeded with periosteal cells. In group II, defects were repaired using untreated polyglycolic acid implants. In group III, the defects were left unrepaired. Rabbits were killed at 4 and 12 weeks postoperatively. Defect sites were then studied histologically, biochemically, and radiographically. In vitro analysis of the cultured periosteal cells indicated an osteoblastic phenotype, with production of osteocalcin upon 1,25(OH)2 vitamin D3 induction. In vivo results at 4 weeks showed islands of bone in the defects repaired with polyglycolic acid implants with periosteal cells (group I), whereas the defects repaired with untreated polyglycolic acid implants (group II) were filled with fibrous tissue. Collagen content was significantly increased in group I compared with group II (2.90 +/- 0.80 microg/mg dry weight versus 0.08 +/- 0.11 microg/mg dry weight, p < 0.006), as was the ash weight (0.58 +/- 0.11 mg/mg dry weight versus 0.35 +/- 0.06 mg/mg dry weight, p < 0.015). At 12 weeks there were large amounts of bone in group I, whereas there were scattered islands of bone in groups II and III. Radiodensitometry demonstrated significantly increased radiodensity of the defect sites in group I, compared with groups II and III (0.740 +/- 0.250 OD/mm2 versus 0.404 +/- 0.100 OD/mm2 and 0.266 +/- 0.150 OD/mm2, respectively, p < 0.05). Bromodeoxyuridine label, as detected by immunofluorescence, was identified in the newly formed bone in group I at both 4 and 12 weeks, confirming the contribution of the cultured periosteal cells to this bone formation. This study thus demonstrates a tissue-engineering approach to the repair of bone defects, which may have clinical applications in craniofacial and orthopedic surgery.

Accelerated repair of cortical bone defects using a synthetic extracellular matrix to deliver human demineralized bone matrix

“…36 The fact that osteoblasts became active so quickly at the site of injury suggests that, at least during early stages of healing, skeletal progenitor cells are recruited from a nearby site such as the bone marrow, the endosteum, periosteum, and/or the vasculature. [37][38][39][40] At later stages, skeletal progenitor cells may also come from the muscle. 41 To understand more completely how Carbylan TM -GSX hydrogels and sponges with or without DBM may influence these putative sources of skeletal progenitor cells, we addressed separately the osteoconductive and the osteoinductive properties of Carbylan TM -GSX hydrogels and sponges with or without DBM.…”

The effect of aging on distraction osteogenesis in the rat

“…This is consistent with the observation that approximately 25% of trabecular bone is resorbed and replaced yearly, compared with only 3% of cortical bone [16]; the slower rate of remodeling of cortical bone would require fewer osteoblastic cell doublings compared to trabecular bone of equivalent age. Further, this suggests that the periosteum could be considered as a preferred donor site for tissue or cell-based therapy for repair of bony defects in older subjects [6,21].…”

Osteoblastic differentiation of periosteum‐derived cells is promoted by the physical contact with the bone matrix in vivo

“…Periosteal cells have been shown to possess the osteogenic potential in vitro (Koshihara et al, 1989;Tenenbaum et al, 1986) and in vivo (Breitbart et al, 1998;Nakahara et al, 1990b). The phenotype of periosteal cells, however, may be altered through being removed from the original site, dissected for cell culture, or suffering from surroundings that are different from in situ.…”

“…Cultured cells derived from the periosteum exhibit osteoblastic differentiation in the presence of ascorbic acid and 1,25 dihydroxy vitamin D3 in vitro (Fang and Hall, 1996;Koshihara et al, 1989;Tenenbaum et al, 1986). Periosteal explants or cells derived from the periosteum loaded in diffusion chambers or combined with porous ceramics showed the osteogenic potential in implantation experiments in vivo (Breitbart et al, 1998;Gally et al, 1994;Jaroma and Ritsila, 1988;Nakahara et al, 1990aNakahara et al, , 1991. The studies in vitro and in vivo described above suggest that periosteal cells contain subsets of progenitor cells that possess the potential to differentiate to osteoblasts.…”