title: In vivo 3D analysis with micro-computed tomography of rat calvaria bone regeneration using periosteal cell sheets fabricated on temperature-responsive culture dishes.
abstract: Transplantable cell sheets containing osteoblasts were fabricated from periostea on temperature-responsive culture dishes. This study demonstrated the time-course of bone regeneration in living small animals. This continuous observation of bone regeneration was achieved by micro-computed tomography (µCT), which assessed the osteogenic capability of periosteal cells without biodegradable scaffolds. Real-time bone regeneration was non-invasively monitored in a rat calvarial bone defect model, using µCT. Three-dimensional (3D) images obtained over time by µCT clearly showed that two different bone regeneration modes, specific to the control and experimental groups, were observed. In the control group, bone was regenerated only from the periphery of the defect edges. In the experimental group, bone regeneration was observed in several small regions within the central portions of the defects that were covered by the transplanted cell sheets. However, bone regeneration observed after periosteal cell sheet transplantation was limited. The results of ALP staining and the time-course observations concluded that periosteal cell sheets contained a small fraction of cells that could differentiate osteoblasts. Fibroblasts in transplanted cell sheets or from around subcutaneous tissues suppressed bone regeneration. The periosteal cell sheets had a capability to produce ectopic regenerated bones. Therefore, to increase the content of osteogenic cells in harvested cell sheets, the enrichment of cells that could produce osteoblasts was expected by the modification of the initial cell preparation and the culture conditions. With further possible improvements, scaffold-free periosteal cell sheet fabricated on temperature-responsive culture dishes will be a valuable method for inducing and accelerating bone regeneration.
authors.affiliation: Department of Oral and Maxillofacial Surgery, School of Medicine, Tokyo Women's Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan.
authors.forename: Hiroto
authors.initials: H
authors.lastname: Uchiyama
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yamato
authors.forename: Ryo
authors.initials: R
authors.lastname: Sasaki
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Sekine
authors.forename: Joseph
authors.initials: J
authors.lastname: Yang
authors.forename: Hideki
authors.initials: H
authors.lastname: Ogiuchi
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Ando
authors.forename: Teruo
authors.initials: T
authors.lastname: Okano
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001861:Bone Regeneration / Q000502:physiology*; D002113:Calcification, Physiologic / Q000502:physiology; D021621:Imaging, Three-Dimensional / Q000379:methods*; D010521:Periosteum / Q000166:cytology / Q000201:enzymology / Q000637:transplantation*; D051381:Rats; D012886:Skull / Q000000981:diagnostic imaging* / Q000502:physiology; D013194:Staining and Labeling; D013696:Temperature; D046509:Tissue Culture Techniques / Q000379:methods*; D055114:X-Ray Microtomography / Q000379:methods*
chemical_list: D000469:Alkaline Phosphatase