Role of fibrillar structure of collagenous carrier in bone sialoprotein-mediated matrix mineralization and osteoblast differentiation.
To investigate the effects of the microstructure of collagenous carriers on the in vivo function of bone sialoprotein (BSP) in mineralization and osteoblast differentiation, we examined the ultrastructure of reconstituted type I collagen (collagen) and heat-denatured collagen (gelatin) and the in vivo responses to purified bone-derived BSP that was implanted with collagen or gelatin into surgically created 8-mm rat calvarial bone defects. Scanning and transmission electron microscopies revealed that the collagen displayed a fine fibrillar structure with interconnecting spaces between the fibrils/fibers, while the gelatin completely lost this unique three-dimensional structure after denaturation. The rates of in vivo release of BSP from the collagen scaffold were significantly lower than those from the gelatin. Collagen-BSP, but not gelatin-BSP, induced early mineral deposition in the matrix of proliferating repair cells in the calvarial defects at approximately 4-7 days after implantation. Expression levels of osteoblast markers, alkaline phosphatase activity and amounts of new bone synthesized in the collagen-BSP treated defects were significantly greater than that in the gelatin-BSP treated defects (p<0.001). The data suggest that the fibrillar microstructure of reconstituted collagen is essential for retaining BSP at a higher concentration within the defects, which enhances BSP-mediated matrix mineralization and osteoblast differentiation during the repair of rat calvarial defects.
Xu
Lan
L
Orthopedic Research Laboratory, Department of Orthopedic Surgery, Harvard Medical School and Children's Hospital, Boston, MA 02115, USA.
Anderson
Andy L
AL
Lu
Qinghua
Q
Wang
Jinxi
J
D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001857:Bone Matrix / Q000166:cytology* / Q000378:metabolism*; D002113:Calcification, Physiologic; D002454:Cell Differentiation; D003094:Collagen / Q000378:metabolism* / Q000648:ultrastructure; D005780:Gelatin / Q000378:metabolism / Q000648:ultrastructure; D058574:Integrin-Binding Sialoprotein; D008855:Microscopy, Electron, Scanning; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012795:Sialoglycoproteins / Q000378:metabolism*; D013997:Time Factors
D000596:Amino Acids; C546873:Ibsp protein, rat; D058574:Integrin-Binding Sialoprotein; D012795:Sialoglycoproteins; D005780:Gelatin; D003094:Collagen