Three-dimensional Reconstruction of Fracture Callus Morphogenesis

Louis C. Gerstenfeld ^1*, Yaser M. Alkhiary ¹, Elizabeth A. Krall ¹, Fred H. Nicholls ¹, Stephanie N. Stapleton ¹, Jennifer L. Fitch ¹, Megan Bauer ¹, Rayyan Kayal ¹, Dana T. Graves ¹, Karl J. Jepsen ¹ and Thomas A. Einhorn ¹

¹ Orthopaedic Research Laboratory, Boston University Medical Center, Boston, Massachusetts (LCG,FHN,SNS,JLF,TAE); Prosthodontics Division, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia (YMA); Department of Health Policy and Health Services Research (EAK) and Department of Periodontology and Oral Biology (MB,RK,DTG), Boston University School of Dental Medicine, Boston, Massachusetts; and Department of Orthopaedics, Mount Sinai School of Medicine, New York, New York (KJJ)

^* To whom correspondence should be addressed. E-mail: lgersten{at}bu.edu .

Submitted on March 3, 2006
Accepted on 11 July 2006

	Abstract

Rat and mouse femur and tibia fracture calluses were collected over various time increments of healing. Serial sections were produced at spatial segments across the fracture callus. Standard histological methods and in situ hybridization to col1a1 and col2a1 mRNAs were used to define areas of cartilage and bone formation as well as tissue areas undergoing remodeling. Computer-assisted reconstructions of the histological sections were used to generate three-dimensional images of the spatial morphogenesis of the fracture calluses. Endochondral bone formation occurred in an asymmetric manner in both the femur and the tibia, with cartilage tissues seen primarily proximal or distal to the fractures in the respective calluses of these bones. Remodeling of the calcified cartilage proceeded from the edges of the callus inward towards the fracture producing an inner supporting trabecular structure over which a thin outer cortical shell forms. These data suggest that the specific developmental mechanisms that control the asymmetric pattern of endochondral bone formation in fracture healing recapitulated the original asymmetry of development of a given bone since femurs and tibia grow predominantly from their respective distal and proximal physis. These data further show that the remodeling of the calcified cartilage produces a trabecular bone structure unique to fracture healing that provides the rapid regain in weight-bearing capacity to the injured bone.

Key Words: fracture repair, bone histomorphometry, growth and development, three-dimensional reconstructions, orthopaedics

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