Micro-computed tomography (CT) based assessment of dental regenerative therapy in the canine mandible model

High-resolution 3D bone-tissue structure measurements may provide information critical to the understanding of the bone regeneration processes and to the bone strength assessment. Tissue engineering studies rely on such nondestructive measurements to monitor bone graft regeneration area. In this study, we measured bone yield, fractal dimension and trabecular thickness through micro-CT slices for different grafts and controls. Eight canines underwent surgery to remove a bone volume (defect) in the canine’s jaw at a total of 44 different locations. We kept 11 defects empty for control and filled the remaining ones with three regenerative materials; NanoGen (NG), a FDA-approved material (n=11), a novel NanoCalcium Sulfate (NCS) material (n=11) and NCS alginate (NCS+alg) material (n=11). After a minimum of four and eight weeks, the canines were sacrificed and the jaw samples were extracted. We used a custombuilt micro-CT system to acquire the data volume and developed software to measure the bone yield, fractal dimension and trabecular thickness. The software used a segmentation algorithm based on histograms derived from volumes of interest indicated by the operator. Using bone yield and fractal dimension as indices we are able to differentiate between the control and regenerative material (p<0.005). Regenerative material NCS showed an average 63.15% bone yield improvement over the control sample, NCS+alg showed 55.55% and NanoGen showed 37.5%. The bone regeneration process and quality of bone were dependent upon the position of defect and time period of healing. This study presents one of the first quantitative comparisons using non-destructive Micro-CT analysis for bone regenerative material in a large animal with a critical defect model. Our results indicate that Micro-CT measurement could be used to monitor invivo bone regeneration studies for greater regenerative process understanding.

[1]  K. Hoffmann,et al.  Self-calibration of a cone-beam micro-CT system. , 2008, Medical physics.

[2]  H Weinans,et al.  Detecting and tracking local changes in the tibiae of individual rats: a novel method to analyse longitudinal in vivo micro-CT data. , 2004, Bone.

[3]  R. Reis,et al.  Evaluation of a starch‐based double layer scaffold for bone regeneration in a rat model , 2014, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[4]  Olivier Gauthier,et al.  In vivo bone regeneration with injectable calcium phosphate biomaterial: a three-dimensional micro-computed tomographic, biomechanical and SEM study. , 2005, Biomaterials.

[5]  Dietmar W Hutmacher,et al.  Current strategies for cell delivery in cartilage and bone regeneration. , 2004, Current opinion in biotechnology.

[6]  Christoph Groden,et al.  Application of micro-CT in small animal imaging. , 2010, Methods.

[7]  Ralph Müller,et al.  Monitoring individual morphological changes over time in ovariectomized rats by in vivo micro-computed tomography. , 2006, Bone.

[8]  P. Delmas,et al.  Bone quality--the material and structural basis of bone strength and fragility. , 2006, The New England journal of medicine.

[9]  L. Feldkamp,et al.  Practical cone-beam algorithm , 1984 .

[10]  J. Adams,et al.  Selecting regions of interest on intraoral radiographs for the prediction of bone mineral density. , 2008, Dento maxillo facial radiology.

[11]  R. Genco,et al.  Synthesis and characterization of nanocrystalline calcium sulfate for use in osseous regeneration , 2011, Biomedical materials.

[12]  Stephen Rudin,et al.  Micro-angiographic detector with fluoroscopic capability , 2002, SPIE Medical Imaging.

[13]  R. G. Richards,et al.  Animal models for implant biomaterial research in bone: a review. , 2007, European cells & materials.

[14]  Byung-Ho Choi,et al.  Critical size defect in the canine mandible. , 2005, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[15]  Felix Eckstein,et al.  Quantitative MRI of cartilage and bone: degenerative changes in osteoarthritis , 2006, NMR in biomedicine.

[16]  Ralph Müller,et al.  Guidelines for assessment of bone microstructure in rodents using micro–computed tomography , 2010, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[17]  J. van den Dolder,et al.  The bone regenerative effect of platelet-rich plasma in combination with an osteoconductive material in rat cranial defects. , 2006, Clinical oral implants research.

[18]  L Kinzl,et al.  Correlation of bone mineral density with strength and microstructural parameters of cortical bone in vitro. , 2002, Bone.

[19]  I. Sánchez,et al.  Fractals in dentistry. , 2011, Journal of dentistry.

[20]  B Bianco,et al.  Computational methods for ultrasonic bone assessment. , 1999, Ultrasound in medicine & biology.

[21]  L. Liebovitch,et al.  A fast algorithm to determine fractal dimensions by box counting , 1989 .

[22]  P. Rüegsegger,et al.  The ability of 3-D structural indices to reflect mechanical aspects of trabecular bone , 1999 .

[23]  H. K. Genant,et al.  Advanced CT bone imaging in osteoporosis , 2008, Rheumatology.

[24]  Sharmila Majumdar,et al.  Microarchitectural deterioration of cortical and trabecular bone: Differing effects of denosumab and alendronate , 2010, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[25]  Gadi Pelled,et al.  Microcomputed tomography–based structural analysis of various bone tissue regeneration models , 2011, Nature Protocols.

[26]  Michael V Swain,et al.  State of the Art of Micro-CT Applications in Dental Research , 2009, International Journal of Oral Science.

[27]  L. Vico,et al.  3D micro-computed tomography of trabecular and cortical bone architecture with application to a rat model of immobilisation osteoporosis , 2000, Medical and Biological Engineering and Computing.

[28]  P. Rüegsegger,et al.  Morphometric analysis of human bone biopsies: a quantitative structural comparison of histological sections and micro-computed tomography. , 1998, Bone.

[29]  Yves Amouriq,et al.  Exploring relationships between fractal dimension and trabecular bone characteristics , 2012, Medical Imaging.

[30]  M. Elsalanty,et al.  Three-Dimensional Evaluation of Mandibular Bone Regenerated By Bone Transport Distraction Osteogenesis , 2011, Calcified Tissue International.

[31]  Daniel R. Bednarek,et al.  Cone-Beam Micro-CT System Based on LabVIEW Software , 2008, Journal of Digital Imaging.