Multimodal analysis of in vivo resorbable CaP bone substitutes by combining histology, SEM, and microcomputed tomography data.

This study introduced and demonstrated a new method to investigate the repair process of bone defects using micro- and macroporous beta-tricalcium phosphate (β-TCP) substitutes. Specifically, the new method combined and aligned histology, SEM, and preimplantation microcomputed tomography (mCT) data to accurately characterize tissue phases found in biopsies, and thus better understand the bone repair process. The results included (a) the exact fraction of ceramic remnants (CR); (b) the fraction of ceramic resorbed and substituted by bone (CSB); and (c) the fraction of ceramic resorbed and not substituted by bone (CNSB). The new method allowed in particular the detection and quantification of mineralized tissues within the 1-10 µm micropores of the ceramic ("micro-bone"). The utility of the new method was demonstrated by applying it on biopsies of two β-tricalcium phosphate bone substitute groups with two differing macropore sizes implanted in an ovine model for 6 weeks. The total bone deposition and ceramic resorption of the two substitute groups, having macropore sizes of 510 and 1220 μm, were 25.1 ± 8.1% and 67.5 ± 3.2%, and 24.4 ± 4.1% and 61.4 ± 6.5% for the group having the larger pore size. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1567-1577, 2018.

[1]  Xiang Li,et al.  Influence of Architecture of β-Tricalcium Phosphate Scaffolds on Biological Performance in Repairing Segmental Bone Defects , 2012, PloS one.

[2]  R. Legeros Calcium Phosphate-Based Osteoinductive Materials , 2009 .

[3]  M Bohner,et al.  Geometric analysis of porous bone substitutes using micro-computed tomography and fuzzy distance transform. , 2010, Acta biomaterialia.

[4]  M. Mastrogiacomo,et al.  Role of scaffold internal structure on in vivo bone formation in macroporous calcium phosphate bioceramics. , 2006, Biomaterials.

[5]  Samantha J. Polak,et al.  The effect of BMP-2 on micro- and macroscale osteointegration of biphasic calcium phosphate scaffolds with multiscale porosity. , 2010, Acta biomaterialia.

[6]  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.

[7]  Alberto Diaspro,et al.  Order versus Disorder: in vivo bone formation within osteoconductive scaffolds , 2012, Scientific Reports.

[8]  P. Buma,et al.  Mechanism of bone incorporation of beta-TCP bone substitute in open wedge tibial osteotomy in patients. , 2005, Biomaterials.

[9]  L. Gibson,et al.  The effect of pore size on cell adhesion in collagen-GAG scaffolds. , 2005, Biomaterials.

[10]  Marcus Abboud,et al.  Comparison of three hydroxyapatite/β-tricalcium phosphate/collagen ceramic scaffolds: an in vivo study. , 2014, Journal of biomedical materials research. Part A.

[11]  Amy J Wagoner Johnson,et al.  Multiscale osteointegration as a new paradigm for the design of calcium phosphate scaffolds for bone regeneration. , 2010, Biomaterials.

[12]  P H Krebsbach,et al.  Engineering craniofacial scaffolds. , 2005, Orthodontics & craniofacial research.

[13]  G. Baroud,et al.  Effect of subvoxel processes on non-destructive characterization of β-tricalcium phosphate bone graft substitutes. , 2011, Acta biomaterialia.

[14]  S. Hofmann,et al.  Effect of grain size and microporosity on the in vivo behaviour of β-tricalcium phosphate scaffolds. , 2014, European cells & materials.

[15]  R M Pilliar,et al.  The optimum pore size for the fixation of porous-surfaced metal implants by the ingrowth of bone. , 1980, Clinical orthopaedics and related research.

[16]  Dietmar W Hutmacher,et al.  Assessment of bone ingrowth into porous biomaterials using MICRO-CT. , 2007, Biomaterials.

[17]  Anke Bernstein,et al.  Characterization and distribution of mechanically competent mineralized tissue in micropores of β-tricalcium phosphate bone substitutes , 2017 .

[18]  L. Podaropoulos,et al.  Bone regeneration using beta-tricalcium phosphate in a calcium sulfate matrix. , 2009, The Journal of oral implantology.

[19]  G H van Lenthe,et al.  Synthesis and characterization of porous beta-tricalcium phosphate blocks. , 2005, Biomaterials.

[20]  Xing‐dong Zhang,et al.  Tissue responses of calcium phosphate cement: a study in dogs. , 2000, Biomaterials.

[21]  Salvatore Candido,et al.  Automated segmentation of micro-CT images of bone formation in calcium phosphate scaffolds , 2012, Comput. Medical Imaging Graph..

[22]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[23]  L. Mathieu,et al.  Augmentation of bone defect healing using a new biocomposite scaffold: an in vivo study in sheep. , 2010, Acta biomaterialia.

[24]  M Bohner,et al.  Commentary: Deciphering the link between architecture and biological response of a bone graft substitute. , 2011, Acta biomaterialia.

[25]  R. Haddad,et al.  Optimum pore size for bone cement fixation. , 1987, Clinical orthopaedics and related research.

[26]  Ralph Müller,et al.  Nondestructive micro-computed tomography for biological imaging and quantification of scaffold-bone interaction in vivo. , 2007, Biomaterials.

[27]  G. Baroud,et al.  A novel method for segmenting and aligning the pre- and post-implantation scaffolds of resorbable calcium-phosphate bone substitutes. , 2017, Acta biomaterialia.

[28]  Clemens A van Blitterswijk,et al.  Bone regeneration: molecular and cellular interactions with calcium phosphate ceramics , 2006, International journal of nanomedicine.

[29]  Elisabeth H Burger,et al.  Localisation of osteogenic and osteoclastic cells in porous beta-tricalcium phosphate particles used for human maxillary sinus floor elevation. , 2005, Biomaterials.

[30]  Samantha J. Polak,et al.  Analysis of the roles of microporosity and BMP-2 on multiple measures of bone regeneration and healing in calcium phosphate scaffolds. , 2011, Acta biomaterialia.

[31]  Ralph Müller,et al.  In vivo behavior of calcium phosphate scaffolds with four different pore sizes. , 2006, Biomaterials.

[32]  Anja K. Stalder,et al.  Combined use of micro computed tomography and histology to evaluate the regenerative capacity of bone grafting materials , 2014 .

[33]  Bastian Brand,et al.  Biocompatibility and resorption of a brushite calcium phosphate cement. , 2005, Biomaterials.

[34]  Jing Li,et al.  Clinical evaluation of β-TCP in the treatment of lacunar bone defects: a prospective, randomized controlled study. , 2013, Materials science & engineering. C, Materials for biological applications.

[35]  M. Bohner,et al.  Resorbable biomaterials as bone graft substitutes , 2010 .

[36]  K. Draenert,et al.  Osseointegration of hydroxyapatite and remodeling‐resorption of tricalciumphosphate ceramics , 2013, Microscopy research and technique.

[37]  P. Rüegsegger,et al.  A new method for the model‐independent assessment of thickness in three‐dimensional images , 1997 .