Long-Bone-Regeneration Process in a Sheep Animal Model, Using Hydroxyapatite Ceramics Prepared by Tape-Casting Method

This study was designed to investigate the effects of hydroxyapatite (HA) ceramic implants (HA cylinders, perforated HA plates, and nonperforated HA plates) on the healing of bone defects, addressing biocompatibility, biodegradability, osteoconductivity, osteoinductivity, and osteointegration with the surrounding bone tissue. The HA ceramic implants were prepared using the tape-casting method, which allows for shape variation in samples after packing HA paste into 3D-printed plastic forms. In vitro, the distribution and morphology of the MC3T3E1 cells grown on the test discs for 2 and 9 days were visualised with a fluorescent live/dead staining assay. The growth of the cell population was clearly visible on the entire ceramic surfaces and very good osteoblastic cell adhesion and proliferation was observed, with no dead cells detected. A sheep animal model was used to perform in vivo experiments with bone defects created on the metatarsal bones, where histological and immunohistochemical tissue analysis as well as X-ray and CT images were applied. After 6 months, all implants showed excellent biocompatibility with the surrounding bone tissue with no observed signs of inflammatory reaction. The histomorphological findings revealed bone growth immediately over and around the implants, indicating the excellent osteoconductivity of the HA ceramic implants. A number of islands of bone tissue were observed towards the centres of the HA cylinders. The highest degree of biodegradation, bioresorption, and new bone formation was observed in the group in which perforated HA plates were applied. The results of this study suggest that HA cylinders and HA plates may provide a promising material for the functional long-bone-defect reconstruction and further research.

[1]  Yuer Zhang,et al.  Biomaterial-based strategy for bone tumor therapy and bone defect regeneration: An innovative application option , 2022, Frontiers in Materials.

[2]  Wen‐Cheng Chen,et al.  Morphological Changes, Antibacterial Activity, and Cytotoxicity Characterization of Hydrothermally Synthesized Metal Ions-Incorporated Nanoapatites for Biomedical Application , 2022, Pharmaceuticals.

[3]  Yilong Cheng,et al.  The Osteoinductivity of Calcium Phosphate-Based Biomaterials: A Tight Interaction With Bone Healing , 2022, Frontiers in Bioengineering and Biotechnology.

[4]  Xing‐dong Zhang,et al.  Optimal regenerative repair of large segmental bone defect in a goat model with osteoinductive calcium phosphate bioceramic implants , 2021, Bioactive materials.

[5]  M. Marcacci,et al.  Bone Regeneration in Load-Bearing Segmental Defects, Guided by Biomorphic, Hierarchically Structured Apatitic Scaffold , 2021, Frontiers in Bioengineering and Biotechnology.

[6]  D. Ribeiro,et al.  In vitro and in vivo biological performance of hydroxyapatite from fish waste , 2021, Journal of Materials Science: Materials in Medicine.

[7]  N. Kamboj,et al.  Ionic substituted hydroxyapatite for bone regeneration applications: A review , 2021 .

[8]  M. Y. Bajuri,et al.  Tissue-Engineered Hydroxyapatite Bone Scaffold Impregnated with Osteoprogenitor Cells Promotes Bone Regeneration in Sheep Model , 2021, Tissue Engineering and Regenerative Medicine.

[9]  Xiubo Zhao,et al.  Electrospun Icariin-Loaded Core-Shell Collagen, Polycaprolactone, Hydroxyapatite Composite Scaffolds for the Repair of Rabbit Tibia Bone Defects , 2020, International journal of nanomedicine.

[10]  E. Potier,et al.  Custom-made macroporous bioceramic implants based on triply-periodic minimal surfaces for bone defects in load-bearing sites. , 2020, Acta biomaterialia.

[11]  S. R. Gavinho,et al.  Calcium Phosphate Cements in Tissue Engineering , 2020, Contemporary Topics about Phosphorus in Biology and Materials.

[12]  H. Yoshikawa,et al.  Bone regeneration with hydroxyapatite-based biomaterials , 2019, Emergent Materials.

[13]  Callinca Paolla Gomes Machado,et al.  Evaluation of strontium-containing hydroxyapatite as bone substitute in sheep tibiae. , 2019, Brazilian Journal of Implantology and Health Sciences.

[14]  C. Elias,et al.  Nanosized hydroxyapatite and β-tricalcium phosphate composite: Physico-chemical, cytotoxicity, morphological properties and in vivo trial , 2019, Scientific Reports.

[15]  M. Ansari.  Bone tissue regeneration: biology, strategies and interface studies , 2019, Progress in Biomaterials.

[16]  Huawei Qu,et al.  Biomaterials for bone tissue engineering scaffolds: a review , 2019, RSC advances.

[17]  M. Trunec,et al.  Structure degradation and strength changes of sintered calcium phosphate bone scaffolds with different phase structures during simulated biodegradation in vitro. , 2019, Materials science & engineering. C, Materials for biological applications.

[18]  Yue-Wern Huang,et al.  Evaluation of Open Hollow Hydroxyapatite Microsphere on Bone Regeneration in Rat Calvarial Defects , 2019, bioRxiv.

[19]  Nathaniel S. Hwang,et al.  Bioactive calcium phosphate materials and applications in bone regeneration , 2019, Biomaterials Research.

[20]  Jianhua Li,et al.  A method to visually observe the degradation-diffusion-reconstruction behavior of hydroxyapatite in the bone repair process. , 2019, Acta biomaterialia.

[21]  A. W. Wagoner Johnson,et al.  Mineralization in micropores of calcium phosphate scaffolds. , 2019, Acta biomaterialia.

[22]  F. Luyten,et al.  Bone regeneration strategies: Engineered scaffolds, bioactive molecules and stem cells current stage and future perspectives. , 2018, Biomaterials.

[23]  H. Larjava,et al.  Hydoxyapatite/beta-tricalcium phosphate biphasic ceramics as regenerative material for the repair of complex bone defects. , 2018, Journal of biomedical materials research. Part B, Applied biomaterials.

[24]  M. Ginebra,et al.  Bioceramics and bone healing , 2018, EFORT open reviews.

[25]  M. Barbeck,et al.  Purification processes of xenogeneic bone substitutes and their impact on tissue reactions and regeneration , 2018, The International journal of artificial organs.

[26]  Yufeng Zheng,et al.  In vitro and in vivo studies on zinc-hydroxyapatite composites as novel biodegradable metal matrix composite for orthopedic applications. , 2018, Acta biomaterialia.

[27]  G. Kerckhofs,et al.  The Impact of Type 2 Diabetes on Bone Fracture Healing , 2018, Front. Endocrinol..

[28]  M. Salehi,et al.  Regeneration of sciatic nerve crush injury by a hydroxyapatite nanoparticle-containing collagen type I hydrogel , 2018, The Journal of Physiological Sciences.

[29]  K. Yeung,et al.  Bone grafts and biomaterials substitutes for bone defect repair: A review , 2017, Bioactive materials.

[30]  N. Baldini,et al.  Focus Ion Beam/Scanning Electron Microscopy Characterization of Osteoclastic Resorption of Calcium Phosphate Substrates. , 2017, Tissue engineering. Part C, Methods.

[31]  K. Aoki,et al.  Physico-Chemical, In Vitro, and In Vivo Evaluation of a 3D Unidirectional Porous Hydroxyapatite Scaffold for Bone Regeneration , 2017, Materials.

[32]  Cuie Wen,et al.  Bioactive Materials , 2017 .

[33]  V. Kattimani,et al.  Hydroxyapatite—Past, Present, and Future in Bone Regeneration , 2016 .

[34]  S. Goodman,et al.  Inflammation, fracture and bone repair. , 2016, Bone.

[35]  Pamela Habibovic,et al.  Calcium phosphates in biomedical applications: materials for the future? , 2016 .

[36]  H. Petite,et al.  Comparative study of the osteogenic ability of four different ceramic constructs in an ectopic large animal model , 2016, Journal of tissue engineering and regenerative medicine.

[37]  Mark A. Lee,et al.  Bone Grafting: Sourcing, Timing, Strategies, and Alternatives , 2015, Journal of orthopaedic trauma.

[38]  C. V. van Blitterswijk,et al.  Elucidating the individual effects of calcium and phosphate ions on hMSCs by using composite materials. , 2015, Acta biomaterialia.

[39]  Y. Seo,et al.  Biologic properties of nano-hydroxyapatite: An in vivo study of calvarial defects, ectopic bone formation and bone implantation. , 2015, Bio-medical materials and engineering.

[40]  G. Logroscino,et al.  Bone substitutes in orthopaedic surgery: from basic science to clinical practice , 2014, Journal of Materials Science: Materials in Medicine.

[41]  E. Sacher,et al.  Nanoscale surface characterization of biphasic calcium phosphate, with comparisons to calcium hydroxyapatite and β-tricalcium phosphate bioceramics. , 2014, Journal of colloid and interface science.

[42]  J. Granjeiro,et al.  Comparative In Vivo Study of Biocompatibility of Apatites Incorporated with 1% Zinc or Lead Ions versus Stoichiometric Hydroxyapatite , 2014 .

[43]  I. Martin,et al.  Reconstruction of Extensive Long-Bone Defects in Sheep Using Porous Hydroxyapatite Sponges , 2014, Calcified Tissue International.

[44]  A. Piattelli,et al.  Blood vessels are concentrated within the implant surface concavities: a histologic study in rabbit tibia , 2014, Odontology.

[45]  Sergey V. Dorozhkin,et al.  Calcium Orthophosphate-Based Bioceramics , 2013, Materials.

[46]  L. Donahue,et al.  The influence of genetic factors on the osteoinductive potential of calcium phosphate ceramics in mice. , 2012, Biomaterials.

[47]  Melba Navarro,et al.  Control of microenvironmental cues with a smart biomaterial composite promotes endothelial progenitor cell angiogenesis. , 2012, European cells & materials.

[48]  J. Hua,et al.  Comparison of mesenchymal stem cell proliferation and differentiation between biomimetic and electrochemical coatings on different topographic surfaces , 2012, Journal of Materials Science: Materials in Medicine.

[49]  Jaebeom Lee,et al.  Nanoscale hydroxyapatite particles for bone tissue engineering. , 2011, Acta biomaterialia.

[50]  Huipin Yuan,et al.  BIOMATERIALS : CURRENT KNOWLEDGE OF PROPERTIES , EXPERIMENTAL MODELS AND BIOLOGICAL MECHANISMS , 2011 .

[51]  Huipin Yuan,et al.  Osteoinductive ceramics as a synthetic alternative to autologous bone grafting , 2010, Proceedings of the National Academy of Sciences.

[52]  X. Lu,et al.  Osteoinduction of hydroxyapatite/beta-tricalcium phosphate bioceramics in mice with a fractured fibula. , 2010, Acta biomaterialia.

[53]  Y. Leng,et al.  Study of hydroxyapatite osteoinductivity with an osteogenic differentiation of mesenchymal stem cells. , 2009, Journal of biomedical materials research. Part A.

[54]  R. Legeros,et al.  Calcium phosphate-based osteoinductive materials. , 2008, Chemical reviews.

[55]  Hyoun‐Ee Kim,et al.  Phase conversion of tricalcium phosphate into Ca-deficient apatite during sintering of hydroxyapatite-tricalcium phosphate biphasic ceramics. , 2008, Journal of biomedical materials research. Part B, Applied biomaterials.

[56]  Masanori Nakasu,et al.  Development of superporous hydroxyapatites and their examination with a culture of primary rat osteoblasts. , 2007, Journal of biomedical materials research. Part A.

[57]  V. Bousson,et al.  Long‐bone critical‐size defects treated with tissue‐engineered grafts: A study on sheep , 2007, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[58]  Alberto J Ambard,et al.  Calcium phosphate cement: review of mechanical and biological properties. , 2006, Journal of prosthodontics : official journal of the American College of Prosthodontists.

[59]  J. C. Zhu,et al.  In vivo study on biocompatibility and bonding strength of hydroxyapatite-20vol%Ti composite with bone tissues in the rabbit. , 2006, Bio-medical materials and engineering.

[60]  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.

[61]  V. Bousson,et al.  De novo reconstruction of functional bone by tissue engineering in the metatarsal sheep model. , 2005, Tissue engineering.

[62]  Karin A. Hing,et al.  Bioceramic Bone Graft Substitutes: Influence of Porosity and Chemistry , 2005 .

[63]  J. Camilli,et al.  The use of hydroxyapatite and autogenous cancellous bone grafts to repair bone defects in rats. , 2005, International journal of oral and maxillofacial surgery.

[64]  PhD Hideki Yoshikawa MD,et al.  Bone tissue engineering with porous hydroxyapatite ceramics , 2005, Journal of Artificial Organs.

[65]  K. Bachus,et al.  Determining relevance of a weight-bearing ovine model for bone ingrowth assessment. , 2004, Journal of biomedical materials research. Part A.

[66]  Arun K Gosain,et al.  A 1-year study of osteoinduction in hydroxyapatite-derived biomaterials in an adult sheep model: part I. , 2002, Plastic and reconstructive surgery.

[67]  G Pradal,et al.  Cellular mechanisms of calcium phosphate ceramic degradation. , 1999, Histology and histopathology.

[68]  C J Damien,et al.  Bone graft and bone graft substitutes: a review of current technology and applications. , 1991, Journal of applied biomaterials : an official journal of the Society for Biomaterials.

[69]  P. Eggli,et al.  Porous hydroxyapatite and tricalcium phosphate cylinders with two different pore size ranges implanted in the cancellous bone of rabbits. A comparative histomorphometric and histologic study of bony ingrowth and implant substitution. , 1988, Clinical orthopaedics and related research.

[70]  C. Klein,et al.  Biodegradation behavior of various calcium phosphate materials in bone tissue. , 1983, Journal of biomedical materials research.