Animal-bone derived hydroxyapatite in biomedical applications

Abstract The use of animal bone-derived bone for producing bone grafting and other types of materials with biomedical applications has been a fundamental part of biomaterials science since the 1950s, when Kiel bone was first developed and used. Those early experiences paved the way for the large volume of materials and clinical research that were to follow involving animal bone-derived bone. This chapter deals with the (vertebral) species of animal bone sourced for biomedical materials that was and still is principally bovine bone but that has, in recent times, branched out to include equine, porcine, cervine, ovine, fish, ostrich, and duck-head-sourced bone. The rationales for using animal-derived bone are explored and then the processing, characterization, and dependence of the ultimate phase of hydroxyapatite obtained on the process used to extract it from bone sources are discussed. The scare in Europe and other countries from the emergence of transmissible spongiform encephalopathies in cattle herds approximately 20 years ago and the link to variant Creutzfeldt–Jakob disease (v-CJD) in humans have impelled the need (as dictated by regulatory requirements, notably in Europe) to process animal-origin products, particularly bovine bone using methods that eliminate the possibility of transmission to humans of v-CJD from use of bovine-derived implants. These methods are discussed along with a brief word on the European Commission regulations that make it mandatory for manufacturers of such products to ensure their safety when used in vivo. Finally, the chapter will close with descriptions of some commercially supplied animal bone-derived bone graft materials available on the current market and the surgical studies that have used them. In addition, a selection of other clinical studies involving noncommercially sourced animal bone-derived biomedical materials will be covered. In general, the chapter will serve to show the fundamental reach of these products in the field of biomaterials science and practice.

[1]  J. Auyeung,et al.  Outcome of subtalar fusion using bovine cancellous bone graft: a retrospective case series. , 2011, The Journal of foot and ankle surgery : official publication of the American College of Foot and Ankle Surgeons.

[2]  J. Freitas,et al.  Effect of the calcination temperature on the composition and microstructure of hydroxyapatite derived from human and animal bone , 2010 .

[3]  M. Swiontkowski,et al.  Bone-graft substitutes , 1999, The Lancet.

[4]  T. Keaveny,et al.  Characterization of the mechanical and ultrastructural properties of heat-treated cortical bone for use as a bone substitute. , 1999, Journal of biomedical materials research.

[5]  Besim Ben-Nissan,et al.  Biomimetics and Bioceramics , 2004 .

[6]  N. Haas,et al.  Osseous integration of hydroxyapatite grafts in metaphyseal bone defects of the proximal tibia (CT-study). , 2002, Acta chirurgiae orthopaedicae et traumatologiae Cechoslovaca.

[7]  S. Caputi,et al.  Histological and ultrastructural evaluation of bone around Bio-Oss particles in sinus augmentation. , 2007, Oral diseases.

[8]  María Vallet-Regí,et al.  Ceramics for medical applications , 2001 .

[9]  K. Haberko,et al.  Effect of Sintering Atmosphere on the Selected Properties of the Natural Origin Hydroxyapatite Materials , 2010 .

[10]  A. Worth,et al.  The evaluation of processed cancellous bovine bone as a bone graft substitute. , 2005, Clinical oral implants research.

[11]  M. Lombardi,et al.  Gelcast Components Having Controlled Porosity Features Obtained from a Natural Hydroxyapatite Powder , 2009 .

[12]  B. Nies,et al.  Chemical and physicochemical characterization of porous hydroxyapatite ceramics made of natural bone. , 2000, Biomaterials.

[13]  A. Doostmohammadi,et al.  A comparative physico-chemical study of bioactive glass and bone-derived hydroxyapatite , 2011 .

[14]  R. Moses,et al.  The putative collagen binding peptide hastens periodontal ligament cell attachment to bone replacement graft materials. , 2001, Journal of periodontology.

[15]  Sergey V. Dorozhkin,et al.  Calcium Orthophosphates as Bioceramics: State of the Art , 2010, Journal of functional biomaterials.

[16]  Muzafar A. Kanjwal,et al.  Synthesis and characterization of bovine femur bone hydroxyapatite containing silver nanoparticles for the biomedical applications , 2011 .

[17]  L. Gotzen,et al.  [Biomechanical evaluation of biointegrable suture anchors composed of bovine compact bone in a pull-to-failure test in porcine tibial head specimens]. , 2006, Zeitschrift fur Orthopadie und ihre Grenzgebiete.

[18]  P. Herbison,et al.  Long term follow up of bone derived hydroxyapatite orbital implants , 2002, The British journal of ophthalmology.

[19]  J. Lindhe,et al.  The effect of a fibrin glue on the integration of Bio-Oss with bone tissue. A experimental study in labrador dogs. , 2002, Journal of clinical periodontology.

[20]  C. D. de Klein,et al.  Intensification of grassland and forage use: driving forces and constraints , 2014, Crop and Pasture Science.

[21]  M. Barbeck,et al.  Potential lack of "standardized" processing techniques for production of allogeneic and xenogeneic bone blocks for application in humans. , 2014, Acta biomaterialia.

[22]  M Epple,et al.  A thorough physicochemical characterisation of 14 calcium phosphate-based bone substitution materials in comparison to natural bone. , 2004, Biomaterials.

[23]  E. Sherry,et al.  Comparison of in vitro biocompatibility of NanoBone(®) and BioOss(®) for human osteoblasts. , 2011, Clinical Oral Implants Research.

[24]  B. Ben-Nissan,et al.  A Comparative Study of Thai and Australian Crocodile Bone for Use as a Potential Biomaterial , 2006 .

[25]  N. Hadrup,et al.  Oral toxicity of silver ions, silver nanoparticles and colloidal silver--a review. , 2014, Regulatory toxicology and pharmacology : RTP.

[26]  A. Molteno,et al.  "Physiological" orbital implant. , 1973, The British journal of ophthalmology.

[27]  G. Raghoebar,et al.  Applicability of equine hydroxyapatite collagen (eHAC) bone blocks for lateral augmentation of the alveolar crest. A histological and histomorphometric analysis in rats. , 2011, International journal of oral and maxillofacial surgery.

[28]  K. Haberko,et al.  Behaviour of bone origin hydroxyapatite at elevated temperatures and in O2 and CO2 atmospheres , 2009 .

[29]  S. Ramakrishna,et al.  Porous bovine hydroxyapatite for drug delivery. , 2008, Journal of applied biomaterials & biomechanics : JABB.

[30]  R. Cohen,et al.  Phenotypic characterization of mononuclear inflammatory cells following equine hydroxyapatite/collagen block grafting in rats , 2012, Biomedical materials.

[31]  D. Taylor Bovine spongiform encephalopathy--the beginning of the end? , 1996, The British veterinary journal.

[32]  S. H. Keshel,et al.  Clinical, Cosmetic and Investigational Dentistry Dovepress the Comparative Effectiveness of Demineralized Bone Matrix, Beta-tricalcium Phosphate, and Bovine-derived Anorganic Bone Matrix on Inflammation and Bone Formation Using a Paired Calvarial Defect Model in Rats , 2022 .

[33]  A. Bigham,et al.  Xenogenic demineralized bone matrix and fresh autogenous cortical bone effects on experimental bone healing: radiological, histopathological and biomechanical evaluation , 2008, Journal of Orthopaedics and Traumatology.

[34]  P Zioupos,et al.  The fracture toughness of cancellous bone. , 2009, Journal of biomechanics.

[35]  Jin-Woo Park,et al.  Increased new bone formation with a surface magnesium-incorporated deproteinized porcine bone substitute in rabbit calvarial defects. , 2012, Journal of biomedical materials research. Part A.

[36]  G. Rujijanagul,et al.  Synthesis and characterization of nanocrystalline hydroxyapatite from natural bovine bone , 2008 .

[37]  K. Haberko,et al.  Natural hydroxyapatite - Its behaviour during heat treatment , 2006 .

[38]  J. Ackerman,et al.  A Comparison of the Physical and Chemical Differences Between Cancellous and Cortical Bovine Bone Mineral at Two Ages , 2008, Calcified Tissue International.

[39]  R. Elliot,et al.  Failed operative treatment in two cases of pseudarthrosis of the clavicle using internal fixation and bovine cancellous xenograft (Tutobone). , 2011, Journal of pediatric orthopedics. Part B.

[40]  S. Meyer,et al.  Histological osseointegration of Tutobone®: first results in human , 2008, Archives of Orthopaedic and Trauma Surgery.

[41]  H. Redl,et al.  Simultaneous in vivo comparison of bone substitutes in a guided bone regeneration model. , 2008, Biomaterials.

[42]  M. Elder,et al.  Bone implants after enucleation. , 1991, Australian and New Zealand journal of ophthalmology.

[43]  A. Piattelli,et al.  Osseointegration in a sinus augmented with bovine porous bone mineral: histological results in an implant retrieved 4 years after insertion. A case report. , 2004, Journal of periodontology.

[44]  F. A. Sheikh,et al.  Physiochemical characterizations of hydroxyapatite extracted from bovine bones by three different methods: extraction of biologically desirable Hap , 2008 .

[45]  Fulin Chen,et al.  A novel hydroxyapatite ceramic bone substitute transformed by ostrich cancellous bone: characterization and evaluations of bone regeneration activity. , 2011, Journal of biomedical materials research. Part B, Applied biomaterials.

[46]  E. Firth,et al.  Combined xeno/auto-grafting of a benign osteolytic lesion in a dog, using a novel bovine cancellous bone biomaterial , 2007, New Zealand veterinary journal.

[47]  J. Wiltfang,et al.  Ectopic bone formation with the help of growth factor bFGF. , 1996, Journal of cranio-maxillo-facial surgery : official publication of the European Association for Cranio-Maxillo-Facial Surgery.

[48]  P. Revell,et al.  Coralline hydroxyapatite bone graft substitute: A review of experimental studies and biomedical applications. , 2004, Journal of applied biomaterials & biomechanics : JABB.

[49]  Stefan Schultze-Mosgau,et al.  Histologic findings in sinus augmentation with autogenous bone chips versus a bovine bone substitute. , 2003, The International journal of oral & maxillofacial implants.

[50]  J. Klawitter,et al.  History of ceramic orthopedic implants , 1972 .

[51]  B. Oesch,et al.  Analysis of the risk of transmitting bovine spongiform encephalopathy through bone grafts derived from bovine bone. , 2001, Biomaterials.

[52]  Jordan M Katz,et al.  Incorporation and Immunogenicity of Cleaned Bovine Bone in a Sheep Model , 2009, Journal of biomaterials applications.

[53]  N. Lang,et al.  The effect of a deproteinized bovine bone mineral on bone regeneration around titanium dental implants. , 1998, Clinical oral implants research.

[54]  Y. Leng,et al.  Infrared spectroscopic characterization of carbonated apatite: a combined experimental and computational study. , 2014, Journal of biomedical materials research. Part A.

[55]  M. Wolkewitz,et al.  Evaluation of Guided Bone Regeneration around Oral Implants over Different Healing Times Using Two Different Bovine Bone Materials: A Randomized, Controlled Clinical and Histological Investigation. , 2015, Clinical implant dentistry and related research.

[56]  E. Marcantonio,et al.  Comparison of biomaterial implants in the dental socket: histological analysis in dogs. , 2010, Clinical implant dentistry and related research.

[57]  W. Winkelmann,et al.  Lack of toxicological side-effects in silver-coated megaprostheses in humans. , 2007, Biomaterials.

[58]  H. Kim,et al.  Characterisation of bioresourced hydroxyapatite containing silver nanoparticles , 2012 .

[59]  K. Donath,et al.  BIO-OSS--a resorbable bone substitute? , 1998, Journal of long-term effects of medical implants.

[60]  P. Brown,et al.  Novel methods for disinfection of prion-contaminated medical devices , 2004, The Lancet.

[61]  Moustafa N. Aboushelib,et al.  Influence of Material Properties on Rate of Resorption of Two Bone Graft Materials after Sinus Lift Using Radiographic Assessment , 2012, International journal of dentistry.

[62]  Yanmin Zhao,et al.  Biological Assessment of Composite Materials Based on Poly-L-lactide and Bovine Bone , 2013 .

[63]  K. Kim,et al.  Preliminary evaluation of bone graft substitute produced by bone of duck beak , 2014 .

[64]  Su-Hyang Kim,et al.  Chemical, structural properties, and osteoconductive effectiveness of bone block derived from porcine cancellous bone. , 2004, Journal of biomedical materials research. Part B, Applied biomaterials.

[65]  Mamoru Mitsuishi,et al.  Specifications for machining the bovine cortical bone in relation to its microstructure. , 2009, Journal of biomechanics.

[66]  G. McMurray The evaluation of Kiel bone in spinal fusions. , 1982, The Journal of bone and joint surgery. British volume.

[67]  H. C. Killey,et al.  The response of the rabbit to implants of processed calf bone [boplant]. , 1968, Archives of oral biology.

[68]  A. Palmieri,et al.  Genetic effects of anorganic bovine bone (Bio-Oss) on osteoblast-like MG63 cells. , 2006, Archives of oral biology.

[69]  A. Chamberlain,et al.  Bone mineral change during experimental heating: an X-ray scattering investigation. , 2003, Biomaterials.

[70]  I. Anderson,et al.  The processing and characterization of animal-derived bone to yield materials with biomedical applications. Part III: material and mechanical properties of fresh and processed bovine cancellous bone , 2000, Journal of materials science. Materials in medicine.

[71]  D. Kim,et al.  Vertical ridge augmentation using an equine block infused with recombinant human platelet-derived growth factor-BB: a histologic study in a canine model. , 2009, The International journal of periodontics & restorative dentistry.

[72]  K. Becker,et al.  Osseous integration of bovine hydroxyapatite ceramic in metaphyseal bone defects of the distal radius. , 2000, The Journal of hand surgery.

[73]  K. Simkiss Bone and biomineralization , 1975 .

[74]  M. Murata,et al.  Osteoinduction by Functionally Graded Apatites of Bovine Origin Loaded with Bone Morphogenetic Protein-2 , 2005 .

[75]  D. Qiu,et al.  Probing the calcium and sodium local environment in bones and teeth using multinuclear solid state NMR and X-ray absorption spectroscopy. , 2010, Physical chemistry chemical physics : PCCP.

[76]  B. Wielage,et al.  The novel use of waste animal bone from New Zealand agricultural sources as a feedstock for forming plasma sprayed hydroxyapatite coatings on biomedical implant materials. , 2004, Journal of applied biomaterials & biomechanics : JABB.

[77]  X. Dereka,et al.  Effect of rhBMP-7 combined with two bone grafts on human periodontal ligament cell differentiation , 2009, Growth factors.

[78]  H. C. Killey,et al.  The response of the rabbit to implants of processed bovine bone (Kiel bone) and the effects of varying the relationship between implant and host bone. , 1970, Archives of oral biology.

[79]  K. S. Chen,et al.  Preparation of a biphasic porous bioceramic by heating bovine cancellous bone with Na4P2O7.10H2O addition. , 1999, Biomaterials.

[80]  D. Manning,et al.  Structural and chemical changes of thermally treated bone apatite , 2007 .

[81]  M. Mucalo,et al.  The processing and characterization of animal-derived bone to yield materials with biomedical applications. Part II: milled bone powders, reprecipitated hydroxyapatite and the potential uses of these materials , 2000, Journal of materials science. Materials in medicine.

[82]  I. Tsesis,et al.  Effect of guided tissue regeneration on newly formed bone and cementum in periapical tissue healing after endodontic surgery: an in vivo study in the cat. , 2012, Journal of endodontics.

[83]  A. Icaro Cornaglia,et al.  Ten-year follow-up in a maxillary sinus augmentation using anorganic bovine bone (Bio-Oss). A case report with histomorphometric evaluation. , 2003, Clinical oral implants research.

[84]  A. Bigham,et al.  Experimental bone defect healing with xenogenic demineralized bone matrix and bovine fetal growth plate as a new xenograft: radiological, histopathological and biomechanical evaluation , 2009, Cell and Tissue Banking.

[85]  Simin Li,et al.  Variability and anisotropy of mechanical behavior of cortical bone in tension and compression. , 2013, Journal of The Mechanical Behavior of Biomedical Materials.

[86]  M. Mucalo,et al.  Vacuum-assisted infiltration of chitosan or polycaprolactone as a structural reinforcement for sintered cancellous bovine bone graft. , 2012, Journal of biomedical materials research. Part A.

[87]  E. Kusrini,et al.  Characterization of x-ray diffraction and electron spin resonance: Effects of sintering time and temperature on bovine hydroxyapatite , 2012 .

[88]  N. Hunter,et al.  Transmissible spongiform encephalopathies: transmission, mechanism of disease, and persistence. , 1998, Current opinion in microbiology.

[89]  M. Pietruska A comparative study on the use of Bio-Oss and enamel matrix derivative (Emdogain) in the treatment of periodontal bone defects. , 2001, European journal of oral sciences.

[90]  W. Neuman,et al.  THE CHEMICAL DYNAMICS OF BONE MINERAL , 1959 .

[91]  I. Oladele Development of Bone Ash and Bone Particulate Reinforced Polyester Composites for Biomedical Applications , 2013 .

[92]  G. Martins,et al.  Physicochemical characterization of biomaterials commonly used in dentistry as bone substitutes--comparison with human bone. , 2010, Journal of biomedical materials research. Part B, Applied biomaterials.

[93]  M. Mucalo,et al.  The processing and characterization of animal-derived bone to yield materials with biomedical applications Part 1: Modifiable porous implants from bovine condyle cancellous bone and characterization of bone materials as a function of processing , 2000, Journal of materials science. Materials in medicine.

[94]  T. Kumar,et al.  Heat-deproteinated xenogeneic bone from slaughterhouse waste: Physico-chemical properties , 2003 .

[95]  A. Scarano,et al.  Bone reactions to anorganic bovine bone (Bio-Oss) used in sinus augmentation procedures: a histologic long-term report of 20 cases in humans. , 1999, The International journal of oral & maxillofacial implants.

[96]  S. Ramakrishna,et al.  Nanoporous hydroxy-carbonate apatite scaffold made of natural bone , 2006 .

[97]  D. Benke,et al.  Protein-chemical analysis of Bio-Oss bone substitute and evidence on its carbonate content. , 2001, Biomaterials.

[98]  W. Nakkiew,et al.  Preparation and Characterisation of Cattle Bone Based Hydroxyapatite Implant Coatings Using Electrostatic Spray Deposition , 2013 .

[99]  C. Bourauel,et al.  Bone substitute material composition and morphology differentially modulate calcium and phosphate release through osteoclast-like cells. , 2014, International journal of oral and maxillofacial surgery.

[100]  S. Ramesh,et al.  Properties of hydroxyapatite produced by annealing of bovine bone , 2007 .

[101]  G. Bakalim,et al.  Kiel bone in the surgical treatment of tibial condylar fractures. , 1972, Acta orthopaedica Scandinavica.

[102]  Hyung-Sup Kim,et al.  Preparation of a novel anorganic bovine bone xenograft with enhanced bioactivity and osteoconductivity. , 2013, Journal of biomedical materials research. Part B, Applied biomaterials.

[103]  Iain A Anderson,et al.  Orthogonal cutting of cancellous bone with application to the harvesting of bone autograft. , 2008, Medical engineering & physics.

[104]  F. Oktar,et al.  Plasma-sprayed bovine hydroxyapatite coatings , 2004 .

[105]  Virgílio M Roriz,et al.  Treatment of Class III furcation defects with expanded polytetrafluoroethylene membrane associated or not with anorganic bone matrix/synthetic cell-binding peptide: a histologic and histomorphometric study in dogs. , 2006, Journal of periodontology.

[106]  New studies on the heat resistance of hamster-adapted scrapie agent: threshold survival after ashing at 600 degrees C suggests an inorganic template of replication. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[107]  J. Kolmas,et al.  Solid-state NMR and IR characterization of commercial xenogeneic biomaterials used as bone substitutes. , 2012, Journal of pharmaceutical and biomedical analysis.

[108]  A BAUERMEISTER,et al.  A method of bone maceration; results in animal experiments. , 1957, The Journal of bone and joint surgery. American volume.

[109]  A. Sola,et al.  Characterization and in vitro-bioactivity of natural hydroxyapatite based bio-glass―ceramics synthesized by thermal plasma processing , 2010 .

[110]  S. Caputi,et al.  Maxillary Sinus Augmentation With Different Biomaterials: A Comparative Histologic and Histomorphometric Study in Man , 2006, Implant dentistry.

[111]  E. Elvevoll,et al.  The Emerging Importance of Dietary Lipids, Quantity and Quality, in the Global Disease Burden: the Potential of Aquatic Resources , 2001, Nutrition and health.

[112]  Mohd Hamdi,et al.  The influence of sintering temperature on the properties of compacted bovine hydroxyapatite , 2009 .

[113]  R. Aspden,et al.  Effect of the Proportion of Organic Material in Bone on Thermal Decomposition of Bone Mineral: An Investigation of a Variety of Bones from Different Species Using Thermogravimetric Analysis coupled to Mass Spectrometry, High-Temperature X-ray Diffraction, and Fourier Transform Infrared Spectroscopy , 2004, Calcified Tissue International.

[114]  P. Giannoudis,et al.  Reconstruction of iliac crest with bovine cancellous allograft after bone graft harvest for symphysis pubis arthrodesis , 2012, International Orthopaedics.

[115]  Michael T. Wilson,et al.  In vitro antibacterial efficacy of tetracycline hydrochloride adsorbed onto Bio-Oss bone graft. , 2010, Journal of biomedical materials research. Part B, Applied biomaterials.

[116]  Jui-Sheng Sun,et al.  Preparation of ?TCP/HAP biphasic ceramics with natural bone structure by heating bovine cancellous bone with the addition of (NH4)2HPO4 , 2000 .

[117]  M. U. Helber,et al.  [Metaphyseal defect substitute: hydroxylapatite ceramic. Results of a 3 to 4 year follow up]. , 2000, Der Unfallchirurg.

[118]  Yi‐Cheng Huang,et al.  Hydroxyapatite extracted from fish scale: Effects on MG63 osteoblast-like cells , 2011 .

[119]  S. Tröster,et al.  [Hydroxyapatite ceramics in clinical application. Histological findings in 23 patients]. , 1997, European Journal of Trauma.

[120]  M. Lombardi,et al.  Processing of a natural hydroxyapatite powder: From powder optimization to porous bodies development , 2011 .

[121]  S. Fulle,et al.  Functional assay, expression of growth factors and proteins modulating bone-arrangement in human osteoblasts seeded on an anorganic bovine bone biomaterial. , 2010, European cells & materials.

[122]  M. Hamonic,et al.  Bone grafting with Boplant. Results in thirty-three cases. , 1968, The Journal of bone and joint surgery. British volume.

[123]  Z. Wzorek,et al.  The influence of calcination parameters on free calcium oxide content in natural hydroxyapatite , 2012 .

[124]  A. Worth,et al.  Comparison between a novel bovine xenoimplant and autogenous cancellous bone graft in tibial tuberosity advancement. , 2012, Veterinary surgery : VS.

[125]  A. U. Daniels,et al.  Preparation, chemistry and physical properties of bone-derived hydroxyapatite particles having a negative zeta potential , 2012 .

[126]  W. Bonfield,et al.  Biomechanical assessment of bone ingrowth in porous hydroxyapatite , 1997, Journal of materials science. Materials in medicine.

[127]  M. Połomska,et al.  FT NIR Raman studies on γ-irradiated bone , 2007 .

[128]  F. Lusquiños,et al.  Biological hydroxyapatite obtained from fish bones , 2012 .

[129]  M. Khil,et al.  Extraction of pure natural hydroxyapatite from the bovine bones bio waste by three different methods , 2009 .

[130]  P. Biskupski,et al.  Radiation sterilized bone response to dynamic loading. , 2012, Materials science & engineering. C, Materials for biological applications.

[131]  M. Bosetti,et al.  Evaluation of bioresorbable implants from bovine bone: In vitro preliminary observations. , 2008, Journal of applied biomaterials & biomechanics : JABB.

[132]  J. Fages,et al.  Histological evaluation of xenogeneic bone treated by supercritical CO2 implanted into sheep , 1995 .

[133]  A. A. Muraev,et al.  Synthetic materials used for the substitution of bone defects , 2013 .

[134]  M. Horton,et al.  Human osteoclast formation and activity on a xenogenous bone mineral. , 2009, Journal of biomedical materials research. Part A.

[135]  K. Urban,et al.  In vivo behaviour of low-temperature calcium-deficient hydroxyapatite: comparison with deproteinised bovine bone , 2011, International Orthopaedics.

[136]  E. Huffman,et al.  Determination of trace organic carbon and nitrogen in the presence of carbonates in anorganic bovine bone graft materials , 2003 .

[137]  R Z LeGeros,et al.  A Raman and infrared spectroscopic investigation of biological hydroxyapatite. , 1990, Journal of inorganic biochemistry.

[138]  R E Horch,et al.  Evaluation of processed bovine cancellous bone matrix seeded with syngenic osteoblasts in a critical size calvarial defect rat model , 2006, Journal of cellular and molecular medicine.

[139]  C. Seebach,et al.  Comparison of six bone-graft substitutes regarding to cell seeding efficiency, metabolism and growth behaviour of human mesenchymal stem cells (MSC) in vitro. , 2010, Injury.

[140]  C. Ulrich,et al.  Metaphysärer Defektersatz mit Hydrosylapatitkeramik 3- bis 4-Jahresnachuntersuchungs-Ergebnisse , 2000, Der Unfallchirurg.

[141]  A. Rakowska,et al.  Chemical and microstructural characterization of natural hydroxyapatite derived from pig bones , 2008 .

[142]  Leszek Kubisz,et al.  Differential scanning calorimetry and temperature dependence of electric conductivity in studies on denaturation process of bone collagen , 2005 .

[143]  C. Rey Calcium Phosphates for Medical Applications , 1998 .

[144]  Vilmos Vécsei,et al.  Combination of anorganic bovine‐derived hydroxyapatite with binding peptide does not enhance bone healing in a critical‐size defect in a rabbit model , 2008, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.