Conversion of natural marine skeletons as scaffolds for bone tissue engineering

Abstract

Marine CaCO₃ skeletons have tailored architectures created by nature, which give them structural support and other functions. For example, seashells have dense lamellar structures, while coral, cuttlebone and sea urchin spines have interconnected porous structures. In our experiments, seashells, coral and cuttlebone were hydrothermally converted to hydroxyapatite (HAP), and sea urchin spines were converted to Mg-substituted tricalcium phosphate, while maintaining their original structures. Partially converted shell samples have mechanical strength, which is close to that of compact human bone. After implantation of converted shell and spine samples in rat femoral defects for 6 weeks, there was newly formed bone growth up to and around the implants. Some new bone was found to migrate through the pores of converted spine samples and grow inward. These results show good bioactivity and osteoconductivity of the implants, indicating the converted shell and spine samples can be used as bone defect fillers. The interconnected porous HAP scaffolds from converted coral or cuttlebone that have pore size larger than 100 μm likely support infiltration of bone cells and vessels, and finally encourage new bone ingrowth.

Keywords

seashell / sea urchin spine / coral / cuttlebone / hydrothermal conversion / hydroxyapatite (HAP) / β-tricalcium phosphate (β-TCP)

References

[1] Laurencin C T, Attawia M, Borden M D. Advancements in tissue engineered bone substitutes. Current Opinion in Orthopaedics , 1999, 10(6): 445–451
[2] Enneking W. Transplanting allografts. Journal of the American College of Surgeons , 2005, 201(1): 5–6
[3] LeGeros R Z. Biodegradation and bioresorption of calcium phosphate ceramics. Clinical Materials , 1993, 14(1): 65–88
[4] Karageorgiou V, Kaplan D. Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials , 2005, 26(27): 5474–5491
[5] Hing K A. Bioceramic bone graft substitutes: Influence of porosity and chemistry. International Journal of Applied Ceramic Technology , 2005, 2(3): 184–199
[6] LeGeros R Z. Properties of osteoconductive biomaterials: calcium phosphates. Clinical Orthopaedics and Related Research , 2002, 395(395): 81–98
[7] Vallet-Regí M, González-Calbet J M. Calcium phosphates as substitution of bone tissues. Progress in Solid State Chemistry , 2004, 32(1-2): 1–31
[8] Aoki H, Kato K, Ebihara M, . Studies on the application of apatite to dental materials. (I) Apatite ceramics. Shika Rikogaku Zasshi , 1976, 17(39): 200–205
[9] Barney V C, Levin M P, Adams D F. Bioceramic implants in surgical periodontal defects. A comparison study. Journal of Periodontology , 1986, 57(12): 764–770
[10] Cranin A N, Tobin G P, Gelbman J. Applications of hydroxylapatite in oral and maxillofacial surgery. Part II: Ridge augmentation and repair of major oral defects. Compendium , 1987, 8(5): 334–335 , 337 -338 , 340
[11] Denissen H W, de Groot K. Immediate dental root implants from synthetic dense calcium hydroxylapatite. The Journal of Prosthetic Dentistry , 1979, 42(5): 551–556
[12] Ellinger R F, Nery E B, Lynch K L. Histological assessment of periodontal osseous defects following implantation of hydroxyapatite and biphasic calcium phosphate ceramics: a case report. The International Journal of Periodontics & Restorative Dentistry , 1986, 6(3): 22–33
[13] Albee F H, Morrison H A R O L D F. Studies in bone growth: triple calcium phosphate as a stimulus to osteogenesis. Annals of Surgery , 1920, 71(1): 32–39
[14] Daculsi G, Passuti N, Martin S, . Macroporous calcium phosphate ceramic for long bone surgery in humans and dogs. Clinical and histological study. Journal of Biomedical Materials Research , 1990, 24(3): 379–396
[15] Sun L, Berndt C C, Gross K A, . Material fundamentals and clinical performance of plasma-sprayed hydroxyapatite coatings: a review. Journal of Biomedical Materials Research , 2001, 58(5): 570–592
[16] Hing K A, Revell P A, Smith N, . Effect of silicon level on rate, quality and progression of bone healing within silicate-substituted porous hydroxyapatite scaffolds. Biomaterials , 2006, 27(29): 5014–5026
[17] Kondo N, Ogose A, Tokunaga K, . Bone formation and resorption of highly purified beta-tricalcium phosphate in the rat femoral condyle. Biomaterials , 2005, 26(28): 5600–5608
[18] Hasegawa S, Ishii S, Tamura J, . A 5-7 year in vivo study of high-strength hydroxyapatite/poly(l-lactide) composite rods for the internal fixation of bone fractures. Biomaterials , 2006, 27(8): 1327–1332
[19] Ohura K, Bohner M, Hardouin P, . Resorption of, and bone formation from, new β-tricalcium phosphate-monocalcium phosphate cements: An in vivo study. Journal of Biomedical Materials Research , 1996, 30(2): 193–200
[20] Tanaka T, Kumagae Y, Saito M, . Bone formation and resorption in patients after implantation of β-tricalcium phosphate blocks with 60% and 75% porosity in opening-wedge high tibial osteotomy. Journal of Biomedical Materials Research Part B: Applied Biomaterials , 2008, 86B(2): 453–459
[21] Okuda T, Ioku K, Yonezawa I, . The effect of the microstructure of beta-tricalcium phosphate on the metabolism of subsequently formed bone tissue. Biomaterials , 2007, 28(16): 2612–2621
[22] Yoshikawa H, Tamai N, Murase T, . Interconnected porous hydroxyapatite ceramics for bone tissue engineering. Journal of the Royal Society Interface , 2009, 6(Suppl 3): S341–S348
[23] Bose S, Roy M, Bandyopadhyay A. Recent advances in bone tissue engineering scaffolds. Trends in Biotechnology , 2012, 30(10): 546–554
[24] Suchanek W, Yoshimura M. Processing and properties of hydroxyapatite-based biomaterials for use as hard tissue replacement implants. Journal of Materials Research , 1998, 13(01): 94–117
[25] Kaplan D L. Mollusc shell structures: novel design strategies for synthetic materials. Current Opinion in Solid State and Materials Science , 1998, 3(3): 232–236
[26] Belcher A M, Wu X H, Christensen R J, . Control of crystal phase switching and orientation by soluble mollusc-shell proteins. Nature , 1996, 381(6577): 56–58
[27] Sudo S, Fujikawa T, Nagakura T, . Structures of mollusc shell framework proteins. Nature , 1997, 387(6633): 563–564
[28] Vecchio K S, Zhang X, Massie J B, . Conversion of bulk seashells to biocompatible hydroxyapatite for bone implants. Acta Biomaterialia , 2007, 3(6): 910–918
[29] Roy D M, Linnehan S K. Hydroxyapatite formed from coral skeletal carbonate by hydrothermal exchange. Nature , 1974, 247(5438): 220–222
[30] Birchall J D, Thomas N L. On the architecture and function of cuttlefish bone. Journal of Materials Science , 1983, 18(7): 2081–2086
[31] Kim B-S, Kim J S, Sung H-M, . Cellular attachment and osteoblast differentiation of mesenchymal stem cells on natural cuttlefish bone. Journal of Biomedical Materials Research Part A , 2012, 100A(7): 1673–1679
[32] Vecchio K S, Zhang X, Massie J B, . Conversion of sea urchin spines to Mg-substituted tricalcium phosphate for bone implants. Acta Biomaterialia , 2007, 3(5): 785–793
[33] Lin A Y M, Meyers M A, Vecchio K S. Mechanical properties and structure of Strombus gigas, Tridacna gigas, and Haliotis rufescens sea shells: A comparative study. Materials Science and Engineering C , 2006, 26(8): 1380–1389
[34] Camprasse S, Camprasse G, Pouzol M, . Artificial dental root made of natural calcium carbonate (bioracine). Clinical Materials , 1990, 5(2-4): 235–250
[35] Lamghari M, Almeida M J, Berland S, . Stimulation of bone marrow cells and bone formation by nacre: in vivo and in vitro studies. Bone , 1999, 25(2 Supplement 1): 91S–94S
[36] Lopez E, Vidal B, Berland S, . Demonstration of the capacity of nacre to induce bone formation by human osteoblasts maintained in vitro. Tissue and Cell , 1992, 24(5): 667–679
[37] Atlan G, Delattre O, Berland S, . Interface between bone and nacre implants in sheep. Biomaterials , 1999, 20(11): 1017–1022
[38] Zhang X, Vecchio K S. Creation of dense hydroxyapatite (synthetic bone) by hydrothermal conversion of seashells. Materials Science and Engineering C , 2006, 26(8): 1445–1450
[39] Menig R, Meyers M H, Meyers M A, . Quasi-static and dynamic mechanical response of Strombus gigas (conch) shells. Materials Science and Engineering A , 2001, 297(1-2): 203–211
[40] Hou D F, Zhou G S, Zheng M. Conch shell structure and its effect on mechanical behaviors. Biomaterials , 2004, 25(4): 751–756
[41] Kamat S, Su X, Ballarini R, . Structural basis for the fracture toughness of the shell of the conch Strombus gigas. Nature , 2000, 405(6790): 1036–1040
[42] Weber J N, White E W, Lebiedzik J. New porous biomaterials by replication of echinoderm skeletal microstructures. Nature , 1971, 233(5318): 337–339
[43] Rocha J H, Lemos A F, Agathopoulos S, . Hydrothermal growth of hydroxyapatite scaffolds from aragonitic cuttlefish bones. Journal of Biomedical Materials Research Part A , 2006, 77A(1): 160–168
[44] LeGeros R Z, Gatti A M, Kijkowska R, . Mg-substituted tricalcium phosphates: formation and properties. Key Engineering Materials , 2004, 254-256: 127–130
[45] Zhang X, Takahashi T, Vecchio K S. Development of bioresorbable Mg-substituted tricalcium phosphate scaffolds for bone tissue engineering. Materials Science and Engineering C , 2009, 29(6): 2003–2010
[46] Zhang X, Jiang F, Groth T, . Preparation, characterization and mechanical performance of dense β-TCP ceramics with/without magnesium substitution. Journal of Materials Science: Materials in Medicine , 2008, 19(9): 3063–3070
[47] Rezwan K, Chen Q Z, Blaker J J, . Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. Biomaterials , 2006, 27(18): 3413–3431
[48] Wasielewski R C, Sheridan K C, Lubbers M A. Coralline hydroxyapatite in complex acetabular reconstruction. Orthopedics , 2008, 31(4): 367
[49] Knackstedt M A, Arns C H, Senden T J, . Structure and properties of clinical coralline implants measured via 3D imaging and analysis. Biomaterials , 2006, 27(13): 2776–2786
[50] Hsu C-J, Chou W-Y, Teng H-P, . Coralline hydroxyapatite and laminectomy-derived bone as adjuvant graft material for lumbar posterolateral fusion. Journal of Neurosurgery: Spine , 2005, 3(4): 271–275
[51] Ivankovic H, Gallego Ferrer G, Tkalcec E, . Preparation of highly porous hydroxyapatite ceramics from cuttlefish bone. Advances in Science and Technology , 2006, 49: 142–147
[52] Zhang X. Preparation and characterization of calcium phosphate ceramics and composites as bone substitutes. Dissertation for the Doctoral Degree . San Diego, USA: University of California, 2007
[53] Klawitter J J, Bagwell J G, Weinstein A M, . An evaluation of bone growth into porous high density polyethylene. Journal of Biomedical Materials Research , 1976, 10(2): 311–323
[54] Klawitter J J, Hulbert S F. Application of porous ceramics for the attachment of load bearing internal orthopedic applications. Journal of Biomedical Materials Research Part A , 1971, 5(6): 161–229
[55] Sherrard K M. Cuttlebone morphology limits habitat depth in eleven species of Sepia (Cephalopoda: Sepiidae). The Biological Bulletin , 2000, 198(3): 404–414
[56] Denton E J, Gilpin-Brown J B. The buoyancy of the cuttlefish, Sepia officinalis (L.). Journal of the Marine Biological Association of the United Kingdom , 1961, 41(2): 319–342
[57] Walsh P J, Buchanan F J, Dring M, . Low-pressure synthesis and characterisation of hydroxyapatite derived from mineralise red algae. Chemical Engineering Journal , 2008, 137(1): 173–179
[58] Felicio-Fernandes G, Laranjeira M C M. Calcium phosphate biomaterials from marine algae. Hydrothermal synthesis and characterisation. Quimica Nova , 2000, 23(4): 441–446
[59] dePaula S M, Huila M F, Araki K, . Confocal Raman and electronic microscopy studies on the topotactic conversion of calcium carbonate from Pomacea lineate shells into hydroxyapatite bioceramic materials in phosphate media. Micron , 2010, 41(8): 983–989
[60] Cunningham E, Dunne N, Walker G, . Hydroxyapatite bone substitutes developed via replication of natural marine sponges. Journal of Materials Science: Materials in Medicine , 2010, 21(8): 2255–2261