An improved collagen scaffold for skeletal regeneration
Serafim M. Oliveira
Department of Mechanical Engineering, ESTG-Escola Superior de Tecnologia e Gestão, 3504-510 Viseu, Portugal
Divisao de Biomateriais, INEB-Instituto de Engenharia Biomédica, 3500 Porto, Portugal
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010
Search for more papers by this authorRushali A. Ringshia
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010
Search for more papers by this authorRacquel Z. Legeros
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010
Search for more papers by this authorElizabeth Clark
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010
Search for more papers by this authorMichael J. Yost
Department of Surgery, University of South Carolina, Columbia, South Carolina 29208
Search for more papers by this authorLouis Terracio
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010
Search for more papers by this authorCorresponding Author
Cristina C. Teixeira
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010Search for more papers by this authorSerafim M. Oliveira
Department of Mechanical Engineering, ESTG-Escola Superior de Tecnologia e Gestão, 3504-510 Viseu, Portugal
Divisao de Biomateriais, INEB-Instituto de Engenharia Biomédica, 3500 Porto, Portugal
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010
Search for more papers by this authorRushali A. Ringshia
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010
Search for more papers by this authorRacquel Z. Legeros
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010
Search for more papers by this authorElizabeth Clark
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010
Search for more papers by this authorMichael J. Yost
Department of Surgery, University of South Carolina, Columbia, South Carolina 29208
Search for more papers by this authorLouis Terracio
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010
Search for more papers by this authorCorresponding Author
Cristina C. Teixeira
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010
Department of Basic Science and Craniofacial Biology/Department of Biomaterials & Biomimetics, New York University College of Dentistry, New York, New York 10010Search for more papers by this authorAbstract
Bone repair and regeneration is one of the most extensively studied areas in the field of tissue engineering. All of the current tissue engineering approaches to create bone focus on intramembranous ossification, ignoring the other mechanism of bone formation, endochondral ossification. We propose to create a transient cartilage template in vitro, which could serve as an intermediate for bone formation by the endochondral mechanism once implanted in vivo. The goals of the study are (1) to prepare and characterize type I collagen sponges as a scaffold for the cartilage template, and (2) to establish a method of culturing chondrocytes in type I collagen sponges and induce cell maturation. Collagen sponges were generated from a 1% solution of type I collagen using a freeze/dry technique followed by UV light crosslinking. Chondrocytes isolated from two locations in chick embryo sterna were cultured in these sponges and treated with retinoic acid to induce chondrocyte maturation and extracellular matrix deposition. Material strength testing as well as microscopic and biochemical analyzes were conducted to evaluate the properties of sponges and cell behavior during the culture period. We found that our collagen sponges presented improved stiffness and supported chondrocyte attachment and proliferation. Cells underwent maturation, depositing an abundant extracellular matrix throughout the scaffold, expressing high levels of type X collagen, type I collagen and alkaline phosphatase. These results demonstrate that we have created a transient cartilage template with potential to direct endochondral bone formation after implantation. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res 2010
References
- 1 Pountos I,Jones E,Tzioupis C,Mcgonagle D,Giannoudis PV. Growing bone and cartilage. The role of mesenchymal stem cells. J Bone Joint Surg Br 2006; 88: 421–426.
- 2 Newman AP. Articular cartilage repair. Am J Sports Med 1998; 26: 309–324.
- 3 Tomihisa K,Tomoo M,Toshitaka T,Tomoyuki S. New bone formation around porous hydroxyapatite wedge implanted in opening wedge high tibial osteotomy in patients with osteoarthritis. Biomaterials 2001; 22: 1579–1582.
- 4 Weiss P,Layrolle P,Clergeau LP,Enckel B,Pilet P,Amouriq Y,Daculsi G,Giumelli B. The safety and efficacy of an injectable bone substitute in dental sockets demonstrated in a human clinical trial. Biomaterials 2007; 28: 3295–3305.
- 5 Verlaan JJ,Oner FC,Dhert WJ. Anterior spinal column augmentation with injectable bone cements. Biomaterials 2006; 27: 290–301.
- 6 Zelzer E,Olsen BR. The genetic basis for skeletal diseases. Nature 2003; 423: 343–348.
- 7 Ivaska KK,Gerdhem P,Åkesson K,Garnero P,Obrant KJ. Effect of fracture on bone turnover markers: A longitudinal study comparing marker levels before and after injury in 113 elderly women. J Bone Miner Res 2007; 22: 1155–1164.
- 8 Hunziker EB. Articular cartilage repair: Basic science and clinical progress. A review of the current status and prospects. Osteoarthritis Cartilage 2002; 10: 432–463.
- 9 Curran SJ,Chen R,Curran JM,Hunt JA. Expansion of human chondrocytes in an intermittent stirred flow bioreactor, using modified biodegradable microspheres. Tissue Eng 2005; 11: 1312–1322.
- 10 Vinatier C,Magne D,Moreau A,Gauthier O,Malard O,Vignes-Colombeix C,Daculsi G,Weiss P,Guicheux J. Engineering cartilage with human nasal chondrocytes and a silanized hydroxypropyl methylcellulose hydrogel. J Biomed Mater Res Part A 2007; 80: 66–74.
- 11 Pound JC,Green DW,Chaudhuri JB,Mann S,Roach HI,Oreffo RO. Strategies to promote chondrogenesis and osteogenesis from human bone marrow cells and articular chondrocytes encapsulated in polysaccharide templates. Tissue Eng 2006; 12: 2789–2799.
- 12 Kasten P,Vogel J,Luginbuhl R,Niemeyer P,Tonak M,Lorenz H,Helbig L,Weiss S,Fellenberg J,Leo A. Ectopic bone formation associated with mesenchymal stem cells in a resorbable calcium deficient hydroxyapatite carrier. Biomaterials 2005; 26: 5879–5889.
- 13 Lee JY,Musgrave D,Pelinkovic D,Fukushima K,Cummins J,Usas A,Robbins P,Fu FH,Huard J. Effect of bone morphogenetic protein-2-expressing muscle-derived cells on healing of critical-sized bone defects in mice. J Bone Joint Surg Am 2001; 83: 1032–1039.
- 14 Sumanasinghe RD,Osborne JA,Loboa EG. Mesenchymal stem cell-seeded collagen matrices for bone repair: Effects of cyclic tensile strain, cell density, and media conditions on matrix contraction in vitro. J Biomed Mater Res Part A 2008; 88: 778–786.
- 15 Alsberg E,Anderson KW,Albeiruti A,Rowley JA,Mooney DJ. Engineering growing tissues. Proc Natl Acad Sci USA 2002; 99: 12025–12030.
- 16 Arpornmaeklong P,Suwatwirote N,Pripatnanont P,Oungbho K. Growth and differentiation of mouse osteoblasts on chitosan-collagen sponges. Int J Oral Maxillacial Surgery 2007; 36: 328–337.
- 17 Lode A,Bernhardt A,Gelinsky M. Cultivation of human bone marrow stromal cells on three-dimensional scaffolds of mineralized collagen: influence of seeding density on colonization, proliferation and osteogenic differentiation. J Tissue Eng Regenerative Med 2008; 2: 400–407.
- 18 Balooch M,Habelitz S,Kinney JH,Marshall SJ,Marshall GW. Mechanical properties of mineralized collagen fibrils as influenced by demineralization. J Struct Biol 2008; 162: 404–410.
- 19 Gelse K,Poschl E,Aigner T. Collagens-structure, function, and biosynthesis. Adv Drug Deliv Rev 2003; 55: 1531–1546.
- 20 Berglund JD,Mohseni MM,Nerem RM,Sambanis A. A biological hybrid model for collagen-based tissue engineered vascular constructs. Biomaterials 2003; 24: 1241–1254.
- 21 Chvapil M. Collagen sponge: Theory and practice of medical applications. J Biomed Mater Res 1977; 11: 721–741.
- 22 Chvapil M,Kronenthal L,Van Winkle W. Medical and surgical applications of collagen. Int Rev Connect Tissue Res 1973; 6: 1–61.
- 23 Matton G,Anseeuw A,De Keyser F. The history of injectable biomaterials and the biology of collagen. Aesthetic Plast Surg 1985; 9: 133–140.
- 24 Radhika M,Babu M,Sehgal PK. Cellular proliferation on desamidated collagen matrices. Comp Biochem Physiol C Pharmacol Toxicol Endocrinol 1999; 124: 131–139.
- 25 Elliott JT,Woodward JT,Langenbach KJ,Tona A,Jones PL,Plant AL. Vascular smooth muscle cell response on thin films of collagen. Matrix Biol 2005; 24: 489–502.
- 26 Song J,Rolfe BE,Hayward IP,Campbell GR,Campbell JH. Effects of collagen gel configuration on behavior of vascular smooth muscle cells in vitro: Association with vascular morphogenesis. In Vitro Cell Dev Biol Anim 2000; 36: 600–610.
- 27 Weinberg CB,Bell E. A blood vessel model constructed from collagen and cultured vascular cells. Science 1986; 231: 397–400.
- 28 Reid GG,Gorham SD,Lackie JM. The attachment, spreading and growth of baby hamster-kidney cells on collagen, chemically modified collagen and collagen-composite substrata. J Mater Sci-Mater Med 1993; 4: 201–209.
- 29 Michalopoulos G,Pitot HC. Primary culture of parenchymal liver cells on collagen membranes. Morphological and biochemical observations. Exp Cell Res 1975; 94: 70–78.
- 30 Winder SJ,Turvey A,Forsyth IA. Characteristics of ruminant mammary epithelial cells grown in primary culture in serum-free medium. J Dairy Res 1992; 59: 491–498.
- 31 Sato T,Chen G,Ushida T,Ishii T,Ochiai N,Tateishi T. Tissue-engineered cartilage by in vivo culturing of chondrocytes in PLGA-collagen hybrid sponge. Mater Sci Engineering: C 2001; 17: 83–89.
- 32 Takahiro O,Keizo T,Yosuke H,Takashi U,Tamotsu T,Tetsuya T. Effect of type I and type II collagen sponges as 3D scaffolds for hyaline cartilage-like tissue regeneration on phenotypic control of seeded chondrocytes in vitro. Mater Sci Eng C 2004; 24: 407–411.
- 33 Fuchs JR,Kaviani A,Oh JT,Lavan D,Udagawa T,Jennings RW,Wilson JM,Fauza DO. Diaphragmatic reconstruction with autologous tendon engineered from mesenchymal amniocytes. J Pediatr Surg 2004; 39: 834–838.
- 34 Kall S,Nöth U,Reimers K,Choi CYU,Muehlberger T,Allmeling C,Jahn S,Heymer A,Vogt PM. In vitro fabrication of tendon substitutes using human mesenchymal stem cells and a collagen type i gel. Handchir Mikrochir Plast Chir 2004; 4: 205–211.
10.1055/s-2004-815745 Google Scholar
- 35 Domaschke H,Gelinsky M,Burmeister B,Fleig R,Hanke T,Reinstorf A,Pompe W,Rosen-Wolff A. In vitro ossification and remodeling of mineralized collagen i scaffolds. Tissue Eng 2006; 12: 949–958.
- 36 Rodrigues CVM,Serricella P,Linhares ABR,Guerdes RM,Borojevic R,Rossi MA,Duarte MEL,Farina M. Characterization of a bovine collagen-hydroxyapatite composite scaffold for bone tissue engineering. Biomaterials 2003; 24: 4987–4997.
- 37 Cooperman L,Michaeli D. The immunogenicity of injectable collagen. I. A 1-year prospective study. J Am Acad Dermatol 1984; 10: 638–646.
- 38 Delustro F,Condell RA,Nguyen MA,Mcpherson JM. A comparative study of the biologic and immunologic response to medical devices derived from dermal collagen. J Biomed Mater Res 1986; 20: 109–120.
- 39 Sandberg M,Vurio E. Localization of types I, II, and III collagen mRNAs in developing human skeletal tissues by in situ hybridization. J Cell Biol 1987; 104: 1077–1084.
- 40 Delustro F,Smith ST,Sundsmo J,Salem G,Kincaid S,Ellingsworth L. Reaction to injectable collagen: Results in animal models and clinical use. Plast Reconstr Surg 1987; 79: 581–594.
- 41 Lynn AK,Yannas IV,Bonfield W. Antigenicity and immunogenicity of collagen. J Biomed Mater Res Part B: Appl Biomaterials 2004; 71: 343–354.
- 42 Mizuno M,Shindo M,Kobayashi D,Tsuruga E,Amemiya A,Kuboki Y. Osteogenesis by bone marrow stromal cells maintained on type I collagen matrix gels in vivo. Bone 1997; 20: 101–107.
- 43 Wambach BA,Cheung H,Josephson GD. Cartilage tissue engineering using thyroid chondrocytes on a type i collagen matrix. Laryngoscope 2000; 110: 2008–2011.
- 44 Rajpurohit R,Koch CJ,Tao Z,Teixeira CM,Shapiro IM. Adaptation of chondrocytes to low oxygen tension: relationship between hypoxia and cellular metabolism. J Cell Physiol 1996; 168: 424–432.
10.1002/(SICI)1097-4652(199608)168:2<424::AID-JCP21>3.0.CO;2-1 CASPubMedWeb of Science®Google Scholar
- 45 Schipani E,Ryan HE,Didrickson S,Kobayashi T,Knight M,Johnson RS. Hypoxia in cartilage: HIF-1 alpha is essential for chondrocyte growth arrest and survival. Genes Dev 2001; 15: 2865–2876.
- 46 Maes C,Carmeliet P,Moermans K,Stockmans I,Smets N,Collen D,Bouillon R,Carmeliet G. Impaired angiogenesis and endochondral bone formation in mice lacking the vascular endothelial growth factor isoforms VEGF164 and VEGF188. Mech Dev 2002; 111: 61–73.
- 47 Petersen W,Tsokos M,Pufe T. Expression of VEGF121 and VEGF165 in hypertrophic chondrocytes of the human growth plate and epiphyseal cartilage. J Anat 2002; 201: 153–157.
- 48 Oliveira SM,Amaral IF,Barbosa MA,Teixeira CC. Engineering endochondral bone: In vitro studies. Tissue Eng 2009; 15: 625–634.
- 49 Teixeira CC,Nemelivsky Y,Karkia C,Legeros RZ. Biphasic calcium phosphate: A scaffold for growth plate chondrocyte maturation. Tissue Eng 2006; 12: 2283–2289.
- 50 Oliveira SM,Turner G,Mijares D,Amaral IF,Barbosa MA,Teixeira CC. Engineering endochondral bone: In vivo studies. Tissue Eng 2009; 15: 635–643.
- 51 Hiraoka Y,Kimura Y,Ueda H,Tabata Y. Fabrication and biocompatibility of collagen sponge reinforced with poly(glycolic acid) fiber. Tissue Eng 2003; 9: 1101–1112.
- 52 Yost MJ,Baicu CF,Stonerock CE,Terracio L. A novel tubular scaffold for cardiovascular tissue engineering. Tissue Eng 2004; 10: 273–284.
- 53 Mow VC,Kuei SC,Lai WM,Armstrong CG. Biphasic creep and stress relaxation of articular cartilage in compression? Theory and experiments. J Biomech Eng 1980; 102: 73–84.
- 54 Iwamoto M,Shapiro IM,Yagami K,Boskey AL,Leboy PS,Adams SL,Pacifici M. Retinoic acid induces rapid mineralization and expression of mineralization-related genes in chondrocytes. Exp Cell Res 1993; 207: 413–420.
- 55 Teixeira CC,Hatori M,Leboy PS,Pacifici M,Shapiro IM. A rapid and ultrasensitive method for measurement of DNA, calcium and protein content, and alkaline phosphatase activity of chondrocyte cultures. Calcif Tissue Int 1995; 56: 252–256.
- 56 Leboy PS,Vaias L,Uschmann B,Golub E,Adams SL,Pacifici M. Ascorbic acid induces alkaline phosphatase, type X collagen, and calcium deposition in cultured chick chondrocytes. J Biol Chem 1989; 264: 17281–17286.
- 57 Whyte MP. Hypophosphatasia and the role of alkaline phosphatase in skeletal mineralization. Endocr Rev 1994; 15: 439–461.
- 58 Wang W,Kirsch T. Retinoic acid stimulates annexin-mediated growth plate chondrocyte mineralization. J Cell Biol 2002; 157: 1061–1069.
- 59 Chen D,Zhao M,Mundy GR. Bone morphogenetic proteins. Growth Factors 2004; 22: 233–241.
- 60 Gerstenfeld LC,Shapiro FD. Expression of bone-specific genes by hypertrophic chondrocytes: Implications of the complex functions of the hypertrophic chondrocyte during endochondral bone development. J Cell Biochem 1996; 62: 1–9.
- 61 Bianco P,Cancedda FD,Riminucci M,Cancedda R. Bone formation via cartilage models: The “borderline” chondrocyte. Matrix Biol 1998; 17: 185–192.
- 62 De Bernard B,Bianco P,Bonucci E,Costantini M,Lunazzi GC,Martinuzzi P,Modricky C,Moro L,Panfili E,Pollesello P. Biochemical and immunohistochemical evidence that in cartilage an alkaline phosphatase is a Ca 2”—Binding glycoprotein. J Cell Biol 1986; 103: 1615–1623.
- 63 Galotto M,Campanile G,Robino G,Cancedda FD,Bianco P,Cancedda R. Hypertrophic chondrocytes undergo further differentiation to osteoblast-like cells and participate in the initial bone formation in developing chick embryo. J Bone Miner Res 1994; 9: 1239–1249.
- 64 Mark MP,Butler WT,Prince CW,Finkelman RD,Ruch JV. Developmental expression of 44-kDa bone phosphoprotein (osteopontin) and bone gamma-carboxyglutamic acid (Gla)-containing protein (osteocalcin) in calcifying tissues of rat. Differentiation 1988; 37: 123–136.
- 65 Sims CD,Butler PE,Casanova R,Lee BT,Randolph MA,Lee WP,Vacanti CA,Yaremchuk MJ. Injectable cartilage using polyethylene oxide polymer substrates. Plast Reconstr Surg 1996; 98: 843–850.
- 66 Woodard JC,Donovan GA,Fisher LW. Pathogenesis of vitamin (A and D)-induced premature growth-plate closure in calves. Bone 1997; 27: 171–182.
- 67 Liao E,Yaszemski M,Krebsbach P,Hollister S. Tissue-engineered cartilage constructs using composite hyaluronic acid/collagen I hydrogels and designed poly(propylene fumarate) scaffolds. Tissue Eng 2007; 13: 537–550.
- 68 Gerstenfeld LC,Cruceta J,Shea CM,Sampath K,Barnes GL,Einhorn TA. Chondrocytes provide morphogenic signals that selectively induce osteogenic differentiation of mesenchymal stem cells. J Bone Miner Res 2002; 17: 221–230.