Background The basic strategy to construct tissue engineered bone graft (TEBG) is to combine osteoblastic cells with three dimensional (3D) scaffold. Based on this strategy, we proposed the “Totally Vitalized TEBG” (TV-TEBG) which was characterized by abundant and homogenously distributed cells with enhanced cell proliferation and differentiation and further investigated its biological performance in repairing segmental bone defect. Methods In this study, we constructed the TV-TEBG with the combination of customized flow perfusion seeding/culture system and β-tricalcium phosphate (β-TCP) scaffold fabricated by Rapid Prototyping (RP) technique. We systemically compared three kinds of TEBG constructed by perfusion seeding and perfusion culture (PSPC) method, static seeding and perfusion culture (SSPC) method, and static seeding and static culture (SSSC) method for their in vitro performance and bone defect healing efficacy with a rabbit model. Results Our study has demonstrated that TEBG constructed by PSPC method exhibited better biological properties with higher daily D-glucose consumption, increased cell proliferation and differentiation, and better cell distribution, indicating the successful construction of TV-TEBG. After implanted into rabbit radius defects for 12 weeks, PSPC group exerted higher X-ray score close to autograft, much greater mechanical property evidenced by the biomechanical testing and significantly higher new bone formation as shown by histological analysis compared with the other two groups, and eventually obtained favorable healing efficacy of the segmental bone defect that was the closest to autograft transplantation. Conclusion This study demonstrated the feasibility of TV-TEBG construction with combination of perfusion seeding, perfusion culture and RP technique which exerted excellent biological properties. The application of TV-TEBG may become a preferred candidate for segmental bone defect repair in orthopedic and maxillofacial fields.
References
[1]
Kruyt MC, de Bruijn JD, Wilson CE, Oner FC, van Blitterswijk CA, et al. (2003) Viable osteogenic cells are obligatory for tissue-engineered ectopic bone formation in goats. Tissue Eng 9: 327–336. doi: 10.1089/107632703764664792
[2]
Zhang ZY, Teoh SH, Hui JH, Fisk NM, Choolani M, et al. (2012) The potential of human fetal mesenchymal stem cells for off-the-shelf bone tissue engineering application. Biomaterials 33: 2656–2672. doi: 10.1016/j.biomaterials.2011.12.025
[3]
Khan Y, Yaszemski MJ, Mikos AG, Laurencin CT (2008) Tissue engineering of bone: material and matrix considerations. The Journal of bone and joint surgery American volume 90 Suppl 136–42. doi: 10.2106/jbjs.g.01260
[4]
Ge Z, Baguenard S, Lim LY, Wee A, Khor E (2004) Hydroxyapatite-chitin materials as potential tissue engineered bone substitutes. Biomaterials 25: 1049–1058. doi: 10.1016/s0142-9612(03)00612-4
[5]
Vunjak-Novakovic G, Obradovic B, Martin I, Bursac PM, Langer R, et al. (1998) Dynamic cell seeding of polymer scaffolds for cartilage tissue engineering. Biotechnol Prog 14: 193–202. doi: 10.1021/bp970120j
[6]
Burg KJ, Delnomdedieu M, Beiler RJ, Culberson CR, Greene KG, et al. (2002) Application of magnetic resonance microscopy to tissue engineering: a polylactide model. J Biomed Mater Res 61: 380–390. doi: 10.1002/jbm.10146
[7]
Xie Y, Yang ST, Kniss DA (2001) Three-dimensional cell-scaffold constructs promote efficient gene transfection: implications for cell-based gene therapy. Tissue Eng 7: 585–598. doi: 10.1089/107632701753213200
[8]
van den Dolder J, Spauwen PH, Jansen JA (2003) Evaluation of various seeding techniques for culturing osteogenic cells on titanium fiber mesh. Tissue Eng 9: 315–325. doi: 10.1089/107632703764664783
[9]
van den Dolder J, Bancroft GN, Sikavitsas VI, Spauwen PH, Jansen JA, et al. (2003) Flow perfusion culture of marrow stromal osteoblasts in titanium fiber mesh. Journal of biomedical materials research Part A 64: 235–241. doi: 10.1002/jbm.a.10365
[10]
Sikavitsas VI, Bancroft GN, Lemoine JJ, Liebschner MA, Dauner M, et al. (2005) Flow perfusion enhances the calcified matrix deposition of marrow stromal cells in biodegradable nonwoven fiber mesh scaffolds. Ann Biomed Eng 33: 63–70. doi: 10.1007/s10439-005-8963-x
[11]
Holtorf HL, Sheffield TL, Ambrose CG, Jansen JA, Mikos AG (2005) Flow perfusion culture of marrow stromal cells seeded on porous biphasic calcium phosphate ceramics. Ann Biomed Eng 33: 1238–1248. doi: 10.1007/s10439-005-5536-y
[12]
Sikavitsas VI, Bancroft GN, Holtorf HL, Jansen JA, Mikos AG (2003) Mineralized matrix deposition by marrow stromal osteoblasts in 3D perfusion culture increases with increasing fluid shear forces. Proceedings of the National Academy of Sciences of the United States of America 100: 14683–14688. doi: 10.1073/pnas.2434367100
[13]
Leong KF, Cheah CM, Chua CK (2003) Solid freeform fabrication of three-dimensional scaffolds for engineering replacement tissues and organs. Biomaterials 24: 2363–2378. doi: 10.1016/s0142-9612(03)00030-9
[14]
Ma PX, Choi JW (2001) Biodegradable polymer scaffolds with well-defined interconnected spherical pore network. Tissue Eng 7: 23–33. doi: 10.1089/107632701300003269
[15]
Murphy WL, Dennis RG, Kileny JL, Mooney DJ (2002) Salt fusion: an approach to improve pore interconnectivity within tissue engineering scaffolds. Tissue Eng 8: 43–52. doi: 10.1089/107632702753503045
[16]
Shea LD, Wang D, Franceschi RT, Mooney DJ (2000) Engineered bone development from a pre-osteoblast cell line on three-dimensional scaffolds. Tissue Eng 6: 605–617. doi: 10.1089/10763270050199550
[17]
Holtorf HL, Datta N, Jansen JA, Mikos AG (2005) Scaffold mesh size affects the osteoblastic differentiation of seeded marrow stromal cells cultured in a flow perfusion bioreactor. Journal of biomedical materials research Part A 74: 171–180. doi: 10.1002/jbm.a.30330
[18]
Gomes ME, Holtorf HL, Reis RL, Mikos AG (2006) Influence of the porosity of starch-based fiber mesh scaffolds on the proliferation and osteogenic differentiation of bone marrow stromal cells cultured in a flow perfusion bioreactor. Tissue Eng 12: 801–809. doi: 10.1089/ten.2006.12.801
[19]
Manjubala I, Woesz A, Pilz C, Rumpler M, Fratzl-Zelman N, et al. (2005) Biomimetic mineral-organic composite scaffolds with controlled internal architecture. J Mater Sci Mater Med 16: 1111–1119. doi: 10.1007/s10856-005-4715-6
[20]
Yu X, Botchwey EA, Levine EM, Pollack SR, Laurencin CT (2004) Bioreactor-based bone tissue engineering: the influence of dynamic flow on osteoblast phenotypic expression and matrix mineralization. Proceedings of the National Academy of Sciences of the United States of America 101: 11203–11208. doi: 10.1073/pnas.0402532101
[21]
Song K, Wang H, Zhang B, Lim M, Liu Y, et al. (2013) Numerical simulation of fluid field and in vitro three-dimensional fabrication of tissue-engineered bones in a rotating bioreactor and in vivo implantation for repairing segmental bone defects. Cell Stress Chaperones 18: 193–201. doi: 10.1007/s12192-012-0370-2
[22]
Raimondi MT, Boschetti F, Falcone L, Fiore GB, Remuzzi A, et al. (2002) Mechanobiology of engineered cartilage cultured under a quantified fluid-dynamic environment. Biomech Model Mechanobiol 1: 69–82. doi: 10.1007/s10237-002-0007-y
[23]
Alvarez-Barreto JF, Sikavitsas VI (2007) Improved mesenchymal stem cell seeding on RGD-modified poly(L-lactic acid) scaffolds using flow perfusion. Macromol Biosci 7: 579–588. doi: 10.1002/mabi.200600280
[24]
Leclerc E, David B, Griscom L, Lepioufle B, Fujii T, et al. (2006) Study of osteoblastic cells in a microfluidic environment. Biomaterials 27: 586–595. doi: 10.1016/j.biomaterials.2005.06.002
[25]
Janssen FW, Hofland I, van Oorschot A, Oostra J, Peters H, et al. (2006) Online measurement of oxygen consumption by goat bone marrow stromal cells in a combined cell-seeding and proliferation perfusion bioreactor. Journal of Biomedical Materials Research Part A 79A: 338–348. doi: 10.1002/jbm.a.30794
[26]
Schumacher M, Uhl F, Detsch R, Deisinger U, Ziegler G (2010) Static and dynamic cultivation of bone marrow stromal cells on biphasic calcium phosphate scaffolds derived from an indirect rapid prototyping technique. Journal of Materials Science: Materials in Medicine 21: 3039–3048. doi: 10.1007/s10856-010-4153-y
[27]
Wang L, Hu YY, Wang Z, Li X, Li DC, et al. (2009) Flow perfusion culture of human fetal bone cells in large beta-tricalcium phosphate scaffold with controlled architecture. Journal of biomedical materials research Part A 91: 102–113. doi: 10.1002/jbm.a.32189
[28]
Chu TM, Halloran JW, Hollister SJ, Feinberg SE (2001) Hydroxyapatite implants with designed internal architecture. Journal of materials science Materials in medicine 12: 471–478. doi: 10.1023/a:1011203226053
[29]
Declercq H, Van den Vreken N, De Maeyer E, Verbeeck R, Schacht E, et al. (2004) Isolation, proliferation and differentiation of osteoblastic cells to study cell/biomaterial interactions: comparison of different isolation techniques and source. Biomaterials 25: 757–768. doi: 10.1016/s0142-9612(03)00580-5
[30]
Schmitz JP, Hollinger JO (1986) The critical size defect as an experimental model for craniomandibulofacial nonunions. Clinical orthopaedics and related research: 299–308.
[31]
Yang CY, Simmons DJ, Lozano R (1994) The healing of grafts combining freeze-dried and demineralized allogeneic bone in rabbits. Clinical orthopaedics and related research: 286–295.
[32]
Zhao F, Ma T (2005) Perfusion bioreactor system for human mesenchymal stem cell tissue engineering: Dynamic cell seeding and construct development. Biotechnology and bioengineering 91: 482–493. doi: 10.1002/bit.20532
[33]
Goldstein AS, Juarez TM, Helmke CD, Gustin MC, Mikos AG (2001) Effect of convection on osteoblastic cell growth and function in biodegradable polymer foam scaffolds. Biomaterials 22: 1279–1288. doi: 10.1016/s0142-9612(00)00280-5
[34]
Alvarez-Barreto JF, Linehan SM, Shambaugh RL, Sikavitsas VI (2007) Flow perfusion improves seeding of tissue engineering scaffolds with different architectures. Ann Biomed Eng 35: 429–442. doi: 10.1007/s10439-006-9244-z
[35]
Haynesworth SE, Goshima J, Goldberg VM, Caplan AI (1992) Characterization of cells with osteogenic potential from human marrow. Bone 13: 81–88. doi: 10.1016/8756-3282(92)90364-3
[36]
Olivares AL, Marsal E, Planell JA, Lacroix D (2009) Finite element study of scaffold architecture design and culture conditions for tissue engineering. Biomaterials 30: 6142–6149. doi: 10.1016/j.biomaterials.2009.07.041
[37]
Pountos I, Jones E, Tzioupis C, McGonagle D, Giannoudis PV (2006) Growing bone and cartilage. The role of mesenchymal stem cells. J Bone Joint Surg Br 88: 421–426. doi: 10.1302/0301-620x.88b4.17060
[38]
McGarry JG, Klein-Nulend J, Mullender MG, Prendergast PJ (2005) A comparison of strain and fluid shear stress in stimulating bone cell responses—a computational and experimental study. FASEB journal: official publication of the Federation of American Societies for Experimental Biology 19: 482–484.
[39]
Hillsley MV, Frangos JA (1994) Bone tissue engineering: the role of interstitial fluid flow. Biotechnology and bioengineering 43: 573–581. doi: 10.1002/bit.260430706
[40]
Sikavitsas VI, Temenoff JS, Mikos AG (2001) Biomaterials and bone mechanotransduction. Biomaterials 22: 2581–2593. doi: 10.1016/s0142-9612(01)00002-3
[41]
Minamide A, Yoshida M, Kawakami M, Yamasaki S, Kojima H, et al. (2005) The use of cultured bone marrow cells in type I collagen gel and porous hydroxyapatite for posterolateral lumbar spine fusion. Spine 30: 1134–1138. doi: 10.1097/01.brs.0000162394.75425.04
[42]
Pieri F, Lucarelli E, Corinaldesi G, Aldini NN, Fini M, et al. (2010) Dose-dependent effect of adipose-derived adult stem cells on vertical bone regeneration in rabbit calvarium. Biomaterials 31: 3527–3535. doi: 10.1016/j.biomaterials.2010.01.066
[43]
Nair MB, Varma HK, Menon KV, Shenoy SJ, John A (2009) Tissue regeneration and repair of goat segmental femur defect with bioactive triphasic ceramic-coated hydroxyapatite scaffold. J Biomed Mater Res A 91: 855–865. doi: 10.1002/jbm.a.32239
[44]
Dutta Roy T, Simon JL, Ricci JL, Rekow ED, Thompson VP, et al. (2003) Performance of hydroxyapatite bone repair scaffolds created via three-dimensional fabrication techniques. Journal of biomedical materials research Part A 67: 1228–1237. doi: 10.1002/jbm.a.20034
[45]
Simon JL, Roy TD, Parsons JR, Rekow ED, Thompson VP, et al. (2003) Engineered cellular response to scaffold architecture in a rabbit trephine defect. Journal of biomedical materials research Part A 66: 275–282. doi: 10.1002/jbm.a.10569
[46]
Rouwkema J, Rivron NC, van Blitterswijk CA (2008) Vascularization in tissue engineering. Trends Biotechnol 26: 434–441. doi: 10.1016/j.tibtech.2008.04.009
[47]
Tan H, Yang B, Duan X, Wang F, Zhang Y, et al. (2009) The promotion of the vascularization of decalcified bone matrix in vivo by rabbit bone marrow mononuclear cell-derived endothelial cells. Biomaterials 30: 3560–3566. doi: 10.1016/j.biomaterials.2009.03.029
[48]
Wang L, Fan H, Zhang ZY, Lou AJ, Pei GX, et al. (2010) Osteogenesis and angiogenesis of tissue-engineered bone constructed by prevascularized beta-tricalcium phosphate scaffold and mesenchymal stem cells. Biomaterials 31: 9452–9461. doi: 10.1016/j.biomaterials.2010.08.036
[49]
Karageorgiou V, Kaplan D (2005) Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials 26: 5474–5491. doi: 10.1016/j.biomaterials.2005.02.002
