Biocompatibility and biodegradation studies of PCL/β-TCP bone tissue scaffold fabricated by structural porogen method

Abstract

Three-dimensional printer (3DP) (Z-Corp) is a solid freeform fabrication system capable of generating sub-millimeter physical features required for tissue engineering scaffolds. By using plaster composite materials, 3DP can fabricate a universal porogen which can be injected with a wide range of high melting temperature biomaterials. Here we report results toward the manufacture of either pure polycaprolactone (PCL) or homogeneous composites of 90/10 or 80/20 (w/w) PCL/beta-tricalcium phosphate (β-TCP) by injection molding into plaster composite porogens fabricated by 3DP. The resolution of printed plaster porogens and produced scaffolds was studied by scanning electron microscopy. Cytotoxicity test on scaffold extracts and biocompatibility test on the scaffolds as a matrix supporting murine osteoblast (7F2) and endothelial hybridoma (EAhy 926) cells growth for up to 4 days showed that the porogens removal process had only negligible effects on cell proliferation. The biodegradation tests of pure PCL and PCL/β-TCP composites were performed in DMEM with 10 % (v/v) FBS for up to 6 weeks. The PCL/β-TCP composites show faster degradation rate than that of pure PCL due to the addition of β-TCP, and the strength of 80/20 PCL/β-TCP composite is still suitable for human cancellous bone healing support after 6 weeks degradation. Combining precisely controlled porogen fabrication structure, good biocompatibility, and suitable mechanical properties after biodegradation, PCL/β-TCP scaffolds fabricated by 3DP porogen method provide essential capability for bone tissue engineering.

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References

  1. 1.

    Williams D. Benefit and risk in tissue engineering. Mater Today. 2004;7:24–9.

    Article  CAS  Google Scholar 

  2. 2.

    Kneser U, Schaefer DJ, Polykandriotis E, Horch RE. Tissue engineering of bone: the reconstructive surgeon’s point of view. J Cell Mol Med. 2006;10:7–19.

    Article  CAS  Google Scholar 

  3. 3.

    Burg KJL, Porter S, Kellam JF. Biomaterial developments for bone tissue engineering. Biomaterials. 2000;21:2347–59.

    Article  CAS  Google Scholar 

  4. 4.

    Thomson RC, Mikos AG, Beahm E, Lemon JC, Satterfield WC, Aufdemorte TB, et al. Guided tissue fabrication from periosteum using preformed biodegradable polymer scaffolds. Biomaterials. 1999;20:2007–18.

    Article  CAS  Google Scholar 

  5. 5.

    Ma PX. Biomimetic materials for tissue engineering. Adv Drug Deliv Rev. 2008;60:184–98.

    Article  CAS  Google Scholar 

  6. 6.

    Zhang QW, Mochalin VN, Neitzel I, Knoke IY, Han JJ, Klug CA, et al. Fluorescent PLLA-nanodiamond composites for bone tissue engineering. Biomaterials. 2011;32:87–94.

    Article  Google Scholar 

  7. 7.

    Neumann M, Epple M. Composites of calcium phosphate and polymers as bone substitution materials. Eur J Trauma. 2006;32:125–31.

    Article  Google Scholar 

  8. 8.

    Shin M, Yoshimoto H, Vacanti JP. In vivo bone tissue engineering using mesenchymal stem cells on a novel electrospun nanofibrous scaffold. Tissue Eng. 2004;10:33–41.

    Article  CAS  Google Scholar 

  9. 9.

    Rohner D, Hutmacher DW, Cheng TK, Oberholzer M, Hammer B. In vivo efficacy of bone-marrow-coated polycaprolactone scaffolds for the reconstruction of orbital defects in the pig. J Biomed Mater Res B Appl Biomater. 2003;66B:574–80.

    Article  CAS  Google Scholar 

  10. 10.

    Ruhe PQ, Hedberg EL, Padron NT, Spauwen PHM, Jansen JA, Mikos AG. rhBMP-2 release from injectable poly(dl-lactic-co-glycolic acid)/calcium-phosphate cement composites. J Bone Joint Surg Am. 2003;85A:75–81.

    Google Scholar 

  11. 11.

    Xu HHK, Quinn JB, Takagi S, Chow LC. Synergistic reinforcement of in situ hardening calcium phosphate composite scaffold for bone tissue engineering. Biomaterials. 2004;25:1029–37.

    Article  CAS  Google Scholar 

  12. 12.

    Barralet JE, Grover L, Gaunt T, Wright AJ, Gibson IR. Preparation of macroporous calcium phosphate cement tissue engineering scaffold. Biomaterials. 2002;23:3063–72.

    Article  CAS  Google Scholar 

  13. 13.

    Yang Q, Chen L, Shen X, Tan Z. Preparation of polycaprolactone tissue engineering scaffolds by improved solvent casting/particulate leaching method. J Macromol Sci Part B Phys. 2006;45:1171–81.

    Article  Google Scholar 

  14. 14.

    Heijkants R, Van Tienen TG, De Groot JH, Pennings AJ, Buma P, Veth RPH, et al. Preparation of a polyurethane scaffold for tissue engineering made by a combination of salt leaching and freeze-drying of dioxane. J Mater Sci. 2006;41:2423–8.

    Article  CAS  Google Scholar 

  15. 15.

    Mikos (AGH, TX), Sarakinos, Georgios (Boston, MA), Vacanti, Joseph P. (Winchester, MA), Langer, Robert S. (Newton, MA), Cima, Linda G. (Lexington, MA). Biocompatible polymer membranes and methods of preparation of three-dimensional membrane structures. United States: Massachusetts Institute of Technology (Cambridge, MA), Children’s Medical Center Corporation (Boston, MA); 1996.

  16. 16.

    Fukuhira Y, Kitazono E, Hayashi T, Kaneko H, Tanaka M, Shimomura M, et al. Biodegradable honeycomb-patterned film composed of poly(lactic acid) and dioleoylphosphatidylethanolamine. Biomaterials. 2006;27:1797–802.

    Article  CAS  Google Scholar 

  17. 17.

    Cima LG, Vacanti JP, Vacanti C, Ingber D, Mooney D, Langer R. Tissue engineering by cell transplantation using degradable polymer substrates. J Biomech Eng Trans ASME. 1991;113:143–51.

    Article  CAS  Google Scholar 

  18. 18.

    Zhang W, Yao D, Zhang Q, Zhou JG, Lelkes PI. Fabrication of interconnected microporous biomaterials with high hydroxyapatite nanoparticle loading. Biofabrication. 2010;2.

  19. 19.

    Manjubala I, Woesz A, Pilz C, Rumpler M, Fratzl-Zelman N, Roschger P, et al. Biomimetic mineral-organic composite scaffolds with controlled internal architecture. J Mater Sci Mater Med. 2005;16:1111–9.

    Article  CAS  Google Scholar 

  20. 20.

    Hollister SJ, Maddox RD, Taboas JM. Optimal design and fabrication of scaffolds to mimic tissue properties and satisfy biological constraints. Biomaterials. 2002;23:4095–103.

    Article  CAS  Google Scholar 

  21. 21.

    Roy TD, Simon JL, Ricci JL, Rekow ED, Thompson VP, Parsons JR. Performance of degradable composite bone repair products made via three-dimensional fabrication techniques. J Biomed Mater Res A. 2003;66A:283–91.

    Article  CAS  Google Scholar 

  22. 22.

    Taboas JM, Maddox RD, Krebsbach PH, Hollister SJ. Indirect solid free form fabrication of local and global porous, biomimetic and composite 3D polymer–ceramic scaffolds. Biomaterials. 2003;24:181–94.

    Article  CAS  Google Scholar 

  23. 23.

    Lu L, Zhang Q, Wootton D, Lelkes PI, Zhou J. A novel sucrose porogen-based solid freeform fabrication system for bone scaffold manufacturing. Rapid Prototyping J. 2010;16:365–76.

    Article  Google Scholar 

  24. 24.

    Mooney DJ, Baldwin DF, Suh NP, Vacanti LP, Langer R. Novel approach to fabricate porous sponges of poly(D,L-lactic-co-glycolic acid) without the use of organic solvents. Biomaterials. 1996;17:1417–22.

    Article  CAS  Google Scholar 

  25. 25.

    Mondrinos MJ, Dembzynski R, Lu L, Byrapogu VKC, Wootton DM, Lelkes PI, et al. Porogen-based solid freeform fabrication of polycaprolactone-calcium phosphate scaffolds for tissue engineering. Biomaterials. 2006;27:4399–408.

    Article  CAS  Google Scholar 

  26. 26.

    Boyan BD, Hummert TW, Dean DD, Schwartz Z. Role of material surfaces in regulating bone and cartilage cell response. Biomaterials. 1996;17:137–46.

    Article  CAS  Google Scholar 

  27. 27.

    Geissler U, Hempel U, Wolf C, Scharnweber D, Worch H, Wenzel KW. Collagen type I-coating of Ti6Al4V promotes adhesion of osteoblasts. J Biomed Mater Res. 2000;51:752–60.

    Article  CAS  Google Scholar 

  28. 28.

    Andrianarivo AG, Robinson JA, Mann KG, Tracy RP. Growth on type-I collagen promotes expression of the osteoblastic phenotype in human osteosarcoma MG-63 cells. J Cell Physiol. 1992;153:256–65.

    Article  CAS  Google Scholar 

  29. 29.

    Schantz JT, Hutmacher DW, Ng KW, Khor HL, Lim TC, Teoh SH. Evaluation of a tissue-engineered membrane-cell construct for guided bone regeneration. Int J Oral Maxillofac Implants. 2002;17:161–74.

    Google Scholar 

  30. 30.

    Woodward SC, Brewer PS, Moatamed F, Schindler A, Pitt CG. The Intracellular degradation of poly(epsilon-caprolactone). J Biomed Mater Res. 1985;19:437–44.

    Article  CAS  Google Scholar 

  31. 31.

    Ali SAM, Zhong SP, Doherty PJ, Williams DF. Mechanisms of polymer degradation in implantable devices. 1. Poly(caprolactone). Biomaterials. 1993;14:648–56.

    Article  CAS  Google Scholar 

  32. 32.

    Kim HW, Knowles JC, Kim HE. Development of hydroxyapatite bone scaffold for controlled drug release via poly(epsilon-caprolactone) and hydroxyapatite hybrid coatings. J Biomed Mater Res B Appl Biomater. 2004;70B:240–9.

    Article  CAS  Google Scholar 

  33. 33.

    Timmer MD, Ambrose CG, Mikos AG. In vitro degradation of polymeric networks of poly(propylene fumarate) and the crosslinking macromer poly(propylene fumarate)-diacrylate. Biomaterials. 2003;24:571–7.

    Article  CAS  Google Scholar 

  34. 34.

    Peter SJ, Nolley JA, Widmer MS, Merwin JE, Yaszemski MJ, Yasko AW, et al. In vitro degradation of a poly(propylene fumarate)/beta-tricalcium phosphate composite orthopaedic scaffold. Tissue Eng. 1997;3:207–15.

    Article  CAS  Google Scholar 

  35. 35.

    Looney MA, Park JB. Molecular and mechanical property changes during aging of bone cement in vitro and in vivo. J Biomed Mater Res. 1986;20:555–63.

    Article  CAS  Google Scholar 

  36. 36.

    Misch CE, Qu ZM, Bidez MW. Mechanical properties of trabecular bone in the human mandible: implications for dental implant treatment planning and. Surgical placement. J Oral Maxillofac Surg. 1999;57:700–6.

    Article  CAS  Google Scholar 

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Acknowledgments

We gratefully thank National Science Foundation (NSF) for its financial support (DMI–0300405, CMMI-0700139 and CMMI-0925348). Additionally, the authors are grateful to Dr. Wei Sun for providing access to 3D printer for this study. We also would like to thank the laboratory of Dr. Giuseppe Palmese for assistance with GPC degradation tests and the laboratory of Dr. Boris Polyak for providing the access to plate reader. The Centralized Research Facility (CRF) of the College of Engineering, Drexel University provided access to electron microscopes used in this work.

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Correspondence to Jack Zhou.

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Lu, L., Zhang, Q., Wootton, D. et al. Biocompatibility and biodegradation studies of PCL/β-TCP bone tissue scaffold fabricated by structural porogen method. J Mater Sci: Mater Med 23, 2217–2226 (2012). https://doi.org/10.1007/s10856-012-4695-2

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Keywords

  • Injection Molding
  • Solid Freeform Fabrication
  • Bone Scaffold
  • Scaffold Fabrication
  • Human Cancellous Bone