Characterization of Additive Manufactured Scaffolds

  • Giuseppe CriscentiEmail author
  • Carmelo De Maria
  • Giovanni Vozzi
  • Lorenzo Moroni
Reference work entry
Part of the Reference Series in Biomedical Engineering book series (RSBE)


At the increasing pace with which additive manufacturing technologies are advancing, it is possible nowadays to fabricate a variety of three-dimensional (3D) scaffolds with controlled structural and architectural properties. Examples span from metal cellular solids, which find application as prosthetic devices, to bioprinted constructs holding the promise to regenerate tissues and organs. These 3D porous constructs can display a variety of physicochemical and mechanical properties depending on the used material and on the design of the pore network to be created. To determine how these properties change with changing the scaffold’s design criteria, a plethora of characterization methods are applied in the biofabrication field. In this chapter, we review the most common techniques used to characterize such fabricated scaffolds by additive manufacturing technologies.



This project/research has been made possible with the support of the Dutch Province of Limburg.


  1. Abramowitch SD, Woo SL (2004) An improved method to analyze the stress relaxation of ligaments following a finite ramp time based on the quasi-linear viscoelastic theory. J Biomech Eng 126(1):92–97. Scholar
  2. Barsoukov E (2005) Impedance spectroscopy: theory, experiment, and applications (edited by J Ross Macdonald), 2nd. edn isbn:978-0-471-64749-2.Google Scholar
  3. Bay BK, Smith TS, Fyhrie DP, Saad M (1999) Digital volume correlation: three-dimensional strain mapping using X-ray tomography. Exp Mech 39(3):217–226CrossRefGoogle Scholar
  4. Binnig G, Quate CF, Gerber C (1986) Atomic-Force Microscope. Phys Rev Lett 56(9):930–933CrossRefPubMedGoogle Scholar
  5. Boffito M, Bernardi E, Sartori S, Ciardelli G, Sassi MP (2015) A mechanical characterization of polymer scaffolds and films at the macroscale and nanoscale. J Biomed Mater Res A 103:162–169CrossRefPubMedGoogle Scholar
  6. Carrabba M, De Maria C, Oikawa A, Reni C, Rodriguez-Arabaolaza I, Spencer H, Slater S, Avolio E, Dang Z, Spinetti G, Madeddu P, Vozzi G (2016) Design, fabrication and perivascular implantation of bioactive scaffolds engineered with human adventitial progenitor cells for stimulation of arteriogenesis in peripheral ischemia. Biofabrication 8(1):015020CrossRefPubMedGoogle Scholar
  7. Castilho M, Dias M, Vorndran E, Gbureck U, Fernandes P, Pires I, Gouveia B, Armés H, Pires E, Rodrigues J (2014) Application of a 3D printed customized implant for canine cruciate ligament treatment by tibial tuberosity advancement. Biofabrication 6(2):025005CrossRefPubMedGoogle Scholar
  8. Cheah CM, Chua CK, Leong KF, Chua SW (2003) Development of a tissue engineering scaffold structure library for rapid prototyping. Part 1: investigation and classification. Int J Adv Manuf Technol 21(4):291–301CrossRefGoogle Scholar
  9. Clarke AR (2002) Microscopy techniques for materials science. CRC Press (electronic resource)CrossRefGoogle Scholar
  10. de Gennes PG (1985) Wetting: statics and dynamics. Rev Mod Phys 57:827–863CrossRefGoogle Scholar
  11. De Maria C, Giusti S, Mazzei D, Crawford A, Ahluwalia A (2011) Squeeze pressure bioreactor: a hydrodynamic bioreactor for noncontact stimulation of cartilage constructs. Tissue Eng Part C Methods 17(7):757–764CrossRefPubMedGoogle Scholar
  12. De Maria C, De Acutis A, Vozzi G (2015) Indirect rapid prototyping for tissue engineering. In: Essential of 3D biofabrication and translation. Elsevier. isbn:978-0-12-800972-7Google Scholar
  13. Della Volpe C, Brugnara M (2006) About the possibility of experimentally measuring an equilibrium contact angle and its theoretical and practical consequences. Contact Angle, Wettability and Adhesion 4:79–100Google Scholar
  14. Denk W, Strickler JH, Webb WW (1990) Two-photon laser scanning fluorescence microscopy. Science 248(4951):73–76CrossRefPubMedGoogle Scholar
  15. Dias MR, Fernandes PR, Guedes JM, Hollister SJ (2012) Permeability analysis of scaffolds for bone tissue engineering. J Biomech 45:938–944CrossRefPubMedGoogle Scholar
  16. Doube M, Kłosowski MM, Arganda-Carreras I, Cordeliéres F, Dougherty RP, Jackson J, Schmid B, Hutchinson JR, Shefelbine SJ (2010) BoneJ: free and extensible bone image analysis in ImageJ. Bone 47:1076–1079. Scholar
  17. EC Regulation No 1394/2007 on advanced therapy medicinal products and amending Directive 2001/83/EC and Regulation (EC) No 726/2004Google Scholar
  18. Egerton RF (2005) Physical principles of electron microscopy: an introduction to TEM, SEM, and AEM. Springer, New YorkCrossRefGoogle Scholar
  19. Ehret R, Baumann W, Brischwein M, Schwinde A, Stegbauer K, Wolf B (1997) Monitoring of cellular behaviour by impedance measurements on interdigitated electrode structures. Biosens Bioelectron 12(1):29–41CrossRefPubMedGoogle Scholar
  20. Eshraghi S, Das S (2010) Mechanical and microstructural properties of polycaprolactone scaffolds with one-dimensional, two-dimensional, and three-dimensional orthogonally oriented porous architectures produced by selective laser sintering. Acta Biomater 6(7):2467–2476CrossRefPubMedPubMedCentralGoogle Scholar
  21. Fujie T, Desii A, Ventrelli L, Mazzolai B, Mattoli V (2012) Inkjet printing of protein microarrays on freestanding polymeric nanofilms for spatio-selective cell culture environment. Biomed Microdevices 14(6):1069–1076CrossRefPubMedGoogle Scholar
  22. Fung YC, Perrone N, Anliker M (1972) Stress strain history relations of soft tissues in simple elongation. In: Biomechanics: its foundations and objectives. Prentice Hall, Englewood Cliffs, pp 181–207Google Scholar
  23. Ge Z, Yang F, Goh JCH, Ramakrishna S, Lee EH (2006) Biomaterials and scaffolds for ligament tissue engineering. J Biomed Mater Res A 77:639–652CrossRefPubMedGoogle Scholar
  24. Giaever I, Keese CR (1984) Monitoring fibroblast behavior in tissue culture with an applied electric field. Proc Natl Acad Sci USA 81(12):3761–3764CrossRefPubMedGoogle Scholar
  25. Giaever I, Keese CR (1991) Micromotion of mammalian cells measured electrically. Proc Natl Acad Sci USA 88(17):7896–7900CrossRefPubMedGoogle Scholar
  26. Giessibl FJ, Trafas BM (1994) Piezoresistive cantilevers utilized for scanning tunneling and scanning force microscope in ultrahigh vacuum. Rev Sci Instrum 65(6):1923CrossRefGoogle Scholar
  27. Gilbert PM, Havenstrite KL, Magnusson KE, Sacco A, Leonardi NA, Kraft P, Nguyen NK, Thrun S, Lutolf MP, Blau HM (2010) Substrate elasticity regulates skeletal muscle stem cell self-renewal in culture. Science 329(5995):1078–1081CrossRefPubMedPubMedCentralGoogle Scholar
  28. Goldstein J (2003) Scanning electron microscopy and x-ray microanalysis. Kluwer Adacemic/Plenum Pulbishers, New YorkCrossRefGoogle Scholar
  29. Groll J, Boland T, Blunk T, Burdick JA, Cho DW, Dalton PD, Derby B, Forgacs G, Li Q, Mironov VA, Moroni L, Nakamura M, Shu W, Takeuchi S, Vozzi G, Woodfield TB, Xu T, Yoo JJ, Malda J (2016) Biofabrication: reappraising the definition of an evolving field. Biofabrication 8(1):013001CrossRefGoogle Scholar
  30. Guillemot F, Mironov V, Nakamura M (2010) Bioprinting is coming of age: report from the international conference on bioprinting and biofabrication in Bordeaux (3B'09). Biofabrication 2:010201CrossRefPubMedGoogle Scholar
  31. Ho ST, Hutmacher DW (2006) A comparison of micro CT with other techniques used in the characterization of scaffolds. Biomaterials 27(8):1362–1376CrossRefPubMedGoogle Scholar
  32. Hoque ME, Hutmacher DW, Feng W, Li S, Huang MH, Vert M, Wong YS (2005) Fabrication using a rapid prototyping system and in vitro characterization of PEG-PCL-PLA scaffolds for tissue engineering. J Biomater Sci Polym Ed 16(12):1595–1610CrossRefPubMedGoogle Scholar
  33. Huang J, Pan X, Li S, Peng X, Xiong C, Fang J (2011) A digital volume correlation technique for 3-D deformation measurements of soft gels. Int J Appl Mech 3(2):335–354CrossRefGoogle Scholar
  34. Hutmacher D, Woodfield T, Dalton P, Lewis J (2008) Scaffold design and fabrication in tissue engineering. In: Tissue engineering, pp 403–454. isbn:978-0-12-370869-4CrossRefGoogle Scholar
  35. Ibáñez L, Schroeder W, Ng L, Cates J, Consortium TIS, Hamming R (2003) The ITK software guide. Kitware, Inc., New YorkGoogle Scholar
  36. Kemppainen J (2008) Mechanically stable solid free form fabricated scaffolds with permeability optimized for cartilage tissue engineering. Dissertation, University of Michigan, USAGoogle Scholar
  37. Kemppainen J, Hollister S (2010) Differential effects of designed scaffold permeability on chondrogenesis by chondroyctes and bone marrow stromal cells. Biomaterials 31:279–287CrossRefPubMedGoogle Scholar
  38. Kubitscheck U (2013) Fluorescence microscopy: from principles to biological applications. Wiley-Blackwell, Weinheim. isbn:978-3-527CrossRefGoogle Scholar
  39. Li J, Mak A (2005) Hydraulic permeability of polyglycolic acid scaffolds as a function of biomaterial degradation. J Biomater Appl 19:253–266CrossRefPubMedGoogle Scholar
  40. Loh QL, Choong C (2013) Three-dimensional scaffolds for tissue engineering applications: role of porosity and pore size. Tissue Eng Part B Rev 19(6):485–502CrossRefPubMedPubMedCentralGoogle Scholar
  41. Malda J, Woodfield TB, van der Vloodt F, Wilson C, Martens DE, Tramper J, van Blitterswijk CA, Riesle J (2005) The effect of PEGT/PBT scaffold architecture on the composition of tissue engineered cartilage. Biomaterials 26(1):63–72CrossRefPubMedGoogle Scholar
  42. Malda J, Visser J, Melchels FP, Jüngst T, Hennink WE, Dhert WJ, Groll J, Hutmacher DW (2013) 25th anniversary article: engineering hydrogels for biofabrication. Adv Mater 25(36):5011–5028CrossRefGoogle Scholar
  43. Masaeli E, Morshed M, Nasr-Esfahani MH, Sadri S, Hilderink J, van Apeldoorn A, van Blitterswijk CA, Moroni L (2013) Fabrication, characterization and cellular compatibility of poly(hydroxy alkanoate) composite nanofibrous scaffolds for nerve tissue engineering. PLoS One 8(2):e57157CrossRefPubMedPubMedCentralGoogle Scholar
  44. Mattioli-Belmonte M, De Maria C, Vitale-Brovarone C, Baino F, Dicarlo M, Vozzi G (2015) Pressure-activated microsyringe (PAM) fabrication of bioactive glass-poly(lactic-co-glycolic acid) composite scaffolds for bone tissue regeneration. J Tissue Eng Regen Med.
  45. Menard KP (1999) DMA: introduction to the technique, its applications and theory. CRC Press, Boca RatonGoogle Scholar
  46. Moroni L, de Wijn JR, van Blitterswijk CA (2006) 3D fiber-deposited scaffolds for tissue engineering: influence of pores geometry and architecture on dynamic mechanical properties. Biomaterials 27:974–985CrossRefPubMedGoogle Scholar
  47. Nadeem D, Kiamehr M, Yang X, Su B (2013) Fabrication and in vitro evaluation of a sponge-like bioactive-glass/gelatin composite scaffold for bone tissue engineering. Mater Sci Eng C Mater Biol Appl 33(5):2669–2678CrossRefPubMedGoogle Scholar
  48. Nitzsche H, Metz H, Lochmann A, Bernstein A, Hause G, Groth T, Mäder K (2009) Characterization of scaffolds for tissue engineering by benchtop-magnetic resonance imaging. Tissue Eng Part C Methods 15(3):513–521. Scholar
  49. Oest ME, Dupont KM, Kong HJ, Mooney DJ, Guldberg RE (2007) Quantitative assessment of scaffold and growth factor-mediated repair of critically sized bone defects. J Orthop Res 25(7):941–950CrossRefPubMedGoogle Scholar
  50. Orsi G, De Maria C, Montemurro F, Chauhan VM, Aylott JW, Vozzi G (2015) Combining inkjet printing and sol-gel chemistry for making pH-sensitive surfaces. Curr Top Med Chem 15:271–278CrossRefPubMedGoogle Scholar
  51. Panetta J, Zhou Q, Malomo L, Pietroni N, Cignoni P, Zorin D (2015) Elastic textures for additive fabrication. ACM Trans on Graphics – Siggraph 34(4):12Google Scholar
  52. Park CH, Rios HF, Jin Q, Sugai JV, Padial-Molina M, Taut AD, Flanagan CL, Hollister SJ, Giannobile WV (2012) Tissue engineering bone-ligament complexes using fiber-guiding scaffolds. Biomaterials 33(1):137–145CrossRefPubMedGoogle Scholar
  53. Rainer A, Giannitelli SM, Accoto D, De Porcellinis S, Guglielmelli E, Trombetta M (2012) Load-adaptive scaffold architecturing: a bioinspired approach to the design of porous additively manufactured scaffolds with optimized mechanical properties. Ann Biomed Eng 40(4):966–975CrossRefPubMedGoogle Scholar
  54. Ratner BD, Hoffman AS, Schoen FJ, Lemons JE (2013) Biomaterials science, 3rd edn. Elsevier Academic Press, San Diego/London. ISBN:978-0-12-374626-9CrossRefGoogle Scholar
  55. Shirazi RN, Ronan W, Rochev Y, McHugh P (2016) Modelling the degradation and elastic properties of poly(lactic-co-glycolic acid) films and regular open-cell tissue engineering scaffolds. J Mech Behav Biomed Mater 54:48–59CrossRefPubMedGoogle Scholar
  56. Sudarmadji N, Tan JY, Leong KF, Chua CK, Loh YT (2011) Investigation of the mechanical properties and porosity relationships in selective laser-sintered polyhedral for functionally graded scaffolds. Acta Biomater 7(2):530–537CrossRefPubMedGoogle Scholar
  57. Sutton MA, Orteu JJ, Schreier HW (2009) Image correlation for shape, motion and deformation measurements. Springer, New York. isbn:978-0-387-78746-6Google Scholar
  58. Tirella A, Ahluwalia A (2012) The impact of fabrication parameters and substrate stiffness in direct writing of living constructs. Biotechnol Prog 28(5):1315–1320CrossRefPubMedGoogle Scholar
  59. Tirella A, Vozzi F, Vozzi G, Ahluwalia A (2011) PAM2 (piston assisted microsyringe): a new rapid prototyping technique for biofabrication of cell incorporated scaffolds. Tissue Eng Part C Methods 17(2):229–237CrossRefPubMedGoogle Scholar
  60. Tomlins P (2015) Characterization and design of tissue scaffold. Woodhead Publishing, Cambridge, UK/Waltham/Kidlington. ISBN:9781782420958Google Scholar
  61. Urciuolo A, Quarta M, Morbidoni V, Gattazzo F, Molon S, Grumati P, Montemurro F, Tedesco FS, Blaauw B, Cossu G, Vozzi G, Rando TA, Bonaldo P (2013) Collagen VI regulates satellite cell self-renewal and muscle regeneration. Nat Commun 4:1964CrossRefPubMedPubMedCentralGoogle Scholar
  62. von Burkersroda F, Schedl L, Göpferich A (2002) Why degradable polymers undergo surface erosion or bulk erosion. Biomaterials 23(21):4221–4231CrossRefGoogle Scholar
  63. Whulanza Y, Ucciferri N, Domenici C, Vozzi G, Ahluwalia A (2011) Sensing scaffolds to monitor cellular activity using impedance measurements. Biosens Bioelectron 26(7):3303–3308CrossRefPubMedGoogle Scholar
  64. Whulanza Y, Battini E, Vannozzi L, Vomero M, Ahluwalia A, Vozzi G (2013) Electrical and mechanical characterisation of single wall carbon nanotubes based composites for tissue engineering applications. J Nanosci Nanotechnol 13(1):188–197CrossRefPubMedGoogle Scholar
  65. Zein I, Hutmacher DW, Tan KC, Teoh SH (2002) Fused deposition modeling of novel scaffold architectures for tissue engineering applications. Biomaterials 23(4):1169–1185CrossRefPubMedGoogle Scholar
  66. Zhang Z, Jones D, Yue S, Lee PD, Jones JR, Sutcliffe CJ, Jones E (2013) Hierarchical tailoring of strut architecture to control permeability of additive manufactured titanium implants. Mater Sci Eng C Mater Biol Appl 33(7):4055–4062CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  • Giuseppe Criscenti
    • 1
    • 2
    Email author
  • Carmelo De Maria
    • 2
  • Giovanni Vozzi
    • 2
  • Lorenzo Moroni
    • 1
  1. 1.Department of Complex Tissue Regeneration, MERLN Institute for Technology-Inspired Regenerative MedicineMaastricht UniversityMaastrichtThe Netherlands
  2. 2.Research Center “E. Piaggio”University of PisaPisaItaly

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