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Dry versus hydrated collagen scaffolds: are dry states representative of hydrated states?

  • Tomáš Suchý
  • Monika Šupová
  • Martin Bartoš
  • Radek Sedláček
  • Marco Piola
  • Monica Soncini
  • Gianfranco Beniamino Fiore
  • Pavla Sauerová
  • Marie Hubálek  Kalbáčová
Biomaterials Synthesis and Characterization Original Research
Part of the following topical collections:
  1. Biomaterials Synthesis and Characterization

Abstract

Collagen composite scaffolds have been used for a number of studies in tissue engineering. The hydration of such highly porous and hydrophilic structures may influence mechanical behaviour and porosity due to swelling. The differences in physical properties following hydration would represent a significant limiting factor for the seeding, growth and differentiation of cells in vitro and the overall applicability of such hydrophilic materials in vivo. Scaffolds based on collagen matrix, poly(DL-lactide) nanofibers, calcium phosphate particles and sodium hyaluronate with 8 different material compositions were characterised in the dry and hydrated states using X-ray microcomputed tomography, compression tests, hydraulic permeability measurement, degradation tests and infrared spectrometry. Hydration, simulating the conditions of cell seeding and cultivation up to 48 h and 576 h, was found to exert a minor effect on the morphological parameters and permeability. Conversely, hydration had a major statistically significant effect on the mechanical behaviour of all the tested scaffolds. The elastic modulus and compressive strength of all the scaffolds decreased by ~95%. The quantitative results provided confirm the importance of analysing scaffolds in the hydrated rather than the dry state since the former more precisely simulates the real environment for which such materials are designed.

Notes

Acknowledgements

This study was supported by a grant project awarded by the Ministry of Health of the Czech Republic (15-25813A). This publication is the result of the implementation of the “Technological development of post-doc programmes” project, registration number CZ.1.05/41.00/16.0346, supported by the Research and Development for Innovations Operational Programme (RDIOP), co-financed by European regional development funds and the state budget of the Czech Republic. The project was also supported by the “Progres Q29/1LF, Ministry of Education, Youth and Sports of the Czech Republic” and GAUK no. 400215. We gratefully acknowledge the financial support provided for our work by the long-term conceptual development research organisation under project no. RVO: 67985891. Special thanks go to Darren Ireland for the language revision of the English manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10856_2017_6024_MOESM1_ESM.tif (19.3 mb)
Supplementary Information

References

  1. 1.
    Jungreuthmayer C, Jaasma MJ, Al-Munajjed AA, Zanghellini J, Kelly DJ, O’Brien FJ. Deformation simulation of cells seeded on a collagen-GAG scaffold in a flow perfusion bioreactor using a sequential 3D CFD-elastostatics model. Med Eng Phys. 2009;31:420–7.CrossRefGoogle Scholar
  2. 2.
    Doillon CJ, Whyne CF, Brandwein S, Silver FH. Collagen-based wound dressings: control of the pore structure and morphology. J Biomed Mater Res. 1986;20:1219–28.CrossRefGoogle Scholar
  3. 3.
    O’Brien FJ, Harley BA, Yannas IV, Gibson L. Influence of freezing rate on pore structure in freeze-dried collagen-GAG scaffolds. Biomaterials. 2004;25:1077–86.CrossRefGoogle Scholar
  4. 4.
    Zhu Y, Wu H, Sun S, Zhou T, Wu J, Wan Y. Designed composites for mimicking compressive mechanical properties of articular cartilage matrix. J Mech Behav Biomed Mater. 2014;36:32–46.CrossRefGoogle Scholar
  5. 5.
    Zhu Y, Wan Y, Zhang J, Yin D, Cheng W. Manufacture of layered collagen/chitosan-polycaprolactone scaffolds with biomimetic microarchitecture. Colloid Surf B. 2014;113:352–60.CrossRefGoogle Scholar
  6. 6.
    Gorczyca G, Tylingo R, Szweda P, Augustin E, Sadowska M, Milewski S. Preparation and characterization of genipin cross-linked porous chitosan–collagen–gelatin scaffolds using chitosan–CO2 solution. Carbohyd Polym. 2014;102:901–11.CrossRefGoogle Scholar
  7. 7.
    Elsayed Y, Lekakou C, Labeed F, Tomlins P. Fabrication and characterisation of biomimetic, electrospun gelatin fibre scaffolds for tunica media-equivalent, tissue engineered vascular grafts. Mat Sci Eng C. 2016;61:473–83.CrossRefGoogle Scholar
  8. 8.
    Davidenko N, Campbell JJ, Thian ES, Watson CJ, Cameron RE. Collagen–hyaluronic acid scaffolds for adipose tissue engineering. Acta Biomater. 2010;6:3957–68.CrossRefGoogle Scholar
  9. 9.
    Offeddu GS, Ashworth JC, Cameron RE, Oyen ML. Structural determinants of hydration, mechanics and fluid flow in freeze-dried collagen scaffolds. Acta Biomater. 2016;41:193–203.CrossRefGoogle Scholar
  10. 10.
    Varley MC, Neelakantan S, Clyne TW, Dean J, Brooks RA, Markaki AE. Cell structure, stiffness and permeability of freeze-dried collagen scaffolds in dry and hydrated states. Acta Biomater. 2016;33:166–75.CrossRefGoogle Scholar
  11. 11.
    Beachley V, Wen X. Fabrication of nanofiber reinforced protein structures for tissue engineering. Mat Sci Eng C. 2009;29:2448–53.CrossRefGoogle Scholar
  12. 12.
    Davidenko N, Schuster CF, Bax DV, Raynal N, Farndale RW, Best SM, Cameron RE. Control of cross-linking for tailoring collagen-based scaffolds stability and mechanics. Acta Biomater. 2015;25:131–42.CrossRefGoogle Scholar
  13. 13.
    Xingang W, Qiyin L, Xinlei H, Lie M, Chuangang Y, Yurong Z, Huafeng S, Chunmao H, Changyou G. Fabrication and characterization of poly(L-lactide-co-glycolide)knitted mesh-reinforced collagen–chitosan hybrid scaffolds for dermal tissue engineering. J Mech Behav Biomed Mater. 2012;8:204–15.CrossRefGoogle Scholar
  14. 14.
    Kane RJ, Weiss-Bilka HE, Meagher MJ, Liu Y, Gargac JA, Niebur GL, Wagner DR, Roeder RK. Hydroxyapatite reinforced collagen scaffolds with improved architecture and mechanical properties. Acta Biomater. 2015;17:16–25.CrossRefGoogle Scholar
  15. 15.
    Yin D, Wu H, Liu C, Zhang J, Zhou T, Wu J, Wan Y. Fabrication of composition-graded collagen/chitosan–polylactide scaffolds with gradient architecture and properties. React Funct Polym. 2014;83:98–106.CrossRefGoogle Scholar
  16. 16.
    Jose MV, Thomas V, Dean DR, Nyairo E. Fabrication and characterization of aligned nanofibrous PLGA/Collagen blends as bone tissue scaffolds. Polymer. 2009;50:3778–85.CrossRefGoogle Scholar
  17. 17.
    Arahira T, Todo M. Variation of mechanical behaviour of β-TCP/collagen two phase composite scaffold with mesenchymal stem cell in vitro. J Mech Behav Biomed Mater. 2016;61:464–74.CrossRefGoogle Scholar
  18. 18.
    Elango J, Zhang J, Bao B, Palaniyandi K, Wang S, Wu W, Robinson JS. Rheological, biocompatibility and osteogenesis assessment of fish collagen scaffold for bone tissue engineering. Int J Biol Macromol. 2016;91:51–59.CrossRefGoogle Scholar
  19. 19.
    Tylingo R, Gorczyca G, Mania S, Szweda P, Milewski S. Preparation and characterization of porous scaffolds from chitosan-collagen-gelatin composite. React Funct Polym. 2016;103:131–40.CrossRefGoogle Scholar
  20. 20.
    Parenteau-Bareil R, Gauvin R, Cliche S, Gariépy C, Germain L, Berthod F. Comparative study of bovine, porcine and avian collagens for the production of a tissue engineered dermis. Acta Biomater. 2011;7:3757–65.CrossRefGoogle Scholar
  21. 21.
    Arora A, Kothari A, Katti DS. Pore orientation mediated control of mechanical behavior of scaffolds and its application in cartilage-mimetic scaffold design. J Mech Behav Biomed Mater. 2015;51:169–83.CrossRefGoogle Scholar
  22. 22.
    Ghorbani F, Nojehdehian H, Zamanian A. Physicochemical and mechanical properties of freeze cast hydroxyapatite-gelatin scaffolds with dexamethasone loaded PLGA microspheres for hard tissue engineering applications. Mater Sci Eng C. 2016;69:208–20.CrossRefGoogle Scholar
  23. 23.
    Jeevithan E, Jeya Shakila R, Varatharajakumar A, Jeyasekaran G, Sukumar D. Physico-functional and mechanical properties of chitosan and calcium salts incorporated fish gelatin scaffolds. Int J Biol Macromol. 2013;60:262–7.CrossRefGoogle Scholar
  24. 24.
    Kim MS, Kim GH. Electrohydrodynamic direct printing of PCL/collagen fibrous scaffolds with a core/shell structure for tissue engineering applications. Chem Eng J. 2015;279:317–26.CrossRefGoogle Scholar
  25. 25.
    Muthukumar T, Aravinthan A, Sharmila J, Kim NS, Kim J-H. Collagen/chitosan porous bone tissue engineering composite scaffold incorporated with Ginseng compound K. Carbohyd Polym. 2016;152:566–74.CrossRefGoogle Scholar
  26. 26.
    Cao H, Chen M-M, Liu Y, Liu Y-Y, Huang Y-Q, Wang J-H, Chen J-D, Zhang Q-Q. Fish collagen-based scaffold containing PLGA microspheres forcontrolled growth factor delivery in skin tissue engineering. Colloid Surf B. 2015;136:1098–106.CrossRefGoogle Scholar
  27. 27.
    Suchý T, Šupová M, Sauerová P, Verdánová M, Sucharda Z, Rýglová Š, Žaloudková M, Sedláček R, Hubálek Kalbáčová M. The effects of different cross-linking conditions on collagen-based nanocomposite scaffolds—an in vitro evaluation using mesenchymal stem cells. Biomed Mater. 2015;10:065008.CrossRefGoogle Scholar
  28. 28.
    Murugan R, Ramakrishna S, Panduranga Rao K. Nanoporous hydroxy-carbonate apatite scaffold made of natural bone. Mater Lett. 2006;60:2844–7.CrossRefGoogle Scholar
  29. 29.
    ISO, ISO13314: 2011 Mechanical testing of metals—ductility testing—compression test for porous and cellular metals, 2011.Google Scholar
  30. 30.
    Yavari SA, Wauthle R, van der Stok J, Riemslag AC, Janssen M, Mulier M, Kruth JP, Schrooten J, Weinans H, Zadpoor AA. Fatigue behaviour of porous biomaterials manufactured using selective laser melting. Mater Sci Eng C. 2013;33:4849–58.CrossRefGoogle Scholar
  31. 31.
    Ahmadi SM, Campoli G, Yavari SA, Sajadi B, Wauthlé R, Schrooten J, Weinans H, Zadpoor A. Mechanical behavior of regular open-cell porous biomaterials made of diamond lattice unit cells. J Mech Behav Biomed. 2014;34:106–15.CrossRefGoogle Scholar
  32. 32.
    Bobe K, Willbold E, Morgenthal I, Andersen O, Studnitzky T, Nellesen J, Tillmann W, Vogt C, Vano K, Witte F. In vitro and in vivo evaluation of biodegradable, open-porous scaffolds made of sintered magnesium W4 short fibres. Acta Biomater. 2013;9:8611–23.CrossRefGoogle Scholar
  33. 33.
    Piola M, Soncini M, Cantini M, Sadr N, Ferrario G, Fiore GB. Design and functional testing of a multichamber perfusion platform for three-dimensional scaffolds. Sci World J. 2013;2013:123974.CrossRefGoogle Scholar
  34. 34.
    Kozłowska J, Sionkowska A. Effects of different crosslinking methods on the properties of collagen–calcium phosphate composite materials. Int J Biol Macromol. 2015;74:397–403.CrossRefGoogle Scholar
  35. 35.
    Kemppainen JM, Hollister SJ. Differential effects of designed scaffold permeability on chondrogenesis by chondrocytes and bone marrow stromal cells. Biomaterials. 2010;31:279–87.CrossRefGoogle Scholar
  36. 36.
    O’Brien FJ, Harley BA, Waller MA, Yannas IV, Gibson LJ. The effect of pore size on permeability and cell attachment in collagen scaffolds for tissue engineering. Technol Health Care. 2007;15:3–17.Google Scholar
  37. 37.
    Wang Y, Tomlins PE, Coombes AG, Rides M. On the determination of Darcy permeability coefficients for a microporous tissue scaffold. Tissue Eng C. 2010;16:281–9.CrossRefGoogle Scholar
  38. 38.
    Villa MM, Wang L, Huang J, Rowe DW, Wei M. Bone tissue engineering with a collagen-hydroxyapatite scaffold and culture expanded bone marrow stromal cells. J Biomed Mater Res B. 2015;103:243–53.CrossRefGoogle Scholar
  39. 39.
    Grimm WJ, Williams MJ. Measurements of permeability in human calcaneal trabecular bone. J Biomech. 1997;30:743–5.CrossRefGoogle Scholar
  40. 40.
    Wen D, Androjna C, Vasanji A, Belovich J, Midura RJ. Lipids and collagen matrix restrict the hydraulic permeability within the porous compartment of adult cortical bone. Ann Biomed Eng. 2010;38:558–69.CrossRefGoogle Scholar
  41. 41.
    Gautieri A, Vesentini S, Redaelli A, Buehler MJ. Hierarchical structure and nanomechanics of collagen microfibrils from the atomistic scale up. Nano Lett. 2011;11:757–66.CrossRefGoogle Scholar
  42. 42.
    Fratzl P. Collagen: structure and mechanics. Springer: New York, 2008.Google Scholar
  43. 43.
    Venugopal J, Ramakrishna S. Applications of polymer nanofibers in biomedicine and biotechnology. Appl Biochem Biotechnol. 2005;125:147–58.CrossRefGoogle Scholar
  44. 44.
    Rampichová M, Chvojka J, Buzgo M, Prosecká E, Mikeš P, Vysloužilová L, Tvrdík D, Kochová P, Gregor T, Lukáš D, Amler E. Elastic three-dimensional poly (ε-caprolactone) nanofibre scaffold enhances migration, proliferation and osteogenic differentiation of mesenchymal stem cells. Cell Prolif. 2013;46:23–37.CrossRefGoogle Scholar
  45. 45.
    Engler AJ, Sen S, Sweeney HL, Discher DE. Matrix elasticity directs stem cell lineage specification. Cell. 2006;126:677–89.CrossRefGoogle Scholar
  46. 46.
    Kasten P, Beyen I, Niemeyer P, Luginbühl R, Bohner M, Richter W. Porosity and pore size of β-tricalcium phosphate scaffold can influence protein production and osteogenic differentiation of human mesenchymal stem cells: An in vitro and in vivo study. Acta Biomater. 2008;4:1904–15.CrossRefGoogle Scholar
  47. 47.
    Walters NJ, Gentleman E. Evolving insights in cell–matrix interactions: elucidating how non-soluble properties of the extracellular niche direct stem cell fate. Acta Biomater. 2015;11:3–16.CrossRefGoogle Scholar
  48. 48.
    Anselme K, Ploux L, Ponche A. Cell/material interfaces: influence of surface chemistry and surface topography on cell adhesion. J Adhes Sci Technol. 2012;24:831–52.CrossRefGoogle Scholar
  49. 49.
    Dutta P, Hajra S, Chattoraj DK. Binding of water and solute to protein-mixture and protein-coated alumina. Indian J Biochem Biophys. 1997;34:449–60.Google Scholar
  50. 50.
    Feng B, Chen J, Zhang X. Interaction of calcium and phosphate in apatite coating on titanium with serum albumin. Biomaterials. 2002;23:2499–507.CrossRefGoogle Scholar
  51. 51.
    Akin FA, Zreiqat H, Jordan S, Wijesundara MBJ, Hanley L. Preparation and analysis of macroporous TiO2 films on Ti surfaces for bone–tissue implants. J Biomed Mater Res. 2001;57:588–96.CrossRefGoogle Scholar
  52. 52.
    Mygind T, Stiehler M, Baatrup A, Li H, Zou X, Flyvbjerg A, Kassem M, Bünger C. Mesenchymal stem cell ingrowth and differentiation on coralline hydroxyapatite scaffolds. Biomaterials. 2007;28:1036–47.CrossRefGoogle Scholar
  53. 53.
    Cooper DML, Matyas JR, Katzenberg MA, Hallgrimsson B. Comparison of microcomputed tomographic and microradiographic measurements of cortical bone porosity. Calcif Tissue Int. 2004;74:437–47.CrossRefGoogle Scholar
  54. 54.
    Keaveny TM, Morgan EF, Niebur GL, Yeh OC. Biomechanics of trabecular bone. Annu Rev Biomed Eng. 2001;3:307–33.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Tomáš Suchý
    • 1
    • 2
  • Monika Šupová
    • 1
  • Martin Bartoš
    • 3
  • Radek Sedláček
    • 2
  • Marco Piola
    • 4
  • Monica Soncini
    • 4
  • Gianfranco Beniamino Fiore
    • 4
  • Pavla Sauerová
    • 5
    • 6
  • Marie Hubálek  Kalbáčová
    • 5
    • 6
  1. 1.Department of Composites and Carbon Materials, Institute of Rock Structure and MechanicsAcademy of Sciences of the Czech RepublicPrague 8Czech Republic
  2. 2.Laboratory of Biomechanics, Department of Mechanics, Biomechanics and Mechatronics, Faculty of Mechanical EngineeringCzech Technical University in PraguePrague 6Czech Republic
  3. 3.Department of Stomatology, First Faculty of MedicineCharles University and General University Hospital in PraguePrague 2Czech Republic
  4. 4.Dipartimento di ElettronicaInformazione e Bioingegneria, Politecnico di MilanoMilanoItaly
  5. 5.Biomedical Centre, Faculty of Medicine in PilsenCharles UniversityPilsenCzech Republic
  6. 6.Institute of Inherited Metabolic Disorders, 1st Faculty of MedicineCharles University in PraguePrague 2Czech Republic

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