OBJECTIVES: We prepared 3D poly (ε-caprolactone) (PCL) nanofibre scaffolds and tested their use for seeding, proliferation, differentiation and migration of mesenchymal stem cell (MSCs). MATERIALS AND METHODS: 3D nanofibres were prepared using a special collector for common electrospinning; simultaneously, a 2D PCL nanofibre layer was prepared using a classic plain collector. Both scaffolds were seeded with MSCs and biologically tested. MSC adhesion, migration, proliferation and osteogenic differentiation were investigated. RESULTS: The 3D PCL scaffold was characterized by having better biomechanical properties, namely greater elasticity and resistance against stress and strain, thus this scaffold will be able to find broad applications in tissue engineering. Clearly, while nanofibre layers of the 2D scaffold prevented MSCs from migrating through the conformation, cells infiltrated freely through the 3D scaffold. MSC adhesion to the 3D nanofibre PCL layer was also statistically more common than to the 2D scaffold (P < 0.05), and proliferation and viability of MSCs 2 or 3 weeks post-seeding, were also greater on the 3D scaffold. In addition, the 3D PCL scaffold was also characterized by displaying enhanced MSC osteogenic differentiation. CONCLUSIONS: We draw the conclusion that all positive effects observed using the 3D PCL nanofibre scaffold are related to the larger fibre surface area available to the cells. Thus, the proposed 3D structure of the nanofibre layer will find a wide array of applications in tissue engineering and regenerative medicine.

Laboratory of Tissue Engineering Institute of Experimental Medicine Academy of Science of the Czech Republic Videnska 1083 142 40 Prague Czech Republic

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Langer R, Vacanti J (1993) Tissue engineering. Science 260, 920–926. PubMed

Hutmacher DW, Schantz JT, Lam CX, Tan KC, Lim TC (2007) State of the art and future directions of scaffold‐based bone engineering from a biomaterials perspective. J. Tissue Eng. Regen. Med. 1, 245–260. Epub 2007/11/27. PubMed

Karageorgiou V, Kaplan D (2005) Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials 26, 5474–5491. Epub 2005/04/30. PubMed

Lowery JL, Datta N, Rutledge GC (2010) Effect of fiber diameter, pore size and seeding method on growth of human dermal fibroblasts in electrospun poly(epsilon‐caprolactone) fibrous mats. Biomaterials 31, 491–504. Epub 2009/10/14. PubMed

Baker BM, Gee AO, Metter RB, Nathan AS, Marklein RA, Burdick JA PubMed PMC

Guimaraes A, Martins A, Pinho ED, Faria S, Reis RL, Neves NM (2010) Solving cell infiltration limitations of electrospun nanofiber meshes for tissue engineering applications. Nanomedicine (Lond). 5, 539–554. Epub 2010/06/10. PubMed

Nam J, Huang Y, Agarwal S, Lannutti J (2007) Improved cellular infiltration in electrospun fiber via engineered porosity. Tissue Eng. 13, 2249–2257. Epub 2007/06/01. PubMed PMC

Simonet M, Schneider OD, Neuenschwander P, Stark WJ (2007) Ultraporous 3D polymer meshes by low‐temperature electrospinning: use of ice crystals as a removable void template. Polym. Eng. Sci. 47, 2020–2026.

Sundararaghavan HG, Metter RB, Burdick JA (2010) Electrospun fibrous scaffolds with multiscale and photopatterned porosity. Macromol. Biosci. 10, 265–270. Epub 2009/12/17. PubMed PMC

Liao S, Li B, Ma Y, Wei H, Chan C, Ramakrishna S (2006) Biomimetic electrospun nanofibers for tissue regeneration. Biomed Mater. 1(3), 45–53. PubMed

Li WJ, Laurencin CT, Caterson EJ, Tuan RS, Ko FK (2002) Electrospun nanofibrous structure: a novel scaffold for tissue engineering. J. Biomed. Mater. Res. 60, 613–621. Epub 2002/04/12. PubMed

Liang D, Hsiao BS, Chu B (2007) Functional electrospun nanofibrous scaffolds for biomedical applications. Adv. Drug Deliv. Rev. 59, 1392–1412. PubMed PMC

Noh HK, Lee SW, Kim JM, Oh JE, Kim KH, Chung CP PubMed

Park WH, Min BM, Lee G, Kim SH, Nam YS, Lee TS (2004) Electrospinning of silk fibroin nanofibers and its effect on the adhesion and spreading of normal human keratinocytes and fibroblasts PubMed

Rho KS, Jeong L, Lee G, Seo BM, Park YJ, Hong SD PubMed

Pham QP, Sharma U, Mikos AG (2006) Electrospun poly(epsilon‐caprolactone) microfiber and multilayer nanofiber/microfiber scaffolds: characterization of scaffolds and measurement of cellular infiltration. Biomacromolecules 7, 2796–2805. Epub 2006/10/10. PubMed

Eichhorn SJ, Sampson WW (2005) Statistical geometry of pores and statistics of porous nanofibrous assemblies. J. R. Soc. Interface 2, 309–318. Epub 2006/07/20. PubMed PMC

Blakeney BA, Tambralli A, Anderson JM, Andukuri A, Lim DJ, Dean DR PubMed PMC

Badami AS, Kreke MR, Thompson MS, Riffle JS, Goldstein AS (2006) Effect of fiber diameter on spreading, proliferation, and differentiation of osteoblastic cells on electrospun poly(lactic acid) substrates. Biomaterials 27, 596–606. Epub 2005/07/19. PubMed

Schofer MD, Fuchs‐Winkelmann S, Grabedunkel C, Wack C, Dersch R, Rudisile M PubMed PMC

Norris SC, Humpolickova J, Amler E, Huranova M, Buzgo M, Machan R PubMed

Yasuda K, Inoue S, Tabata Y (2004) Influence of culture method on the proliferation and osteogenic differentiation of human adipo‐stromal cells in nonwoven fabrics. Tissue Eng. 10, 1587–1596. Epub 2004/12/14. PubMed

Bashur CA, Dahlgren LA, Goldstein AS (2006) Effect of fiber diameter and orientation on fibroblast morphology and proliferation on electrospun poly(D,L‐lactic‐co‐glycolic acid) meshes. Biomaterials 27, 5681–5688. Epub 2006/08/18. PubMed

Riboldi SA, Sadr N, Pigini L, Neuenschwander P, Simonet M, Mognol P PubMed

Vaquette C, Cooper‐White JJ (2011) Increasing electrospun scaffold pore size with tailored collectors for improved cell penetration. Acta Biomater. 7, 2544–2557. Epub 2011/03/05. PubMed

Milleret V, Simona B, Neuenschwander P, Hall H (2011) Tuning electrospinning parameters for production of 3D‐fiber‐fleeces with increased porosity for soft tissue engineering applications. Eur Cell Mater. 21, 286–303. Epub 2011/03/25. PubMed

Li WJ, Tuli R, Huang X, Laquerriere P, Tuan RS (2005) Multilineage differentiation of human mesenchymal stem cells in a three‐dimensional nanofibrous scaffold. Biomaterials 26, 5158–5166. Epub 2005/03/29. PubMed

Li WJ, Tuli R, Okafor C, Derfoul A, Danielson KG, Hall DJ PubMed

Li WJ, Danielson KG, Alexander PG, Tuan RS (2003) Biological response of chondrocytes cultured in three‐dimensional nanofibrous poly(epsilon‐caprolactone) scaffolds. J Biomed Mater Res A. 67, 1105–1114. Epub 2003/11/19. PubMed

Lim CT, Tan EPS, Ng SY (2008) Effects of crystalline morphology on the tensile properties of electrospun polymer nanofibers. Appl. Phys. Lett. 92, 141908 (3 pages).

Prosecka E, Buzgo M, Rampichova M, Kocourek T, Kochova P, Vyslouzilova L PubMed PMC

Mickova A, Buzgo M, Benada O, Rampichova M, Fisar Z, Filova E PubMed

Tvrdik D, Povysil C, Svatosova J, Dundr P (2005) Molecular diagnosis of synovial sircoma: RT‐PCR detection of SYT‐SSX1/2 fusion transcripts in paraffin‐embedded tissue. Med. Sci. Monit. 11, Mt1–Mt7. PubMed

Rampichova M, Kostakova E, Filova E, Prosecka E, Plencner M, Ocheretna L PubMed

Zhu X, Cui W, Li X, Jin Y (2008) Electrospun fibrous mats with high porosity as potential scaffolds for skin tissue engineering. Biomacromolecules 9, 1795–1801. Epub 2008/06/27. PubMed

Zussman E (2003) Formation of nanofiber crossbars in electrospinning. Appl. Phys. Lett. 82, 973.

Li D, Wang Y, Xia Y (2003) Electrospinning of polymeric and ceramic nanofibers as uniaxially aligned arrays. Nano Lett. 3, 1167–1171.

Li D, Wang Y, Xia Y (2004) Electrospinning nanofibers as uniaxially aligned arrays and layer‐by‐layer stacked films. Adv Mater (Weinheim, Ger). 16, 361–366.

Li D, Ouyang G, McCann JT, Xia Y (2005) Collecting electrospun nanofibers with patterned electrodes. Nano Lett. 5, 913–916. Epub 2005/05/12. PubMed

Walonen PK, Moutos FT, Kusanagi A, Moretti MG, Diekman BO, Welter JF PubMed PMC

Wise JK, Yarin AL, Megaridis CM, Cho M (2009) Chondrogenic differentiation of human mesenchymal stem cells on oriented nanofibrous scaffolds: engineering the superficial zone of articular cartilage. Tissue Eng. Part A 15, 913–921. PubMed PMC

Mountziaris PM, Tzouanas SN, Mikos AG (2010) Dose effect of tumor necrosis factor‐alpha on PubMed PMC

Zhu L, Skoultchi AI (2001) Coordinating cell proliferation and differentiation. Curr. Opin. Genet. Dev. 11, 91–97. PubMed

Conlon I, Raff M (1999) Size control in animal development. Cell 96, 235–244. PubMed

Xia K, Xue H, Dong D, Zhu S, Wang J PubMed PMC

Brown G, Hughes PJ, Michell RH (2003) Cell differentiation and proliferation‐simultaneous but independent? Exper Cell Res. 291, 282–288. PubMed

Lode A, Bernhardt A, Gelinsky M (2008) 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 Regen Med. 2, 400–407. PubMed

Fisher JP, Kim K, Dean D, Lu AQ, Mikos AG (2011) Early osteogenic signal expression of rat bone marrow stromal cells is influenced by both hydroxyapatite nanoparticle content and initial cell seeding density in biodegradable nanocomposite scaffolds. Acta Biomater. 7, 1249–1264. PubMed PMC

Grayson WL, Bhumiratana S, Cannizzaro C, Chao PH, Lennon DP, Caplan AI PubMed PMC

McBeath R, Pirone DM, Nelson CM, Bhadriraju K, Chen CS (2004) Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev. Cell 6, 483–495. PubMed

Optimizing PCL/PLGA Scaffold Biocompatibility Using Gelatin from Bovine, Porcine, and Fish Origin

Dynamic creep properties of a novel nanofiber hernia mesh in abdominal wall repair

Osteoinductive 3D scaffolds prepared by blend centrifugal spinning for long-term delivery of osteogenic supplements

A polypropylene mesh modified with poly-ε-caprolactone nanofibers in hernia repair: large animal experiment

Composite 3D printed scaffold with structured electrospun nanofibers promotes chondrocyte adhesion and infiltration

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

Osteogenic differentiation of 3D cultured mesenchymal stem cells induced by bioactive peptides

Platelet-functionalized three-dimensional poly-ε-caprolactone fibrous scaffold prepared using centrifugal spinning for delivery of growth factors

Highly efficient mesenchymal stem cell proliferation on poly-ε-caprolactone nanofibers with embedded magnetic nanoparticles

Significant improvement of biocompatibility of polypropylene mesh for incisional hernia repair by using poly-ε-caprolactone nanofibers functionalized with thrombocyte-rich solution

Abdominal closure reinforcement by using polypropylene mesh functionalized with poly-ε-caprolactone nanofibers and growth factors for prevention of incisional hernia formation

Cell penetration to nanofibrous scaffolds: Forcespinning®, an alternative approach for fabricating 3D nanofibers