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Bioprinting predifferentiated adipose-derived mesenchymal stem cell spheroids with methacrylated gelatin ink for adipose tissue engineering

  • Tissue Engineering Constructs and Cell Substrates
  • Original Research
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Abstract

The increasing number of mastectomies results in a greater demand for breast reconstruction characterized by simplicity and a low complication profile. Reconstructive surgeons are investigating tissue engineering (TE) strategies to overcome the current surgical drawbacks. 3D bioprinting is the rising technique for the fabrication of large tissue constructs which provides a potential solution for unmet clinical needs in breast reconstruction building on decades of experience in autologous fat grafting, adipose-derived mesenchymal stem cell (ASC) biology and TE. A scaffold was bioprinted using encapsulated ASC spheroids in methacrylated gelatin ink (GelMA). Uniform ASC spheroids with an ideal geometry and diameter for bioprinting were formed, using a high-throughput non-adhesive agarose microwell system. ASC spheroids in adipogenic differentiation medium (ADM) were evaluated through live/dead staining, histology (HE, Oil Red O), TEM and RT-qPCR. Viable spheroids were obtained for up to 14 days post-printing and showed multilocular microvacuoles and successful differentiation toward mature adipocytes shown by gene expression analysis. Moreover, spheroids were able to assemble at random in GelMA, creating a macrotissue. Combining the advantage of microtissues to self-assemble and the controlled organization by bioprinting technologies, these ASC spheroids can be useful as building blocks for the engineering of soft tissue implants.

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References

  1. WHO. Cancer fact sheets [internet]. 2017 [cited 5 January 2018]. Available from: http://globocan.iarc.fr/Pages/fact_sheets_cancer.aspx.

  2. Alaofi RK, Nassif MO, Al-Hajeili MR. Prophylactic mastectomy for the prevention of breast cancer: Review of the literature. Avicenna J Med. 2018;8:67–77.

    Google Scholar 

  3. Fracon S, et al. Patient satisfaction after breast reconstruction: implants vs. autologous tissues. Acta Chir Plast. 2018;59:120–8.

    CAS  Google Scholar 

  4. Blondeel PN. One hundred free DIEP flap breast reconstructions: a personal experience. Br J Plast Surg. 1999;52:104–11.

    CAS  Google Scholar 

  5. Critchley AC, et al. Current controversies in breast cancer surgery. Clin Oncol (R Coll Radiol). 2013;25:101–8.

    CAS  Google Scholar 

  6. O’Halloran NA, et al. Hydrogels in adipose tissue engineering - potential application in post-mastectomy breast regeneration. J Tissue Eng Regen Med. 2018;12:2234–47.

    Google Scholar 

  7. Zhang YS, et al. 3D bioprinting for tissue and organ fabrication. Ann Biomed Eng. 2016;45:148–63.

    CAS  Google Scholar 

  8. Guiro K, Arinzeh TL. Bioengineering models for breast cancer research. Breast Cancer. 2015;9:57–70.

    CAS  Google Scholar 

  9. Laschke MW, Menger MD. Life is 3D: boosting spheroid function for tissue engineering. Trends Biotechnol. 2017;35:133–44.

    CAS  Google Scholar 

  10. Jakab K, et al. Tissue engineering by self-assembly of cells printed into topologically defined structures. Tissue Eng Part A. 2008;14:413–21.

    CAS  Google Scholar 

  11. Steinberg MS. Differential adhesion in morphogenesis: a modern view. Curr Opin Genet Dev. 2007;17:281–6.

    CAS  Google Scholar 

  12. Nichol JW, Khademhosseini A. Modular tissue engineering: engineering biological tissues from the bottom up. Soft Matter. 2009;5:1312–9.

    CAS  Google Scholar 

  13. Dean DM, et al. Rods, tori, and honeycombs: the directed self-assembly of microtissues with prescribed microscale geometries. FASEB J. 2007;21:4005–12.

    CAS  Google Scholar 

  14. Jakab K, et al. Tissue engineering by self-assembly and bio-printing of living cells. Biofabrication. 2010;2:022001.

    Google Scholar 

  15. Jakab K, et al. Relating cell and tissue mechanics: implications and applications. Dev Dyn. 2008;237:2438–49.

    Google Scholar 

  16. Miyagawa Y, et al. A microfabricated scaffold induces the spheroid formation of human bone marrow-derived mesenchymal progenitor cells and promotes efficient adipogenic differentiation. Tissue Eng Part A. 2011;17:513–21.

    CAS  Google Scholar 

  17. Turner PA, et al. Adipogenic differentiation of human adipose-derived stem cells grown as spheroids. Process Biochem. 2017;59:312–20.

    CAS  Google Scholar 

  18. Cheng NC, Wang S, Young TH. The influence of spheroid formation of human adipose-derived stem cells on chitosan films on stemness and differentiation capabilities. Biomaterials. 2012;33:1748–58.

    CAS  Google Scholar 

  19. Kapur SK, et al. Human adipose stem cells maintain proliferative, synthetic and multipotential properties when suspension cultured as self-assembling spheroids. Biofabrication. 2012;4:025004.

    CAS  Google Scholar 

  20. Murphy SV, Atala A. 3D bioprinting of tissues and organs. Nat Biotechnol. 2014;32:773–85.

    CAS  Google Scholar 

  21. Wang X, Liu C. 3D bioprinting of adipose-derived stem cells for organ manufacturing. Adv Exp Med Biol. 2018;1078:3–14.

    CAS  Google Scholar 

  22. Chang CC, et al. Direct-write bioprinting three-dimensional biohybrid systems for future regenerative therapies. J Biomed Mater Res B Appl Biomater. 2011;98:160–70.

    Google Scholar 

  23. Kang HW, et al. A 3D bioprinting system to produce human-scale tissue constructs with structural integrity. Nat Biotechnol. 2016;34:312–9.

    CAS  Google Scholar 

  24. Kolesky DB, et al. 3D bioprinting of vascularized, heterogeneous cell-laden tissue constructs. Adv Mater. 2014;26:3124–30.

    CAS  Google Scholar 

  25. Panwar A, Tan LP. Current status of bioinks for micro-extrusion-based 3D bioprinting. Molecules. 2016;21:685.

  26. Vashi AV, et al. Adipose tissue engineering based on the controlled release of fibroblast growth factor-2 in a collagen matrix. Tissue Eng. 2006;12:3035–43.

    CAS  Google Scholar 

  27. Yao R, et al. Injectable cell/hydrogel microspheres induce the formation of fat lobule-like microtissues and vascularized adipose tissue regeneration. Biofabrication. 2012;4:045003.

    Google Scholar 

  28. Van Vlierberghe S, Dubruel P, Schacht E. Biopolymer-based hydrogels as scaffolds for tissue engineering applications: a review. Biomacromolecules. 2011;12:1387–408.

    Google Scholar 

  29. Tan H, et al. Injectable in situ forming biodegradable chitosan-hyaluronic acid based hydrogels for cartilage tissue engineering. Biomaterials. 2009;30:2499–506.

    CAS  Google Scholar 

  30. Wang Z, et al. A simple and high-resolution stereolithography-based 3D bioprinting system using visible light crosslinkable bioinks. Biofabrication. 2015;7:045009.

    Google Scholar 

  31. Billiet T, et al. The 3D printing of gelatin methacrylamide cell-laden tissue-engineered constructs with high cell viability. Biomaterials. 2014;35:49–62.

    CAS  Google Scholar 

  32. Occhetta P, et al. VA-086 methacrylate gelatine photopolymerizable hydrogels: a parametric study for highly biocompatible 3D cell embedding. J Biomed Mater Res A. 2015;103:2109–17.

    CAS  Google Scholar 

  33. Clevenger TN, et al. Cell-mediated remodeling of biomimetic encapsulating hydrogels triggered by adipogenic differentiation of adipose stem cells. J Tissue Eng. 2016;7:2041731416670482.

    Google Scholar 

  34. Huber B, et al. Methacrylated gelatin and mature adipocytes are promising components for adipose tissue engineering. J Biomater Appl. 2016;30:699–710.

    CAS  Google Scholar 

  35. Declercq HA, et al. Bone grafts engineered from human adipose-derived stem cells in dynamic 3D-environments. Biomaterials. 2013;34:1004–17.

    CAS  Google Scholar 

  36. Declercq H, et al. Isolation, proliferation and differentiation of osteoblastic cells to study cell/biomaterial interactions: comparison of different isolation techniques and source. Biomaterials. 2004;25:757–68.

    CAS  Google Scholar 

  37. Gevaert E, et al. High throughput micro-well generation of hepatocyte micro-aggregates for tissue engineering. PLoS ONE. 2014;9:e105171.

    Google Scholar 

  38. Berneel E, et al. Redifferentiation of high-throughput generated fibrochondrocyte micro-aggregates: impact of low oxygen tension. Cells Tissues Organs. 2016;202:369–81.

    CAS  Google Scholar 

  39. De Moor L, et al. High-throughput fabrication of vascularized spheroids for bioprinting. Biofabrication. 2018;10:035009.

    Google Scholar 

  40. Declercq HA, et al. Calcification as an indicator of osteoinductive capacity of biomaterials in osteoblastic cell cultures. Biomaterials. 2005;26:4964–74.

    CAS  Google Scholar 

  41. Rosen ED, et al. Transcriptional regulation of adipogenesis. Genes Dev. 2000;14:1293–307.

    CAS  Google Scholar 

  42. Fasshauer M, et al. Serum levels of the adipokine adipocyte fatty acid-binding protein are increased in preeclampsia. Am J Hypertens. 2008;21:582–6.

    CAS  Google Scholar 

  43. Mironov V, et al. Organ printing: tissue spheroids as building blocks. Biomaterials. 2009;30:2164–74.

    CAS  Google Scholar 

  44. Hurtley S. Spatial cell biology. Location, location, location. Introduction. Science. 2009;326:1205.

    CAS  Google Scholar 

  45. Kim SJ, et al. Hydrogels with an embossed surface: an all-in-one platform for mass production and culture of human adipose-derived stem cell spheroids. Biomaterials. 2018;188:198–212.

    Google Scholar 

  46. Pati F, et al. Biomimetic 3D tissue printing for soft tissue regeneration. Biomaterials. 2015;62:164–75.

    CAS  Google Scholar 

  47. Kayabolen A, et al. Native extracellular matrix/fibroin hydrogels for adipose tissue engineering with enhanced vascularization. Biomed Mater. 2017;12:035007.

    Google Scholar 

  48. Gungor-Ozkerim PS, et al. Bioinks for 3D bioprinting: an overview. Biomater Sci. 2018;6:915–46.

    CAS  Google Scholar 

  49. Wang Z, et al. An ultrafast hydrogel photocrosslinking method for direct laser bioprinting. RSC Adv. 2016;6:21099–104.

    CAS  Google Scholar 

  50. Zhu J, Marchant RE. Design properties of hydrogel tissue-engineering scaffolds. Expert Rev Med Devices. 2011;8:607–26.

    Google Scholar 

  51. Rouillard AD, et al. Methods for photocrosslinking alginate hydrogel scaffolds with high cell viability. Tissue Eng Part C Methods. 2011;17:173–9.

    CAS  Google Scholar 

  52. Lin RZ, et al. Transdermal regulation of vascular network bioengineering using a photopolymerizable methacrylated gelatin hydrogel. Biomaterials. 2013;34:6785–96.

    CAS  Google Scholar 

  53. Zhao Y, et al. Three-dimensional printing of Hela cells for cervical tumor model in vitro. Biofabrication. 2014;6:035001.

    Google Scholar 

  54. Eto H, et al. Characterization of structure and cellular components of aspirated and excised adipose tissue. Plast Reconstr Surg. 2009;124:1087–97.

    CAS  Google Scholar 

  55. Chhaya MP, et al. Sustained regeneration of high-volume adipose tissue for breast reconstruction using computer aided design and biomanufacturing. Biomaterials. 2015;52:551–60.

    CAS  Google Scholar 

  56. Moldovan NI, Hibino N, Nakayama K. Principles of the Kenzan method for robotic cell spheroid-based three-dimensional bioprinting. Tissue Eng Part B Rev. 2017;23:237–44.

    CAS  Google Scholar 

  57. Kim JH, et al. Therapeutic angiogenesis of three-dimensionally cultured adipose-derived stem cells in rat infarcted hearts. Cytotherapy. 2013;15:542–56.

    CAS  Google Scholar 

  58. Jain RK, et al. Engineering vascularized tissue. Nat Biotechnol. 2005;23:821–3.

    CAS  Google Scholar 

  59. Liu J, et al. Monitoring nutrient transport in tissue-engineered grafts. J Tissue Eng Regen Med. 2015;9:952–60.

    CAS  Google Scholar 

  60. Hoch E, Tovar GE, Borchers K. Bioprinting of artificial blood vessels: current approaches towards a demanding goal. Eur J Cardiothorac Surg. 2014;46:767–78.

    Google Scholar 

  61. Nunes SS, et al. Generation of a functional liver tissue mimic using adipose stromal vascular fraction cell-derived vasculatures. Sci Rep. 2013;3:2141.

    CAS  Google Scholar 

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Correspondence to Julien Colle.

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Colle, J., Blondeel, P., De Bruyne, A. et al. Bioprinting predifferentiated adipose-derived mesenchymal stem cell spheroids with methacrylated gelatin ink for adipose tissue engineering. J Mater Sci: Mater Med 31, 36 (2020). https://doi.org/10.1007/s10856-020-06374-w

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  • DOI: https://doi.org/10.1007/s10856-020-06374-w

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