Skip to main content

Preparation and Endothelialization of Multi-level Vessel-like Network in Enzymated Gelatin Scaffolds

Abstract

Loss of function of large tissues is an urgent clinical problem. Although the artificial microfluidic network fabricated in large tissue- engineered constructs has great promise, it is still difficult to develop an efficient vessel-like design to meet the requirements of the biomimetic vascular network for tissue engineering applications. In this study, we used a facile approach to fabricate a branched and multi-level vessel-like network in a large muscle scaffolds by combining stereolithography (SL) technology and enzymatic crosslinking mechanism. The morphology of microchannel cross-sections was characterized using micro-computed tomography. The square cross-sections were gradually changed to a seamless circular microfluidic network, which is similar to the natural blood vessel. In the different micro-channels, the velocity greatly affected the attachment and spread of Human Umbilical Vein Endothelial Cell (HUVEC)-Green Fluorescent Protein (GFP). Our study demonstrated that the branched and multi-level microchannel network simulates biomimetic microenvironments to promote endothelialization. The gelatin scaffolds in the circular vessel-like networks will likely support myoblast and surrounding tissue for clinical use.

This is a preview of subscription content, access via your institution.

References

  1. Hurd S, Bhati N, Walker A, Kasukonis B, Wolchok J C. Development of a biological scaffold engineered using the extracellular matrix secreted by skeletal muscle cells. Biomaterials, 2015, 49, 9–17.

    Article  Google Scholar 

  2. Valentin J E, Freytes D O, Grasman J M, Pesyna C, Freund J, Gilbert T W, Badylak S F. Oxygen diffusivity of biologic and synthetic scaffold materials for tissue engineering. Journal of Biomedical Materials Research Part A, 2009, 91, 1010–1017.

    Article  Google Scholar 

  3. Mandru M, Ionescu C, Chirita M. Modelling mechanical properties in native and biomimetically formed vascular grafts. Journal of Bionic Engineering, 2009, 6, 371–377.

    Article  Google Scholar 

  4. Cokelet G R, Soave R, Pugh G, Rathbun L. Fabrication of in vitro microvascular blood flow systems by photolithography. Microvascular Research, 1993, 46, 394–400.

    Article  Google Scholar 

  5. He J K, Chen R M, Lu Y J, Zhan L, Liu Y X, Jin Z M. Fabrication of circular microfluidic network in enzymatically- crosslinked gelatin hydrogel. Materials Science & Engineering C-Materials for Biological Applications, 2016, 59, 53–60.

    Article  Google Scholar 

  6. Choi J S, Piao Y, Seo T S. Fabrication of a circular PDMS microchannel for constructing a three-dimensional endothelial cell layer. Bioprocess and Biosystems Engineering, 2013, 36, 1871–1878.

    Article  Google Scholar 

  7. Choi J S, Piao Y, Seo T S. Circumferential alignment of vascular smooth muscle cells in a circular microfluidic channel. Biomaterials, 2014, 35, 63–70.

    Article  Google Scholar 

  8. Huang Z C, Li X, Martinsgreen M M, Liu Y X. Microfabrication of cylindrical microfluidic channel networks for microvascular research. Biomedical Microdevices, 2012, 14, 873–883.

    Article  Google Scholar 

  9. Wong C C, Agarwal A, Balasubramanian N, Kwong D L. Fabrication of self-sealed circular nano/microfluidic channels in glass substrates. Nanotechnology, 2007, 18, 135304.

    Article  Google Scholar 

  10. Jana S, Leung M, Chang J L, Zhang M Q. Effect of nanoand micro-scale topological features on alignment of muscle cells and commitment of myogenic differentiation. Biofabrication, 2014, 6, 035012.

    Article  Google Scholar 

  11. Chen S W, Nakamoto T, Kawazoe N, Chen G P. Engineering multi-layered skeletal muscle tissue by using 3D microgrooved collagen scaffolds. Biomaterials, 2015, 73, 23–31.

    Article  Google Scholar 

  12. Hosseini V, Kollmannsberger P, Ahadian S, Ostrovidov S, Kaji H, Vogel V, Khademhosseini A. Fiber-assisted molding (FAM) of surfaces with tunable curvature to guide cell alignment and complex tissue architecture. Small, 2014, 10, 4851–4857.

    Article  Google Scholar 

  13. Fiddes L K, Raz N, Srigunapalan S, Tumarkan E, Simmons C A, Wheeler A R, Kumacheva E. A circular cross-section PDMS microfluidics system for replication of cardiovascular flow conditions. Biomaterials, 2010, 31, 3459–3464.

    Article  Google Scholar 

  14. Kolesky D B, Truby R L, Gladman A S, Busbee T A, Homan K A, Lewis J A. 3D bioprinting of vascularized, heterogeneous cell-laden tissue constructs. Advanced materials, 2014, 26, 3124–3130.

    Article  Google Scholar 

  15. Kubis H P, Scheibe R J, Decker B, Hufendiek K, Hanke N, Gros G, Meissner J D. Primary skeletal muscle cells cultured on gelatin bead microcarriers develop structural and biochemical features characteristic of adult skeletal muscle. Cell Biology International, 2016, 40, 364–374.

    Article  Google Scholar 

  16. Baniasadi H, Mashayekhan S, Fadaoddini S, Haghirsharifzamini Y. Design, fabrication and characterization of oxidized alginate-gelatin hydrogels for muscle tissue engineering applications. Journal of Biomaterials Applications, 2016, 31, 152–161.

    Article  Google Scholar 

  17. Hajiabbas M, Mashayekhan S, Nazaripouya A, Naji M, Hunkeler D, Zeleti S R, Sharifiaghdas F. Chitosan-gelatin sheets as scaffolds for muscle tissue engineering. Artificial Cells, Nanomedicine, and Biotechnology, 2015, 43, 124–132.

    Article  Google Scholar 

  18. Dong G R, Lian Q, Yang L X, Mao W, Liu S Y, Xu C. Design and fabrication of vascular network for muscle tissue engineering. Advances in Engineering Research, 2016, 75, 613–621.

    Google Scholar 

  19. Murray C D. The physiological principle of minimum work: I. The vascular system and the cost of blood volume. Proceedings of the National Academy of Sciences, 1926, 12, 207–214.

    Google Scholar 

  20. Zamir M. Optimality principles in arterial branching. Journal of Theoretical Biology, 1976, 62, 227–251.

    Article  Google Scholar 

  21. Kroll M H, Hellums J D, Mcintire L V, Schafer A I, Moake J L. Platelets and shear stress. Blood, 1996, 88, 1525–1541.

    Google Scholar 

  22. Lowe G D O. Virchow’s triad revisited: Abnormal flow. Pathophysiol Haemost Thromb, 2003, 33, 455–457.

    Article  Google Scholar 

  23. Hoganson D M, Pryor H I, Spool I D, Burns O H, Gilmore J R, Vacanti J P. Principles of biomimetic vascular network design applied to a tissue-engineered liver scaffold. Tissue Engineering Part A, 2010, 16, 1469–1477.

    Article  Google Scholar 

  24. Camp J P, Shuler S M L. Fabrication of a multiple-diameter branched network of microvascular channels with semicircular cross-sections using xenon difluoride etching. Biomedical Microdevices, 2008, 10, 179–186.

    Article  Google Scholar 

  25. Huh D, Matthews B D, Mammoto A, Montoya-Zavala M, Hsin H Y, Ingber D E. Reconstituting organ-level lung functions on a chip. Science, 2010, 328, 1662–1668.

    Article  Google Scholar 

  26. Shah G, Costello B J. Soft tissue regeneration incorporating 3-dimensional biomimetic scaffolds. Oral & Maxillofacial Surgery Clinics, 2017, 29, 9–18.

    Article  Google Scholar 

  27. Badhe R V, Bijukumar D, Chejara D R, Mabrouk M, Choonara Y E, Kumar P, du Toit L C, Kondiah P P D, Pillay V. A composite chitosan-gelatin bi-layered, biomimetic macroporous scaffold for blood vessel tissue engineering. Carbohydrate Polymers, 2017, 157, 1215–1225.

    Article  Google Scholar 

  28. Mansouri N, Bagheri S. The influence of topography on tissue engineering perspective. Materials Science & Engineering C–Materials for Biological Applications, 2016, 61, 906–921.

    Article  Google Scholar 

  29. Visone R, Gilardi M, Marsano A, Rasponi M, Bersini S, Moretti M. Cardiac meets skeletal: What’s new in microfluidic models for muscle tissue engineering. Molecules, 2016, 21, 1128.

    Article  Google Scholar 

  30. Jeong K Y, Ji S L, Paik D H, Jung W K, Choi S W. Fabrication of cell-penetrable microfibrous matrices with a highly porous structure using a simple fluidic device for tissue engineering. Materials Letters, 2016, 168, 116–120.

    Article  Google Scholar 

  31. Agarwal P, Choi J K, Huang H S, Zhao S T, Dumbleton J, Li J R, He X M. A biomimetic core-shell platform for miniaturized 3D cell and tissue engineering. Particle & Particle Systems Characterization, 2015, 32, 809–816.

    Article  Google Scholar 

  32. Ren L, Qian Z H, Ren L Q. Biomechanics of musculoskeletal system and its biomimetic implications: A review. Journal of Bionic Engineering, 2014, 11, 159–175.

    Article  Google Scholar 

  33. Ostrovidov S, Ahadian S, Ramon-Azcon J, Hosseini V, Fujie T, Parthiban S P, Shiku H, Matsue T, Kaji H, Ramalingam M, Bae H, Khademhosseini A. Three-dimensional co-culture of C2C12/PC12 cells improves skeletal muscle tissue formation and function. Journal of Tissue Engineering and Regenerative Medicine, 2017, 11, 582–595.

    Article  Google Scholar 

  34. 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. Materials Science & Engineering C–Materials for Biological Applications, 2016, 61, 473–483.

    Article  Google Scholar 

  35. Williams C, Xie A W, Yamato M, Okano T, Wong J Y. Stacking of aligned cell sheets for layer-by-layer control of complex tissue structure. Biomaterials, 2011, 32, 5625–5632.

    Article  Google Scholar 

  36. Syed H, Unnikrishnan V U, Olcmen S. Characteristics of time-varying intracranial pressure on blood flow through cerebral artery: A fluid-structure interaction approach. Proceedings of the Institution of Mechanical Engineers, Part H, 2016, 230, 111–121.

    Article  Google Scholar 

  37. Hudlicka O, Brown M D. Adaptation of skeletal muscle microvasculature to increased or decreased blood flow: Role of shear stress, nitric oxide and vascular endothelial growth factor. Journal of Vascular Research, 2009, 46, 504–512.

    Article  Google Scholar 

  38. Issa B, Moore R J, Bowtell R W, Baker P N, Johnson I R, Worthington B S, Gowland P A. Quantification of blood velocity and flow rates in the uterine vessels using echo planar imaging at 0.5 Tesla. Journal of Magnetic Resonance Imaging, 2010. 31, 921–927.

    Article  Google Scholar 

Download references

Acknowledgment

This work was supported by National Natural Science Foundation of China (Grant No. 51375371) and the High-Tech Projects of China (Grant Nos. 2015AA020303 and 2015AA042503).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Qin Lian.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Dong, G., Lian, Q., Yang, L. et al. Preparation and Endothelialization of Multi-level Vessel-like Network in Enzymated Gelatin Scaffolds. J Bionic Eng 15, 673–681 (2018). https://doi.org/10.1007/s42235-018-0055-3

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s42235-018-0055-3

Keywords

  • large-size scaffolds
  • vascular network
  • enzymatically gelatin hydrogel
  • endothelialization
  • muscle tissue engineering