The Use of Alginate to Inhibit Mineralization for Eventual Vascular Development

Abstract

Bone is a complex tissue with a complex architecture and numerous types of cells. Bone can contain two different architectures (cortical and trabecular) and several cell types (osteoblasts, osteoclasts, osteocytes, neurons, and stem cells to name a few). Nutrient transport is extremely important for a tissue with such complexity in design and cellular population. Therefore, when considering tissue engineering or regenerative engineering, options for bone vascularization must be considered. Along with placement of the vasculature, conditions for vascular development should be of the utmost importance. Optimizing the tissue-engineered scaffold for vascular development can often go overlooked as researchers focus on bone growth and scaffold strength. In this study, we look at scaffold conditions that are best for vascular development in bone scaffolding. Specifically, we investigate how the presence of crystalized calcium phosphate on a scaffold surface affects the ability of vascular endothelial cells to proliferate and differentiate for the eventual production of blood vessels. In addition, we use alginate as a way to maintain a “vascular friendly” surface inside of a mineralized scaffold. The results show that vascular endothelial cells prefer a non-mineralized scaffold and that the presence of alginate on surfaces during mineralization of a scaffold protects that surface from mineralization, making it capable of producing vasculature from seeded vascular endothelial cells.

Lay Summary

A piece of bone contains many structures, tissues, and cell types. In order to support them all, a bone has an extensive network of blood vessels. This makes the formation of useable blood vessels important for tissue engineering. In this study, we investigate how calcium phosphate formation affects blood vessels inside of bone scaffolds. Specifically, we investigate how calcium phosphate on a scaffold affects blood vessel formation in bone tissue engineering scaffolds. In addition, we use alginate to enhance vessel formation in the scaffolds. The results show that vessel formation is optimal without calcium phosphate on the scaffolds.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

References

  1. 1.

    Who are candidates for preventation and treatment for osteoporosis?. Osteoporos Int, 1997. 7(1): p. 7.

  2. 2.

    Kanis JA, et al. Long-term risk of osteoporotic fracture in Malmo. Osteoporos Int. 2000;11(8):669–74.

    CAS  Google Scholar 

  3. 3.

    Melton LJ, et al. Bone density and fractures risk in men. J Bone Miner Res. 1998;13(12):1915–23.

    Google Scholar 

  4. 4.

    CR P. Bone repair techniques, bone graft, and bone graft substitutes. Clin Orthop Relat Res. 1999;360:71–86.

    Google Scholar 

  5. 5.

    Shin M, Abukawa H, Troulis MJ, Vacanti JP. Development of a biodegradable scaffold with interconnected pores by heat fusion and its application to bone tissue engineering. J Biomed Mater Res A. 2008;84(3):702–9.

    Google Scholar 

  6. 6.

    Melton LJ, et al. Perspective. How many women have osteoporosis. J Bone Miner Res. 1992;7(9):10005–1010.

    Google Scholar 

  7. 7.

    Randell A, et al. Direct clinical and welfare costs of osteoporotic fractures in elderly men and women. Osteoporos Int. 1995;5(6):427–32.

    CAS  Google Scholar 

  8. 8.

    Wang X, Puram S. The toughness of cortical bone and its relationship with age. Ann Biomed Eng. 2004;32(1):123–35.

    Google Scholar 

  9. 9.

    An YH, Draughn RA. Mechanical testing of bone and the bone-implant interface. Boca Raton: CRC Press; 2000.

    Google Scholar 

  10. 10.

    Bose S, Roy M, Bandyopadhyay A. Recent advances in bone tissue engineering scaffolds. Trends Biotechnol. 2012;30(10):546–54.

    CAS  Google Scholar 

  11. 11.

    Krishnan L, Willett NJ, Guldberg RE. Vascularization strategies for bone regeneration. Ann Biomed Eng. 2014;42(2):432–44.

    Google Scholar 

  12. 12.

    Liu X, et al. Vascularized bone tissue formation induced by fiber-reinforced scaffolds cultured with osteoblasts and endothelial cells. Biomed Res Int. 2013;2013:854917.

    Google Scholar 

  13. 13.

    Fedorovich N, et al. The role of endothelial progenitor cells in prevascularized bone tissue engineering: development of heterogeneous constructs. Tissue Eng A. 2010;16(7):2355–67.

    CAS  Google Scholar 

  14. 14.

    Mongiat, M., et al., Extracellular matrix, a hard player in angiogenesis. Int J Mol Sci, 2016. 17(11).

  15. 15.

    Ausprunk DH, Folkman J. Migration and proliferation of endothelial cells in preformed and newly formed blood vessels during tumor angiogenesis. Microvasc Res. 1977;14(1):53–65.

    CAS  Google Scholar 

  16. 16.

    Dejana E, Languino LR, Polentarutti N, Balconi G, Ryckewaert JJ, Larrieu MJ, et al. Interaction between fibrinogen and cultured endothelial cells. Induction of migration and specific binding. J Clin Invest. 1985;75(1):11–8.

    CAS  Google Scholar 

  17. 17.

    Tolsma SS, Volpert OV, Good DJ, Frazier WA, Polverini PJ, Bouck N. Peptides derived from two separate domains of the matrix protein thrombospondin-1 have anti-angiogenic activity. J Cell Biol. 1993;122(2):497–511.

    CAS  Google Scholar 

  18. 18.

    Kyriakides TR, Zhu YH, Yang Z, Bornstein P. The distribution of the matricellular protein thrombospondin 2 in tissues of embryonic and adult mice. J Histochem Cytochem. 1998;46(9):1007–15.

    CAS  Google Scholar 

  19. 19.

    Kyriakides TR, Zhu YH, Smith LT, Bain SD, Yang Z, Lin MT, et al. Mice that lack thrombospondin 2 display connective tissue abnormalities that are associated with disordered collagen fibrillogenesis, an increased vascular density, and a bleeding diathesis. J Cell Biol. 1998;140(2):419–30.

    CAS  Google Scholar 

  20. 20.

    Kohn JC, Zhou DW, Bordeleau F, Zhou AL, Mason BN, Mitchell MJ, et al. Cooperative effects of matrix stiffness and fluid shear stress on endothelial cell behavior. Biophys J. 2015;108(3):471–8.

    CAS  Google Scholar 

  21. 21.

    Wu W, Allen R, Gao J, Wang Y. Artificial niche combining elastomeric substrate and platelets guides vascular differentiation of bone marrow mononuclear cells. Tissue Eng Part A. 2011;17(15–16):1979–92.

    CAS  Google Scholar 

  22. 22.

    Sack KD, Teran M, Nugent MA. Extracellular matrix stiffness controls VEGF signaling and processing in endothelial cells. J Cell Physiol. 2016;231(9):2026–39.

    CAS  Google Scholar 

  23. 23.

    Andric T, Wright LD, Freeman JW. Rapid mineralization of electrospun scaffolds for bone tissue engineering. J Biomater Sci Polym Ed. 2011;22(11):1535–50.

    CAS  Google Scholar 

  24. 24.

    Andric T, Wright LD, Taylor BL, Freeman JW. Fabrication and characterization of three-dimensional electrospun scaffolds for bone tissue engineering. J Biomed Mater Res A. 2012;100(8):2097–105.

    Google Scholar 

  25. 25.

    Andric T, Taylor BL, Whittington AR, Freeman JW. Fabrication and characterization of three-dimensional electrospun scaffolds for bone tissue engineering. Regen Eng Transl Med. 2015;1(1–4):32–41.

    Google Scholar 

  26. 26.

    Taylor BL, Limaye A, Yarborough J, Freeman JW. Investigating processing techniques for bovine gelatin electrospun scaffolds for bone tissue regeneration. J Biomed Mater Res B Appl Biomater. 2017;105(5):1131–40.

    CAS  Google Scholar 

  27. 27.

    Andric, T., Taylor B.L., Degen K.E., Whittington A.R., Freeman J.W., Fabrication and characterization of three dimensional electrospun cortical bone scaffolds. Nanomater Environ, 2014. 2(1).

  28. 28.

    Wright LD, Young RT, Andric T, Freeman JW. Fabrication and mechanical characterization of 3D electrospun scaffolds for tissue engineering. Biomed Mater. 2010;5(5):055006.

    CAS  Google Scholar 

  29. 29.

    Moffa M, Polini A, Sciancalepore AG, Persano L, Mele E, Passione LG, et al. Microvascular endothelial cell spreading and proliferation on nanofibrous scaffolds by polymer blends with enhanced wettability. Soft Matter. 2013;9(23):5529–39.

    CAS  Google Scholar 

  30. 30.

    Patel PP, Buckley C, Taylor BL, Sahyoun CC, Patel SD, Mont AJ, et al. Mechanical and biological evaluation of a hydroxyapatite-reinforced scaffold for bone regeneration. J Biomed Mater Res A. 2019;107(4):732–41.

    CAS  Google Scholar 

  31. 31.

    Browe DP, Wood C, Sze MT, White KA, Scott T, Olabisi RM, et al. Characterization and optimization of actuating poly(ethylene glycol) diacrylate/acrylic acid hydrogels as artificial muscles. Polymer. 2017;117:331–41.

    CAS  Google Scholar 

  32. 32.

    Shi X, Zhou K, Huang F, Wang C. Interaction of hydroxyapatite nanoparticles with endothelial cells: internalization and inhibition of angiogenesis in vitro through the PI3K/Akt pathway. Int J Nanomedicine. 2017;12:5781–95.

    CAS  Google Scholar 

  33. 33.

    Rao RR, Ceccarelli J, Vigen ML, Gudur M, Singh R, Deng CX, et al. Effects of hydroxyapatite on endothelial network formation in collagen/fibrin composite hydrogels in vitro and in vivo. Acta Biomater. 2014;10(7):3091–7.

    CAS  Google Scholar 

  34. 34.

    Nemati S, Rezabakhsh A, Khoshfetrat AB, Nourazarian A, Biray Avci Ç, Goker Bagca B, et al. Alginate-gelatin encapsulation of human endothelial cells promoted angiogenesis in in vivo and in vitro milieu. Biotechnol Bioeng. 2017;114(12):2920–30.

    CAS  Google Scholar 

  35. 35.

    Saha K, Keung AJ, Irwin EF, Li Y, Little L, Schaffer DV, et al. Substrate modulus directs neural stem cell behavior. Biophys J. 2008;95(9):4426–38.

    CAS  Google Scholar 

  36. 36.

    Eroshenko N, et al. Effect of substrate stiffness on early human embryonic stem cell differentiation. J Biol Eng. 2013;7(1):7.

    CAS  Google Scholar 

  37. 37.

    Evans ND, et al. Substrate stiffness affects early differentiation events in embryonic stem cells. Eur Cell Mater. 2009;18:1–13 discussion 13-4.

    CAS  Google Scholar 

  38. 38.

    Charrier EE, Pogoda K, Wells RG, Janmey PA. Control of cell morphology and differentiation by substrates with independently tunable elasticity and viscous dissipation. Nat Commun. 2018;9(1):449.

    Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Joseph W. Freeman.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Ai, X., Pellegrini, M. & Freeman, J.W. The Use of Alginate to Inhibit Mineralization for Eventual Vascular Development. Regen. Eng. Transl. Med. 6, 154–163 (2020). https://doi.org/10.1007/s40883-019-00104-7

Download citation

Keywords

  • Vascularization
  • Inhibition
  • Mineralization
  • Alginate
  • Calcium phosphate
  • Bone tissue engineering