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Embryonic Mesenchymal Multipotent Cell Differentiation on Electrospun Biodegradable Poly(ester amide) Scaffolds for Model Vascular Tissue Fabrication

Abstract

Vascular differentiation of stem cells and matrix component production on electrospun tubular scaffolds is desirable to engineer blood vessels. The mouse embryonic multipotent mesenchymal progenitor cell line (10T1/2) provides an excellent tool for tissue engineering since it shares similar differentiation characteristics with mesenchymal stem cells. Although 10T1/2 cells have been widely studied in the context of skeletal tissue engineering, their differentiation to smooth muscle lineage is less known. In this study, we fabricated tubular electrospun poly(ester amide) (PEA) fibers from l-phenylalanine-derived biodegradable biomaterials and investigated cell-scaffold interactions as well as their differentiation into vascular smooth muscle cell and subsequent elastin expression. PEA scaffolds fabricated under different collector speeds did not have an impact on the fiber directionality/orientation. 10T1/2 cytocompatibility and proliferation studies showed that PEA fibres were not cytotoxic and were able to support proliferation for 14 days. Furthermore, cells were observed infiltrating the fibrous scaffolds despite the small pore sizes (~ 5 µm). Vascular differentiation studies of 10T1/2 cells using qPCR, Western blot, and immunostaining showed a TGFβ1-induced upregulation of vascular smooth muscle cell (VSMC)-specific markers smooth muscle alpha-actin (SM-α–actin) and smooth muscle myosin heavy chain (SM-MHC). Differentiated 10T1/2 cells produced both elastin and fibrillin-1 suggesting the potential of fibrous PEA scaffolds to fabricate model vascular tissues.

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Funding

Funding was provided by Natural Sciences and Engineering Research Council of Canada (Grant No. RGPIN-2018-06310).

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Correspondence to Kibret Mequanint.

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Kiros, S., Lin, S., Xing, M. et al. Embryonic Mesenchymal Multipotent Cell Differentiation on Electrospun Biodegradable Poly(ester amide) Scaffolds for Model Vascular Tissue Fabrication. Ann Biomed Eng 48, 980–991 (2020). https://doi.org/10.1007/s10439-019-02276-3

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Keywords

  • Poly(ester amide)s
  • 10T1/2 cell differentiation
  • Tubular electrospinning
  • Vascular smooth muscle cells
  • Elastin
  • Fibrillin-1