Avidity-controlled hydrogels for injectable co-delivery of induced pluripotent stem cell-derived endothelial cells and growth factors

Widya Mulyasasmita; Lei Cai; Ruby E Dewi; Arshi Jha; Sabrina D Ullmann; Richard H Luong; Ngan F Huang; Sarah C Heilshorn

doi:10.1016/j.jconrel.2014.05.015

Avidity-controlled hydrogels for injectable co-delivery of induced pluripotent stem cell-derived endothelial cells and growth factors

J Control Release. 2014 Oct 10:191:71-81. doi: 10.1016/j.jconrel.2014.05.015. Epub 2014 May 18.

Authors

Widya Mulyasasmita¹, Lei Cai², Ruby E Dewi³, Arshi Jha⁴, Sabrina D Ullmann⁵, Richard H Luong⁶, Ngan F Huang⁷, Sarah C Heilshorn⁸

Affiliations

¹ Department of Bioengineering, Stanford University, Stanford, CA, USA.
² Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA; Stanford Cardiovascular Institute, Stanford University, Stanford, CA, USA.
³ Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA.
⁴ Stanford Cardiovascular Institute, Stanford University, Stanford, CA, USA; Veterans Affairs Palo Alto Health Care System, Palo Alto, CA, USA.
⁵ Department of Chemistry, RWTH Aachen University, Aachen, Germany.
⁶ Department of Comparative Medicine, Stanford University, Stanford, CA, USA.
⁷ Stanford Cardiovascular Institute, Stanford University, Stanford, CA, USA; Veterans Affairs Palo Alto Health Care System, Palo Alto, CA, USA; Division of Cardiovascular Medicine, Stanford University, Stanford, CA, USA.
⁸ Department of Bioengineering, Stanford University, Stanford, CA, USA; Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA; Stanford Cardiovascular Institute, Stanford University, Stanford, CA, USA. Electronic address: heilshorn@stanford.edu.

Abstract

To translate recent advances in induced pluripotent stem cell biology to clinical regenerative medicine therapies, new strategies to control the co-delivery of cells and growth factors are needed. Building on our previous work designing Mixing-Induced Two-Component Hydrogels (MITCHs) from engineered proteins, here we develop protein-polyethylene glycol (PEG) hybrid hydrogels, MITCH-PEG, which form physical gels upon mixing for cell and growth factor co-delivery. MITCH-PEG is a mixture of C7, which is a linear, engineered protein containing seven repeats of the CC43 WW peptide domain (C), and 8-arm star-shaped PEG conjugated with either one or two repeats of a proline-rich peptide to each arm (P1 or P2, respectively). Both 20kDa and 40kDa star-shaped PEG variants were investigated, and all four PEG-peptide variants were able to undergo a sol-gel phase transition when mixed with the linear C7 protein at constant physiological conditions due to noncovalent hetero-dimerization between the C and P domains. Due to the dynamic nature of the C-P physical crosslinks, all four gels were observed to be reversibly shear-thinning and self-healing. The P2 variants exhibited higher storage moduli than the P1 variants, demonstrating the ability to tune the hydrogel bulk properties through a biomimetic peptide-avidity strategy. The 20kDa PEG variants exhibited slower release of encapsulated vascular endothelial growth factor (VEGF), due to a decrease in hydrogel mesh size relative to the 40kDa variants. Human induced pluripotent stem cell-derived endothelial cells (hiPSC-ECs) adopted a well-spread morphology within three-dimensional MITCH-PEG cultures, and MITCH-PEG provided significant protection from cell damage during ejection through a fine-gauge syringe needle. In a mouse hindlimb ischemia model of peripheral arterial disease, MITCH-PEG co-delivery of hiPSC-ECs and VEGF was found to reduce inflammation and promote muscle tissue regeneration compared to a saline control.

Keywords: Endothelial cell; Hydrogel; Protein engineering; VEGF; iPSC.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Cell Shape
Cells, Cultured
Chemistry, Pharmaceutical
Delayed-Action Preparations
Disease Models, Animal
Elastic Modulus
Endothelial Progenitor Cells / metabolism
Endothelial Progenitor Cells / transplantation*
Hindlimb
Humans
Hydrogels
Induced Pluripotent Stem Cells / metabolism
Induced Pluripotent Stem Cells / transplantation*
Injections, Intramuscular
Ischemia / pathology
Ischemia / physiopathology
Ischemia / therapy*
Kinetics
Male
Mice, Inbred NOD
Mice, SCID
Molecular Weight
Muscle, Skeletal / blood supply*
Muscle, Skeletal / pathology
Muscle, Skeletal / physiopathology
Necrosis
Polyethylene Glycols / chemistry*
Protein Binding
Recombinant Proteins / chemistry*
Regeneration / drug effects
Solubility
Technology, Pharmaceutical / methods
Tissue Scaffolds*
Vascular Endothelial Growth Factor A / administration & dosage*
Vascular Endothelial Growth Factor A / chemistry
Viscosity

Substances

Delayed-Action Preparations
Hydrogels
Recombinant Proteins
VEGFA protein, human
Vascular Endothelial Growth Factor A
Polyethylene Glycols

Abstract

Publication types

MeSH terms

Substances

Grants and funding