Avidity-controlled hydrogels for injectable co-delivery of induced pluripotent stem cell-derived endothelial cells and growth factors.
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.
Department of Bioengineering, Stanford University, Stanford, CA, USA.
Widya
W
Mulyasasmita
Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA; Stanford Cardiovascular Institute, Stanford University, Stanford, CA, USA.
Lei
L
Cai
Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA.
Ruby E
RE
Dewi
Stanford Cardiovascular Institute, Stanford University, Stanford, CA, USA; Veterans Affairs Palo Alto Health Care System, Palo Alto, CA, USA.
Arshi
A
Jha
Department of Chemistry, RWTH Aachen University, Aachen, Germany.
Sabrina D
SD
Ullmann
Department of Comparative Medicine, Stanford University, Stanford, CA, USA.
Richard H
RH
Luong
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.
Ngan F
NF
Huang
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.
Sarah C
SC
Heilshorn
D000818:Animals; D048430:Cell Shape; D002478:Cells, Cultured; D002626:Chemistry, Pharmaceutical; D003692:Delayed-Action Preparations; D004195:Disease Models, Animal; D055119:Elastic Modulus; D066026:Endothelial Progenitor Cells / Q000378:metabolism / Q000637:transplantation*; D006614:Hindlimb; D006801:Humans; D020100:Hydrogels; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000637:transplantation*; D007273:Injections, Intramuscular; D007511:Ischemia / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D007700:Kinetics; D008297:Male; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008970:Molecular Weight; D018482:Muscle, Skeletal / Q000098:blood supply* / Q000473:pathology / Q000503:physiopathology; D009336:Necrosis; D011092:Polyethylene Glycols / Q000737:chemistry*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry*; D012038:Regeneration / Q000187:drug effects; D012995:Solubility; D013678:Technology, Pharmaceutical / Q000379:methods; D054457:Tissue Scaffolds; D042461:Vascular Endothelial Growth Factor A / Q000008:administration & dosage* / Q000737:chemistry; D014783:Viscosity
D003692:Delayed-Action Preparations; D020100:Hydrogels; D011994:Recombinant Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D011092:Polyethylene Glycols
Endothelial cell; Hydrogel; Protein engineering; VEGF; iPSC