title: Optimized Repopulation of Tendon Hydrogel: Synergistic Effects of Growth Factor Combinations and Adipose-Derived Stem Cells.
abstract: Background:Tendon-derived extracellular matrix (ECM) hydrogel has been shown to augment tendon healing in vivo. We hypothesized that reseeding of the gel with adipose-derived stem cells (ASCs) could further assist repopulation of the gel and that combinations of growth factors (GFs) would improve the survival of these cells after reseeding.Methods:A tendon-specific ECM solution was supplemented with varying concentrations of basic fibroblast growth factor (bFGF), insulin-like growth factor-1 (IGF-1), and platelet-derived growth factor-BB (PDGF-BB). Gels were then seeded with ASCs transfected with a green fluorescent protein/luciferin construct. Cell proliferation was determined using the MTT assay and histology, and GF and ASC augmented gels were injected into the back of Sprague Dawley rats. Bioluminescence of seeded gels was continuously followed after reseeding, and cell counts were performed after the gels were explanted at 14 days.Results:Synergistic effects of the GFs were seen, and an optimal combination was determined to be 10 ng/mL bFGF, 100 ng/mL IGF-1, and 100 ng/mL PDGF-BB (2.8-fold increase;P< .05). In vivo bioluminescence showed an improved initial survival of cells in gels supplemented with the optimal concentration of GF compared with the control group (10.6-fold increase at 8 days;P< .05). Cell counts of explants showed a dramatic endogenous repopulation of gels supplemented by GF + ASCs compared with both gels with GF but no ASCs (7.6-fold increase) and gels with ASCs but no GF (1.6-fold increase).Conclusion:Synergistic effects of GFs can be used to improve cellular proliferation of ASCs seeded to a tendon ECM gel. Reseeding with ASCs stimulates endogenous repopulation of the gel in vivo and may be used to further augment tendon healing.
authors.affiliation: VA Palo Alto Health Care System, Livermore, CA, USA; Linköping University, Sweden.
authors.forename: Simon
authors.initials: S
authors.lastname: Farnebo
authors.affiliation: Stanford University, CA, USA.
authors.forename: Lovisa
authors.initials: L
authors.lastname: Farnebo
authors.affiliation: VA Palo Alto Health Care System, Livermore, CA, USA; Stanford University, CA, USA.
authors.forename: Maxwell
authors.initials: M
authors.lastname: Kim
authors.affiliation: VA Palo Alto Health Care System, Livermore, CA, USA; Stanford University, CA, USA.
authors.forename: Colin
authors.initials: C
authors.lastname: Woon
authors.affiliation: VA Palo Alto Health Care System, Livermore, CA, USA; Stanford University, CA, USA.
authors.forename: Hung
authors.initials: H
authors.lastname: Pham
authors.affiliation: VA Palo Alto Health Care System, Livermore, CA, USA; Stanford University, CA, USA.
authors.forename: James
authors.initials: J
authors.lastname: Chang
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D005109:Extracellular Matrix / Q000637:transplantation*; D048091:Guided Tissue Regeneration / Q000379:methods*; D020100:Hydrogels; D036341:Intercellular Signaling Peptides and Proteins / Q000008:administration & dosage / Q000494:pharmacology*; D017207:Rats, Sprague-Dawley; D033581:Stem Cell Transplantation / Q000379:methods*; D013710:Tendons / Q000166:cytology*; D023822:Tissue Engineering; D054457:Tissue Scaffolds
chemical_list: D020100:Hydrogels; D036341:Intercellular Signaling Peptides and Proteins
keywords: ECM; flexor tendon; hydrogel; reconstructive hand surgery; tissue engineering