title: Capillary force seeding of hydrogels for adipose-derived stem cell delivery in wounds.
abstract: Effective skin regeneration therapies require a successful interface between progenitor cells and biocompatible delivery systems. We previously demonstrated the efficiency of a biomimetic pullulan-collagen hydrogel scaffold for improving bone marrow-derived mesenchymal stem cell survival within ischemic skin wounds by creating a "stem cell niche" that enhances regenerative cytokine secretion. Adipose-derived mesenchymal stem cells (ASCs) represent an even more appealing source of stem cells because of their abundance and accessibility, and in this study we explored the utility of ASCs for hydrogel-based therapies. To optimize hydrogel cell seeding, a rapid, capillary force-based approach was developed and compared with previously established cell seeding methods. ASC viability and functionality following capillary hydrogel seeding were then analyzed in vitro and in vivo. In these experiments, ASCs were seeded more efficiently by capillary force than by traditional methods and remained viable and functional in this niche for up to 14 days. Additionally, hydrogel seeding of ASCs resulted in the enhanced expression of multiple stemness and angiogenesis-related genes, including Oct4, Vegf, Mcp-1, and Sdf-1. Moving in vivo, hydrogel delivery improved ASC survival, and application of both murine and human ASC-seeded hydrogels to splinted murine wounds resulted in accelerated wound closure and increased vascularity when compared with control wounds treated with unseeded hydrogels. In conclusion, capillary seeding of ASCs within a pullulan-collagen hydrogel bioscaffold provides a convenient and simple way to deliver therapeutic cells to wound environments. Moreover, ASC-seeded constructs display a significant potential to accelerate wound healing that can be easily translated to a clinical setting.
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA.
authors.forename: Ravi K
authors.initials: RK
authors.lastname: Garg
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Rennert
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA.
authors.forename: Dominik
authors.initials: D
authors.lastname: Duscher
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Sorkin
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA.
authors.forename: Revanth
authors.initials: R
authors.lastname: Kosaraju
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA.
authors.forename: Lauren J
authors.initials: LJ
authors.lastname: Auerbach
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA.
authors.forename: James
authors.initials: J
authors.lastname: Lennon
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Chung
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA.
authors.forename: Kevin
authors.initials: K
authors.lastname: Paik
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA.
authors.forename: Johannes
authors.initials: J
authors.lastname: Nimpf
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA.
authors.forename: Jayakumar
authors.initials: J
authors.lastname: Rajadas
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Longaker
authors.affiliation: Hagey Laboratory, Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, California, USA; Max F. Perutz Laboratories, Department of Medical Biochemistry and Molecular Biology, Medical University of Vienna, Vienna, Austria; Biomaterials and Advanced Drug Delivery Center, Stanford University, Stanford, California, USA ggurtner@stanford.edu.
authors.forename: Geoffrey C
authors.initials: GC
authors.lastname: Gurtner
mesh_terms: D017667:Adipocytes / Q000166:cytology*; D000818:Animals; D015153:Blotting, Western; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D004195:Disease Models, Animal; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D020100:Hydrogels; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008855:Microscopy, Electron, Scanning; D060888:Real-Time Polymerase Chain Reaction; D012867:Skin / Q000293:injuries; D023822:Tissue Engineering / Q000379:methods*; D014945:Wound Healing
chemical_list: D020100:Hydrogels
keywords: Adipose stem cells; Angiogenesis; Cellular therapy; Cytokines; Seeding