Enzymatic regulation of functional vascular networks using gelatin hydrogels.
To manufacture tissue engineering-based functional tissues, scaffold materials that can be sufficiently vascularized to mimic the functionality and complexity of native tissues are needed. Currently, vascular network bioengineering is largely carried out using natural hydrogels as embedding scaffolds, but most natural hydrogels have poor mechanical stability and durability, factors that critically limit their widespread use. In this study, we examined the suitability of gelatin-phenolic hydroxyl (gelatin-Ph) hydrogels that can be enzymatically crosslinked, allowing tuning of the storage modulus and the proteolytic degradation rate, for use as injectable hydrogels to support the human progenitor cell-based formation of a stable and mature vascular network. Porcine gelatin-Ph hydrogels were found to be cytocompatible with human blood-derived endothelial colony-forming cells and white adipose tissue-derived mesenchymal stem cells, resulting in >87% viability, and cell proliferation and spreading could be modulated by using hydrogels with different proteolytic degradability and stiffness. In addition, gelatin was extracted from mouse dermis and murine gelatin-Ph hydrogels were prepared. Importantly, implantation of human cell-laden porcine or murine gelatin-Ph hydrogels into immunodeficient mice resulted in the rapid formation of functional anastomoses between the bioengineered human vascular network and the mouse vasculature. Furthermore, the degree of enzymatic crosslinking of the gelatin-Ph hydrogels could be used to modulate cell behavior and the extent of vascular network formation in vivo. Our report details a technique for the synthesis of gelatin-Ph hydrogels from allogeneic or xenogeneic dermal skin and suggests that these hydrogels can be used for biomedical applications that require the formation of microvascular networks, including the development of complex engineered tissues.
Department of Applied Science, National Hsinchu University of Education, Hsinchu 30014, Taiwan, ROC.
Chia-Hui
CH
Chuang
Department of Cardiac Surgery, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA.
Ruei-Zeng
RZ
Lin
Department of Applied Science, National Hsinchu University of Education, Hsinchu 30014, Taiwan, ROC.
Han-Wen
HW
Tien
Department of Applied Science, National Hsinchu University of Education, Hsinchu 30014, Taiwan, ROC.
Ya-Chun
YC
Chu
Material and Chemical Research Laboratories, Industrial Technology Research Institute, Hsinchu 31040, Taiwan, ROC.
Yen-Cheng
YC
Li
Department of Cardiac Surgery, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA.
Juan M
JM
Melero-Martin
Department of Applied Science, National Hsinchu University of Education, Hsinchu 30014, Taiwan, ROC. Electronic address: yingchiehchen@gmail.com.
Ying-Chieh
YC
Chen
D001672:Biocompatible Materials / Q000138:chemical synthesis; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D003432:Cross-Linking Reagents / Q000737:chemistry; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005780:Gelatin / Q000737:chemistry*; D006735:Horseradish Peroxidase / Q000737:chemistry; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D008422:Materials Testing; D055806:Microvessels / Q000502:physiology*; D018919:Neovascularization, Physiologic / Q000502:physiology*; D010636:Phenols / Q000737:chemistry; D054457:Tissue Scaffolds
D001672:Biocompatible Materials; D003432:Cross-Linking Reagents; D020100:Hydrogels; D010636:Phenols; D005780:Gelatin; D006735:Horseradish Peroxidase
Gelatin hydrogels; In vivo; Tissue engineering; Vascular graft; Vascularization