Surasak Kasetsiriku1, Dettachai Ketpun1, Yon Jin Chuah2, Yannapol Sriphutkiat1, Dong-An Wang2, Yufeng Zhou3. 1. School of Mechanical and Aerospace Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore, 639798, Singapore. 2. School of Chemical and Biomedical Engineering, Nanyang Technological University, 62 Nanyang Drive, Singapore, 637459, Singapore. 3. School of Mechanical and Aerospace Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore, 639798, Singapore. yfzhou@ntu.edu.sg.
Abstract
BACKGROUND: Surface modification is used to modify the biomaterials for the regulation of cell culture using different approaches, such as chemical graft and mechanical treatment. However, those conventional methodologies often require precise fabrication in a high resolution involving either high cost or laborious steps to remove chemical residues that are toxic to the cells. METHODS: A novel and simple method was proposed and evaluated to rapidly generate surface ceases on the gelatin methacrylate (gelMA) surface using the heating-hydration process. Human umbilical vein endothelial cells (HUVECs) were cultured on the gelMA surface. The surface binding was characterized using the RGD (Arg-Gly-Asp) antibodies and cell adhesion pattern captured by scanning electron microscopy. The effect of the heating-hydration parameters on the creasing formation was investigated. The morphology of HUVECs cultured on such micropatterned gelMA was characterized and compared. RESULTS: It is found that the hydration solution, gelMA mixture, and hydration rate are the major factors that influence the cracking sizes in the range from 20 to 120 µm which resulted in capillary-like patterns on the gelMA surface. Low concentration of gelMA, high water concentration of cooling agent, and slow hydration rate result in the long creases, and heating of at least 60 min is required for complete dehydration. Strong fluorescence was around the creases with RGD-staining. Consequently, micropatterned gelMA demonstrated good biocompatibility with endothelial cells with more than 95% cell viability and continuous cell proliferation throughout 2 weeks as well as a good trace of neovascular formation. In comparison, normal gelMA surface did not exhibit RGD-fluorescent signals, and the cultured HUVECs on it were rounded with no spreading for network formation. CONCLUSION: The heating-hydration approach can successfully and easily produce the micropatterned gelMA that allows rapid and effective vascularization to potentially improve the functionalities of the tissue-engineered construct.
BACKGROUND: Surface modification is used to modify the biomaterials for the regulation of cell culture using different approaches, such as chemical graft and mechanical treatment. However, those conventional methodologies often require precise fabrication in a high resolution involving either high cost or laborious steps to remove chemical residues that are toxic to the cells. METHODS: A novel and simple method was proposed and evaluated to rapidly generate surface ceases on the gelatin methacrylate (gelMA) surface using the heating-hydration process. Human umbilical vein endothelial cells (HUVECs) were cultured on the gelMA surface. The surface binding was characterized using the RGD (Arg-Gly-Asp) antibodies and cell adhesion pattern captured by scanning electron microscopy. The effect of the heating-hydration parameters on the creasing formation was investigated. The morphology of HUVECs cultured on such micropatterned gelMA was characterized and compared. RESULTS: It is found that the hydration solution, gelMA mixture, and hydration rate are the major factors that influence the cracking sizes in the range from 20 to 120 µm which resulted in capillary-like patterns on the gelMA surface. Low concentration of gelMA, high water concentration of cooling agent, and slow hydration rate result in the long creases, and heating of at least 60 min is required for complete dehydration. Strong fluorescence was around the creases with RGD-staining. Consequently, micropatterned gelMA demonstrated good biocompatibility with endothelial cells with more than 95% cell viability and continuous cell proliferation throughout 2 weeks as well as a good trace of neovascular formation. In comparison, normal gelMA surface did not exhibit RGD-fluorescent signals, and the cultured HUVECs on it were rounded with no spreading for network formation. CONCLUSION: The heating-hydration approach can successfully and easily produce the micropatterned gelMA that allows rapid and effective vascularization to potentially improve the functionalities of the tissue-engineered construct.
Authors: Jinhwan Yoon; Pei Bian; Jungwook Kim; Thomas J McCarthy; Ryan C Hayward Journal: Angew Chem Int Ed Engl Date: 2012-06-12 Impact factor: 15.336
Authors: Molly M Stevens; Robert P Marini; Dirk Schaefer; Joshua Aronson; Robert Langer; V Prasad Shastri Journal: Proc Natl Acad Sci U S A Date: 2005-07-29 Impact factor: 11.205
Authors: Jason W Nichol; Sandeep T Koshy; Hojae Bae; Chang M Hwang; Seda Yamanlar; Ali Khademhosseini Journal: Biomaterials Date: 2010-04-24 Impact factor: 12.479
Authors: Philippe E Van den Steen; Bénédicte Dubois; Inge Nelissen; Pauline M Rudd; Raymond A Dwek; Ghislain Opdenakker Journal: Crit Rev Biochem Mol Biol Date: 2002-12 Impact factor: 8.250