Preparation Of Fish College Peptide-hyaluronic Acid Hydrogel And Its Biocompatibility Of Hyaluronic Acid Hydrogel Biocompatibility

Object: To apply the collagen peptide derived from the fish and hyaluronic acid to the synthetize of the hydrogel and to find a tissue engineering scaffold material with good biocompatibility,safety and convenience.Methods: The precursors of the hydrogels are synthesized by means of adding acrylate to the collagen peptide and hyaluronic acid and hydrogels are formed by irradiation of 365 nm ultraviolet and catalyze of photoinitiator I2965.Human umbilical cord-derived mesenchymal stem cells were embed in the hydrogels during the process of the formation and were cultured.The size of the voids inside the gels made by different concentrations were observed by scanning electron microscope.The survival status of the cells cultured on the surface of the gels and cultured in the gels were identified by dyeing of Fluorescein diacetate.Precursors were dissolved in the medium and the cells were cultured in it.And the proliferation rate were examined by the staining of Ed U.The cells were cultured and the relationship between the Proliferation rate and the concentration of the precursors of the gels were examined by the staining of Ed U too.The proliferation rate of the cells cultured in gels were measured on different days by detection of MTT and the profiles of proliferation of cells were drawled.The cytoskeleton protein β-acting were stained by immunofluorescence to observe the morphology and outline of the cells in the gel.Results: The scanning electron microscope showed that porosity of gels were decreased with the increases of the concentration of gels.Fluorescein diacetate staining and immunostaining of β-acting showed that the cells cultured on the surface and inside of the gels were both survived well.The cells cultured on the surface and in the materials were in spherical ship at the initial stage of culture.But with the increase of the culture time,a small number of cells in the gels began turn into fusiform or polygonal.Test of Ed U staining showed that the Precursors we synthesized had no adverse effect on the cells in the dissolved state.The result of the Ed U staining of three tips of hydrogels we used in culture were formed by three different concentrations of acrylic-collagen and same concentration of acrylic-hyaluronic acid showed that there were most positively stained cells in the groupof 30 g / L of acrylic-collagen and 10 g / L of acrylic-hyaluronic acid,and the group of 50 g / L of acrylic-collagen showed lower than that and the group of 100 g / L of acryliccollagen were lowest of all.MTT assay showed that the proliferation of 2D-cultured cells maintained only to the 6th day of our culture,but the proliferation of the cells cultured in the gels maintained through all 8 days of our culture.Conclusions: The hydrogels formed by collagen peptide and hyaluronic acid chemical modified by acrylic acid is a suitable material for mesenchymal stem cells to grow.Compared with the two-dimensional cultured cells,the cells cultured in the gel can maintain their proliferation much longer,and 30 g / L of acrylic-collagen and 10 g / L of acrylichyaluronic acid can produce most suitable gels for cells to grow.The cells inside the gels are spherical in the early stages of our culture,but with the extension of the culture time,some of the cells on the edges of gels appear in the shape of a shuttle and that may suggesting that may the material can be attached by cells.However,further experiments are needed to prove that why fusiform cells appeared only in the edge of the gel.