Establishment of in vitro models of denatured collagen
Department of Burns and Plastic Surgery, Shanghai Institute of Burns, Shanghai Research Center of Wound Repair, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
Abstract：Objective To investigate influence of different temperatures on secondary structure of type Ⅰ collagen, determine the proper temperature for collagen denaturation, observe changes of collagen fibre arrangement and three dimensional collagen gel properties after thermal denaturation, compare morphological variation of fibroblasts seeded in mediums with denatured collagen and therefore establish a standardized culture model with denatured collagen in vitro. Methods Changes of the secondary structure of type Ⅰ collagen was measured by circular dichroism spectrameter after the collagen solution had been treated with different temperatures. Changes of the fibre structure after collagen denaturation were observed by scanning probe microscope. Different three-dimensional collagen gels were prepared and break moduli of the collagen gels were determined by gas pressure machine. The denatured collagens were coated on culture plate or mixed into three-dimensional collagen gels. Morphology of the fibroblasts seeded in different culture conditions was detected using the inverted phase contrast microscope and the light microscope. Results At 50℃, the secondary structure of type Ⅰ collagen changed dramatically. Collagen fibers were aggregated when denatured collagens were coated on culture plate. Break moduli of three-dimensional collagen gels containing denatured collagens had distinct decrease. Morphology of the fibroblasts seeded in mediums with denatured collagen were varied significantly. Conclusion Type Ⅰ collagen reveals an obvious change of its secondary structure at 50 ℃. Break moduli of three-dimensional collagen gels containing the denatured collagens present a great degree of decrease. Coating and three-dimensional collagen gel models containing type Ⅰ denatured collagen present different influences on morphology of fibroblasts and thus can be taken as in vitro models to investigate effect of the denatured collagen on bioactivity of cells.
SU Rongjia,WANG Zhiyong,LIU Yingkai et al. Establishment of in vitro models of denatured collagen[J]. CHINESE JOURNAL OF TRAUMA, 2013, 29(4): 353-358.
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