Stability of a biological tissue fixed with a naturally occurring crosslinking agent (genipin).
The study was undertaken to investigate the stability of a biological tissue fixed with a naturally occurring crosslinking agent (genipin) at distinct elapsed storage durations. The glutaraldehyde-fixed counterpart was used as a control. Porcine pericardia procured from a slaughterhouse were used as raw materials. After fixation, the fixed tissues were sterilized in a graded series of ethanol solutions and thoroughly rinsed in phosphate buffered saline for 1 day, and then stored in a jar containing sterilized water. The samples were taken out and tested for their stability during the durations of 1day through 6 months after storage. The stability of each study group was tested by measuring its tensile strength, free-amino-group content, and denaturation temperature. Additionally, the cytotoxicity of each test sample and its corresponding storage solution were investigated in vitro using 3T3 fibroblasts. The results were examined using a microscope and 3-(4,5-dimethylthiazol-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. It was found that the stability of the genipin-fixed tissue during storage was superior to its glutaraldehyde-fixed counterpart. The differences in stability between the genipin- and glutaraldehyde-fixed tissues during storage may be caused by their differences in crosslinking structure. There was no apparent cytotoxicity for both the genipin-fixed tissue and its corresponding storage solution throughout the entire course of the study, whereas significant cytotoxicity was observed for both the glutaraldehyde-fixed tissue and its storage solution. However, the cytotoxicity of the glutaraldehyde-fixed tissue decreased with increasing elapsed storage duration, whereas that of its corresponding storage solution increased. This suggested that the toxic residues remaining in the glutaraldehyde-fixed tissue leached out slowly into its corresponding storage solution during the course of storage.
Sung
H W
HW
Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan 30013. hwsung@che.nthu.edu.tw
Liang
I L
IL
Chen
C N
CN
Huang
R N
RN
Liang
H F
HF
D016475:3T3 Cells; D000818:Animals; D003432:Cross-Linking Reagents; D057889:Iridoid Glycosides; D039823:Iridoids; D051379:Mice; D010496:Pericardium; D011714:Pyrans; D013552:Swine; D016707:Tissue Fixation; D014021:Tissue Preservation
D003432:Cross-Linking Reagents; D057889:Iridoid Glycosides; D039823:Iridoids; D011714:Pyrans; C007834:genipin