摘要
目的研究混合醇改性对戊二醛牛心包细胞毒性的影响;并验证0.3%戊二醛保存对改性后的戊二醛牛心包细胞毒性有无影响。方法取新鲜牛心包心前区部分,分三组处理:A组:用0.625%戊二醛固定两周后用0.3%戊二醛保存;B组:牛心包用0.625%戊二醛固定二周后,用混合醇后处理并保存;C组:将B组心包在混合醇中处理2个月后,转入0.3%戊二醛中保存。D组:用玻片作为对照组。将三种心包切成同样大小(2×2cm2),用生理盐水分别漂洗10min3次。然后分别种植同样数目的(4×104)人胚肺成纤维细胞,培养7d后,将细胞洗下来,0.4%台盼兰染色,计算每片心包上的活细胞数。同时对心包片细胞作光镜及扫描电镜(SEM)观察。结果4个组活细胞计数结果:三组心包片上的活细胞数分别为:A组(戊二醛牛心包)(:5.83±2.04)×103;B组(混合醇改性戊二醛牛心包)(:16.67±2.04)×103;C组(0.3%戊二醛保存的混合醇改性牛心包):(18.75±1.12)×103。与对照组([176.25±18.15)×103]相比,三种心包均对细胞增殖有不同程度的抑制。混合醇改性后(B、C组)与未改性戊二醛牛心包(A组)的活细胞数有显著性差异(P<0.001),而B组与C组无显著性差异(P>0.05);光镜细胞形态学观察:对照组细胞生长良好,且形态正常,无死亡细胞。细胞计数时见A、B、C三组均有死亡细胞,尤以A组最多;扫描电镜观察:A组细胞较少,且多为圆球形细胞,大部分为裸露的胶原纤维;B、C组均有较多的细胞覆盖生长,细胞展开,可见细胞分裂相。结论经混合醇处理后的戊二醛牛心包细胞毒性明显降低,但仍有细胞毒性;混合醇改性后的戊二醛牛心包保存在0.3%戊二醛溶液中,其细胞毒性无明显改变。
[Objective] To research that post-treated with polydiol affect the cytotoxicity of glutaraldehyde tanned bovine pericardium （G-tBP）, and test that post-treated G-tBP preserved in 0.3% glutaraldehyde dose not affect its cytotoxicity. [Methods] Fresh bovine pericardium after the fai had been trimmed away, then washed with normal saline, and processed with different preservatives. Three groups of variouly preserved tissue samples were used： Group A： Pericardium tanned with 0.625% glutaraldehyde （GA） for two weeks, then preserved in 0.3% GA; Group B： treated as group A, then post-treated with polydiol for two months; Gorup C： treated as group B, then preserved in 0.3% glutaraldehyde; Group D： A sheet of glass was used as a control. Each group 6 specimens. The tissue was cut into same size （2 cm×2 cm）, then washed with NS for three times 10 minutes period. Same amount （4×10^4） of human embryo lung （HEL） fibroblast were seeding on the smooth surface of pericardium and glass sheet. After cultured for 7 days, living cells on each specimens were counted, and light microscopy and scanning electron microscopy （SEM） was examined. [Result] The amount of living cells in Group A were （5.83±2.04）×10^3; and in group B： （16.67±2.04）×10^3; Group C （18.75±1.12）×10^3; Group D（176.25±18.15）×10^3. Group A vs Group B, C, P〈0.001; Group D vs Group A, B, C, P〈0.001; Group B vs Group C, P〉0.05. Light microscopy showed the cells of group D were growing well, and no dead cells. There were some dead cells in other three groups, especially in group A. SEM showed there were only a few cells on the surface of pericardium in group A, most of the collagen fibrils were naked. There were much more cells on the surface of pericardium in group B and C. [Conclusions] After glutaraldehyde tanned bovine pericardium post treated with polydiol, its cytotoxicity reduced significantly. There is no significant cytotoxicity changes when polydiol post treated glutaraldehyde tanned bovine pericardium was preserved in 0.3% glutaraldehyde.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2005年第18期2760-2763,共4页
China Journal of Modern Medicine
基金
温州医学院课题经费资助:基金编号2004A016
关键词
混合醇
牛心包
戊二醛
细胞毒性
Polydiol
Bovine pericardium
Glutaraldehyde
Cytotoxicity
