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L-谷氨酸预防戊二醛固定的牛心包钙化:大鼠实验研究

L-glutamic acid in the prevention of the calcification of bovine pericardial fixed in glutaraldehyde: study in rats.

作者信息

Ferreira Andrea Dumsch de Aragon, Costa Francisco Diniz Affonso da, Santos Eduardo Antonio Andrade dos, Sardeto Evandro Antonio, Gomes Carlos Henrique Gori, Collatusso Claudinei, Lopes Sérgio Augusto Veiga, Perruzzo Angela, Costa Iseu de Santo Elias Affonso da

机构信息

Pontifical Catholic University of Paraná, Curitiba, PR, Brazil.

出版信息

Rev Bras Cir Cardiovasc. 2007 Jul-Sep;22(3):303-9. doi: 10.1590/s0102-76382007000300006.

Abstract

OBJECTIVE

To evaluate the efficiency of L-glutamic acid to prevent calcification of glutaraldehyde bovine pericardium implanted in rats' subcutaneous tissues.

METHODS

Fifty four Wistar rats were divided in six groups according to the type of the bovine pericardium implanted. At first, all pericardia were initially cross-linked with 0.5% glutaraldehyd (GDA) fixative for 72 h. In Group I, after the initial fixation, the pericardia were preserved in 0.2% GDA fixative until the implantation, whereas in Group II they were stocked in Paraben solution. In Groups III and IV, after the initial fixation in 0.5% GDA fixative, the pericardia were treated with 8% L-glutamic acid at pH 7.4 and 3.5, respectively, being subsequently stocked in Paraben solution. Groups V and VI were similar to III and IV, except for the concentration of L-Glutamic acid which was 0.8%. Explantation was done at 15, 30, and 60 days, and the specimens submitted to histological analysis with Hematoxylin and eosin (HE) and Von Kossa stains, besides calcium quantification with atomic spectrofotometry.

RESULTS

Microscopic analysis demonstrated severe and progressive calcification in groups I, II, and III, whereas in groups IV, V, and VI calcification, when present, was mild and focal. Spectrofotomety confirmed these findings, revealing calcium contents of 1.93 microg/mg of tissue at 60 days in the control group. Groups IV and VI showed the least calcium contents (0.063 e 0.066, respectively).

CONCLUSIONS

The use of L-glutamic acid in segments of bovine pericardium with glutaraldehyde fixative was effective in preventing the calcification when implanted in rats' subcutaneous up to 60 days.

摘要

目的

评估L-谷氨酸对预防植入大鼠皮下组织的戊二醛处理牛心包钙化的效果。

方法

54只Wistar大鼠根据植入的牛心包类型分为六组。首先,所有心包均先用0.5%戊二醛(GDA)固定液交联固定72小时。第一组,初始固定后,心包保存在0.2% GDA固定液中直至植入,而第二组心包保存在对羟基苯甲酸酯溶液中。第三组和第四组,在用0.5% GDA固定液初始固定后,心包分别用pH值为7.4和3.5的8% L-谷氨酸处理,随后保存在对羟基苯甲酸酯溶液中。第五组和第六组与第三组和第四组相似,只是L-谷氨酸浓度为0.8%。在第15、30和60天取出标本,除了用原子分光光度法进行钙定量外,标本还用苏木精和伊红(HE)染色及冯·科萨染色进行组织学分析。

结果

显微镜分析显示,第一组、第二组和第三组有严重且进行性的钙化,而第四组、第五组和第六组如有钙化则为轻度且局灶性。分光光度法证实了这些结果,显示对照组在60天时组织钙含量为1.93微克/毫克。第四组和第六组钙含量最低(分别为0.063和0.066)。

结论

戊二醛固定的牛心包片段使用L-谷氨酸在植入大鼠皮下长达60天的时间内可有效预防钙化。

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