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一种基于V79细胞集落形成的生物材料毒理学评价方法。

A method for toxicological evaluation of biomaterials based on colony formation of V79 cells.

作者信息

Kotoura Y, Yamamuro T, Shikata J, Kakutani Y, Kitsugi T, Tanaka H

出版信息

Arch Orthop Trauma Surg (1978). 1985;104(1):15-9. doi: 10.1007/BF00449950.

Abstract

This report describes a method for cytotoxicity screening of biomaterials based on colony formation of V79 cells. For this test, two metals (titanium and nickel), two ceramics (alumina ceramic and tricalcium phosphate), and two types of polymeric material [high density polyethylene (HDP) and polyvinylchloride (PVC)] were used. Each metal and ceramic was cast into a disk and semidisk 49 mm in diameter and 1 to 2 mm thick. The HDP was molded into a petri dish and PVC was used as a thin film. The materials were sterilized by heating or with ethylene oxide and placed in plastic petri dishes, after which 8 ml cell suspension containing 100 cells were added to each dish. After 1 week, the colonies formed on the materials were fixed, stained, and then the number of colonies was counted. Titanium, alumina ceramic, and HDP showed no differences from the controls in terms of colonies. On the disks and the semidisks of nickel and tricalcium phosphate and on the thin disks of PVC, however, no colonies were detected. The V79 cells used in this experiment showed a rapid and logarithmically stable growth curve and such a high rate of colony formation as to form visible noticeable colonies, and were therefore suitable cells for screening test the cytotoxicity of biomaterials. Unlike other previously reported methods of in vitro cytotoxicity testing, this method permits assay of colonies formed from a single cell after proliferation directly on the materials. Moreover, the test with semidisks permits simple screening to assess the cytotoxicity is caused by either the chemical substances or the physical properties of the materials.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

本报告描述了一种基于V79细胞集落形成的生物材料细胞毒性筛选方法。在该试验中,使用了两种金属(钛和镍)、两种陶瓷(氧化铝陶瓷和磷酸三钙)以及两种类型的聚合材料[高密度聚乙烯(HDP)和聚氯乙烯(PVC)]。每种金属和陶瓷被铸造成直径49毫米、厚1至2毫米的圆盘和半圆盘。HDP被模制成培养皿,PVC被用作薄膜。材料通过加热或环氧乙烷灭菌,然后置于塑料培养皿中,之后向每个培养皿中加入8毫升含有100个细胞的细胞悬液。1周后,对材料上形成的集落进行固定、染色,然后计数集落数量。钛、氧化铝陶瓷和HDP在集落方面与对照组无差异。然而,在镍和磷酸三钙的圆盘和半圆盘以及PVC的薄盘上未检测到集落。本实验中使用的V79细胞显示出快速且呈对数稳定的生长曲线,以及如此高的集落形成率以至于能形成可见的明显集落,因此是用于筛选生物材料细胞毒性的合适细胞。与其他先前报道的体外细胞毒性测试方法不同,该方法允许直接在材料上对增殖后由单个细胞形成的集落进行测定。此外,使用半圆盘的试验允许进行简单筛选,以评估细胞毒性是由材料的化学物质还是物理性质引起的。(摘要截断于250字)

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