Does anyone use ISO -5:1999 standerd procedure for in vitro cytotoxicity evelation? I am a new guy here, working with bone tissue scaffolds. i dnot know is that possible to find the protocol here? Thx
Here are some links which may help you -
ISO Link -
ISO 10993-5:1999 Biological evaluation of medical devices -- Part 5: Tests for in vitro cytotoxicity
AAMI Preview Link -
ANSI/AAMI/ISO 10993-5:1999 Biological evaluationof medical devicesPart 5: Tests for cytotoxicity, in vitro methods
This research paper may be of some help to you...
A.J. Salgado, O.P. Coutinho, R.L. Reis. Novel Starch-Based Scaffolds for Bone Tissue Engineering: Cytotoxicity, Cell Culture, and Protein Expression. Tissue Engineering. March 1, 2004, 10(3-4): 465-474. doi:10.1089/107632704323061825.
Starch-based biomaterials and scaffolds have been proposed for several biomedical applications. In the present work new scaffolds based on a 50/50 (wt%) blend of corn starch/ethylene-vinyl alcohol (SEVA-C) were studied. These scaffolds were processed by a melt-based technology, which has been used before with other starch-based materials but never with SEVA-C. Scanning electron microscopy (SEM) observation showed that the developed porous structures were 60% porous with pore size between 200 and 900 μm and a reasonable degree of interconnectivity. Moreover, scaffolds presented a compressive modulus of 117.50 ± 3.7 MPa and a compressive strength of 20.8 ± 2.4 MPa. Cytotoxicity evaluation was performed according to ISO/EN 10993 part 5 guidelines, and revealed that the developed scaffolds were nontoxic and did not inhibit cell growth. Direct contact assays were also carried out by use of a cell line of human osteoblast-like cells (SaOS-2). Cells were seeded (3 × 105 per scaffold) and allowed to grow for 4 weeks at 37°C, in a humidified atmosphere containing 5% CO2. Total protein assay showed that the cells were able to grow for the 4 weeks of the experiment. These data were further confirmed by SEM. Moreover, a cell viability assay (MTS test) demonstrated that cells were perfectly viable after the 4 weeks of culture, showing the adequacy of the developed structure in supporting them. Finally, Western blot analysis revealed that osteopontin was being actively expressed by the cells, which, in association with collagen deposition observed by SEM, seems to indicate that bone extracellular matrix was being deposited. Consequently it is believed that starch-based scaffolds should be considered as an alternative for bone tissue-engineering applications in the near future.
Hope this helps... Good luck!
These info is so much helpful. Thx