fLecture 1 15 nov 2021
ISO Standard 10993-5- cytotoxicity
- Extract testing
- Direct contact testing
- Indirect contact testing
Cytotoxicity – principle of extract testing
Cytotoxicity – Extraction
Material preparation
Cell culture
,Cytotoxicity – MTT assay
Apolar (non-charged) compounds can easily travel through the membrane
Polar compounds can’t move outside the cell
No colour = no metabolic activity
Can be measure by a …….
Diluted in a two x serial dilution series
Analysis of cellular responses by all current cell biological methods (discussed in second
part), here with a confocal microscope
,Use of fluorescence labels
(In)Direct contact testing
Different metal surfaces with cells on it to see what they do
Transparent system, looking at different surfaces, numbers are different time scales
(seconds), different cell behaviour is noticed. w/o is without and w/w is with (fibronectin in
this case).
, Cells leave the surface (in A more cytotoxicity kind of thing because of the surface, that can’t
adhere well)
(In)Direct contact testing
Coil system, with winding
Nitinol Is a nickel titanium alloy
Memory metal (remembers its shape)
Can be applied through a catheter and will retain its shape and close the defect
Intermediate layer – linking polymer make a covalent binding with proteins (fibronectin,
heparin (anti blood clotting), hirudin)
Coil touching the cell – direct contact
But it’s not much
Endothelial cells in the presence of a coil; some distance from the coil
ISO Standard 10993-5- cytotoxicity
- Extract testing
- Direct contact testing
- Indirect contact testing
Cytotoxicity – principle of extract testing
Cytotoxicity – Extraction
Material preparation
Cell culture
,Cytotoxicity – MTT assay
Apolar (non-charged) compounds can easily travel through the membrane
Polar compounds can’t move outside the cell
No colour = no metabolic activity
Can be measure by a …….
Diluted in a two x serial dilution series
Analysis of cellular responses by all current cell biological methods (discussed in second
part), here with a confocal microscope
,Use of fluorescence labels
(In)Direct contact testing
Different metal surfaces with cells on it to see what they do
Transparent system, looking at different surfaces, numbers are different time scales
(seconds), different cell behaviour is noticed. w/o is without and w/w is with (fibronectin in
this case).
, Cells leave the surface (in A more cytotoxicity kind of thing because of the surface, that can’t
adhere well)
(In)Direct contact testing
Coil system, with winding
Nitinol Is a nickel titanium alloy
Memory metal (remembers its shape)
Can be applied through a catheter and will retain its shape and close the defect
Intermediate layer – linking polymer make a covalent binding with proteins (fibronectin,
heparin (anti blood clotting), hirudin)
Coil touching the cell – direct contact
But it’s not much
Endothelial cells in the presence of a coil; some distance from the coil