3 Materials and Methods
4.2 Degradation process of MgAg1 % sticks
4.2.4 Histological parameters
4.2.4.1 Udder tissue incubated with MgAg1% sticks
Both unstained teat tissue (Fig. 24, B) and teat tissues stained with hematoxylin and eosin (H&E) treated with MgAg1% sticks (Fig. 24, D) showed black deposits precipitated on the upper layer, which is more prominent compared to the unstained and stained tissues without treatment (Fig. 24, A & C). This indicates the degradation of MgAg1% sticks during incubation
Fig. 24: The unstained teat tissue treated with MgAg1% sticks (B) and stained teat tissue with hematoxylin and eosin (H&E) treated with MgAg1% sticks (D) showed black deposits precipitated on the upper layer compared to control teat (A, C); magnification = 40x; bar = 50 μm.
C D
A B
Results
4.2.5 Silver concentrations in the degradation medium
The amount of silver ions increased after 25 days of incubation in the degradation medium. The 1 ml volume of degradation medium contains higher concentration of silver than the volume of 3 and 10 ml. The mean concentration of Ag+ ranged from 0.02 -1 mmol/l (Fig. 25)
Fig. 25: Mean silver concentration (mmol/l) in degradatiom medium after incubation of MgAg1% sticks in 1, 3 or 10 ml medium. There is a significant increase of silver concentration in degradation medium after 25 days of incubation (* p< 0.05, n=6).
4.2.6 Magnesium concentrations in the degradation medium
The amount of Mg++ in the degradation medium increased after 10 days up to 25 days of incubation without showing any significant differences between 1, 3 and 10 ml volume of degradation medium. The mean concentration of magnesium ranged from 0.2 - 9 mmol/l (Fig. 26).
Degradation medium
0 0.2 0.4 0.6 0.8 1 1.2 1.4
5 10 15 20 25
Days of incubation
Amount of silver (mmol\l)
1ml 3ml 10ml
*
* *
Results
Fig. 26: Mean Mg++ concentration (mmol/l) in degradation medium after incubation of MgAg 1% sticks in 1, 3 or 10 ml medium. There is an enhanced concentration of magnesium in degradation medium on day 10 up to 25 (* P< 0.05, n=6).
Degradation medium
0 2 4 6 8 10 12 14
5 10 15 20 25
Days of incubation
Amount of Mg (mmol/l)
1 ml 3 ml 10 ml
*
*
*
* *
*
**
*
* *
*
Results
4.3 Biocompatibility tests 4.3.1 Cell viability and proliferation 4.3.1.1 MTS assay
1. Murine cells
a. Incubated with silver ions
Murine fibroblasts (L929 cells) were treated with different concentrations of AgNO3
solution. A significant reduction in the viability of murine fibroblasts of 50 % was observed, when incubated with concentrations equal or above 0.1 mmol/l and more than 75 %, when incubated with concentrations above or equal to 0.3 mmol/l compared to the negative control cells (Fig. 27).
Fig. 27: Cell viability (MTS) of murine fibroblasts as indicated by the mean optical density of formazan after incubation in various concentration of AgNo3. A significant reduction in the viability of murine fibroblasts at 0.1 mmol/l of AgNO3 solution compared to control was observed (* p< 0.05, n= 6).
Raw murine macrophages and primary murine macrophages were treated with different concentration of AgNO3 solution. A significant reduction in the viability of raw macrophages was observed by 30 %, when incubated with concentrations equal or higher than 0.3 mmol/l, while primary macrophages showed a significant reduction in viability by 35 % when incubated with concentrations equal or below 1 mmol/l (Fig.
28).
control
0.00010.0003 0.001 0.003 0.01 0.03 0.1
Concentration of AgNO3 (mmol/l)
L929 cells
Results
Fig. 28: Cell viability (MTS) of raw macrophages and primary macrophages as indicated by the mean optical density of formazan after incubation in various concentration of AgNo3. A significant reduction in the viability in raw macrophages at 0.3 mmol/l AgNo3 solution compared to control, while primary macrophages showed a significant reduction in the viability at 1 mmol/l AgNo3 solution compared to control (* p< 0.05, n= 5).
Raw macrophages
Concentration of AgNO3 (mmol/l)
Optical density
Concentration of AgNO3 (mmol/l)
Optical density
Primary macrophages
Results
b. Incubated with MgAg1% sticks
Murine fibroblasts were treated with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium). A significant reduction in viability of about 75 % was observed, when murine fibroblasts were incubated with volumes equal or below 1 ml of degradation medium (1 ml = 0.5 mmol/l Mg++,0.2 mmol/l Ag+) (Fig. 29).
Fig. 29: Viability test (MTS assay) of murine fibroblasts as indicated by the mean optical density of formazan after treatment with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium). A significant reduction in the viability of murine fibroblasts at 1ml compared to control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3 mmol/l Mg++,0.04 mmol/l Ag+ and 1 ml ≈ 0.5 mmol/l Mg++,0.2 mmol/l Ag+ (* p< 0.05, n= 5).
control 10 ml 3 ml 1 ml 0
1 2 3
*
Different volumes of degradation medium (ml)
Optical density
L929
Results
Both raw murine macrophages and primary murine macrophages did not show a significant reduction in the viability, when treated with degradation medium (MgAg1%
sticks incubated in 1, 3, 10 ml medium) (Fig. 30).
Fig. 30: Cell viability (MTS asaay) of raw macrophages and primary macrophages as indicated by the mean optical density of formazan after treatment withdegradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium). No significant reduction in the viability of raw macrophages and primary macrophages compared to control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3 mmol/l Mg++,0.04 mmol/l Ag+ and 1 ml ≈ 0.5 mmol/l
Different volumes of degradation medium (ml)
Raw macrophages
Different volumes of degradation medium (ml)
Primary macrophages
Results
2. Bovine cells
a. Incubated with silver ions
Primary mammary epithelial cells were treated with different concentrations of AgNO3 solution and showed a slight tendency toward reduction, when incubated with concentrations equal or below 1 mmol/l (Fig. 31).
Fig. 31: Cell viability (MTS assay) of primary mammary epithelial cells as indicated by the mean optical density of formazan after incubation in various concentration of AgNo3. A tendency towards reduction in the viability was observed at 1 mmol/l AgNO3 solution compared to control (* p< 0.05, n= 6).
Primary mammary fibroblasts were treated with different concentrations of AgNO3
solution. A significant reduction in the viability of about 75 % was observed, when incubated with concentrations equal or below 1 mmol/l (Fig. 32)
Fig. 32: Cell viability (MTS) of primary mammary fibroblasts as indicated by the mean optical density of formazan after incubation in various concentration of AgNo3. A significant reduction in the viability at 1 mmol/l of AgNO3 solution compared to control (* p< 0.05, n= 6).
cont
Concentration of AgNO3 (mmol/l)
Optical density
٭
Results
b. Incubated with MgAg1% sticks
Primary mammary epithelial cells were treated with degradation medium (MgAg1%
sticks incubated in 1, 3, 10 ml medium) did not show a reduction in the viability (Fig.
33).
Fig. 33: Cell viability (MTS assay) of primary mammary epithelial cells as indicated by the mean optical density formazan after treatment with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium). No significant reduction in the viability was observed compared with control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3 mmol/l Mg++,0.04 mmol/l Ag+ and 1 ml ≈ 0.5 mmol/l Mg++,0.2 mmol/l Ag+l (n= 5).
Primary mammary fibroblasts were treated with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium), while there was no significant reduction in the viability of primary mammary fibroblasts (Fig. 34).
Fig. 34: Cell viability (MTS assay) primary mammary fibroblasts as indicated by the mean optical density of formazan after treatment with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium). They showed no significant reduction in the viability compared with control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3
Diffe re nt v olume s of de gradation me dium (ml)
control 10 ml 3 ml 1 ml
Different volumes of degradation medium (ml)
Optical density
Primary mammary fibroblasts
Results
4.3.1.2 Neutral red assay 1. Murine cells
a. Incubated with silver ions
Murine fibroblasts were treated with different concentrations of AgNO3 solution. A significant reduction in the viability of murine fibroblasts of 50 %, 70 % and 85 % was observed, when incubated with concentrations equal or higher than 0.1, 0.3 and 1 mmol/l, respectively (Fig. 35).
Fig. 35: Cell viability (Neutral red assay) of murine fibroblasts as indicated by the mean optical density of neutral red after incubation in various concentrations of AgNo3. A significant reduction in the viability of murine fibroblasts at 0.1 mmol/l compared with control was observed, (* p< 0.05, n= 6).
Both raw murine macrophages and primary murine macrophages were treated with different concentrations of AgNO3.A significant reduction in the viability of 60 % and 85 % was observed, when raw macrophages were treated with concentrations above 0.03 and 0.3 mmol/l, respectively (Fig. 36).
L929 cells
Results
Fig. 36: Cell viability (Neutral red assay) of raw macrophages and primary macrophages fibroblasts as indicated by the mean optical density of neutral red after incubation in various concentrations of AgNo3. A significant reduction in the viability of raw macrophages at 0.1 mmol/l of AgNO3 solution was observed compared with control, while primary macrophages showed significant reduction in the viability at 0.3 mmol/l compared with control (* p< 0.05, n= 6).
Concentration of AgNO3 (mmol/l)
Optical density
Concentration of AgNO3 (mmol/l)
Optical density
* *
Primary macrophages
Results
a. Incubated with MgAg1% sticks
Murine fibroblasts did not show a reduction in viability, when treated with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium) (Fig. 37).
Fig. 37: Cell viability (Neutral red assay) of murine fibroblasts as indicated by the mean optical density of neutral red after treatment with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium). No significant reduction in the viability was observed, when compared with control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3 mmol/l Mg++,0.04 mmol/l Ag+ and 1 ml ≈ 0.5 mmol/l Mg++,0.2 mmol/l Ag+ (n= 4).
Con trol
10 ml 3 ml 1 ml 0.30
0.35 0.40 0.45 0.50 0.55
Optical density
Different volumes of degradation medium (ml)
L929 cells
Results
Raw macrophages and primary macrophages did not show a reduction in the viability, when treated with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium/) (Fig. 38).
Fig. 38: Cell viability (Neutral red assay) of raw macrophages and primary macrophages fibroblasts as indicated by the mean optical density of neutral red after treatment with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium). No significant reduction in the viability of raw macrophages and primary macrophages were observed compared to control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3 mmol/l Mg++,0.04 mmol/l Ag+ and 1 ml ≈ 0.5 mmol/l Mg++,0.2 mmol/l Ag+ (n= 4).
Raw macrophages
control 10 ml 3 ml 1 ml 0.0
0.1 0.2 0.3 0.4
Different volumes of degradation medium (ml)
Optical density
Cont
rol 10 ml 3 m
l 1 ml 0.0
0.2 0.4 0.6
Optical denisty
Different volumes of degradation medium (ml)
Primary macrophages
Results
2. Bovine cells
a. Incubated with silver ions
Primary mammary epithelial cells were treated with different concentrations of AgNO3 solution. A significant reduction in the viability of primary mammary epithelium cells of 20 % was observed, when incubated with concentrations equal or below 1 mmol/l (Fig. 39).
Fig. 39: Cell viability (Neutral red assay) of primary mammary epithelial cells fibroblasts as indicated by the mean optical density of neutral red after incubation in various concentrations of AgNo3. A significant reduction in the viability was observed at 1 mmol/l of AgNo3 solution compared to control (
*
p< 0.05, n= 4).Primary mammary fibroblasts were treated with different concentrations of AgNO3 solution. A significant reduction in viability of 80 % was observed, when incubated with concentrations equal or below 1 mmol/l (Fig. 40).
Fig. 40: Cell viability (Neutral red assay) of primary mammary fibroblasts as indicated by the mean optical density of neutral red after incubation in various concentrations of AgNo3. A significant reduction in the viability of primary mammary fibroblasts at 1 mmol/l was observed compared to control (* p< 0.05, n= 6).
Concentration of AgNO3 (mmol/l)
Optical density
Concentration of AgNO3 (mmol/l)
Optical density
Results
b. Incubated with MgAg1% sticks
Primary mammary epithelium cells did not show a reduction in the viability, when treated with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium) (Fig. 41).
Fig. 41: Cell viability (Neutral red assay) of primary mammary epithelium cells as indicated by the mean optical density of neutral red after treatment with degradation medium (MgAg1%
sticks incubated in 1, 3, 10 ml medium). No significant reduction in the viability was observed compared to control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3 mmol/l Mg++, 0.04 mmol/l Ag+ and 1 ml ≈ 0.5 mmol/l Mg++, 0.2 mmol/l Ag+ (n= 4).
Primary mammary fibroblasts did not show a reduction in viability, when treated with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium) (Fig. 42).
Fig. 42: Cell viability (Neutral red assay) of primary mammary fibroblasts as indicated by the mean optical density of neutral red after treatment with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium). No significant reduction in the viability was observed compared with control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3 mmol/l Mg++, 0.04 mmol/l Ag+ and 1 ml ≈ 0.5 mmol/l Mg++, 0.2 mmol/l Ag+ (n= 4).
Different volumes of degradation medium (ml)
Optical density
Different volumes of degradation medium (ml)
Optical density
Results
4.3.1.3 Measuring the amount of protein in the supernatants
Primary mammary epithelium cells showed a reduction in the protein amounts when treated with different concentrations of AgNO3 solution. A significant reduction was observed at concentrations equal or above 0.3 mmol/l. When they were treated with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium), they did not show a significant decrease in the protein amount (Fig. 43)
Fig. 43: Bio-Rad assay of primary mammary epithelial cells. Mean optical density for Coomassive ® Brilliant Blue G-250 dye as an indicator of protein concentration. A significant decrease in the protein amount of primary mammary epithelial cells at concentrations equal or above 0.3 mmol/l was observed. While no significant difference was observed when treated with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium) compared to control (* p< 0.05, n= 4).
Concentrations of AgNO3 (mmol/l)
Optical density
Different volumes of degradation medium (ml)
Optical density
Results
4.3.2 Measurement of metabolic activities
4.3.2.1 Measurement of SDH activity in the supernatant 1. Murine cells
Murine fibroblasts showed an increase in SDH activity, when incubated with different concentrations of AgNO3 solution equal or higher than 0.3 mmol/l, while they showed no significant differences when treated with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium) (Fig. 44).
Fig. 44: Mean SDH activity of murine fibroblasts. A significant increase in the SDH activity of L929 at 0.3 mmol/L was observed compared to control, while no significant difference were observed with different volumes of degradation medium of MgAg1% sticks compared to control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3 mmol/l Mg++,0.04
Different volumes of degradation medium (ml)
SDHOD/g protein
Concentration of AgNO3 (mmol/l) SDHOD/g protein
L929 cells
L929 cells
Results
2. Bovine cells
Primary mammary epithelium cells showed an increase in the SDH activity by 65%, when incubated with concentrations of AgNO3 solution equal or below 1 mmol/l.
Furthermore, they showed the highest SDH activity, when treated with volume equal or below 1 ml degradation medium of MgAg1% sticks (Fig. 45).
Fig. 45: Mean SDH activity of primary mammary epithelial cells. A significant increase in the SDH activity of primary mammary epithelial cells at 1 mmol/l compared to control was observed, in addition, a significant increase was observed at 1 ml of the degradation medium of MgAg1% compared to control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3
Different volumes of degradation medium (ml) SDHOD/g protein
Concentration of AgNO3 (mmol/l) SDHOD/g protein
Primary mammary epithelial cells
Primary mammary epithelial cells
Results
Primary mammary fibroblasts did not show differences in SDH activity, when incubated with different concentrations of AgNO3 solution, while they showed the highest SDH activity, when treated with volume equal or below 1 ml degradation medium of MgAg1% sticks (Fig. 46).
Fig. 46: Mean SDH activity of primary mammary fibroblasts. There is no significant difference of SDH activity of primary mammary fibroblasts compared to control when treated with concentrations of AgNO3 solution, while a significant increase was observed at 1 ml volume of the degradation medium of MgAg1% sticks compared to control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3 mmol/l Mg++,0.04 mmol/l Ag+ and 1 ml ≈
Different volumes of degradation medium (ml) SDHOD/g protein
Concentration of AgNO3 (mmol/l) SDHOD/g protein
Results
4.3.2.2 Measurement of PK activity in supernatant 1. Murine cells
Murine fibroblasts cells showed an increase in the PK activity by about 60 %, when incubated with concentrations equal or higher than 0.3 mmol/l, while they showed no significant difference, when incubated with degradation medium (MgAg1% sticks incubated in 1, 3, 10 ml medium) (Fig. 47).
Fig. 47: Mean PK activity of murine fibroblasts. A significant increase in the PK activity of L929 at 0.3 mmol/l compared to control was observed, while no significant difference was observed with different volumes of degradation medium of MgAg1% compared to control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3 mmol/l Mg++,0.04 mmol/l Ag+ and 1 ml ≈ 0.5 mmol/l Mg++,0.2 mmol/l Ag+ (* p< 0.05, n= 6).
control 10 ml 3 ml 1 ml
0.0 0.2 0.4 0.6
Different volumes of degradation medium (ml) PKOD/g protein
L929 cells control
0.0001 0.0003
0.001
0.003 0.01 0.03 0.1 0.3 1 0.0
0.1 0.2 0.3 0.4
0.5
* *
Concentration of AgNO3 (mmol/l) PKOD/g protein
Results
2. Bovine cells
Primary mammary epithelium cells showed an increase in the PK activity by about 60
%, when incubated with concentrations equal or below 1 mmol/l. Furthermore, they showed the highest PK activity when incubated with 1 ml volume of the degradation medium of MgAg1% (Fig. 48).
Fig. 48: Mean PK activity of primary mammary epithelial cells. A significant increase in the PK activity of primary mammary epithelium at 1 mmol/l compared to control, in addition to a significant increase was observed at 1 ml of the degradation medium of MgAg1% sticks compared to control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3
Concentration of AgNO3 (mmo/l)
PKOD/g protein
Different volumes of degradation medium (ml)
PKOD/g protein
Primary mammary epithelial cells
Results
Primary mammary fibroblasts did not show significant difference of PK activity, when incubated with different concentrations of AgNO3 solution, while they showed highest PK activity, when incubated with 1 ml of degradation medium of MgAg1% sticks (Fig. 49).
Fig. 49: Mean PK activity of primary mammary fibroblasts. No significant increase in the PK activity of primary mammary fibroblasts was observed, when treated with different concentrations of AgNO3 solution compared to control. In addition, a significant increase was observed at 1 ml of the degradation medium of MgAg1% sticks compared to control regarding that, 10 ml ≈ 0.2 mmol/l Mg++, 0.02 mmol/l Ag+, 3 ml ≈ 0.3 mmol/l Mg++,0.04 mmol/l
Different concentration of AgNO3 (mmol/l) PKOD/g protein
Results
4.3.2.3 Succinate staining of tissue and cells 4.3.2.3.1 Teat tissues
After succinate staining, the teat tissues, which were incubated with MgAg1% sticks, showed a higher SDH activity presented by a violet or dark blue diformazan from nitrozolium blue reduction through succinate oxidation (Fig. 50 B) compared to the untreated teat (Fig. 50 A). Furthermore, after staining with succinate and H&E, higher SDH activity was also observed in the teat tissues which were incubated with MgAg 1% sticks (Fig. 50 D) compared to the untreated teat (Fig. 50 C).
Fig. 50: Tissue sections of untreated teat (A, C) and the treated teat with MgAg1% sticks for 5-6 hours (B, D). The treated teat stained by succinate stain (B) and succinate stain / H&E (D) showed a dense dark blue colour compared to untreated teat; magnification = 40x; bar = 50 μm.
A B
D
C
Results
4.3.2.3.2 Primary mammary epithelial cells
Primary mammary epithelial cells showed a higher SDH activity compared to control, when incubated with higher concentrations of AgNO3 solution at 1 mmol/l (Fig. 51).
Fig. 51: Light microscopic pictures of primary mammary epithelium cells stained with succinate and nitrozolium blue. Primary mammary epithelium cells treated with 1 mmol/l of AgNO3 showed darker blue colour (B) compared to untreated control cells (A) which showed less colour; magnification = 20x; bar = 100 μm.
Primary mammary epithelial cells stained with succinate and nitrozolium blue did not show significant differences, when incubated with either different concentrations of AgNO3 or different volumes of degradation medium of MgAg1% (Fig. 52).
Fig. 52: Mean SDH activity of primary mammary epithelial cells treated with succinate and nitrozolium blue. There is no significant difference in incubated primary epithelium cells with different concentration of AgNO3 and degradation medium of MgAg1% sticks (n= 4).
con
Different volumes of degradation medium (ml)
Primary mammary epithelial cells
A B
Results
4.3.3 Biomarkers of inflammatory reactions
4.3.3.1 Measurement of IL-1 beta in the culture medium supernatant
Primary macrophages, raw macrophages and L929 did not show indications of an enhance of IL-1 beta release after incubation with different concentrations of AgNO3
solution or different volumes of degradation medium MgAg1%, although there was a significant response for stimulation with LPS (Fig. 53).
Fig. 53. Detection of IL-1 beta of primary macrophage, raw macrophage and L929 cells.
primary macrophage, raw macrophage and L929 cells treated with AgNO3 and degradation medium of MgAg1% showed no significant difference of IL-1 beta release compared to control (* p< 0.05, n= 4).
Different volumes of degradation medium (ml)
control
Different volumes of degradation medium (ml) control
Concentration of AgNO3 (mmol/l)
pg/ml
Different volumes of degradation medium (ml) control
Concentration of AgNO3 (mmol/l)
pg/ml
Concentration of AgNO3 (mmol/l)
pg/ml
Results
4.3.3.2 Measurement of IL- 6 in the culture medium supernatant
Primary macrophages and raw macrophages did not show indication of an enhance of IL-6 release after incubation with different concentrations of silver nitrate or different volumes of degradation medium, although there was a significant response for stimulation with LPS (Fig. 54).
Fig. 54: Detection of IL-6 of primary macrophage, raw macrophage and L929 cells. Primary macrophage, raw macrophage and L929 cells treated with AgNO3 and MgAg1% showed no significant difference of IL-6 release compared to control (* p< 0.05, n= 4).
raw macrophages
Concentration of AgNO3 (mmol/l)
pg/ml
Different volumes of degradatiom medium (ml)
pg/ml
Different volumes of degradatiom medium (ml)
L929 cells
Different volumes of degradation medium (ml)
*
Concentration of AgNO3 (mmol/l) pg/ml raw macrophage
Concentration of AgNO3 (mmol/l) pg/ml raw macrophage