• Keine Ergebnisse gefunden

3 Materials and Methods

4.2 Degradation process of MgAg1 % sticks

4.2.1 Isolated perfused udder

4.2.1.2 Measuring glucose consumption in the perfused udder

The glucose consumption during the perfusation process ranged from 45-77 mg/dl on both sides of the udder. Glucose concentration did not show a significant difference on both sides over time (Fig. 18).

Fig. 18: Mean glucose concentration (mg/dl) in the perfusate of perfused udder. There is no significant difference of glucose consumptions lactate production of left and right sides of perfused udder, n=6.

0 2 4 6

0 50 100

150 Left side

Right side

mg/dl

Time (h)

0 2 4 6

0.0 0.2 0.4

0.6 Left side

Right side

mg/ml

Time (h)

Results

4.2.1.3 Measuring lactate dehydrogenase enzyme (LDH) activity in the perfused udder

The lactate dehydrogenase production during the perfusation process ranged from 1000 -1750 mU/l on both sides of the udder. LDH concentration did not show a significant difference on both sides over time (Fig. 19).

Fig. 19: Mean LDH concentration in the perfusate of perfused udder (mU/l). There is no significant difference of (LDH) production of left and right sides of perfused udder, n=6.

4.2.2 Degradation of MgAg1% sticks in bovine udder

The MgAg1% sticks degraded in bovine udder after being incubated for 5-6 hours in the teat of isolated bovine udder (Fig. 20).

Fig. 20:MgAg1% sticks. The stick before incubation in the teat of bovine udder (A), while (B) showed the stick after the incubation.

0 2 4 6 8

0 1000 2000

3000 Left side

Right side

Time (h)

mU/l

A B

Results

The initial and the final weight of MgAg1% sticks incubated in the teat of isolated bovine udder for 5-6 hours for 7 separated experiments was listed in (Tab. 8). The mean percentage of weigh loss during incubation of MgAg1% sticks was about 3 % of the initial weight (Fig. 21).

Tab. 8: The initial and final weight of MgAg 1% sticks incubated in the teat of bovine udder for 5-6 hours (n=7)

Trials 1 2 3 4 5 6 7

Initial weight/mg 56.06 53.27 53.97 54.43 54.63 53.3 52.6 Final weight/mg

after 6 h

51.43 52.06 53.56 51.33 53.87 53.23 51.93

Fig. 21: Mean weight of MgAg1% sticks before and after incubation in the teat of bovine udder. There is a significant decrease of weight of MgAg1% sticks incubated 6 h in the teat of bovine udder (* P< 0.05, n= 7).

Initial we ight

Weig ht after loss 45

50 55 60

*

Weight in mg

Results

4.2.3 Degradation of MgAg1% sticks in secretion samples from cows at dry off Period

MgAg 1% stick was cutted into three parts nearly of the same weight.The initial and final weight of MgAg1% sticks after incubation in secretion from dry cows for 21 days was listed in (Tab. 9). The mean loss percentage for 6 separated experiments after incubation was 60 % of the initial weight, regarding that the stick of the highest weight was completely degradable after 21 days.

Tab. 9: The initial and final weight of MgAg1% sticks incubated in dry secretion for 21 days (n=6)

Trials 1 2 3 4 5 6

Initial weight 15.7 mg 14.0 mg 10.3 mg 9.5 mg 10.1 mg 8.6 mg Final weight Completely

degraded

3.8 mg 6.6 mg 4.0 mg 4.7 mg 5.3 mg

The mean final weigh of 6 separated experiments of MgAg1% sticks, after incubation in dry secretion for 21 days, revealed a significant difference compared to the initial weight (Fig. 22).

Fig. 22: Mean weight of MgAg1% sticks before and after incubation in dry off period secretion. There is a significant decrease of weight of MgAg1% sticks incubated 21 days in dry secretion (* P< 0.05, n=6).

initia l we

igh t

Final weight 0

5 10 15 20

weight in mg

*

Results

The MgAg1% sticks degraded after incubation in dry off period secretion for 21 days compared to the initial stick (Fig. 23).

Fig. 23: Cutted MgAg1% sticks. The stick before incubation (A). The stick after the 21 days of incubation in dry off period secretion (B).

A B

Results

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

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