DEFINICIÓN DEL TEMA DE INVESTIGACIÓN Y JUSTIFICACIÓN DEL MISMO
2.5 Norma del Reglamento del ACI 318S-08 Establecida Para el Diseño de Sistemas de Anclaje al Concreto: Apéndice D
Following 8 and 16 weeks of supplementation with Se supplemented with vitamin C and vitamin E (AS7), decrease in levels of LPO and 8-OHdG were found in supplemented Groups two and three (Group two+AS and Group three+AS) when compared with non-supplemented Groups two and three (Group two–AS and Group three–AS). Increases in TAS, vitamin C, vitamin E and Se levels, GPx activities were observed in Group two+AS and Group three+AS compared with Group two–AS and Group three–AS (Figures 4.1a–4.8b).
The ABDEFS wound healing assessing tools revealed reduction in score of supplemented diabetic foot ulcer and increase in score of non-supplemented diabetic foot ulcer subjects (Figure 4.9).
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Table 4.9. Selenium, Vitamins C and E (S7) supplementation on oxidative stress indices and antioxidant status in supplemented and non-supplemented rats
Paramete rs/Groups
Group A Group B Group C Group D Group E
Before Supple mentati on
After Supple mentati on
*%
differ ences
Before Supple mentati on
After Supple mentati on
*%
differ ences
Before Supple mentati on
After Supple mentati on
*%
differ ences
Before Supple mentati on
After Supple mentati on
*%
differ ences
Before Supple mentati on
After Supple mentati on
*%
differ ences 4–HNE
(μg/ml) 11.979
± 0.763
11.630
± 0.467
–2.91 12.114
± 0.666
11.521
± 1.080
–4.90 12.789
± 0.763
11.665
± 0.274
–8.79 12.241
± 0.045
10.956
± 0.117 – 10.50
13.517
± 0.124
10.772
± 0.133 – 20.31 LPO
(μmol/l)
102.761
± 2.062
95.063
± 1.302
–7.49 123.984
± 10.879
110.085
± 3.820 – 11.21
162.721
± 30.753
155.065
± 14.591
–4.70 138.161
± 3.075
108.203
± 6.238 – 21.68
186.541
± 1.831
142.810
± 23.663
– 23.44 8–OHdG
(ng/ml)
3.559 ± 0.310
3.095 ± 0.618
– 13.04
5.139 ± 0.495
4.041 ± 0.018
– 21.37
5.612 ± 0.159
4.782 ± 0.090
– 14.79
5.541 ± 0.619
3.956 ± 0.032
– 28.60
6.697 ± 0.029
4.516 ± 0.103
– 32.57 SOD
(U/g Hb)
5058.42 4 ± 289.245
5498.72 7 ± 46.130
8.70 4294.20 4 ± 447.366
4680.32 9 ± 86.175
8.99 4555.47 2 ± 26.055
5031.92 2 ± 28.509
10.46 4105.10 6 ± 70.416
4922.42 3 ± 180.827
19.91 4432.79 5 ± 95.389
5211.57 4 ± 590.057
17.57
GPx (U/g Hb)
1068.98 1 ± 7.887
1080.73 2 ± 42.510
1.10 1137.62 1 ± 38.723
1176.58 9 ± 70.291
3.43 1099.56 2 ± 9.899
1140.26 5 ± 151.378
3.70 1184.10 6 ± 114.002
1177.00 0 ± 18.859
–0.60 1109.00 5 ± 62.543
1141.36 5 ± 28.878
2.92
TAS (mmol/l)
0.726 ± 0.095
0.742 ± 0.168
2.20 0.667 ± 0.325
0.707 ± 0.129
6.00 0.550 ± 0.028
0.584 ± 0.058
6.18 0.620 ± 0.063
0.707 ± 0.127
14.03 0.516 ± 0.045
0.585 ± 0.153
13.37 Vitamin
C (mg/dl)
0.069 ± 0.010
0.078 ± 0.014
13.04 0.071 ± 0.022
0.088 ± 0.008
23.94 0.067 ± 0.011
0.084 ± 0.009
25.37 0.065 ± 0.003
0.067 ± 0.007
3.08 0.061 ± 0.011
0.075 ± 0.014
22.95 Vitamin
E (mg/dl)
0.789 ± 0.094
0.932 ± 0.088
18.12 0.555 ± 0.020
0.665 ± 0.093
19.82 0.537 ± 0.081
0.741 ± 0.147
37.99 0.527 ± 0.026
0.715 ± 0.181
35.67 0.518 ± 0.007
0.835 ± 0.060
61.20 Se
(μmol/l)
0.230 ± 0.074
0.236 ± 0.010
2.61 0.227 ± 0.034
0.231 ± 0.014
1.76 0.199 ± 0.023
0.221 ± 0.016
11.06 0.224 ± 0.012
0.230 ± 0.013
2.68 0.194 ± 0.005
0.219 ± 0.010
12.89 Results are expressed as Mean ± S.D
*supplemented Groups A, B, C, D and E were compared with non-supplemented Groups A, B, C, D and E
% = Percentage increases and decreases where negative (–) values denotes percentage decreases and positive (+) values percentage increases Group A = non-diabetic Control rats without ulcer
Group B = Diabetic Rats without Ulcer Group C = Diabetic Rats with Ulcer
Group D = Glibenclamide treated Diabetic Rats without Ulcer Group E = Glibenclamide treated Diabetic Rats with Ulcer
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Table 4.10. The Friedman significant table of rats results of supplemented Groups A, B, C, D and E compared with supplemented groups in all the supplementations (S1 to S7)
Groups 4-HNE
(µg/ml)
LPO (µmol/l) 8-OHdG
(ng/ml)
SOD (U/g Hb) GPx (U/g Hb) TAS (mmol/l) Vitamin C (mg/dl)
Vitamin E (mg/dl)
Se (µmol/l)
Fried-man
P-value
Fried-man
P-value
Fried-man
P-value
Fried-man
P-value
Fried-man
P-value
Fried-man
P-value
Fried-man
P-value
Fried-man
P-value
Fried-man
P-value
†Before Supplementation
A, B, & C 1.733NS 0.420 8.000* 0.018 6.000 0.050 6.500* 0.039 8.000* 0.018 6.500* 0.039 0.500NS 0.779 6.500* 0.039 1.500NS 0.472 A, D & E 6.500* 0.039 8.000* 0.018 8.000* 0.018 8.000* 0.018 3.500NS 0.174 4.500NS 0.105 0.933NS 0.627 6.000 0.050 4.500NS 0.105
#After Supplementation
A1, B1 & C1 2.000NS 0.368 8.000* 0.018 8.000* 0.018 6.500* 0.039 0.500NS 0.779 6.000 0.050 0.000NS 1.000 8.000* 0.018 2.000NS 0.368 A1, D1 & E1 6.000 0.050 8.000* 0.018 6.000 0.050 3.500NS 0.174 0.500NS 0.779 2.800NS 0.247 2.000NS 0.368 3.500NS 0.174 2.000NS 0.368 A2, B2 & C2 8.000* 0.018 8.000* 0.018 8.000* 0.018 3.500NS 0.174 4.500NS 0.105 8.000* 0.018 2.000NS 0.368 8.000* 0.018 1.733NS 0.420 A2, D2 & E2 8.000* 0.018 8.000* 0.018 8.000* 0.018 6.500* 0.039 6.500* 0.039 8.000* 0.018 0.500NS 0.779 4.500NS 0.105 0.500NS 0.779 A3, B3 & C3 2.000NS 0.368 8.000* 0.018 6.500* 0.039 8.000* 0.018 2.800NS 0.247 8.000* 0.018 3.500NS 0.174 6.000 0.050 0.400NS 0.819 A3, D3 & E3 1.500NS 0.472 8.000* 0.018 6.500* 0.039 4.500NS 0.105 6.000 0.050 3.500NS 0.174 8.000* 0.018 6.500* 0.039 3.500NS 0.174 A4, B4 & C4 1.500NS 0.472 8.000* 0.018 8.000* 0.018 6.500* 0.039 6.500* 0.039 8.000* 0.018 0.500NS 0.779 8.000* 0.018 0.933NS 0.627 A4, D4 & E4 3.500NS 0.174 8.000* 0.018 6.500* 0.039 2.000NS 0.368 6.500* 0.039 6.500* 0.039 3.500NS 0.174 8.000* 0.018 0.500NS 0.779 A5, B5 & C5 6.500* 0.039 6.500* 0.039 8.000* 0.018 8.000* 0.018 8.000* 0.018 2.000NS 0.368 1.200NS 0.549 0.500NS 0.779 3.500NS 0.174 A5, D5 & E5 8.000* 0.018 4.500NS 0.105 8.000* 0.018 8.000* 0.018 3.500NS 0.174 1.500NS 0.472 0.500NS 0.779 0.500NS 0.779 0.000NS 1.000 A6, B6 & C6 6.500* 0.039 0.500NS 0.779 6.000 0.050 8.000* 0.018 4.500NS 0.105 4.500NS 0.105 1.500NS 0.472 1.500NS 0.472 0.400NS 0.819 A6, D6 & E6 6.500* 0.039 8.000* 0.018 6.500* 0.039 3.500NS 0.174 4.500NS 0.105 6.000 0.050 1.500NS 0.472 6.000 0.050 0.933NS 0.627 A7, B7 & C7 0.000NS 1.000 8.000* 0.018 8.000* 0.018 8.000* 0.018 2.000NS 0.368 3.500NS 0.174 2.286NS 0.319 6.500* 0.039 2.364NS 0.307 A7, D7 & E7 3.500NS 0.174 8.000* 0.018 8.000* 0.018 2.000NS 0.368 6.500* 0.039 2.000NS 0.368 2.286NS 0.319 6.000 0.050 3.000NS 0.223
†Non-supplemented Groups B and C and Groups D and E were compared with non-supplemented Group A at the beginning of the study.
#Supplemented Groups B and C and Groups D and E were compared with Supplemented Group A.
Not significant = NSP > 0.05 *Significant at p < 0.05
Group A = non-diabetic control rats without ulcer Group B = Diabetic rats without ulcer Group C = Diabetic Rats with Ulcer Group D = Glibenclamide treated diabetic rats without ulcer Group E = Glibenclamide treated diabetic rats with ulcer
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A. Rat Tissue with Fibroblast B. Rat Tissue with Fibroblast 2
C. Rat Tissue with Fibroblast 3 D. Rat Tissue with Fibroblast 4
E. Normal Tissue of Rat F. Normal Tissue of Rat
Figure 4.03a. Healing process in Wistar rats as depicted by the presence of fibroblast and granulated Tissues (A – F)
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G. Rat Tissue with granulation cells 1 H. Rat Tissue with granulation cells 2
I. Rat Tissue with granulation cells 3 J. Rat Tissue with granulation cells 4
K. Normal Tissue of Rat L. Normal Tissue of Rat
Figure 4.03b. Healing process in Wistar rats as depicted by the presence of fibroblast and granulated Tissues (G – L)
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Table 4.11. The comparison of parameters at 8 and 16 weeks of antioxidant supplementation with baseline parameters in Group two and Group three using percentage differences
Before Antioxidant Supplementation
8 and 16 weeks Antioxidant
Supplementation in DM with and without DFU
8 and 16 weeks of non – Antioxidant Supplementation in DM with and without
DFU Parameters *Group
two (%) 0 week
n = 56
*Group three (%)
0 week n = 70
¶Group two (%) 8 week
†n = 25
¶Group2 (%) 16week
†n =25
¶Group three (%)
8 week
†n = 25
¶Group three (%)
16week
†n = 25
¶Group2 (%) 8 week
†n = 25
¶Group2 (%) 16 week
†n = 25
¶Group three
(%) 8 week
†n = 25
¶Group three
(%) 16 week
†n = 25
BMI(Kg/m2) +20.80 +13.74 –5.27 –8.18 –4.55 –6.58 –3.51 –6.73 –3.84 –6.07
FPG (mg/dl) +138.90 +155.13 –5.78 –7.70 –4.79 –9.61 –5.90 –12.94 –4.71 –10.34
HbA1c (%) +89.22 +111.76 –13.53 –18.79 –5.13 –9.44 –10.10 –11.76 –5.75 –8.06
LPO (µmol/l) +42.60 +76.60 –7.43 –10.54 –12.55 –22.53 +4.20 +7.63 +12.44 +23.10
8-OHdG(ng/ml) +46.72 +56.76 –6.71 –10.19 –10.49 –22.54 +14.34 +17.28 +12.46 +26.81
SOD (U/g Hb) –13.99 –3.27 +2.53 +3.56 –3.03 –0.43 –3.61 –5.66 –6.14 –11.29
GPx (U/g Hb) +68.66 +160.28 +5.30 +6.02 –4.77 +17.79 –1.60 –3.27 +5.66 +2.87
TAS (mmol/l) –53.52 –52.82 +7.81 +12.50 +4.55 +9.09 –4.41 –8.82 –7.25 –11.59
VitaminC(mg/dl) –20.00 –30.00 +44.93 +60.87 +6.00 +20.00 –8.57 –15.71 –13.24 –14.71 VitaminE(mg/dl) –20.00 –22.73 +19.76 +26.67 +22.03 +31.53 +5.41 +8.47 –4.69 –15.43
Se (µmol/l) –39.51 –40.74 +11.11 +18.67 +11.02 +19.54 –6.95 –13.68 –12.14 –16.96
ABDEFS – – – – –13.21 –24.81 – – +5.96 +14.04
*The percentage increases and decreases of Groups two and three were computed using Group one as reference at baseline.
†n = Number of subjects that completed the study
¶The percentage increases and decreases of supplemented and non-supplemented Groups two and three at 8 and 16 weeks were compared with Groups two and three at baseline respectively.
n = Number of subjects
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Figure 4.1a. Selenium, vitamin C and vitamin E supplementation effects on lipid peroxide (LPO) levels at baseline, 8 and 16 weeks in DM subjects without diabetic foot ulcer (Group two)
Group two + AS = Diabetes mellitus subjects without diabetic foot ulcer subjects administered with antioxidant supplementation
Group two – AS = Diabetes mellitus subjects without diabetic foot ulcer subjects not administered with antioxidant supplementation
At baseline, a non-significant change was observed when the LPO values of supplemented Group two was compared with the value of non-supplemented group. The supplementation at 8 and 16 weeks resulted in non-significant decreases of LPO levels in supplemented group compared with non-supplemented group.
44.40
41.10 39.72
45.19 ns
47.09 ns
48.64 ns
0 30 60
baseline 8 weeks 16 weeks
Values of LPO (µmol/L)
Group two + AS Group two – AS
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Figure 4.1b. Selenium, vitamins C and E supplementation effects on lipid peroxide (LPO) levels at baseline, 8 and 16 weeks in DM subjects with diabetic foot ulcer (Group three)
Group three + AS = Diabetes mellitus subjects with diabetic foot ulcer subjects administered with antioxidant supplementation
Group three – AS = Diabetes mellitus subjects with diabetic foot ulcer not administered with antioxidant supplementation
In this group, when LPO of supplemented group was compared with non-supplemented group at baseline, a non-significant change in levels was observed. Following, 8 and 16 weeks of supplementation a respectively non-significant and significant reduction of this parameter in the supplemented group compared with the non-supplemented group was revealed.
54.49
48.27 48.67
54.67 ns
59.85 ns
62.31
**
0 40 80
baseline 8 weeks 16 weeks
Values of LPO (µmol/L)
Group three + AS Group three – AS
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Figure 4.2a. Selenium, vitamins C and E supplementation effects on
8-hydroxy-2-deoxyguanosine (8-OHdG) levels at baseline, 8 and 16 weeks in DM subjects without diabetic foot ulcer (Group two)
Group two + AS = Diabetes mellitus subjects without diabetic foot ulcer administered with antioxidant supplementation
Group two – AS = Diabetes mellitus subjects without diabetic foot ulcer not administered with antioxidant supplementation
In this group, there is no significant change in the value of 8-OHdG between the supplemented and non-supplemented of Group two at baseline. Following 8 and 16 weeks of antioxidant supplementation, decreases in values that are non-significant and significant respectively were found when supplemented group were compared with the non-supplemented group.
46.51
43.39 41.77
47.00 ns
53.74 ns
55.12
**
0 40 80
baseline 8 weeks 16 weeks
Values of 8-OHdG (ng/mL)
Group two + AS Group two – AS
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Figure 4.2b. Selenium, vitamins C and E supplementation effects on 8-
hydroxy-2-deoxyguanosine (8-OHdG) levels at baseline, 8 and 16 weeks in DM subjects with diabetic foot ulcer (Group three)
Group three + AS = Diabetes mellitus subjects with diabetic foot ulcer administered with antioxidant supplementation
Group three – AS =Diabetes mellitus subjects with diabetic foot ulcer not administered with antioxidant supplementation
At baseline, 8-OHdG value in non-supplemented diabetic foot ulcer group was found to be lower than the value obtained in the supplemented group (p<0.001). A non-significant decrease at 8 weeks and a significant reduction at 16 weeks (p<0.001) of antioxidant micronutrients supplementation were observed in the supplemented diabetic foot ulcer group when compared with the non-supplemented group
53.55
47.93
41.48 46.37
*
52.15 ns
58.80
*
0 40 80
baseline 8 weeks 16 weeks
Values of 8-OHdG (ng/mL)
Group three + AS Group three – AS
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Figure 4.3a. Selenium, vitamins C and E supplementation effects on
superoxide dismutase (SOD) activities at baseline, 8 and 16 weeks in DM subjects without diabetic foot ulcer (Group two)
Group two + AS = Diabetes mellitus subjects without diabetic foot ulcer administered with antioxidant supplementation
Group two – AS =Diabetes mellitus subjects without diabetic foot ulcer not administered with antioxidant supplementation
The beginning of the study before supplementation, the activities of superoxide dismutase in the supplemented diabetic subjects without ulcer reveal no significant change when compared with the non-supplemented group. Following 8 and 16 weeks of antioxidant supplementation, non-significant increase in activities of this enzyme was found in the supplemented group compared with the non-supplemented group (p>0.05).