Supplementary material

Title: Cord Mesenchymal Stem Cells Derived Extracellular Vesicles Can Safely Ameliorate
the Progression of Chronic Kidney Diseases
Authors: Wael Nassar MD1,2, Mervat El-Ansary M D3, Dina Sabry MD5, Mostafa A Mostafa
MRCP, MD 2, Tarek Fayad MD4, Esam Kotb MSc 2, Mahmoud Temraz MSc 1, Abdel-Naser
Saad MSc 1, Wael Essa MSc 1, and Heba Adel MSc3.
Affiliation: Nephrology department, Sahel Teaching Hospital 1, General Organization of
Teaching Hospitals and Institutes (GOTHI), Cairo. Egypt. Department of Internal medicine and
nephrology, October Six University 2, Cairo. Egypt . Department of clinical pathology and stem
cells 3; department of nephrology 4, Departement of biochemistry 5, Faculty of medicine, Cairo
University, Cairo. Egypt.
Group A
(1) TGF-

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
Mean
SD

Before
3.54
3.12
3.81
4.1
3.25
2.31
3.22
3.46
3.24
3.62
4.21
3.78
3.73
3.55
4.1
4.31
4.77
3.28
4.12
4.12
3.682
0.546006

pg/ml
12
4 weeks 8 weeks
weeks
20.8
31.4
22.8
23.7
32.5
25.7
18.8
22.3
20.8
31.3
24.8
33.3
29.6
29.6
31.6
22.9
27.7
24.9
27.8
22.6
29.2
19.4
21.3
21.4
28.3
20.1
30.3
26.9
23.4
28.9
29.4
20.8
31.4
30.2
23.7
32.5
19.3
18.8
22.3
22.8
31.3
24.8
27.6
29.6
29.6
25.9
22.9
27.7
20.6
27.8
22.6
19.3
19.4
21.3
18.1
28.3
20.1
19.7
26.9
23.4
24.12
25.26
26.23
4.472677 4.318796 4.341246

6 mon
20.2
17.3
18.4
19.2
12.9
14.6
13.5
14.5
17.1
15.9
17.4
14.1
12.8
15.7
13.9
16.2
14.9
15.1
14.1
18.3
15.805
2.14144

9 mon
15.8
16.3
13.5
14.7
12.3
11.4
10.5
15.2
16.3
11.4
10.5
10.6
2.4
11.2
10.9
11.3
10.9
12.2
9.8
13.1
12.015
3.080204

12 mon
6.4
3.5
5.2
9.5
8.6
5.2
7.5
7.4
8.2
3.2
9.8
8.4
8.7
4.3
5.1
6.5
7.9
8.9
8.9
7.2
7.02
2.00121

GroupA
(2) IL-10

pg/ml

12
Before
4 weeks 8 weeks
weeks
6 mon
9 mon
2.5
8.6
9.6
16.2
9.6
4.2
3.92
7.8
8.4
12.7
10.6
4.1
4.9
7.3
9.5
11.7
8.3
5.1
4.2
9.6
7.8
11.7
8.4
5.3
2.7
8.4
9.17
11.9
8.4
6.2
3.9
8.6
8.6
13.3
7.3
4.9
3.8
9.4
9.3
13.7
7.5
7.8
3.4
8.6
9.33
11.8
7.4
4.2
3.1
8.7
9.4
12.4
8.2
5.1
3.34
9.8
8.5
15.1
7.8
5.9
4.71
9.6
16.2
15.2
8.9
5.1
3.7
8.4
9.5
12.7
8.4
7.2
4.75
9.5
10.9
11.5
6.7
6.6
4.3
7.8
9.6
12.7
8.2
4.6
3.74
9.17
11.9
13.2
7.9
2.8
4.21
8.6
10.8
13.5
7.9
5.8
4.6
9.3
13.7
15.1
8.3
4.7
4.7
9.33
13.4
16.3
7.9
4.1
3.27
9.4
12.4
14.7
8.5
4.6
3.33
8.5
9.8
11.6
8.2
4.3
3.8535
8.82
10.39
13.35
8.22
5.13
0.66235 0.687474 2.126596 1.495549 0.788368 1.124659

12 mon
2.98
4.21
4.11
4.38
3.7
4.16
5.28
4.27
5.4
4.32
3.7
4.2
5.33
4.11
3.28
4.1
4.8
3.11
4.2
3.4
4.152
0.66857

Group A
(3) TNF-

pg/ml

12
Before
4 weeks 8 weeks
weeks
6 mon
9 mon.
12 mon.
5.8
4.3
2.9
1.3
1.4
1.5
1.5
3.7
3.1
2.7
1.4
1.6
1.4
1.7
4.2
3.7
2.8
1.2
1.1
1.4
1.5
4.1
3.4
2.4
1.7
1.3
1.3
1.6
3.6
3.2
2.7
1.4
1.4
1.6
1.3
4.7
3.9
2.4
1.1
1.2
1.7
1.6
2.9
2.4
1.8
1.2
1.1
1.5
1.5
5.3
4.1
2.7
1.8
1.5
1.7
1.6
4.2
3.8
2.5
1.7
1.7
1.6
1.6
6.2
4.2
3.1
1.3
1.5
1.7
1.4
3.5
3.1
2.6
1.1
1.3
1.5
1.3
2.6
2.1
2.1
1.5
1.6
1.6
1.5
4.8
3.9
2.5
1.7
1.2
1.9
1.2
5.7
4.2
2.9
1.5
1.4
1.6
1.7
7.4
3.1
2.6
1.2
1.6
1.8
1.3
3.8
3.4
2.4
1.1
1.4
1.7
1.4
6.8
5.8
3.1
1.6
1.7
1.9
1.4
8.4
5.1
2.3
1.3
1.5
1.8
1.1
4.7
3.6
2.2
1.1
1.6
1.9
1.7
5.4
4.2
2.6
1.1
1.3
1.7
1.1
4.89
3.73
2.565
1.365
1.42
1.64
1.45
1.465572 0.842365 0.324889 0.230814 0.180555 0.168523 0.147619
0.00465
0.0842
-0.1

Group A

GFR = 60-15
(4) Bl.
urea

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
Mean
SD

mg/dl

8
12
Before
4 weeks weeks
weeks
6 month 9 month 12 mon.
122
118
70
66
63
68
70
124
110
62
64
68
64
69
125
98
72
66
59
66
71
131
104
61
73
68
63
69
134
95
84
70
63
61
68
122
93
64
62
66
68
66
143
102
72
72
64
61
67
112
88
63
61
58
66
72
152
100
75
84
76
68
71
127
104
86
64
60
63
68
124
107
66
72
62
64
65
113
89
72
63
61
66
73
132
94
74
75
66
60
74
148
101
66
86
69
63
68
121
106
78
66
70
62
66
117
87
79
72
59
60
63
134
89
77
74
67
62
68
116
100
81
63
64
63
69
87
82
73
61
60
66
74
132
103
74
66
65
61
73
125.8
98.5
72.45
74.65
64.4
63.75
69.2
14.15553 8.894114 7.20727 6.906519 4.500292 2.653201 2.976575

Group A
(5) S.
creat
.

mg/dl

Before
4 wks
8 wks
12wks
3.1
2.9
2.8
2.4
2.9
2.4
2.6
2.2
3.5
2.8
2.1
2
3.6
2.7
2.3
2.1
3.8
2.6
2.2
2.3
3.1
2.5
2.1
2.1
4.1
2.9
2.6
3.1
2.4
2.1
2.5
2.1
4.1
3.2
2.5
2.3
3.3
3.1
2.2
2.1
3.2
2.7
2.3
2.1
2.8
2.5
2
2
3.6
2.6
2.4
2.2
3.8
2.9
2.2
2.4
3.2
2.7
2.3
2
2.6
2.2
2.1
2.1
3.7
3.4
2
1.8
4.3
3.6
2.2
2.1
3.8
3.1
2.3
2.1
3.8
2.8
2.2
2.2
3.435
2.785
2.295
2.185
0.493071 0.374552 0.213923 0.251545

6
month 9 month 12 mon.
2.7
2.9
2.8
2.5
2.8
2.7
2.3
2.9
2.4
2.2
2.7
2.2
2
2.3
2.6
2.1
2.1
2.2
2.8
2.4
2.3
2.6
2.2
2.3
2.4
2.6
2.7
2.3
2.2
2.4
2.2
2.4
2.1
2.2
2.3
2.2
1.9
2.3
2.1
2.1
2.2
2.2
2.3
2.3
2.6
2.2
2.3
2.2
2.1
2.2
2.1
1.9
2.1
2.3
2.4
2.2
2.4
2.3
2.6
2.3
2.275
2.4
2.355
0.24252 0.257519 0.210891

Group A
(6) UACR

mg/g

12
Before
4 weeks 8 weeks
weeks
1135
1118
866
580
1527
1243
857
613
1542
1340
857
724
1375
983
640
512
1298
1073
718
589
1273
948
588
531
1722
1318
743
489
1372
1099
689
622
1578
1413
714
579
1677
1226
649
614
1744
1431
693
582
2133
1518
715
513
1673
1261
743
627
1273
993
688
530
1422
895
590
488
1566
1031
739
614
1433
978
613
588
2133
1105
841
592
1537
1008
663
586
1677
1121
755
513
1554.5
1155.1
718.05
574.3
259.2856 179.0031 85.62616 57.60035

6 mon
618
723
815
688
741
783
598
774
683
782
633
683
742
683
584
727
755
672
673
749
705.3
64.6766

9 mon
12 mon
912
873
887
912
830
855
761
770
618
713
624
683
820
795
583
630
915
879
499
538
627
716
754
693
763
790
742
851
628
804
815
792
824
805
730
806
742
837
798
831
743.6
778.65
114.653 89.84057

Group A
(7) eGFR

ml/min

12
Before
4 weeks 8 weeks
weeks
6 mon
9 mon
12 mon
30
38
53
57
55
53
51
32
37
41
44
45
43
40
32
36
44
47
46
44
42
37
37
41
46
47
48
41
26
29
42
44
47
43
41
29
30
40
39
42
44
42
26
28
36
41
40
41
38
31
31
39
49
45
40
40
28
29
34
37
38
40
38
41
41
41
53
47
42
42
29
30
40
44
46
44
41
34
33
43
48
43
41
40
32
34
42
49
46
42
41
37
38
41
53
48
45
42
32
31
46
52
50
47
41
34
33
43
53
47
47
39
26
32
45
51
50
44
43
29
31
41
47
47
45
44
25
28
41
48
43
43
40
25
29
35
43
44
45
41
30.75
32.75
41.4
47.25
45.8
44.05
41.35
4.363425 3.864481 4.083342 5.138861 3.707744 3.068945 2.587056

Group B

GFR =60-15
B
Bl. Urea
Before

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
7
18
19
Mean
SD

mg/dl
After

S. creat.
Before

mg/dl
After

UACR
Before

mg/g
After

122
124
3.1
4.1
1250
876
124
113
2.9
3.3
988
935
125
132
3.5
3.2
948
844
131
148
3.6
2.8
972
844
134
121
3.8
3.6
983
984
122
117
3.1
3.8
851
819
143
134
4.1
3.2
855
957
112
116
2.4
2.6
850
836
152
146
4.1
3.7
1104
953
122
124
3.1
4.1
893
876
124
113
2.9
3.3
988
935
125
132
3.5
3.2
948
844
131
148
3.6
2.8
972
844
134
121
3.8
3.6
983
984
122
117
3.1
3.8
851
819
143
134
4.1
3.2
855
957
112
116
2.4
2.6
850
836
152
146
4.1
3.7
1104
953
124
113
2.9
3.3
988
935
129.4444 127.8889
3.4
3.366667 977.8889 894.2222
12.1461
12.8981 0.572276 0.482183 131.6068 62.79287

Group B
eGFR
Before

ml/min
After

TGF-beta
Before

pg/ml
After

IL-10
Before

pg/ml
After

37
31
9.31
9.12
4.39
4.27
36
35
8.22
7.16
3.11
4.79
29
28
8.95
8.79
4.25
3.88
33
34
9.63
9.28
4.91
4.57
31
31
9.54
8.93
4.88
4.82
28
26
8.97
8.89
4.31
4.13
37
25
6.33
7.28
4.85
4.13
32
31
9.58
9.14
4.32
4.19
37
38
7.41
8.82
4.93
4.76
37
31
9.31
9.12
4.39
4.27
36
35
8.22
7.16
3.11
4.79
29
28
8.95
8.79
4.25
3.88
33
34
9.63
9.28
4.91
4.57
31
31
9.54
8.93
4.88
4.82
28
26
8.97
8.89
4.31
4.13
37
5
6.33
7.28
4.85
4.13
32
31
9.58
9.14
4.32
4.19
37
38
7.41
8.82
4.93
4.76
36
35
8.22
7.16
3.11
4.79
33.33333
31
8.66
8.601111 4.438889 4.393333
3.570714 4.242641 1.137398 0.799835 0.576789 0.347095

Other Biopsies of the treatment group A

(1) CD133 expressed at the renal tubular epithelial cells (cytoplasmic) X 630

(2) CD133 expressed at the renal tubular epithelial cells (cytoplasmic) X 630

(3) CD133 tubular casts in atrophic tubules X100

(4) Ki67 within the inflammatory cells

Other Biopsies of the Control group B

(1) Absent CD133 expression in tubular cells X 100

(2) One Ki67 in tubular epithelial cells of the control group X 400

(3) Scanty Ki67 in tubular epithelial cells of the control group X 100