Jack Kent Cooke research fellowship presentation.

1. Peritoneal Dialysis and
Secondary Renal Function

2. Background/Goals
 End Stage Renal Disease (ESRD)
 Current therapy:
- Peritoneal Dialysis vs. Hemodialysis
- Few kidneys available for transplantation
 Development of a viable renal function
replacement system

3. Methods
Animal model of uremia: Bilateral nephrectomies 45-
60 kg female sheep performed in two surgical
stages.
Stage 1
Left nephrectomy + PD catheters placement (infusion and removal)
Stage 2
Right nephrectomy, lines placed in carotid artery and jugular vein (to
monitor and control hemodynamics).
PD and BREC-d was turned on 24 hrs after second
surgery.
Data was collected up to 10 days.

4. Data Collection
 Hemodynamics:
 Heart Rate
 Mean Arterial Pressure
 Central Venous Pressure
 Respiratory Rate
 Electrolytes :
 K+
 Renal function:
 BUN / CREAT
 PD Fluid Flow Rate

5. Roller BREC-d F-40
pump
F-80 Trilogy
OUT
MC3 Trilogy
pump
Waste PD IN

6. Study Groups
 Control (n=5)
 Acellular BREC-d
 Study - Cellular (n=11): Renal Epithelial Cells
 Lamb BREC-d (n=6)
 Human BREC-d (n=5)

7. Sheep Number BREC-d Cell type Duration of Study
Study 3 Lamb 10 days
5 Lamb 10 days
Groups 6 Lamb 10 days
7 Lamb 7 days
8 Lamb <1 day
9 Lamb 8 days
11 Human <1 day
12 Human 10 days
16 Human 10 days
17 Human 10 days
18 Human 10 days
20 Acellular 2.5 days
21 Acellular 5 days
22 Acellular 10 days
23 Acellular 7 days
24 Acellular 10 days

8. Acellular Control Group (n=5)
Vs.
Cellular Study Group (n=11)

9. Normal Range for sheep (60-120 BPM)

10. Normal Range for Sheep (73-120 mmHg)
Average Mean Arterial Blood Pressure: Acellular vs Cellular
Error bars = SEM
Acellular-BREC (n=5) Cellular-BREC (n=11)
140
120
100
mmHg
80
60
40
20
0
0 24 48 72 96 120 144 168 192 216 240
Experimental Time (Hrs)

11. Average Sheep Central Venous Pressure per Treatment
Group
Error bars = SEM
Acellular-BREC (n=5) Cellular- BREC (n=11)
35
30
25
mmHg
20
15
10
5
0
0 24 48 72 96 120 144 168 192 216 240
Experimental Time (Hrs)

12. Normal Range for sheep (10-20 Breaths/Min)
Average Sheep Respiration Rate per Treatment Group
Error bars = SEM
Acellular-BREC (n=5) Cellular-BREC (n=11)
55
50
45
40
Breaths/min
35
30
25
20
15
10
5
0
0 24 48 72 96 120 144 168 192 216 240
Experimental Time (Hrs)

13. Reference Range 3.5-4.5mEq/L
Avg Acellular K+ Values (n=3)
Avg K+ Before PD Exchange Avg K+ After PD Exchange
8
7
6
5
mEq/L
4
3
2
1
0
0 1 2 3 4 5 6 7 8
Study Day
PD Fluid Exchange Rate: 1/Day

14. Reference Range 3.5-4.5mEq/L
Avg Cellular K+ Values (n=3)
Avg K+before PD Exchange Avg K+ after PD Exchange
7
6
5
mEq/L
4
3
2
1
0
1 2 3 4 5 6 7 8 9 10
Study Days
PD Fluid Exchange Rate: 1/Day

15. Metabolic Waste
BUN (Blood Urea Nitrogen)
Natural bi-product of metabolic function, can
cause damage to tissue if not excreted.
Value greater than 60mg/dl indicates severe
renal impairment
Creatine- Another waste product produce
through metabolism with an average range
of 0.6-1.5 mg/dL

16. Average PD Flow per Treatment Group
Error bars = SEM
Acellular-BREC (n=5) Cellular-BREC (n=11)
140
120
100
mL/min
80
60
40
20
0
0 24 48 72 96 120 144 168 192 216 240
Experimental Time (Hrs)

17. Sheep 18 Metabolic Waste
90
80
70
60
50
mg/dl
40
30
20
10
0
Pre Pre Brecs Brecs Day Brecs Day Brecs Day Final
Surgery 1 Day 0 1 2 3
BUN mg/dL (Reference Range 5.-20.) Creatine mg/dL (Reference Range 0.6-1.5)

18. Comparative BUN and Creatine
Data
 Due to constant recirculation of PD fluid, metabolic
wastes such as blood urea nitrogen (BUN) and creatine
reach elevated levels.
 Recirculation occurs so that renal cells can be kept alive
 Thus PD effectiveness is significantly reduced
 BREC-d does not replace primary functions, therefore no
effect on metabolites.

19. Viability of Renal Cell System
 Oxygen Consumption in vivo
Animal Oxygen Consumption of
Recovered Renal Cells
Sheep 16 49.76 +/- 3.25 mmoles of Oxygen/ min
Sheep 17 35.88+/- 7.86 mmoles of Oxygen/ min
Sheep 18 33.08 +/- 4.18 mmoles of Oxygen/ min
 Average oxygen consumption in vitro is 19.24 mmoles of
Oxygen/ min
 This data illustrates that renal cells can survive in
BREC-d environment.

20. Conclusions
 Cellular BREC-d maintains acceptable MAP.
 Respiration rate elevated
 Discomfort due to dialysate volume
 More PD fluid exchanges necessary to better
control K+; BUN; CREAT
 Renal cells are viable for course of study.

21. Future Research
 More effective PD fluid
 Analyze effects of BREC-d unit for longer period
time (>2 weeks)
 Analyze sheep vs human BREC-d
 Increase frequency of PD fluid exchange
Twice a day (am/pm)