Blood Transfusion, Endothelial Physiopathology, Nitric Oxide, Pulmonary Hypertension
Conditions
Keywords
Blood Transfusion, Autologous, Blood Preservation/adverse effects, Blood Transfusion/adverse effects, Endothelium, Vascular/physiopathology, Hemoglobins, Nitric Oxide, Pulmonary Hypertension
Brief summary
The purpose of this study is to determine whether storage time affects how human body responds to autologous blood transfusion. An autologous blood transfusion is when a person donates blood and then receives that same blood back in the transfusion. We also want to find out if in this situation inhaled nitric oxide can help to prevent the potential reduction of vasodilation capacity. Vasodilation capacity is the ability of the blood vessel to widen when needed.
Detailed description
The objective of this study is to assess effects of the storage of PRBC on pulmonary vasoconstriction measured as increase in pulmonary artery pressure and endothelial function measured as a change in reactive hyperemia index in overweight people with existing endothelial dysfunction at baseline. The present study consists of three different parts, which will be scheduled in a randomized order on the same subject (crossover study). During one phase of the study, 14 healthy human volunteers will donate a unit of Packed Red Blood Cells (PRBC), which will be leukoreduced and stored in Additive Solutions-1 (AS-1), and then transfused back to the subjects after 3 days of storage at 4º C in the MGH Blood Bank (Fresh Blood arm). The second part of the study consists in the collection of another unit of PRBC from the same volunteers which will be transfused back to them after 40 days of storage (Old Blood arm). Finally in the third part, like in the second one, one unit of PRBC will be withdraw and stored for 40 days, but 80 ppm (parts per million by volume) Nitric Oxide in air will be administered together with the transfusion. There will be a 2 weeks interval after each PRBC transfusion. We hypothesize that old red blood cells stored under conventional conditions may trigger a complex, pro-inflammatory, pro-thrombotic and vasoconstriction response. We will compare the response to PRBC stored for 3 days with the response to PRBC stored for 40 days in the same healthy volunteers. We also want to test the hypothesis that inhaled nitric oxide may reverse these adverse effects. We will monitor/measure the following parameters: 1. Pulmonary vasoconstriction by trans-thoracic echocardiography 2. Endothelium-mediated changes in vascular (arterial) tone, measured as reactive hyperemia index.
Interventions
PROCEDURERed blood Cells auto-transfusion
Withdrawal from 14 overweight volunteers of one unit of red blood cells and auto-transfusion after 3 days storage time. The same 14 subjects will be included in every arm of the study.
DRUGInhaled Nitric Oxide (iNO) administration
Subjects will breath iNO (80ppm) for 10 minutes before, during and for one hour after the autologous old-blood transfusion.
Sponsors
Massachusetts General Hospital
Study design
Allocation
RANDOMIZED
Intervention model
CROSSOVER
Primary purpose
BASIC_SCIENCE
Masking
SINGLE (Outcomes Assessor)
Eligibility
Sex/Gender
ALL
Age
18 Years to 60 Years
Healthy volunteers
No
Inclusion criteria
1. Have a photo ID 2. Age\>18 years old (required to provide informed consent) 3. Age \<60 years old 4. Mild impairment of endothelial function, assessed by post ischemic vasodilation (L\_RHI,\<0.7) (38) 5. Body mass index (BMI) \>27 kg/m\^2 and \<40 kg/m\^2 6. Fasting during the days of transfusion. 7. Avoiding intake of high nitrate food (i.e. green leafy vegetables: lettuce, spinach) on the day prior to the study 8. Feel well on the day of blood donation 9. KG within normal limits 10. Normotensive (systolic blood pressure \<140 mmHg and diastolic \<90 mmHg) 11. Normal physical exam and blood test results as indicated by: 1. WBC 4.5-11.0 n x103/μL 2. HGB 12.0-17.5 gm/dl 3. PLT 150-400 n x103/μL 4. Plasma Sodium 135-145 mmol/L 5. Plasma Potassium 3.4-4.8 mmol/L 6. Plasma Chloride 98-108 mmol/L 7. Plasma Carbon Dioxide 23.0-31.9 mmol/L 8. Plasma Urea Nitrogen 8-25 mg/dl 9. Plasma Creatinine 0.60-1.50 mg/dl 10. Plasma Glucose 70-110 mg/dl 11. Transaminase-SGPT 10-55 U/L 12. Transaminase-SGOT 10-40 U/L 13. Total Bilirubin \< 2 mg/dl 14. Fasting plasma glucose \< 110 mg/dl 15. Methemoglobin \< 3%
Exclusion criteria
1. Psychiatric disturbances such as anxiety, depression, schizophrenia requiring pharmacological treatment or hospitalization in the last year 2. Systemic disease with or without any functional limitation 3. Pregnancy determined by urine pregnancy test, detecting presence of human chorionic gonadotropin (hCG), or less than six weeks postpartum 4. Active smoking. Volunteers may be enrolled if they quit smoking for more than 1 year. 5. Excess alcohol use: more than ½ L/day of wine consumption or equivalent 6. Any current use of a medication other than: over-the-counter oral medications, herbal remedies, nutritional supplements, and oral contraceptives. 7. Antibiotic use within 48 hours of blood donation 8. Use of NSAIDS, corticosteroids, aspirin during the past 7 days 9. Dental work within 24 hours prior to the donation 10. Received or donated blood in the last 4 months 11. Have had any forms of cancer with the exceptions of basal cell skin cancer or treatment for in situ uterine cervical cancer 12. Currently enrolled in another research study
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Systolic Pulmonary Artery Pressure | Post-transfusion | Pulmonary vasoconstriction was measured by estimation of Systolic Pulmonary Artery Pressure in millimeter of mercury (mmHg) by trans-thoracic echocardiography |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Endothelial Function: Reactive Hyperemia Index | Post-transfusion | Reactive Hyperemia Index (RHI) measures Endothelial function and is assessed by digital pulse amplitude tonometry and it is a sensitive indicator of endothelial function. RHI is a calculated as a ratio between tested versus contralateral finger dilatation, thus there is no unit measure. |
Other
| Measure | Time frame | Description |
|---|---|---|
| Concentration of Cytokines | Before and after blood transfusion | To assess concentration of plasma cytokines at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion. |
| Activation of Platelets | Before and after blood transfusion | To assess concentration of plasma activation of platelets at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion. |
| Hemolysis | before and after blood transfusion | To assess concentration of plasma Hemoglobin at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion. |
| Changes in Gene Expression | Before and after blood transfusion | To assess concentration of changes in gene expression at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion. |
| Endothelial Function | Before and after blood transfusion | To assess Endothelial function by RHI at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion. |
| Activation of Inflammatory Lipid Mediators | Before and after blood transfusion | To assess concentration of plasma activation of inflammatory lipid mediators at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion. |
| Nitric Oxide Metabolites | Before and after blood transfusion | To assess concentration of plasma Nitric oxide metabolites at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion. |
Countries
United States
Participant flow
Participants by arm
| Arm | Count |
|---|---|
| Fresh Blood-Old Blood-Old Blood + Inhaled Nitric Oxide FRESH BLOOD:
Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 3 days storage time.
OLD BLOOD:
Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 40 days storage time.
OLD BLOOD + inhaled NITRIC OXIDE Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 40 days storage time.
Inhaled Nitric Oxide (iNO) administration: Subjects will breath iNO (80ppm) for 10 minutes before, during and for one hour after the autologous old-blood transfusion. | 2 |
| Fresh Blood-Old Blood + Inhaled Nitric Oxide-Old Blood- FRESH BLOOD:
Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 3 days storage time.
OLD BLOOD:
Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 40 days storage time.
OLD BLOOD + inhaled NITRIC OXIDE Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 40 days storage time.
Inhaled Nitric Oxide (iNO) administration: Subjects will breath iNO (80ppm) for 10 minutes before, during and for one hour after the autologous old-blood transfusion. | 3 |
| Old Blood- Fresh Blood- Old Blood + Inhaled Nitric Oxide FRESH BLOOD:
Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 3 days storage time.
OLD BLOOD:
Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 40 days storage time.
OLD BLOOD + inhaled NITRIC OXIDE Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 40 days storage time.
Inhaled Nitric Oxide (iNO) administration: Subjects will breath iNO (80ppm) for 10 minutes before, during and for one hour after the autologous old-blood transfusion. | 3 |
| Old Blood- Old Blood + Inhaled Nitric Oxide- Fresh Blood FRESH BLOOD:
Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 3 days storage time.
OLD BLOOD:
Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 40 days storage time.
OLD BLOOD + inhaled NITRIC OXIDE Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 40 days storage time.
Inhaled Nitric Oxide (iNO) administration: Subjects will breath iNO (80ppm) for 10 minutes before, during and for one hour after the autologous old-blood transfusion. | 2 |
| Old Blood + Inhaled Nitric Oxide- Fresh Blood- Old Blood FRESH BLOOD:
Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 3 days storage time.
OLD BLOOD:
Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 40 days storage time.
OLD BLOOD + inhaled NITRIC OXIDE Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 40 days storage time.
Inhaled Nitric Oxide (iNO) administration: Subjects will breath iNO (80ppm) for 10 minutes before, during and for one hour after the autologous old-blood transfusion. | 2 |
| Old Blood + Inhaled Nitric Oxide- Old Blood- Fresh Blood FRESH BLOOD:
Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 3 days storage time.
OLD BLOOD:
Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 40 days storage time.
OLD BLOOD + inhaled NITRIC OXIDE Red blood Cells auto-transfusion: Withdrawal from the same overweight volunteers of one unit of red blood cells and auto-transfusion after 40 days storage time.
Inhaled Nitric Oxide (iNO) administration: Subjects will breath iNO (80ppm) for 10 minutes before, during and for one hour after the autologous old-blood transfusion. | 2 |
| Total | 14 |
Baseline characteristics
| Characteristic | Fresh Blood-Old Blood-Old Blood + Inhaled Nitric Oxide | Fresh Blood-Old Blood + Inhaled Nitric Oxide-Old Blood- | Old Blood- Fresh Blood- Old Blood + Inhaled Nitric Oxide | Old Blood- Old Blood + Inhaled Nitric Oxide- Fresh Blood | Old Blood + Inhaled Nitric Oxide- Fresh Blood- Old Blood | Old Blood + Inhaled Nitric Oxide- Old Blood- Fresh Blood | Total |
|---|---|---|---|---|---|---|---|
| Age, Categorical <=18 years | 0 Participants | 0 Participants | 0 Participants | 0 Participants | 0 Participants | 0 Participants | 0 Participants |
| Age, Categorical >=65 years | 0 Participants | 0 Participants | 0 Participants | 0 Participants | 0 Participants | 0 Participants | 0 Participants |
| Age, Categorical Between 18 and 65 years | 2 Participants | 3 Participants | 3 Participants | 2 Participants | 2 Participants | 2 Participants | 14 Participants |
| Age, Continuous | 46 years | 37 years | 46 years | 26.5 years | 51 years | 41 years | 41 years |
| Body Mass Index | 37 kg/m^2 | 33 kg/m^2 | 30 kg/m^2 | 33.5 kg/m^2 | 35.5 kg/m^2 | 33 kg/m^2 | 33 kg/m^2 |
| Region of Enrollment United States | 2 participants | 3 participants | 3 participants | 2 participants | 2 participants | 2 participants | 14 participants |
| Sex: Female, Male Female | 0 Participants | 1 Participants | 1 Participants | 1 Participants | 1 Participants | 1 Participants | 5 Participants |
| Sex: Female, Male Male | 2 Participants | 2 Participants | 2 Participants | 1 Participants | 1 Participants | 1 Participants | 9 Participants |
Adverse events
| Event type | EG000 affected / at risk | EG001 affected / at risk | EG002 affected / at risk |
|---|---|---|---|
| deaths Total, all-cause mortality | 0 / 14 | 0 / 13 | 0 / 12 |
| other Total, other adverse events | 0 / 14 | 0 / 13 | 0 / 12 |
| serious Total, serious adverse events | 0 / 14 | 0 / 13 | 0 / 12 |
Outcome results
Primary
Systolic Pulmonary Artery Pressure
Pulmonary vasoconstriction was measured by estimation of Systolic Pulmonary Artery Pressure in millimeter of mercury (mmHg) by trans-thoracic echocardiography
Time frame: Post-transfusion
Population: In the fresh blood only 9 volunteers estimation of Systolic Pulmonary Artery Pressure was successfully studied by trans-thoracic echocardiography. In the old blood only 11 volunteers and In the old blood + Inhaled Nitric Oxide only 12 volunteers estimation of Systolic Pulmonary Artery Pressure was successfully evaluated.
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| Fresh Blood | Systolic Pulmonary Artery Pressure | 39 mmHg | Standard Error 3 |
| Old Blood | Systolic Pulmonary Artery Pressure | 46 mmHg | Standard Error 3 |
| Old Blood + Inhaled Nitric Oxide | Systolic Pulmonary Artery Pressure | 31 mmHg | Standard Error 2 |
Secondary
Endothelial Function: Reactive Hyperemia Index
Reactive Hyperemia Index (RHI) measures Endothelial function and is assessed by digital pulse amplitude tonometry and it is a sensitive indicator of endothelial function. RHI is a calculated as a ratio between tested versus contralateral finger dilatation, thus there is no unit measure.
Time frame: Post-transfusion
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| Fresh Blood | Endothelial Function: Reactive Hyperemia Index | 0.87 LnRHI | Standard Error 0.06 |
| Old Blood | Endothelial Function: Reactive Hyperemia Index | 0.9 LnRHI | Standard Error 0.06 |
| Old Blood + Inhaled Nitric Oxide | Endothelial Function: Reactive Hyperemia Index | 0.78 LnRHI | Standard Error 0.06 |
Other Pre-specified
Activation of Inflammatory Lipid Mediators
To assess concentration of plasma activation of inflammatory lipid mediators at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion.
Time frame: Before and after blood transfusion
Other Pre-specified
Activation of Platelets
To assess concentration of plasma activation of platelets at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion.
Time frame: Before and after blood transfusion
Other Pre-specified
Changes in Gene Expression
To assess concentration of changes in gene expression at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion.
Time frame: Before and after blood transfusion
Other Pre-specified
Concentration of Cytokines
To assess concentration of plasma cytokines at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion.
Time frame: Before and after blood transfusion
Other Pre-specified
Endothelial Function
To assess Endothelial function by RHI at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion.
Time frame: Before and after blood transfusion
Other Pre-specified
Hemolysis
To assess concentration of plasma Hemoglobin at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion.
Time frame: before and after blood transfusion
Other Pre-specified
Nitric Oxide Metabolites
To assess concentration of plasma Nitric oxide metabolites at baseline, 10 minutes after blood transfusion, 1 hour after blood transfusion, 2 hours after blood transfusion and 4 hours after blood transfusion.
Time frame: Before and after blood transfusion