Clinical Trial Details
— Status: Completed
Administrative data
| NCT number |
NCT04323852 |
| Other study ID # |
13312 |
| Secondary ID |
ISRCTN44896820IR |
| Status |
Completed |
| Phase |
Phase 4
|
| First received |
|
| Last updated |
|
| Start date |
September 1, 2017 |
| Est. completion date |
January 21, 2019 |
Study information
| Verified date |
September 2021 |
| Source |
Shahid Beheshti University of Medical Sciences |
| Contact |
n/a |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Interventional
|
Clinical Trial Summary
Background and study aim:
Heart diseases are among the most common causes of death worldwide. A large proportion of
deaths are caused by heart attacks (myocardial infarction), where blood flow to the heart is
reduced resulting in damage to the heart muscle. If the arteries supplying blood to the heart
start to become blocked, Coronary Artery Bypass Grafting (CABG) surgery is a treatment to
replace the blocked sections of artery can reduce angina (chest pain). However, CABG surgery
has complications, including an increased risk of heart attack. Vitamin D deficiency is
thought to be linked to poorer recovery from heart attack and CABG surgery. This study aims
to investigate if vitamin D supplementation can reduce injury to the heart following CABG
surgery.
Who can participate? Adults with vitamin D deficiency undergoing CABG
What does the study involve? Participants are randomly allocated to one of two groups. Those
in the first group receive vitamin D at 3 doses per day for 3 days before surgery. The second
group will receive a dummy pill (placebo). Both groups will have standard CABG surgery.
What are the possible benefits and risks of participating? Those in the vitamin D group might
benefit from its effects. Vitamin D has few side effects, especially when taken for only a
few days.
Where is the study run from? Shahid Modarres Hospital (Iran)
When is the study starting and how long is it expected to run for? September 2017 to January
2019
Who is funding the study? Deputy of Research of Shahid Beheshti School of Medicine
Who is the main contact? Dr Erfan Tasdighi erfan.tasdighi@gmail.com
Description:
Enrollment started in June 2018 and was completed in December 2018. The inclusion criteria
were as following: the patients referred for elective and isolated Coronary Artery Bypass
Graft (CABG) using Cardiopulmonary Bypass (CPB) with vitamin D deficiency (defined as
25-hydroxyvitamin D [25(OH) D] < 20 ng/mL) and normal kidney function (creatinine <1.5mg/dL).
The exclusion criteria were: recent myocardial infarction, urgent CABG, non-isolated coronary
surgery, redo surgery, malignant disease, presence of acute or chronic inflammatory diseases,
history of vitamin D treatment within previous 6 months, or unwillingness to participate.
Intervention Following informed consent, eligible study participants were randomly assigned
(by using a computer- generated random code) in a 1:1 ratio to receive either placebo or a
total of 450,000 international units (IU) vitamin D3 (three 50,000 IU of vitamin D3 tablet
daily for 3 days) before operation. The placebo group received three inactive medication
tablets daily at the same time point. With the exception of the pharmacists, all the
investigators, patients and the medical team were blinded to the group allocation.
Coronary artery bypass was done in the culprit lesions for both groups by one surgical team.
The standard protocol for general anesthesia, surgical and CPB management were performed for
all patients and have already been described in detail [16].
Outcome measures The primary outcome was the degree of heart apoptosis by measurement of
caspase 2, 3 and 7 activity from right atrial specimen with immunohistochemistry staining,
and the serum level of anti-inflammatory interleukin-10 (IL-10) and insulin- like growth
factor (IGF-1), and N-terminal pro v-type Brain Natriuretic Peptide (nt-pro BNP). The biopsy
from right atrial appendage was taken at the end of surgery after venous cannula removal in a
nontraumatic fashion, kept into formalin and in less than 24h parafinized. Blood samples were
collected at the baseline (T1), before anesthesia induction (T2), at the end of surgery after
protamine reversal (T3) and the first postoperative day (T4) to measure the serum level of
IGF-1, IL-10 and pro BNP. The blood samples were centrifuged at 2500 rpm for 15 min within
one hour after blood sampling, and the serum was stored at -20°C until assayed.
Enzyme-linked immunosorbent assay The concentration of IL-10 was measured by a quantitative
ELISA kit . The concentration of the IGF- 1 was measured by a quantitative ELISA kit .
Serum vitamin D was detected by using the high performance liquid chromatography method . The
pro BNP measurement was done using a commercially available two- site chemiluminescent
immunometric assay .
Immunohistochemistry studies Immunohistochemical staining was performed on 5-micrometer thick
sections. The slides were incubated at 37°C for 24 hours and de-paraffinized in pre-heated
xylene and rehydrated through descending grades of alcohol, washed in distilled water. Heat
induced antigen retrieval was done by microwave oven with citrate buffer (pH 6.0) for
anti-caspase-7 and Ethylenediamine Tetraacetic Acid; buffered solution (Tris-EDTA) (pH=8) for
anti-caspase-2 and 3. endoperoxidase blocking was done by adding hydrogen peroxide on the
sections. The protein block then added for 5 minutes, slides were washed in Tris-Buffered
Saline (TBS). .The primary antibody as anti-caspase-2 antibody, rabbit monoclonal ,
anti-caspase-3 antibody, rabbit monoclonal , anti-caspase-7 antibody, and mouse monoclonal
(clone 7-1-11 , abcam) antibody were added and kept for 30 minutes, washed in TBS. Mouse and
Rabbit Specific horseradish peroxidase/Diaminobenzidine (HRP/DAB) immunohistochemistry (IHC)
Detection Micro-polymer Kit were used and incubated for 15 minutes then washed with
tris-buffered saline (TBS). Diaminobenzidine (DAB) chromogen was added and kept for 5
minutes. Slides washed in distilled water and counter stained with hematoxylin. Sections
containing lymph node tissue were used as positive control. Negative control included primary
antibody replaced with phosphate buffered saline (PBS). Immunostained sections were reviewed
for cytoplasmic expression of anti-caspase-2, anti-caspase-3 and anti-caspase-7. The number
of immune-reactive cells per High Power Field (HPF) (X 400) was counted. For this purpose at
least 10 HPF were assessed and the average of positive cells was recorded.
Sample size and statistical methods The determination of the patient number (30 patients per
group) was based on previous trials investigating the caspase activity in the CABG setting .
Categorical variables were reported as numbers and percentages, whereas mean± standard
deviation was expressed for continuous variables. Repeated measures of analysis of variance
and multiple comparisons using the Bonferroni correction (type I error correction) were
applied for evaluating the change of measured inflammatory markers between the groups over
time. The Kolmogorov-Smirnov test for normality was performed. Continuous variables and
categorical variables were compared between groups using Student's t test (or Mann-Whitney
test for those meeting abnormal distribution) and Chi-square, respectively. All the
statistical analyses were performed using SPSS version 23 (SPSS, Chicago, IL, USA). A p
values <0.05 was considered to be significant.
