Expression of specific matrix proteins in atherosclerotic plaques in patients who underwent carotid artery surgery
| ISRCTN | ISRCTN46536832 |
|---|---|
| DOI | https://doi.org/10.1186/ISRCTN46536832 |
- Submission date
- 23/05/2021
- Registration date
- 25/05/2021
- Last edited
- 16/09/2022
- Recruitment status
- No longer recruiting
- Overall study status
- Completed
- Condition category
- Circulatory System
Plain English summary of protocol
Background and study aims
A stroke is a serious life-threatening medical condition that happens when the blood supply to part of the brain is cut off. Atherosclerosis is a disease in which plaque builds up inside your arteries. Plaque is made up of fat, cholesterol, calcium, and other substances found in the blood. Over time, plaque hardens and narrows your arteries. This limits the flow of oxygen-rich blood to your organs and other parts of your body.
Atherosclerosis is responsible for one-third of all stroke. The rate of death from the first stroke is about 35%, and from the next up to 65%, while as many as 15 - 30% of survivors are permanently incapable of work.
Atherosclerotic plaque formation is a complex process that may be partially mediated by the enzymes matrix metalloproteinase-9 (MMP-9) and cyclooxygenase-2 (COX-2).
Components of a carotid artery (main blood vessel to the head) atherosclerotic plaque can be analyzed by noninvasive preoperative radiological imaging (magnetic resonance imaging, computed tomography and ultrasound), while the expression of matrix metalloproteinase 9 and cyclooxygenase-2, proteins with a role in remodelling of atherosclerotic plaques, can be analyzed immunohistochemically.
The aim of this study is to analyze the characteristics of carotid plaques in symptomatic and asymptomatic patients.
Who can participate?
Participants with an indication for carotid endarterectomy (CEA, a surgical procedure to remove the atheromatous plaque material) of the internal carotid artery will be included in the study.
What does the study involve?
Patients undergoing carotid endarterectomy will be informed of the study and consented to participate.
Preoperatively, participants will undergo a routine clinical examination. Routine blood tests and highly sensitive C-reactive protein will be performed preoperatively.
Participants will undergo preoperative ultrasound-colour Doppler, MRI or MSCT scan of carotid arteries.
Carotid atherosclerotic plaque will be excised during CEA and preserved for analysis. Pathohistological characteristics and expression of MMP-9 and COX-2 proteins in carotid atherosclerotic plaque after carotid endarterectomy will be analysed.
What are the possible benefits and risks of participating?
All study participants could benefit from more close monitoring than would be possible outside of the study.
Defining the association of preoperative radiological characteristics of carotid atherosclerotic plaque with pathohistological features and biological indicators of carotid atherosclerotic plaque is important in clinical practice due to the possible better identification of patients in whom CEA is indicated with the potential predictability of preoperative diagnostic methods.
There are no potential adverse effects
Where is the study run from?
University Hospital Merkur, Zagreb (Croatia)
When is the study starting and how long is it expected to run for?
July 2013 to July 2021
Who is funding the study?
INEL – medicinska tehnika d.o.o., Zagreb (Croatia)
Who is the main contact?
Mr. Davorin Sef
davorin.sef@gmail.com, davorin.sef@nhs.net
Contact information
Scientific
University Hospital Merkur
Zajceva 19
Zagreb
10000
Croatia
 ORCID ID |
0000-0001-5053-3815 |
|---|---|
| Phone | +385 994888552 |
| davorin.sef@nhs.net |
Study information
| Study design | Multicenter single-blinded observational cohort study |
|---|---|
| Primary study design | Observational |
| Secondary study design | Cohort study |
| Study setting(s) | Hospital |
| Study type | Diagnostic |
| Participant information sheet | ISRCTN46536832_PIS.pdf |
| Scientific title | The expression of MMP-9 and COX-2 proteins in the analysis of carotid atherosclerotic plaques in patients with carotid endarterectomy |
| Study objectives | Radiological features of carotid plaque instability analyzed by preoperative noninvasive radiological diagnostics (ultrasound/colour Doppler, MRI and/or MSCT scan) are significantly associated with the expression of MMP-9 and COX-2 proteins in atherosclerotic plaque and are highly prevalent in patients with unstable plaque. |
| Ethics approval(s) | 1. Approved 18/07/2013, University Hospital Merkur Ethics Committee (Zajceva ulica 19, Zagreb, 10000, Croatia; +385 (0)1 2431390; no email provided), ref: none 2. Approved 16/11/2015, Clinic for Cardiovascular Diseases - Magdalena Ethics Committee (Ljudevita Gaja 2, Krapinske Toplice, 49217, Croatia; +385 (0)49 244444; no email provided), ref: none 3. Approved 09/10/2020, University Hospital Centre Rijeka Ethics Committee (Kresimirova 42, Rijeka, 51000, Croatia; +385 (0)51 658808; no email provided), ref: 2170-29-02/1-20-2 |
| Health condition(s) or problem(s) studied | Carotid artery stenosis |
| Intervention | Participants with an indication for carotid endarterectomy (CEA) of the internal carotid artery will be included in the study. The indication for surgery will be set according to the recommendations of multicenter randomized studies (NASCET, ECST, ACAS, ACST and ESVS). Both symptomatic and asymptomatic patients will be enrolled. The identity of participants will be protected by the identification number of the protocol of radiological diagnostic examination and biopsy of carotid atherosclerotic plaque after carotid endarterectomy. Preoperatively, participants will undergo a clinical examination. Routine blood tests and highly sensitive C-reactive protein will be performed preoperatively. Participants will undergo preoperative ultrasound-colour Doppler, MRI or MSCT scan of carotid arteries. The stenosis of carotid artery will be determined according to NASCET criteria, and then the plaque cross-section at the site of greatest narrowing will be analyzed. Carotid atherosclerotic plaque will be qualified as a stable or unstable plaque. The characteristics of carotid atherosclerotic plaque will be determined: degree of carotid artery stenosis, expression and size of lipid nucleus, bleeding, calcification, neovascularization, surface ulceration and plaque thrombosis according to the most recent modified AHA classification. The images will be independently analyzed by two researchers. Carotid atherosclerotic plaque will be excised during CEA respecting the rules of atraumatic technique with minimal sample manipulation. The distal end of the plaque sample will be marked and processed by a standard procedure to obtain pathohistological specimen that includes tissue fixation in 10% buffered formalin, embedding in paraffin blocks, cutting to a thickness of 4 μm, dewaxing and standard hemalaun-eosin staining, staining by a Masson method to detect collagen which will be semiquantitatively analyzed. Immunohistochemical analysis of MMP-9, COX-2 expression will be performed. Primary antibodies will be used in the study: MMP-9 (Leica Mikrosysteme Vertieb GMBH, UK, monoclonal, clone 15W2, 1:50 dilution), COX-2 (Abcam, USA, monoclonal, clone SP21, pre-diluted for use), CD68 PG-M1 (DakoCytomation, Denmark, monoclonal, clone PG-M1, pre-diluted for use), SMA (DakoCytomation, Denmark, monoclonal, clone 1A4, pre-diluted for use), CD34 (DakoCytomation, Denmark, monoclonal, clone QBend 10, pre-diluted for use). |
| Intervention type | Other |
| Primary outcome measure | Measured at a single time point: 1. The characteristics and type of carotid atherosclerotic plaque measured by non-invasive radiological diagnostics (ultrasound/colour Doppler, MRI and/or MSCT scan) before surgery 2. Pathohistological characteristics and type of carotid atherosclerotic plaque measured using patohistological analysis after surgery 3. Expression of MMP-9 and COX-2 proteins in carotid atherosclerotic plaque after carotid endarterectomy measured using immunohistochemistry analysis after surgery |
| Secondary outcome measures | Measured at a single time point: 1. Clinical and laboratory characteristics of patients measured by reviewing patient notes before and after surgery 2. Semiquantitative analysis of macrophages, smooth muscle cells and microvascular density in carotid atherosclerotic plaque and the histological type of bleeding within the plaque measured using patohistological analysis after surgery |
| Overall study start date | 15/07/2013 |
| Completion date | 15/07/2021 |
Eligibility
| Participant type(s) | Patient |
|---|---|
| Age group | Adult |
| Sex | Both |
| Target number of participants | Planned sample size: 30 |
| Total final enrolment | 15 |
| Key inclusion criteria | Participants with an indication for carotid endarterectomy (CEA) of the internal carotid artery |
| Key exclusion criteria | Patients with a contraindication for MRI scan (presence of a ferromagnetic foreign body, claustrophobia) |
| Date of first enrolment | 26/09/2013 |
| Date of final enrolment | 30/06/2021 |
Locations
Countries of recruitment
- Croatia
Study participating centres
Zagreb
10000
Croatia
Krapinske Toplice
49217
Croatia
Rijeka
51000
Croatia
Sponsor information
University/education
Zajceva 19
Zagreb
10000
Croatia
| Phone | +385 (0)1 2431390 |
|---|---|
| vinko.vidjak@gmail.com | |
| Website | http://www.kb-merkur.hr/ |
"ROR" |
https://ror.org/01b6d9h22 |
Funders
Funder type
Industry
No information available
Results and Publications
| Intention to publish date | 01/07/2022 |
|---|---|
| Individual participant data (IPD) Intention to share | Yes |
| IPD sharing plan summary | Available on request |
| Publication and dissemination plan | Planned publication in a high-impact peer-reviewed journal. |
| IPD sharing plan | The datasets generated during and/or analysed during the current study are/will be available upon request from (Dr Davorin Sef, davorin.sef@gmail.com; type of data - database in Microsoft Excel, data will become available following the publication of the study and for 12 months afterwards, by reasonable request data will be shared with medical professionals or researchers, for purpose of descriptive and/or statistical analyses, and by formal request to corresponding author, consent from participants was obtained using only anonymous data, ethical or legal restrictions will be provided by the journal in which the results will be published) |
Study outputs
| Output type | Details | Date created | Date added | Peer reviewed? | Patient-facing? |
|---|---|---|---|---|---|
| Participant information sheet | 01/06/2021 | No | Yes | ||
| Results article | 22/12/2021 | 21/03/2022 | Yes | No | |
| Results article | Immunohistochemical analysis of MMP-9 and COX-2 expression in carotid atherosclerotic plaques | 05/09/2022 | 09/09/2022 | Yes | No |
Additional files
- ISRCTN46536832_PIS.pdf
- Uploaded 01/06/2021
Editorial Notes
09/09/2022: Publication reference added.
21/03/2022: The following changes have been made:
1. Publication reference added.
2. The final enrolment number has been added from the reference.
09/08/2021: The study design was changed from "Observational cohort study" to "Multicenter single-blinded observational cohort study".
01/06/2021: The participant information sheet has been uploaded as an additional file.
25/05/2021: Trial's existence confirmed by University Hospital Centre Rijeka Ethics Committee.
