PuraStat®; Post Market Performance during Vascular Surgery

  • Research type

    Research Study

  • Full title

    A Multi-center, Single Arm Post-market Clinical Study to Confirm Safety and Performance of PuraStat® Absorbable Haemostatic Material for the Management of Bleeding In Vascular Surgery

  • IRAS ID

    217324

  • Contact name

    Carole Robin

  • Contact email

    crobin@puramatrix.com

  • Sponsor organisation

    3-D Matrix Europe SAS

  • Duration of Study in the UK

    0 years, 10 months, 31 days

  • Research summary

    The study is a multi-centre, post-market clinical study to confirm the safety and performance of PuraStat®, an absorbable haemostatic material used for the management of bleeding during vascular surgery.

    The study will primarily assess "time-to-haemostasis" after the application of PuraStat® in patients undergoing elective carotid endarterectomy. In addition, the study will continue to assess the safety profile of this already CE-marked haemostatic material.

    Up to 65 patients, clinically indicated for elective carotid endarterectomy will be invited to participate in the study.

    It is anticipated that up to 6 hospitals across Europe, including 2 NHS Hospitals in the UK, will participate and that patient recruitment will be completed within 5 months.

    Lay Summary of Results

    Abstract
    Anastomotic bleeding in vascular surgery can be difficult to control. Patients, in particular those undergoing carotid surgery, have
    often been started on treatment with dual antiplatelet agents and receive systemic heparinization intraoperatively. The use of
    local hemostatic agents as an adjunct to conventional methods is widely reported. 3-D Matrix’s absorbable hemostatic material
    RADA16 (PuraStat®), is a fully synthetic resorbable hemostatic agent. The aim of this study is to confirm the safety and performance
    of this agent when used to control intraoperative anastomotic bleeding during carotid endarterectomy (CEA). A prospective,
    single-arm, multicenter study involving 65 patients, undergoing CEA, in whom the hemostatic agent was applied to the
    suture line after removal of arterial clamps. Patients were followed up at 24 h, discharge, and one month after surgery. Time
    to hemostasis was measured as the primary endpoint. Secondary endpoints included hemostasis efficacy and safety outcomes,
    blood loss, intraoperative and postoperative administration of blood products, and incidence of reoperation for bleeding. A
    total of 65 cases (51 male and 14 female) undergoing CEA, utilizing patch reconstruction (90. 8%), eversion technique
    (6.1%), and direct closure (3.1%) were analyzed. All patients received dual antiplatelet therapy preoperatively and were administered
    systemic intravenous heparin intraoperatively, as per local protocol. The mean time to hemostasis was 83 s±105 s (95%
    CI: 55-110 s). Primary hemostatic efficacy was 90.8%. The mean volume of product used was 1.7 mL±1.1 mL. Hemostasis was
    achieved with a single application of the product in 49 patients (75.3%). Two patients required a transfusion of blood products
    intraoperatively. There were no blood product transfusions during the postoperative period. The intraoperative mean blood loss
    was 127 mL ±111.4 mL and postoperatively, the total mean drainage volume was 49.0 mL±51.2 mL. The mean duration of surgery
    was 119±35 min, and the mean clamp time was 35 min 12 s±19 min 59 s. In 90.8% of patients, there was no presence of
    hematoma at 24 h postoperatively. Three returned to theatre due to bleeding (2 in the first 24 h), however, none of these cases
    were considered product related. Overall, there were no device-related serious adverse events (SAE) or unanticipated devicerelated
    SAEs reported. Use of the hemostatic agent PuraStat® is associated with a high rate of hemostatic efficacy (90.8%) and a
    short time to hemostasis. The safety of the product for use on vascular anastomoses has been demonstrated.
    Keywords
    hemostasis, self-assembling peptides, RADA16, vascular surgery, carotid endarterectomy, case series
    Date received: 7 July 2022; revised: 10 November 2022; accepted: 23 November 2022.
    1 St George’s Vascular Institute, St George’s Hospital, London, UK
    2 St George’s University of London, Cranmer Terrace, London, UK
    3 Academic Vascular Surgical Unit, Hull Royal Infirmary, Hull, UK
    4 Department of Vascular Surgery, University Hospitals Leuven, Leuven, Belgium
    5 Vascular Surgery Unit, York Teaching Hospital NHS Foundation Trust, York, North Yorkshire, UK
    6 Department of Vascular Surgery, St Mary’s Hospital, Imperial College London Healthcare NHS Trust, London, UK
    Corresponding Author:
    Katherine M. Stenson, St George’s Vascular Institute, St George’s Hospital, Blackshaw Road, London SW17 0QT, UK.
    Email: kstenson@doctors.org.uk
    Creative Commons Non Commercial CC BY-NC: This article is distributed under the terms of the Creative Commons
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    reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access
    page (https://us.sagepub.com/en-us/nam/open-access-at-sage).
    Original Article
    Clinical and Applied
    Thrombosis/Hemostasis
    Volume 28: 1-10
    © The Author(s) 2022
    Article reuse guidelines:
    sagepub.com/journals-permissions
    DOI: 10.1177/10760296221144307
    journals.sagepub.com/home/cat
    Introduction
    Carotid endarterectomy (CEA) is the definitive treatment for
    long-term stroke prevention in patients who have suffered a
    recent cerebral ischemic event with moderate to severe internal
    carotid artery stenosis.1,2 Patients with symptomatic carotid
    stenosis of 50% to 99%, according to the North American
    Symptomatic Carotid Endarterectomy Trial (NASCET) criteria,
    or more than 70% according to the European Carotid Surgery
    Trial criteria, should be assessed and referred urgently for
    CEA.3–5 CEA is a procedure whereby the atherosclerotic
    plaque is removed from the carotid artery via an arteriotomy.
    Despite it demonstrating significant long-term benefits with
    regard to primary and secondary stroke prevention and mortality
    reduction, complications of the operation can provide significant
    perioperative challenges.1,6
    Perioperative bleeding is common in patients undergoingCEA
    and othermajor vascular reconstructions. It iswell-recognized that
    treatment before surgery with dual antiplatelet therapy reduces the
    risk of perioperative stroke. The patient often also receive intraoperative
    anticoagulation to further reduce thromboembolic risk,
    however, these contribute to a significant risk of anastomotic
    bleeding.7,8 These patients often have friable vessels with extensive
    atherosclerotic disease and calcification, further contributing
    to anastomotic bleeding risk.3,9
    Anastomotic bleeding itself can be unpredictable, persistent,
    and life-threatening. Postoperative hematoma after CEA has the
    potential to cause rapid onset of airway obstruction, respiratory
    failure, and emergency reintervention.10,11 In the NASCET
    study postoperative neck hematomas occurred in 7.1% of
    patients after CEA, which was subsequently identified as a statistically
    significant risk factor for perioperative stroke and death
    (14.9% in patients with wound hematoma, compared with 5.9%
    in patients without hematoma).12 More recently, Baracchini
    et al8 reported neck bleeding in 8.2% of 1458 cases undergoing
    eversion CEA.
    Hemostasis may be challenging in peripheral vascular surgery,
    due to the requirement of direct arterial and arterial graft suturing.
    The risk of anastomotic bleeding in vascular surgery may be
    reduced bymeticulous technique, using fine needles, suture material,
    and surgical loupes. The use of antiplatelet agents and systemic
    anticoagulants in the prevention of thrombosis during
    periods of operative vessel occlusion will increase the risk
    despite good surgical technique. Increasingly, hemostatic agents
    are being used as an adjunct to promote hemostasis.13
    Rapid hemostasis during CEA results in shorter operative
    times, decreased transfusion requirement, improved patient
    recovery time, and decreased wound healing time.13 Adjuvant
    hemostatic methods alongside accurate methods of primary
    hemostasis may aid in achieving these outcomes.
    Multiple hemostatic agents are available to the surgeon, with
    the aim of controlling and reducing anastomotic bleeding but, to
    date, a reliable option that is easy to use and shows good risk
    and cost–benefit potential is not widely available. Those hemostatic
    agents can be divided into 3 categories: hemostats, sealants,
    and adhesives.13 Five classes of these agents have been
    historically available: fibrin sealants, bovine collagen and
    thrombin, cyanoacrylate, albumin cross-linked with glutaraldehyde,
    and polyethylene glycol polymer.9 The self-assembling
    peptide, RADA16, an absorbable hemostatic material, was
    developed for use as a hemostat (PuraStat®, 3-D Matrix
    Europe, Caluire and Cuire, France) for controlling intra- and
    postoperative bleeding in diverse surgical procedures (eg,
    bleeding from vascular anastomosis, small vessel hemorrhage
    of the gastrointestinal tract and oozing from capillaries of the
    parenchyma and surrounding tissues of solid organs). It is
    made from a chain of 3 naturally occurring amino acids; arginine,
    alanine, and aspartic acid bonded together and repeated
    4 times to form a 16-amino acid oligo-peptide chain,
    RADA16, as shown in Figure 1. It is a transparent, amphiphilic
    self-assembling peptide (SAP), available in a prefilled syringe
    as a 2.5% aqueous solution. Contact with the fluid of physiological
    pH, such as blood causes the acidic peptide solution to be
    neutralized and, as a result, the peptide molecule, which has a
    β structure, quickly forms a peptide hydrogel. This hydrogel
    coats and adheres to the point of vessel bleeding, forming a
    mechanical barrier that effectively seals the damaged part of
    the vessel. This facilitates coagulation deep into the gel, thus
    aiding hemostasis. The gel remains in situ after surgery and is
    gradually absorbed. The majority will have been absorbed
    within 30 days, but some may remain in place for longer.14
    RADA16 was commercialized under the name of PuraStat®
    following Masuhara et al15 first demonstrating its safety and efficacy
    as a hemostat in cardiac surgery. Since 2014, both pro-active
    and reactive postmarket surveillance data have been collected to
    monitor and record serious adverse events causally related to
    PuraStat®, of which none have been declared as of July 2022.
    Subsequent clinical trials have been carried out since 2014 to
    assess and demonstrate the safety and efficacy of PuraStat® in
    multiple surgical fields, including cardiology, hepatology, gastrointestinal,
    otolaryngology, and endocrine surgery.16–24
    Aim/Hypotheses
    The objective of this postmarket clinical follow-up study is to
    report safety and performance data from patients undergoing
    elective CEA where the SAP, PuraStat®, was used for intraoperative
    hemostasis.
    The primary endpoint was defined as total time-tohemostasis
    (TTH).
    Secondary endpoints evaluated were blood loss, drainage
    volume, ease of use of PuraStat®, and rate and quantity of
    transfusion of blood products. Safety data were also collected
    including reintervention for bleeding, adverse events, postoperative
    complications, and length of hospital stay.
    Methods
    Study Design
    This was a single-arm, prospective, multicenter study involving
    consecutive patients undergoing CEA in whom RADA16, the
    2 Clinical and Applied Thrombosis/Hemostasis
    hemostatic agent was applied to the suture line after the removal
    of arterial clamps. Ethics approval was obtained from all
    centers. The study was registered at HRA under number 17/
    LO/0082 and at ClinicalTrials.gov under the Identifier:
    NCT03103282.
    Patient Selection
    Patients over the age of 18 years, undergoing elective CEA
    either by direct closure (without the use of patch), patch reconstruction,
    or eversion technique were screened for eligibility.
    Those, undergoing elective CEA, requiring the use of
    RADA16 were eligible for inclusion. Those with a known coagulation
    disorder or hypersensitivity to any components of
    RADA16 were excluded. No formal statistical hypothesis was
    conducted to derive the sample size.
    Patient Enrollment
    Routine standard of care regarding the informed consent process
    for CEA was followed independently of any information or discussion
    relating to this study. Study inclusion was first discussed
    at the preoperative visit. Informed consent forms related to the
    study were provided to each site and were signed by the patients.
    Once deemed eligible, patients were assigned a unique identification
    number for the duration of the study.
    Patient demographics, medical history, and their use of antiplatelet
    or anticoagulant drugs were recorded prospectively,
    alongside standard local site preoperative examinations and
    blood tests (full blood count, pregnancy test, prothrombin
    time, fibrinogen or partial thromboplastin time).
    Procedure
    CEA was undertaken by 7 senior surgeons with previous experience
    with different hemostatic agents. CEA was carried out to
    the standard of care of the local site by either patch reconstruction,
    direct closure, or eversion technique, subject to the lead
    surgeon’s discretion. CEA involves the removal of atherosclerotic
    plaque via an arteriotomy in the internal carotid artery
    (ICA). Direct closure involves closing the arteriotomy with
    direct suturing of the ICA, whereas patch reconstruction
    involves the incorporation of a synthetic, bovine pericardial,
    or venous patch as the method of closure. The eversion technique
    involves the disconnection of the ICA from the carotid
    bulb via an arteriotomy. The ICA is then everted to allow for
    the atherosclerotic plaque to be removed before the ICA is
    re-anastomosed to the carotid bulb.25 All patients were on
    dual antiplatelet therapy preoperatively and were administered
    systemic intravenous heparin intraoperatively, as per local site
    protocol.
    Procedural instructions for PuraStat® use were provided to
    the study sites. PuraStat® was supplied in prefilled syringes,
    of either 1, 3, or 5 mL. Primary hemostasis was achieved
    through techniques at the lead surgeon’s discretion, including
    single pressure, and further monofilament sutures, with or
    Figure 1. Constituents of PuraStat®. In contact with the fluid of physiological pH, the β sheet nanofiber network rapidly forms a hydrogel
    creating a mechanical barrier to aid in hemostasis.14
    Stenson et al 3
    without pledgets, for bleeding in between existing sutures.
    Cautery was not used on the suture line itself. RADA16 was
    applied if oozing bleeding from suture lines was present after
    the clamps on the vessel were removed. As much blood as possible
    was removed from the bleeding site following initial
    hemostasis before the application of RADA16. The syringe
    nozzle was held as close to the tissue as possible to allow for
    adequate application in a volume sufficient to fully cover the
    bleeding site. The gel was left undisturbed until complete hemostasis
    occurred. In some cases, more than one application or
    syringe was required to achieve this and there was no
    maximum number of applications defined.
    The primary endpoint was defined as total time-tohemostasis
    (TTH). TTH was measured using a stopwatch
    from the initial application of RADA16 to the bleeding site,
    after vessel clamp release, until all visible bleeding had
    ceased, and complete hemostasis was judged as achieved by
    the operator.
    In the case of multiple applications of RADA16, TTH was
    defined as the time from the very first application. TTH is a
    well-accepted outcome measure for hemostasis and has been
    used in previous postmarket studies evaluating the safety and
    efficacy of PuraStat®.17,18 This should allow adequate comparability
    of the results across the different surgical fields.
    Secondary endpoints evaluated were blood loss, drainage
    volume, ease of use of PuraStat®, and rate and quantity of
    transfusion of blood products. Safety data were also collected
    including reintervention for bleeding, adverse events, postoperative
    complications, and length of hospital stay.
    Surgeon-Assessed Outcomes
    Surgeons were asked to categorize the degree of bleeding following
    anastomosis as being “mild,” “moderate,” or “severe.”
    They were asked to comment on the quality of the vessel
    being treated as “poor,” “moderate,” or “good.” At the completion
    of the procedure, investigators were asked to determine
    hemostasis as “complete” or as “showing presence of bleeding.”
    The ease of use of the PuraStat® application system was
    assessed by the operating surgeons as “excellent,” “good,”
    “fair,” or “poor.”
    Follow-up Protocol
    Patients were followed up at 24 h, at discharge, and at one
    month postoperatively. One-month follow-up was completed
    in person or over the phone. Primary endpoint data were collected
    intraoperatively, whereas secondary endpoints and
    safety data were collected throughout the study follow-up
    period.
    Statistical Analysis
    Statistical analysis was undertaken using SAS System®, Version
    9.4. No replacement of missing data has been performed.
    Descriptive statistics were utilized with continuous variables
    summarized as mean (with 95% confidence intervals [CIs]), standard
    deviation, median, quartiles, and range. The number of
    missing observations has also been summarized. Categorical variables
    were summarized as percentages by categories.
    All statistical analysis was performed on the intention-totreat
    (ITT) population of the study, defined as all the patients
    with signed informed consent forms that were treated with at
    least one application of RADA16.
    Results
    Patient Recruitment and Enrollment
    Eighty-nine patients were enrolled in the study between June
    2017 and July 2019 (72 patients in 4 hospitals in the United
    Kingdom and 17 patients in one hospital in Belgium).
    Twenty-four (27%) of this cohort were excluded from the
    ITT population as RADA16 was not used either because
    patients were ineligible (9), or because there were no active
    bleeding sites after primary hemostasis (9) for other reasons
    not reported by the site (3), for unknown reasons (2), 1
    patient who was eligible but did not have RADA16 applied
    during the procedure. Recruitment and enrolment data are summarized
    in Figure 2.
    In the remaining 65 cases, RAD16 was used during elective
    CEA, and these formed the ITT population. One patient completed
    the study with a major deviation, whereby the site did
    not use a stopwatch to measure the TTH resulting in a perprotocol
    population (PP) of 64 participants. 24-h follow-up
    and discharge data were complete for 100% of patients. 93.8%
    (61) completed the 1-month follow-up visit, of which 83.6%
    (51) were completed within the predefined study window.
    Patient Demographics
    Sixty-five patients underwent CEA with intraoperative use of
    RADA16 and were included in the study. Of these, 51
    (78.5%) of participants were male and 14 (21.5%) females,
    with a mean age of 71.8±8.9 years (range 48-87 years).
    Most of the participants, 60 (92.3%) had a normal physical
    examination at the preoperative consultation, with a mean
    body mass index of 26.3±5.3 kg/m2. 44 (68.8%) had a
    history of smoking and 12 (18.5%) were diabetic. A total of
    47 out of 63 (74.6%, data were missing for 2 patients), were
    deemed at high risk, having an American Society of
    Anesthesiologists (ASA) grade of 3 or more.26 Mean preoperative
    systolic blood pressure was 135.7 ±19.9 mm Hg and diastolic
    blood pressure was 71.5±13.8 mm Hg. Full patient
    characteristics are presented in Table 1.
    Procedure
    Fifty-nine (90.8%) CEAs were performed using patch reconstruction,
    of which most cases, 37 (56.9%), used a bovine pericardial
    patch. The eversion technique was used in 4 (6.1%)
    cases and direct closure (without a patch) was performed in 2
    4 Clinical and Applied Thrombosis/Hemostasis
    (3.1%) of cases. The mean duration of the procedure was 118±
    35 min (range: 61-255 min). The mean carotid artery clamp
    time was 35 min 12 s±19 min 59 s (range: 25 s-68 min),
    with data reported in 59 cases.
    After clamp removal, in 50 cases (76.9%) “rescue” sutures
    were inserted to achieve primary hemostasis before the application
    of RADA16. The quality of vessels at the suture or the
    anastomotic site was reported as “good” in 51 cases (79.7%)
    and “moderate” in 13 cases (20.3%). Data were missing for
    one patient. No patients had suture site vessels deemed as
    “poor” quality.
    The mean total number of applications of PuraStat® was 1.3
    ±0.6, with the mean total amount of product used recorded as
    1.7±1.1 mL.
    Figure 2. Patient recruitment and enrolment flowchart.
    Abbreviations: ITT, Intent To Treat; TTH, Time To Hemostasis; PP, Per Protocol.
    Stenson et al 5
    Bleeding and RADA16 Efficacy
    The degree of bleeding prior to the application of RADA16 was
    reported by the operating surgeon as “mild” in 59 cases (90.8%)
    and “moderate” in 6 (9.2%), with no “severe” bleeding,
    recorded. This classification was subjective and observerdependent.
    RADA16 appeared to be effective in the majority
    of cases.
    In 59 cases (90.8%), investigators describe hemostasis as
    “complete” including 3 cases where it had been obtained by
    combining RADA16 with additional hemostatic action. In 6
    cases (9.2%), the investigator reported ongoing “presence of
    bleeding,” of these, 4 had decreased bleeding compared with
    preapplication levels described by the investigator, and 2
    showed no change in bleeding levels. Further action was
    required to achieve hemostasis using additional rescue sutures
    in 3 cases, compressions in 2, and the use of a fibrillar, surgical
    absorbable hemostat in 1 case. Of the 59 cases that achieved
    hemostasis the number of applications of the product was as
    follows: 49 requiring 1 application, 7 requiring 2 applications,
    and 3 required 3 applications of the product. Overall, 6 cases
    (9.2%) did not achieve hemostasis using RADA16. Full data
    on the bleeding condition after the RADA16 application are
    shown in Table 2.
    Primary Endpoint
    The mean TTH reported was in seconds 83±105 (range:
    4-653 s), with a median time of 52 s. TTH was predictably
    greater in those with moderate bleeding compared with mild,
    307 ±266 s and 66±63 s, respectively, and in those requiring
    additional applications of RAD16. In 49 cases, 1 application
    of RADA16 was used, with a mean TTH of 50±35 s. The
    mean TTH in patients receiving 2 applications was 180±
    79 s, and in those receiving 3 applications was 399±256 s.
    Primary endpoint results for the ITT population are presented
    in Table 3.
    Secondary Endpoints
    Secondary performance data included intraoperative blood loss,
    drainage volume at 24 h postprocedure and at discharge, rate,
    and quantity of transfusion of blood products received (intraoperatively
    at 24 h postoperatively and overall), and ease of use
    of PuraStat® during CEA.
    Table 1. Demographics of the Patient Cohort Included in the
    Statistical Analysis.
    Characteristic
    ITT
    N=65
    (n/N) %, (missing)
    Smoking (44/64) 68.8%, (1)
    Diabetes (12/65) 18.5%, (0)
    Type I (1/12) 8.3%, (0)
    Type II (11/12) 91.7%, (0)
    Diabetes treatment (0)
    Oral treatment (8/12) 66.7%, (0)
    Insulin-dependent (4/12) 33.3%, (0)
    Renal failure (2/65) 3.1%, (0)
    Family History (of CVD) (19/65) 29.2%, (0)
    ASA score (2)
    1 (2/63) 3.2%
    2 (14/63) 22.2%
    3 (42/63) 66.7%
    4 (5/63) 7.9%
    5 (0) 0%
    Other vascular condition (24/65) 36.9%, (0)
    Other medical history (24/65) 36.9%, (0)
    Abbreviations: ITT, Intent To Treat; CVD, Cardiovascular Disease; ASA score,
    American Society of Anesthesiology score.
    Table 3. Total Time-to-Hemostasis, Severity of Bleeding Pre-
    Application, and Number of Applications (ITT Population).
    Study primary endpoint
    ITT population N=65 N Missing
    N (%) or
    mean ±SD
    [95% CI] Range
    Total TTH (sec) 59 0 83±105
    [55-110]
    4-653
    Number of patients per
    range of TTH
    59 0
    [ 4 (sec), 29 (Q25%)] 14 (23.7%)
    [ 29 (Q25%), 53 (Q50%)] 16 (27.1%)
    [ 53 (Q50%), 931
    (Q75%)]
    15 (25.4%)
    [ 91 (Q75%), 653 (Max)] 14 (23.7%)
    TTH per severity of bleeding
    (sec)
    Mild 55 0 66±63 5-401
    Moderate 4 0 307±266 4-63
    TTH per number of
    applications of PuraStat®
    (sec)
    1 49 0 50±35 4-66
    2 7 0 180 ±79 68-286
    3 3 0 399±256 142-653
    Abbreviations: TTH: Time To Hemostasis; sec: seconds.
    Table 2. Condition After Application of PuraStat®.
    Status postapplication (s)
    ITT patients
    n (%)
    N=65
    Missing 0
    Complete hemostasis 59 (90.8%)
    Persistent bleeding 6 (9.2%)
    Severity of persistent bleeding
    compared to before application
    N=6
    Identical 2 (33.3%)
    Decreased 4 (66.7%)
    Worse 0 (0.0%)
    Abbreviation: ITT, Intent To Treat.
    6 Clinical and Applied Thrombosis/Hemostasis
    The mean intraoperative blood loss was 127 ±111.4 mL
    (95% CI: 99.2-154.8; range: 0-600 mL), with data missing for
    one case. The case recording 600 mL of blood loss required
    no specific further action and no adverse event was reported.
    At 24 h, 60 patients (92.3%) had a surgical drain in situ. Five
    patients did not have a drain. The mean drainage volume was
    49.0 ±51.2 mL (range: 0-230 mL) for 52 patients and drainage
    volume data was missing in 13 patients. Two patients (3.1%)
    received blood products intraoperatively, with a mean volume
    of 580 ±113.1 mL received. Neither of these was related to
    bleeding after the RADA16 application. No further blood products
    were received throughout the study period.
    Full data for secondary performance endpoints are presented
    in Table 4.
    Safety Outcomes
    In 3 cases (4.6% [95% CI: 1.0%-12.9%] of the ITT population)
    there was postoperative bleeding requiring a return to the theatre;
    2 within 24 h and one within 15 days. These events were
    resolved without sequelae and evaluated by the surgeons as
    related to the vascular procedure, with no causal relationship to
    RADA16. Adverse events (AEs) were described based on
    System Organ Class (SOC) in the Medical Dictionary for
    Regulatory Activities (MedDRA).27 After 1 month of follow-up,
    24 AEs had been reported in 20 patients. Of these, there were 11
    procedural complications, 6 nervous system disorders (including
    3 strokes), 2 cardiac disorders, and one each of the following: ear
    and labyrinthine, gastrointestinal, general disorders, infection,
    and vascular disorders. Eleven AEs were reported at the time
    of the procedure, 5 within 24-h postprocedure, 3 within 15
    days, and 5 at up to 1 month postprocedure. Seventeen patients
    (26.2%) had AEs related to their CEA, of these, 5 (7.7%) were
    deemed serious adverse events (SAEs). The hematoma was specifically
    reported in 6 cases (9.2%) at 24-h assessment, with one
    of these requiring surgical revisions at 24 h. Two further hematomas
    were recorded within 1 month of CEA but did not
    require additional treatment.
    Overall, 7 AEs in 7 different patients (10.8%) were deemed
    SAEs by the investigators; of these, 3 were classed as stroke (1
    hemorrhagic, 1 intraoperative, and 1 postoperative). The
    remaining 4 SAEs were bradycardia, a hematoma, pneumonia,
    and 1 death. The overall incidence of stroke was 4.6%. No
    SAEs were considered to be related to RADA16.
    Two AEs were considered device-related. Of these, one was
    reported at 24-h postoperatively as a small hematoma that
    required no further action to resolve. The second was reported
    as a failure to achieve hemostasis after 3 applications of
    RADA16, requiring the additional hemostatic effort of compression
    of the carotid vessel with a surgical swab. Neither
    were classed as serious, and no device or product deficiencies
    were reported during the study.
    Table 4. Secondary Performance Endpoints: Blood Loss During Surgery, Drain Insertion and Volume Drained, and Transfusion Products
    Received.
    Secondary endpoints
    ITT population
    N=65 N Missing
    (n/N)% or
    mean±SD [95% CI] Range
    Blood loss assessed during the surgery (mL) 64 1 127±111.4 [99.2-154.8] 0.0-600.0
    Drainage volume (mL) until drain removal
    At 24-h postprocedure 52 13 49 ±51.2 [34.7-63.2] 0.0-230.0
    At discharge 1 64 60
    Rate of transfusion of blood products
    At surgery 65 0
    No 63 (96.9%)
    Yes 2 (3.1%) [0.4%-10.7%]
    At 24-h postoperation 65 0
    No 65 (100%)
    Yes 0 (0.00%)
    At discharge 65 0
    No 65 (100%)
    Yes 0 (0.00%)
    Overall 65 0
    No 63 (96.9%)
    Yes 2 (3.1%) [0.4%-10.7%]
    Quantity of blood products and/or substitutes
    At surgery 2 0 580 ±113.1 500-660
    At 24-h postoperation 0 0
    At discharge 0 0
    Overall 2 0 580 ±113.1 500-660
    Overall ease of use was deemed “good” or “excellent” in 93.8% of the patients (60.0% “excellent”). The ease of preparation, the application system, and the
    application of the gel were graded as “excellent” or “good” in 100%. The most valuable properties of PuraStat®, as recorded by surgeons, were its transparency and
    its overall ease of use.
    Stenson et al 7
    The mean length of hospital stay for the study group was 4.3
    ±11 days (range: 0-85 days). The overall mortality at 1-month
    follow-up post-CEA was 1.5% [95% CI: 0.0%-8.3%].
    Discussion
    CEA is associated with an increased risk of bleeding intra- and
    postoperatively.1,12 This risk can be mitigated using the meticulous
    surgical techniques. In this study, once the anastomosis
    was complete, primary hemostasis was undertaken using
    methods at the lead surgeon’s discretion to ensure the integrity
    of the suture line. Such methods would include simple pressure
    or further monofilament sutures, with or without pledgets, for
    bleeding in between existing sutures.
    Techniques such as additional “rescue” sutures or electrocoagulation
    may be utilized to achieve primary hemostasis.
    Other methods that show hemostatic benefits include intraoperative
    neck flexion as well as simple postoperative direct neck
    pressure.11,28 Despite these options, the risk of bleeding is
    still relatively high, reported as 8.2% by Baracchini et al8 for
    eversion CEA. Increasingly, vascular surgeons are looking at
    hemostatic agents to aid in reducing this risk.13 The mode of
    action of PuraStat® is different from comparable products.
    Self-assembling peptides in contact with the tissues rapidly
    form a hydrogel, which acts as a barrier and blocks the flow
    of blood from the wound.14 PuraStat® has been used successfully
    as a hemostat in cardiovascular, gastrointestinal, otolaryngology,
    and endocrine surgery.
    This study has demonstrated RADA16 (PuraStat®) to be an
    effective hemostatic agent in vascular surgery, successfully
    controlling suture site and anastomotic bleeding in 90.8% of
    cases. This has been achieved without prolonging the length
    of the procedure, while gaining favorable assessment from the
    surgeons in terms of the ease of use, with 93.8% of surgeons
    assessing its overall ease of use as “good” or “excellent.”
    Each of the surgeons operated on approximately the same
    number of patients and therefore there is no bias in this evaluation,
    which despite everything remains obviously subjective.
    Mean TTH, reported in 59 patients, was recorded as 83±
    1058 s and the primary hemostatic efficacy rate was 90.8%.
    This predictably increased with the number of applications of
    PuraStat® required and the amount of bleeding present prior
    to application. Given the very small numbers that did not use
    the patch technique in this study (6% eversion and 3% direct
    closure), it would be difficult to glean any significant differences
    in the incidence of bleeding when comparing the different
    techniques. There is little in the literature to adequately compare
    the incidence of bleeding complications when different techniques
    are used. This is largely related to the fact that bleeding
    is relatively uncommon and so the numbers available to
    compare are too small to draw meaningful conclusions from.
    Gisbert et al29 did not find any difference in bleeding when
    comparing patch closure with eversion in a series of 455
    patients.
    The results are comparable to a study published in 2019 by
    Morshuis et al,17 demonstrating the performance and safety of
    PuraStat® in left ventricular assist device therapy support,
    where complete hemostasis was achieved in 93.1% of surgical
    sites with a mean TTH of 19.38 ±13.01 s. The results also
    concur with the 2018 study by Giritharan et al16 that assessed
    the safety and efficacy of PuraStat® in a range of cardiac surgical
    procedures, reporting that PuraStat® was solely sufficient in
    achieving complete hemostasis in 84% of cases. When compared
    to other hemostatic agents available in cardiovascular
    surgery, TTH achieved with PuraStat® appears favorable.
    Nasso et al30 reported on a gelatin-thrombin-based hemostatic
    agent in cardiac surgery with a mean TTH of 3.8±2.4 min
    and successful hemostasis achieved in 91.8% of patients. Few
    studies on the use of hemostatic agents in CEA exist. Pellenc
    et al31 report preliminary results for a poly(glycerol-sebacate)
    acrylate-based sealant for use in vascular reconstruction,
    achieving complete hemostasis in 84% of cases. It is noteworthy
    that the comparability of previously published data using
    TTH as an outcome measure may be challenging since
    varying definitions exist. Its calculation is also subjective to
    the operator and study populations may differ significantly.
    Stangenberg et al32 reported 1.7% of patients undergoing
    CEA between 2012 and 2013 who received a transfusion of
    blood products before discharge, while 2 patients in our study
    were transfused during surgery and none before discharge
    (3.1%). This should be taken in the context of the study populations,
    with most patients receiving PuraStat® having an ASA
    grade of 3 or more.
    Adverse events were reported in 30.8% of cases in the
    1-month follow-up period, of which only 2 (3.1%) were
    related to the device and neither were serious. The rate of reoperation
    for bleeding was reported as 4.6% (3 cases), comparable
    with results published by Weinrich et al33 who reported a reoperation
    rate of 3.5% after CEA in 565 patients. The mortality
    rate during the study period was 1.5% (1 patient) and was unrelated
    to the device. Kashyap et al34 reported a mortality rate of
    0.7% after CEA in 371 patients.
    Currently available hemostatic agents have the potential to
    enable transmission of viral and prion diseases from human
    blood component-based agents, and systemic inflammatory
    response syndrome to animal-derived agents.35,36 The purely
    synthetic nature of PuraStat® negates these risks. As summarized
    by Sankar et al,14 the completely synthetic nature and
    the transparent aspect of PuraStat® provide unique advantages.
    The transparency of PuraStat®, reported as its best feature in
    70.8% of cases, allows unique visualization of the bleeding
    site after application, meaning potential revisions of primary
    hemostasis can take place intraoperatively instead of requiring
    subsequent re-exploration.14 The application method as a gel
    in a syringe with a nozzle may also enhance precision in a congested
    operating field. In addition, PuraStat® is ready to use in a
    prefilled syringe that is stored at refrigerator temperature and is
    thus immediately ready for use.
    Vyas and Saha13 describe the ideal characteristics of a hemostatic
    agent as being one that could combine biodegradability,
    rapid action of hemostasis, and minimal side effects while preventing
    thrombosis. Lumsden and Heyman9 described their
    8 Clinical and Applied Thrombosis/Hemostasis
    perceived ideal characteristics of a hemostat as “easily applied
    in a controlled fashion, highly predictable in creating hemostasis,
    and nontoxic and must not have an adverse effect on anastomotic
    patency.” This study on CEA has been able to
    demonstrate the safety and rapid efficacy of PuraStat®, alongside
    its inherent ability to be naturally absorbed by the body and
    the favorable opinion of surgeons in terms of its application and
    ease of use.
    Limitations of this study include patients not being stratified
    according to operative technique and general risk factors, but
    the study centers and the cohort of patients were homogenous.
    The proportion of patients with an ASA grade greater than 3
    was significantly high and could well have impacted on the
    results reported. Any further analysis and comparison should
    take this into account. A follow-up study with risk scores stratified
    to evaluate their effects on PuraStat® efficacy and safety
    may be useful.
    TTH, although an accepted outcome measure for hemostasis,
    can be defined in a variety of ways and is a possible
    cofounder. The reporting of “complete hemostasis” is also subjective
    to the observer operating the stopwatch, but evaluation
    of complete hemostasis is common to all surgeons and TTH
    has been widely used to assess the hemostasis efficacy, both
    in the preclinical and clinical setting. However, the development
    of a well-structured definition for TTH would be beneficial.
    A systematic review of hemostasis in vascular surgery to
    identify key outcomes and outcome measures on this topic,
    along with subsequent development of a core outcome set
    could help to provide unified direction in this area of research
    and improve the comparability of studies.
    Conclusion
    This multicentre, postmarket clinical follow-up study was able
    to confirm the safety and efficacy of 3-D Matrix’s PuraStat®
    self-assembling peptide (RADA16) in the management of
    bleeding during elective CEA and opens a promising new perspective
    to manage suture bleeding in carotid endarterectomy.
    PuraStat® provides a safe and effective option to aid hemostasis
    in surgical situations where standard means of hemostasis are
    insufficient or impractical. The efficacy is demonstrated
    through the results achieved with a mean TTH of 83 s and a
    complete hemostasis postapplication in 90.8% of cases. No
    SAEs related to the device, or its application confirm its
    safety, alongside low volumes of intraoperative blood loss
    and the subsequent need for blood transfusion. However, the
    efficacy of long-term hemostasis and reduction of relevant
    hematoma need further evaluation in a larger cohort with a prospective
    two-armed study design.
    Acknowledgments
    All of the authors would like to thank Dr Maurice Bagot d’Arc of
    BluePharm for his invaluable help in preparing the manuscript for
    publication.
    Declaration of Conflicting Interests
    The authors declared no potential conflicts of interest with respect to
    the research, authorship, and/or publication of this article.
    Ethical Approval
    The study was conducted in accordance with the ethical principles that
    have their origins in the Declaration of Helsinki and the requirements
    for medical device investigations as presented in EN/ISO 14155:2011,
    Clinical investigation of medical devices for human subjects—Good
    clinical practice; Annex X of the European Medical Devices
    Directive 93/42/EEC, as amended by Directive 2007/47/EEC,
    MEDDEV 2.7/4 and applicable local regulatory requirements. Ethics
    approval was obtained from all centers. The registration number at
    HRA 17/LO/0082 was granted in March 2017and the study was registered
    under the ClinicalTrials.gov Identifier: NCT03103282.
    Funding
    The authors received no financial support for the research, authorship,
    and/or publication of this article.
    ORCID iD
    Katherine M. Stenson https://orcid.org/0000-0002-1795-4578
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    10 Clinical and Applied Thrombosis/Hemostasis

  • REC name

    London - Surrey Borders Research Ethics Committee

  • REC reference

    17/LO/0082

  • Date of REC Opinion

    14 Mar 2017

  • REC opinion

    Further Information Favourable Opinion