Extended Duration Simulation and Testing of Cellular and Decellularised Heart Valve Roots

Heart valve disease can affect people of all ages, and can be treated by either valve repair or valve replacement surgery. Currently available replacement heart valves, including mechanical prostheses, bioprostheses, autografts and allografts improve patient survival and quality of life, but have limitations. Key limitations include the risk of immunological reaction and the lack of growth potential and regeneration, which is of particular importance in young patients. To address these limitations, low concentration sodium dodecyl sulphate (SDS) decellularised human aortic, human pulmonary, porcine aortic and porcine pulmonary heart valve roots have been developed. Decellularisation of allografts would potentially reduce the risk of immunological reaction, and the development of a decellularised porcine pulmonary heart valve root would potentially provide an option for right ventricular outflow reconstruction in younger patients who have undergone the Ross Procedure. Before moving to clinical trials, the functional performance of decellularised heart valve roots needs to be pre-clinically assessed appropriately to determine mechanical safety. Whilst there are recommended test methods in place for the in vitro functional performance assessment of newly manufactured and modified surgical replacement heart valves, they need to be optimised or replaced with novel methods suitable for decellularised heart valve roots, due to their time dependent viscoelastic properties. The main aim of this research was to optimise in vitro hydrodynamic and biomechanical performance test methods and develop a novel real time fatigue test method for biological heart valve roots. The secondary aim was to apply the developed in vitro test methods to cellular and decellularised (human and porcine) heart valve roots to evaluate the effect of decellularisation, prior to the decellularised heart valve roots being implanted in patients for clinical trials. In collaboration with NHS Blood and Transplant, Tissue and Eye Services, in vitro biomechanical and hydrodynamic performance of decellularised human aortic and pulmonary heart valve roots was evaluated for the first time in this research. This research determined that the hydrodynamic and functional biomechanical performance of human aortic and pulmonary heart valve roots was not affected by decellularisation treatment. Decellularisation, however, significantly altered some of the directional material properties of pulmonary and aortic heart valve root leaflets. To support clinical translation of decellularised porcine pulmonary heart valve roots, material properties of pulmonary heart valve roots was evaluated following 12 months implantation in sheep. In addition, the effect of the processing steps of cryopreservation and decellularisation on the material properties of porcine pulmonary heart valve roots was investigated. Cryopreservation was shown not to alter the material properties of cellular porcine pulmonary heart valve roots, however, decellularisation did have an effect on the material properties of the porcine pulmonary heart valve root wall. Following 12 months implantation in sheep, the decellularised porcine pulmonary heart valve root wall and leaflets showed a trend for decreasing stiffness and strength; becoming more like the cellular ovine, potentially indicating constructive remodelling. A novel method was developed to investigate the real time fatigue of biological heart valve roots, which was then applied to porcine cellular aortic heart valve roots and porcine decellularised aortic heart valve roots at 120 bpm under physiological cyclic pressures for a maximum of 1.2 million cycles. The results showed no fatigue difference between the cellular and decellularised heart valve roots. Overall, a portfolio of in vitro pre-clinical test methods were developed, optimised and applied to assess the hydrodynamic, biomechanical and fatigue performance of biological heart valve roots including decellularised human and porcine heart valve roots. The in vitro pre-clinical test methods developed in this study will lead to the refinement of in vivo large animal studies and revision of international standards; and the data will help in the development of the next generation of replacement biological heart valve roots, such as decellularised heart valve roots

Placental polyp after normal vaginal delivery: a rare diagnostic dilemma

Placental polyp is retained placental tissue within the endometrial cavity, which forms a nidus for inflammation and bleeding. Placental polyp is a rare entity with an incidence of less than 0.25% of all pregnancies as reported. Here, we report a case of 23-year-old P2L2 woman with complaints of intermittent vaginal bleeding since her recent normal vaginal delivery, 1.5 months back. A polypoid mass (51×41 mm) with abundant vascularity was detected as retained products of placenta (RPOC) within the endometrial cavity by imaging studies. A combination of polypoidal mass within the endometrial cavity with normal beta human chorionic gonadotropin (hCG) of <2.0 mIU/ml raising the suspicion of retained products of placenta or trophoblastic neoplasms. After yielding an unsatisfactory biopsy containing only fibrin deposition, total hysterectomy was performed due to profuse bleeding during biopsy. The uterus specimen showed slight globular enlargement with presence of a red-coloured polypoid mass within the endometrial cavity with rough outer surface and fragile consistency. The histological specimen of the protruding lesion, from the exaggerated placental implantation site, showed intermediate trophoblastic cells infiltrated into the myometrium, which might lead to the diagnosis of placental polyp. However, since placental polyp and uterine arteriovenous malformation have similar clinical characteristics, it is important to accurately identify and differentiate between them to ensure optimal treatment therapy. Definite diagnosis is ultimately made by histopathological examination. We report here a case that is suggestive of either a placental polyp or uterine arteriovenous malformation and will discuss the differential diagnoses and treatments for both diseases, based on a literature review

Should the appendix always be removed during surgery for mucinous ovarian tumors?

Background: Appendectomy is performed in all mucinous ovarian tumors (MOT) identified intraoperatively to ensure microscopic metastases from appendix are not missed. Several recent studies suggested that appendectomy should only be performed in cases with a grossly abnormal appendix or with evidence of pseudomyxoma peritonei. Our study aimed to determine the frequency of malignancy in a grossly normal appendix in women undergoing surgery for borderline or malignant MOT.Methods: In a single institution retrospective study, women undergoing surgery for MOT from January 1, 2008 to June 30, 2016 were included. Women with benign MOT, those with a history of either prior appendicectomy or prior gastrointestinal (GI) malignancy were excluded.Results: Of 266 women identified with MOT, 153 with borderline and malignant MOT were included in the study after application of inclusion criteria. The study population comprised of 29 (18.95%) borderline and 124 (81.05%) malignant MOT. Among the borderline MOT, 13/29 had undergone appendectomy. Five (38.46%) had grossly abnormal appendices of whom 1 had mucinous cystadenoma, 3 had borderline mucinous tumor and 1 had mucinous cystadenocarcinoma of the appendix. Histology was normal in all 8 (61.54%) grossly normal appendices. Among the malignant MOT, 80/124 (64.52%) underwent appendicectomy. Nineteen (23.46%) had grossly abnormal appendices and histology was suggestive of adenocarcinoma of appendix. Histology was normal in all 62 (76.54%) macroscopically normal appendices.Conclusions: Present results suggest that appendectomy be performed only for those appendices that are grossly abnormal or associated with pseudomyxoma peritonei at surgery for MO

Implantation of decellularized porcine pulmonary heart valves in sheep - dataset

Data on the in vivo performance of acellular porcine pulmonary heart valve roots in the right ventricular outflow tract of juvenile sheep (sheep weights, ultrasonography analysis). Data on the explanted acellular porcine roots at 1 month, 3 months and 12 months plus data on non-implanted ovine pulmonary roots and implanted ovine pulmonary roots at 12 months including: gross analysis, biomechanical properties, histological analysis, histomorphometric analysis of cellular population using a range of antibody markers, quantitative calcium analysis

Resuscitation with pre-hospital blood products in adults with trauma-related haemorrhagic shock:the RePHILL RCT

Background: The treatment of traumatic haemorrhagic shock has been transformed through better haemorrhage control, use of tranexamic acid and use of blood products. The improved survival seen from these strategies has stimulated an interest in pre-hospital transfusion.Objectives: To determine if the clinical effectiveness of resuscitation with red blood cells and lyophilised plasma was superior to 0.9% saline for improving tissue perfusion and reducing mortality in adults with haemorrhagic shock following major trauma.Design: A multi-centre, allocation concealed, open-label, parallel group, randomised controlled trial (with internal pilot).Setting: The trial was conducted in four civilian pre-hospital critical care services who operated within the National Health Service (NHS) England Major Trauma Networks.Participants: Adults (aged ≥16 years) who had sustained traumatic injuries, were attended by a pre-hospital emergency medical team and were hypotensive (systolic blood pressure &lt;90 mmHg or absence of radial pulse) as a consequence of traumatic haemorrhage were eligible for inclusion. The exclusion criteria were known or apparently &lt;16 years, blood administered on scene prior to the arrival of the RePHILL team, traumatic cardiac arrest where (1) the arrest occurred prior to arrival of the team and/or (2) the primary cause is not hypovolaemia, refusal of blood product administration, known Jehovah’s Witness, pregnancy, isolated head injury without evidence of external haemorrhage, prisoners in the custody of HM Prison and Probation Service.Interventions: Participants were randomised to receive up to either two units each of red blood cells and lyophilised plasma or up to 1 L 0.9% saline. Treatment was administered through the intravenous or intraosseous route.Main outcome measures: The primary outcome was a composite of episode mortality and/or impaired lactate clearance. The secondary outcomes included the individual components of the primary outcome.Results: From 6 December 2016 to 2 January 2021, pre-hospital medical teams randomised 432 participants to red blood cell/lyophilised plasma (n = 209) or 0.9% saline (n = 223) out of a target sample size of 490. Most participants were white (62%), males (82%), median age 38 (interquartile range 26 to 58), involved in a road traffic collision (62%) with severe injuries (median injury severity score 36, interquartile range 25 to 50). Prior to randomisation participants had received on average 430 ml crystalloid fluids and tranexamic acid (90%). The primary outcome occurred in 128/199 (64.3%) of participants randomised to red blood cell/lyophilised plasma and 136/210 (64.8%) randomised to 0.9% saline [adjusted risk difference –0.025% (95% confidence interval –9.0% to 9.0%), p = 0.996]. The event rates for the individual components of the primary outcome, episode mortality and lactate clearance were not statistically different between groups [adjusted average differences −3% (−12% to 7%); p = 0.57 and −5% (−14% to 5%), p = 0.33, respectively].Limitations: Recruitment stopped prematurely due to disruption caused by the COVID-19 pandemic.Future work: Identify the characteristics of patients who may benefit from pre-hospital blood products and whether alternative transfusion regimens are superior to standard care.Conclusions: The trial did not demonstrate that pre-hospital red blood cell/lyophilised plasma resuscitation was superior to 0.9% saline for trauma-related haemorrhagic shock.Trial registration: This trial is registered as ISRCTN62326938.Funding: This award was funded by the National Institute for Health and Care Research (NIHR) Efficacy and Mechanism Evaluation Programme (NIHR award ref: 14/152/14) and is published in full in Efficacy and Mechanism Evaluation; Vol. 11, No. 2. See the NIHR Funding and Awards website for further award information.<br/

Effect of priming interval on reactogenicity, peak immunological response, and waning after homologous and heterologous COVID-19 vaccine schedules: exploratory analyses of Com-COV, a randomised control trial

BackgroundPriming COVID-19 vaccine schedules have been deployed at variable intervals globally, which might influence immune persistence and the relative importance of third-dose booster programmes. Here, we report exploratory analyses from the Com-COV trial, assessing the effect of 4-week versus 12-week priming intervals on reactogenicity and the persistence of immune response up to 6 months after homologous and heterologous priming schedules using the vaccines BNT162b2 (tozinameran, Pfizer/BioNTech) and ChAdOx1 nCoV-19 (AstraZeneca).MethodsCom-COV was a participant-masked, randomised immunogenicity trial. For these exploratory analyses, we used the trial's general cohort, in which adults aged 50 years or older were randomly assigned to four homologous and four heterologous vaccine schedules using BNT162b2 and ChAdOx1 nCoV-19 with 4-week or 12-week priming intervals (eight groups in total). Immunogenicity analyses were done on the intention-to-treat (ITT) population, comprising participants with no evidence of SARS-CoV-2 infection at baseline or for the trial duration, to assess the effect of priming interval on humoral and cellular immune response 28 days and 6 months post-second dose, in addition to the effects on reactogenicity and safety. The Com-COV trial is registered with the ISRCTN registry, 69254139 (EudraCT 2020–005085–33).FindingsBetween Feb 11 and 26, 2021, 730 participants were randomly assigned in the general cohort, with 77–89 per group in the ITT analysis. At 28 days and 6 months post-second dose, the geometric mean concentration of anti-SARS-CoV-2 spike IgG was significantly higher in the 12-week interval groups than in the 4-week groups for homologous schedules. In heterologous schedule groups, we observed a significant difference between intervals only for the BNT162b2–ChAdOx1 nCoV-19 group at 28 days. Pseudotyped virus neutralisation titres were significantly higher in all 12-week interval groups versus 4-week groups, 28 days post-second dose, with geometric mean ratios of 1·4 (95% CI 1·1–1·8) for homologous BNT162b2, 1·5 (1·2–1·9) for ChAdOx1 nCoV-19–BNT162b2, 1·6 (1·3–2·1) for BNT162b2–ChAdOx1 nCoV-19, and 2·4 (1·7–3·2) for homologous ChAdOx1 nCoV-19. At 6 months post-second dose, anti-spike IgG geometric mean concentrations fell to 0·17–0·24 of the 28-day post-second dose value across all eight study groups, with only homologous BNT162b2 showing a slightly slower decay for the 12-week versus 4-week interval in the adjusted analysis. The rank order of schedules by humoral response was unaffected by interval, with homologous BNT162b2 remaining the most immunogenic by antibody response. T-cell responses were reduced in all 12-week priming intervals compared with their 4-week counterparts. 12-week schedules for homologous BNT162b2 and ChAdOx1 nCoV-19–BNT162b2 were up to 80% less reactogenic than 4-week schedules.InterpretationThese data support flexibility in priming interval in all studied COVID-19 vaccine schedules. Longer priming intervals might result in lower reactogenicity in schedules with BNT162b2 as a second dose and higher humoral immunogenicity in homologous schedules, but overall lower T-cell responses across all schedules. Future vaccines using these novel platforms might benefit from schedules with long intervals

Safety and immunogenicity of heterologous versus homologous prime-boost schedules with an adenoviral vectored and mRNA COVID-19 vaccine (Com-COV): a single-blind, randomised, non-inferiority trial

Background: Use of heterologous prime-boost COVID-19 vaccine schedules could facilitate mass COVID-19 immunisation. However, we have previously reported that heterologous schedules incorporating an adenoviral vectored vaccine (ChAdOx1 nCoV-19, AstraZeneca; hereafter referred to as ChAd) and an mRNA vaccine (BNT162b2, Pfizer–BioNTech; hereafter referred to as BNT) at a 4-week interval are more reactogenic than homologous schedules. Here, we report the safety and immunogenicity of heterologous schedules with the ChAd and BNT vaccines. Methods: Com-COV is a participant-blinded, randomised, non-inferiority trial evaluating vaccine safety, reactogenicity, and immunogenicity. Adults aged 50 years and older with no or well controlled comorbidities and no previous SARS-CoV-2 infection by laboratory confirmation were eligible and were recruited at eight sites across the UK. The majority of eligible participants were enrolled into the general cohort (28-day or 84-day prime-boost intervals), who were randomly assigned (1:1:1:1:1:1:1:1) to receive ChAd/ChAd, ChAd/BNT, BNT/BNT, or BNT/ChAd, administered at either 28-day or 84-day prime-boost intervals. A small subset of eligible participants (n=100) were enrolled into an immunology cohort, who had additional blood tests to evaluate immune responses; these participants were randomly assigned (1:1:1:1) to the four schedules (28-day interval only). Participants were masked to the vaccine received but not to the prime-boost interval. The primary endpoint was the geometric mean ratio (GMR) of serum SARS-CoV-2 anti-spike IgG concentration (measured by ELISA) at 28 days after boost, when comparing ChAd/BNT with ChAd/ChAd, and BNT/ChAd with BNT/BNT. The heterologous schedules were considered non-inferior to the approved homologous schedules if the lower limit of the one-sided 97·5% CI of the GMR of these comparisons was greater than 0·63. The primary analysis was done in the per-protocol population, who were seronegative at baseline. Safety analyses were done among participants receiving at least one dose of a study vaccine. The trial is registered with ISRCTN, 69254139. Findings: Between Feb 11 and Feb 26, 2021, 830 participants were enrolled and randomised, including 463 participants with a 28-day prime-boost interval, for whom results are reported here. The mean age of participants was 57·8 years (SD 4·7), with 212 (46%) female participants and 117 (25%) from ethnic minorities. At day 28 post boost, the geometric mean concentration of SARS-CoV-2 anti-spike IgG in ChAd/BNT recipients (12 906 ELU/mL) was non-inferior to that in ChAd/ChAd recipients (1392 ELU/mL), with a GMR of 9·2 (one-sided 97·5% CI 7·5 to ∞). In participants primed with BNT, we did not show non-inferiority of the heterologous schedule (BNT/ChAd, 7133 ELU/mL) against the homologous schedule (BNT/BNT, 14 080 ELU/mL), with a GMR of 0·51 (one-sided 97·5% CI 0·43 to ∞). Four serious adverse events occurred across all groups, none of which were considered to be related to immunisation. Interpretation: Despite the BNT/ChAd regimen not meeting non-inferiority criteria, the SARS-CoV-2 anti-spike IgG concentrations of both heterologous schedules were higher than that of a licensed vaccine schedule (ChAd/ChAd) with proven efficacy against COVID-19 disease and hospitalisation. Along with the higher immunogenicity of ChAd/BNT compared with ChAD/ChAd, these data support flexibility in the use of heterologous prime-boost vaccination using ChAd and BNT COVID-19 vaccines. Funding: UK Vaccine Task Force and National Institute for Health Research