Influence of acute glycaemic level on measures of myocardial infarction in non-diabetic pigs.

OBJECTIVE: Patients with diabetes are at increased risk of experiencing myocardial infarction. The influence of the prevailing plasma glucose level on infarction and mortality after acute ischaemia is however unknown. The aim was to study the effect of the acute plasma glucose level on the myocardial infarction size in a closed-chest pig model. DESIGN: 38 non-diabetic pigs were randomised to hypoglycaemic (1.8-2.2 mmol/l; n = 15), normoglycaemic (5-7 mmol/l; n = 12) or hyperglycaemic glucose clamping (22-23 mmol/l; n = 11). After 30 min within glucose target myocardial infarction was induced for 30 min followed by reperfusion for 120 min. Hereafter the heart was double-stained to delineate infarction from viable tissue within the area at risk. RESULTS: Mean infarction size was 201 ± 35 mm(2) (mean ± SEM) in the hypoglycaemic group, 154 ± 40 mm(2) in the normoglycaemic group and 134 ± 40 mm(2) in the hyperglycaemic group, with no differences in infarction size, infarct/area at risk ratio or troponin T levels between the groups. There was no difference in incidence of ventricular fibrillation or mortality between the groups. CONCLUSION: No statistically significant associations were observed between the acute glycaemic level and measures of myocardial infarction, rates of ventricular fibrillation and subsequent premature death in the setting of acute ischaemia and reperfusion.

Development of left ventricular hypertrophy in a novel porcine model of mitral regurgitation.

OBJECTIVES: We aimed to develop a porcine model for chronic nonischemic mitral regurgitation (MR) to investigate left ventricular (LV) enlargement and eccentric hypertrophy. DESIGN: Nonischemic MR was induced in 30 pigs by open-chest immobilization of the posterior mitral leaflet by transannular traction sutures that where applied in transmyocardial fashion. A sham operated control group (n = 13) was included. Echocardiographic LV size and heart weight assessed at euthanasia were used to evaluate the development of LV enlargement and eccentric hypertrophy after 8 weeks follow-up. RESULTS: Eight pigs died and seven were excluded due to mediastinal infection (n = 2) or failure to produce MR (n = 5). Thus, 28 pigs were included and were divided into three groups: controls (n = 12), mild MR (mMR; n = 10), and moderate to severe MR (sMR; n = 6). The change in LV internal diameter in diastole (LVIDd) from baseline to follow-up was significantly higher in the sMR group compared to that of the control group (P = 0.0017). Furthermore, LV weight was significantly increased in the mMR (P = 0.047) and the sMR (P = 0.0087) groups compared to that of the control group. CONCLUSIONS: A new model for chronic moderate to severe nonischemic MR with development of LV enlargement and eccentric hypertrophy within 8 weeks has been established in pigs.

Surgical relocation of the papillary muscles in functional ischemic mitral regurgitation: what are the forces of the relocation stitches acting on the myocardium?

BACKGROUND AND AIM OF THE STUDY: In patients with chronic functional ischemic mitral regurgitation (FIMR), papillary muscle relocation has the potential to induce reverse left ventricular remodeling. However, in order to optimize function and durability, the forces imposed on the left ventricular myocardium by papillary muscle relocation should be assessed. METHODS: Eight pigs with FIMR were subjected to down-sized ring annuloplasty in combination with relocation of the anterior (5 mm) and posterior (15 mm) papillary muscles towards the respective trigone. Papillary muscle relocation was obtained by a 2-0 expanded polytetrafluoroethylene stitch fixed to the trigone, exteriorized through the myocardium overlying the papillary muscle, and fixed to an epicardial disc. Tension in these stitches was measured at a systolic blood pressure > 80 mmHg using a custom-made sliding caliper with a strain gauge mounted in line. This allowed assessment of the cyclic change from minimal diastolic to maximum systolic papillary muscle relocation stitch tension. RESULTS: Maximum cyclic change in the posterior papillary muscle (PPM) stitch tension was 1.1 N at 15 mm relocation. In comparison, the anterior papillary muscle (APM) tension was increased to a maximum of 1.4 N with only 5 mm relocation. Surprisingly, during each step of isolated PPM relocation, the APM stitch tension increased concomitantly, but in contrast APM relocation did not influence the magnitude of PPM stitch tension. There was no statistically significant difference between cyclic changes in APM and PPM stitch tension at any step of relocation. CONCLUSION: Papillary muscle relocation using stitches attached between epicardial discs and respective trigones induced a cyclic change in papillary muscle relocation stitch tension of 1.1-1.4 N. These values were in the range of normal tension in the mitral valve apparatus, and equivalent to only 19-24% of the total papillary muscle forces. Therefore, this technique does not appear to induce a non-physiologically high cyclic load on the mitral valve complex.

Mitral valve mechanics following posterior leaflet patch augmentation.

BACKGROUND AND AIM OF THE STUDY: Attention towards the optimization of mitral valve repair methods is increasing. Patch augmentation is one strategy used to treat functional ischemic mitral regurgitation (FIMR). The study aim was to investigate the force balance changes in specific chordae tendineae emanating from the posterior papillary muscle in a FIMR-simulated valve, following posterior leaflet patch augmentation. METHODS: Mitral valves were obtained from 12 pigs (body weight 80 kg). An in vitro test set-up simulating the left ventricle was used to hold the valves. The left ventricular pressure was regulated with water to simulate different static pressures during valve closure. A standardized oval pericardial patch (17 x 29 mm) was introduced into the posterior leaflet from mid P2 to the end of the P3 scallop. Dedicated miniature transducers were used to record the forces exerted on the chordae tendineae. Data were acquired before and after 12 mm posterior and 5 mm apical posterior papillary muscle displacement to simulate the effect from one of the main contributors of FIMR, before and after patch augmentation. RESULTS: The effect of displacing the posterior papillary muscle induced tethering on the intermediate chordae tendineae to the posterior leaflet, and resulted in a 39.8% force increase (p = 0.014). Posterior leaflet patch augmentation of the FIMR valve induced a 31.1% force decrease (p = 0.007). There was no difference in force between the healthy and the repaired valve simulations (p = 0.773). CONCLUSION: Posterior leaflet patch augmentation significantly reduced the forces exerted on the intermediate chordae tendineae from the posterior papillary muscle following FIMR simulation. As changes in chordal tension lead to a redistribution of the total stress exerted on the valve, patch augmentation may have an adverse long-term influence on mitral valve function and remodeling.

Recellularization of decellularized mitral heart valves in juvenile pigs.

BACKGROUND AND AIM OF THE STUDY: Glutaraldehyde-preserved bioprosthetic heart valves are non-viable and have a limited durability because of calcification, tissue wear, and inflammation, especially in children. Decellularized porcine heart valves, when treated with deoxycholic acid (DOA), have exhibited complete recellularization and an absence of calcification when implanted into the pulmonary position in juvenile sheep. The study aim was to determine the degree of recellularization and calcification in DOA-treated heart valve prostheses in the mitral position in juvenile pigs. METHODS: A mitral heart valve prosthesis was implanted into each of 17 pigs, and subsequently explanted and fixed in formaldehyde after between five and 26 weeks. A gross pathologic assessment, high-resolution X-ray imaging and histological examination were then performed on each valve. RESULTS: Eight pigs survived the observational period. Five valves had only a slight fibrin deposition and calcification foci within the fibrin deposits. Three valves had severe thrombotic material deposits with disseminated calcification and valve stenosis, and one valve had infective endocarditis. A myofibroblast-like cell ingrowth was observed at different locations of the valve housing in all explanted heart valves, but ingrowth in the basal part of the cusp matrix was limited. In four valve prostheses, endothelial cells covered up to 10% of the cusp surface after six months. Inflammatory cells were observed in large numbers in those valves showing endocarditis and severe thrombosis, but in only limited numbers in the other valves. CONCLUSION: All valves showed the deposition of fibrin and platelet material, in three cases to a severe degree. A limited ingrowth of both endothelial and myofibroblast-like cells was observed in five valves in which calcification was limited to a few commissural foci. The non-endothelialized surface of the decellularized valves makes them very susceptible to platelet and fibrin deposition; however, slow revitalization seems possible.

The effect of haemostatic devices on bone healing 6 months postoperatively in sternotomized pigs.

OBJECTIVES: Bone wax is frequently used to diminish bleeding after sternotomy. Water-soluble polymer wax has been shown to diminish postoperative bleeding and, unlike traditional bone wax, to be absorbed and removed by the organism in an unchanged state. We have previously shown that bone wax impairs early bone healing after sternotomy, whereas polymer wax does not. This difference was observed 6 weeks postoperatively and questions arose as to whether these effects were long term. Therefore, we hypothesized that bone wax impairs bone healing in sternotomized pigs 6 months postoperatively, whereas polymer wax does not. METHODS: Fourteen Landrace/Yorkshire pigs were sternotomized and then randomly assigned to haemostasis by either bone wax (WAX-group) or water-soluble polymer wax (POL-group). After 6 months, the pigs were euthanized and the sternum was removed and prepared for further assessment. Bone fracture strength and bone stiffness were determined using a modified three-point bending test, whereas bone healing was examined by means of quantitative histology. Six pigs died before the end of the study due to failure to thrive, valve prosthesis endocarditis and coronary artery occlusion. RESULTS: The mechanical testing showed no difference between groups with regard to fracture strength [WAX-group versus POL-group; 214.8 (85.5-478.5) vs 203.8 (90.4-478.5) N, P = 0.986] or maximum stiffness [213.0 (81.5-409.5) vs 348.5 (23.3-689.5) N/mm, P = 0.128]. Histology showed predominance of fibroblast-covered surfaces [10.6% (1.8-23.3%) vs 4.1% (0.0-13.0%), P < 0.001] and fibrous tissue volume [45.4% (6.9-82.0%) vs 17.4% (2.9-55.0%), P < 0.001] in animals treated with bone wax. The volume fraction of calcified bone tended to be higher in the POL-group [26.8% (4.3-35.8%) vs 16.7% (1.5-35.8%), P = 0.065]. Granulomas comprised 12.5% (0.0-78.9%) of the volume fraction in the WAX-group compared with 0.0% (0.0-0.0%) in the POL-group (P < 0.001). CONCLUSION: Bone wax and water-soluble polymer wax had similar long-term effects on bone mechanical properties. Histology confirmed our hypothesis and showed a more extensive foreign body reaction in animals treated with bone wax than in those treated with water-soluble polymer wax.

Transapical neochord implantation: is tension of artificial chordae tendineae dependent on the insertion site?

OBJECTIVE: Transapical chordae tendineae replacement is a promising new approach for mitral leaflet prolapse. However, animal studies have raised concerns that the tension of the transapically fixated artificial neochordae might be greater than the tension in the neochordae attached to papillary muscle tips, thereby reducing repair durability. METHODS: In eight 80-kg pigs, the primary anterior leaflet chordae were replaced by a 5-0 polytetrafluoroethylene neochord using a miniature in-line force transducer. The neochord was attached first to the anterior papillary muscle and, on a second cardiac bypass, transapically to the left ventricle apex. Occlusion of the inferior vena cava was performed to examine the effect of left ventricle pressure changes on neochord tension to adjust the crude data to 95 mm Hg. The maximum slope of the chordal tension curve was calculated to compare curve patterns. The data are presented as the mean ± standard deviation. RESULTS: The following tension was measured in the neochordae during papillary muscle and transapical fixation, respectively: peak tension (crude, 0.39 ± 0.32 vs 0.50 ± 0.25 N, P = .17; adjusted, 0.41 ± 0.30 vs 0.46 ± 0.27 N, P = .22), mid-systolic tension (crude, 0.19 ± 0.12 vs 0.19 ± 0.15 N, P = .96; adjusted, 0.28 ± 0.16 vs 0.19 ± 0.11 N, P = .12). There was a significantly lower maximum slope (dF/dtmax) of the neochord tension curves after papillary muscle fixation compared with transapical fixation (7.4 ± 6.9 vs 10.3 ± 7.7 N/s, P = .028). CONCLUSIONS: Overall, the chordal insertion site had little influence on the tension in the artificial neochordae compared with the interindividual variation. However, abnormal tension fluctuations in the transapically fixated neochordae might predispose to leaflet tears and early repair failure.

Mitral valve annular downsizing forces: implications for annuloplasty device development.

OBJECTIVE: Mitral valve repair with annulus downsizing is a popular surgical procedure for functional mitral regurgitation. We investigated the effects of externally applied downsizing on the observed in-plane forces and valvular dimensions. METHODS: Five animals were included in an acute porcine study. Three traction sutures were anchored at the right fibrous trigone (T) and suspended across the annulus for externalization at the P1, P2, and P3 annular segments. The annulus was downsized with the sutures in controlled increments while measuring the tension force in the sutures. Downsizing percentages ranged from a 2% to 32% reduction of the T-P distances. Sonomicrometry was used to measure the resulting valvular dimensions. RESULTS: No difference in force was found between the P1, P2, and P3 segments across all levels of downsizing. The peak forces at 32% downsizing were 1.2 ± 0.9 N, 1.5 ± 1.0 N, and 0.8 ± 0.2 N for the T-P1, T-P2, and T-P3 segments, respectively. The maximum total suture forces in the mitral plane during downsizing increased from 0.12 ± 0.03 N to 3.5 ± 1.3 N (P < .005). Sonomicrometry showed a decrease in the systolic thickening of the posterior myocardial wall at the annular level with annular downsizing (0%-32%) from 5 ± 3 mm to 1 ± 1 mm (P < .05). CONCLUSIONS: Segmental mitral valve annulus downsizing increased in-plane traction suture forces and has a significant influence on the in-plane biomechanics. These results have implications for device design in terms of mechanical strength requirements and can be used to supplement boundary conditions for computational left heart models.

Enhancing coronary Wave Intensity Analysis robustness by high order central finite differences.

BACKGROUND: Coronary Wave Intensity Analysis (cWIA) is a technique capable of separating the effects of proximal arterial haemodynamics from cardiac mechanics. Studies have identified WIA-derived indices that are closely correlated with several disease processes and predictive of functional recovery following myocardial infarction. The cWIA clinical application has, however, been limited by technical challenges including a lack of standardization across different studies and the derived indices' sensitivity to the processing parameters. Specifically, a critical step in WIA is the noise removal for evaluation of derivatives of the acquired signals, typically performed by applying a Savitzky-Golay filter, to reduce the high frequency acquisition noise. METHODS: The impact of the filter parameter selection on cWIA output, and on the derived clinical metrics (integral areas and peaks of the major waves), is first analysed. The sensitivity analysis is performed either by using the filter as a differentiator to calculate the signals' time derivative or by applying the filter to smooth the ensemble-averaged waveforms. Furthermore, the power-spectrum of the ensemble-averaged waveforms contains little high-frequency components, which motivated us to propose an alternative approach to compute the time derivatives of the acquired waveforms using a central finite difference scheme. RESULTS AND CONCLUSION: The cWIA output and consequently the derived clinical metrics are significantly affected by the filter parameters, irrespective of its use as a smoothing filter or a differentiator. The proposed approach is parameter-free and, when applied to the 10 in-vivo human datasets and the 50 in-vivo animal datasets, enhances the cWIA robustness by significantly reducing the outcome variability (by 60%).

Significance of force transfer in mitral valve-left ventricular interaction: in vivo assessment.

OBJECTIVE: The objective of this study was to assess the combined force transfer from the papillary muscle tips to the mitral valve through the chordae tendineae in vivo, and thereby quantify the force transmitted through the papillary-chordal complex to augment left ventricular ejection. METHODS: In an acute porcine model (n = 8), force transfer between papillary muscles and the mitral valve was recorded on the anterior and posterior papillary muscle tip using dedicated force transducers. Ultrasound sonomicrometry was utilized to record and calculate left ventricular long-axis shortening and mitral annular geometry. The closing force acting on the mitral valve leaflets was calculated as mitral annular area multiplied by the transmitral pressure difference throughout systole. Mitral valve competence was verified before measurements with color Doppler ultrasound. RESULTS: Peak force in the anterior and posterior papillary muscle was 5.9 ± 0.6 N and 5.8 ± 0.7 N (mean ± standard error of the mean), respectively, and peak closing force was 6.8 ± 0.3 N all at a transmitral pressure of 90 mm Hg. Peak rate of left ventricular contraction coincided with peak papillary muscle force. CONCLUSIONS: This study is the first to assess the magnitude and time course of the longitudinal force transmitted through the papillary-chordal complex to the left ventricular wall during ejection. The study also demonstrates a significant force transfer to the closing force acting on the mitral valve leaflets that constitutes an essential component of valvular-ventricular interaction to enhance left ventricular systolic pump performance. The magnitude of the combined papillary muscle force component emphasizes the crucial role of preserving mitral valve-left ventricular continuity in mitral valve surgery.

Plasma proANP and SDMA and microRNAs are associated with chronic mitral regurgitation in a pig model.

OBJECTIVE: NON-ISCHEMIC MITRAL REGURGITATION (MR) IS PRIMARILY CAUSED BY MYXOMATOUS MITRAL VALVE (MV) DISEASE LEADING TO ADAPTIVE REMODELING, ENLARGEMENT, AND DYSFUNCTION OF THE LEFT VENTRICLE. THE AIM OF THIS STUDY WAS TO EXAMINE THE REGULATION OF PLASMA MARKERS AND SEVERAL CARDIAC KEY GENES IN A NOVEL PORCINE MODEL OF NON-ISCHEMIC MR. METHODS AND RESULTS: Twenty-eight production pigs (Sus scrofa) were randomized to experimental MR or sham surgery controls. MR was induced by external suture(s) through the posterior MV leaflet and quantified using echocardiography. The experimental group was subdivided into mild MR (mMR, MR=20-50%, n=10) and moderate/severe MR (sMR, MR >50%, n=6) and compared with controls (CON, MR ≤10%, n=12). Eight weeks postoperatively, follow-up examinations were performed followed by killing. Circulating concentrations of pro-atrial natriuretic peptide (proANP), l-arginine, asymmetric dimethylarginine, and symmetric dimethylarginine (SDMA) were measured. MV, anterior papillary muscle, and left ventricular free wall tissues were collected to quantify mRNA expression of eNOS (NOS3), iNOS (NOS2), MMP9, MMP14, ANP (NPPA), BNP (NPPB), and TGFB1, 2, and 3 and five microRNAs by quantitative real-time PCR. Pigs with sMR displayed markedly increased plasma proANP and SDMA concentrations compared with both controls and mMR (P<0.05). The expression of all genes examined differed significantly between the three localizations in the heart. miR-21 and miR-133a were differently expressed among the experimental groups (P<0.05). CONCLUSIONS: Plasma proANP and SDMA levels and tissue expression of miR-21 and miR-133a are associated with severity of chronic MR in an experimental porcine model.

Comparative study of lung sealants in a porcine ex vivo model.

BACKGROUND: Lung sealants are often used to prevent alveolar air leaks after lung resection surgery. Some sealants have shown to be effective in clinical studies, but extensive comparative evaluation has not yet been conducted. We aimed to evaluate different sealant burst pressures in an ex vivo model mimicking air leakage after lung resection. METHODS: Fifty-four porcine lungs comprised the study material. Six different sealants were evaluated: Bioglue (V-Tech, Roskilde, Denmark), TachoSil (Nycomed, Roskilde, Denmark), Tisseel (Baxter, Allerød, Denmark), Evicel (OMRIX biopharmaceuticals S.A, Rhode-St-Genèse, Belgium), TissuePatchDural (Vingmed, Roskilde, Denmark), and Pleuraseal (Covidien, Copenhagen, Denmark). After creating a standardized pleural defect, each lung was randomized into 1 of the 6 treatment groups (n= 9). Each lung was ventilated with incremental airway pressure. Air leakage was assessed after each increment. If leakage occurred, the burst pressure was recorded. RESULTS: The Evicel fibrin sealant and Tisseel fibrin sealant exhibited significantly lower burst pressures compared with the Bioglue, TachoSil, and Pleuraseal (p < 0.05). Bioglue had the highest median burst pressure (55 cm H(2)O) followed by TachoSil (35 cm H(2)O), PleuraSeal (35 cm H(2)O), TissuePatchDural (25 cm H(2)O), Evicel (15 cm H(2)O), and Tisseel (15 cm H(2)O). CONCLUSIONS: This model has shown to be feasible in determining and comparing the burst pressures of different lung sealants. Further studies are needed to determine responses in living tissue and burst pressure over time in vivo.

Degenerative processes in bioprosthetic mitral valves in juvenile pigs.

BACKGROUND: Glutaraldehyde-treated bioprosthetic heart valves are commonly used for replacement of diseased heart valves. However, calcification and wear limit their durability, and the development of new and improved bioprosthetic valve designs is needed and must be evaluated in a reliable animal model. We studied glutaraldehyde-treated valves 6 months after implantation to evaluate bioprosthetic valve complications in the mitral position in juvenile pigs. MATERIALS: The study material comprised eight, 5-month old, 60-kg pigs. All pigs received a size 27, glutaraldehyde-treated, stented, Carpentier-Edwards S.A.V. mitral valve prosthesis. After six months, echocardiography was performed, and the valves explanted for gross examination, high resolution X-ray, and histological evaluation. RESULTS: Five pigs survived the follow-up period. Preexplant echocardiography revealed a median peak and mean velocity of 1.61 m/s (range: 1.17-2.00) and 1.20 (SD = ±0.25), respectively, and a median peak and mean pressure difference of 10.42 mmHg (range: 5.83-16.55) and 6.51 mmHg (SD = ±2.57), respectively. Gross examination showed minor thrombotic depositions at two commissures in two valves and at all three commissures in three valves. High resolution X-ray imaging revealed different degrees of calcification in all explanted valves, primarily in the commissural and belly areas. In all valves, histological evaluation demonstrated various degrees of fibrous sheath formation, limited immunological infiltration, and no overgrowth of host endothelium. CONCLUSIONS: Bioprosthetic glutaraldehyde-treated mitral valves can be implanted into the mitral position in pigs and function after 6 months. Echocardiographic data, calcification, and histological examinations were comparable to results obtained in sheep models and human demonstrating the suitability of the porcine model.