Evaluation of pliable bioresorbable, elastomeric aortic valve prostheses in sheep during 12 months post implantation.

Pliable microfibrous, bioresorbable elastomeric heart valve prostheses are investigated in search of sustainable heart valve replacement. These cell-free implants recruit cells and trigger tissue formation on the valves in situ. Our aim is to investigate the behaviour of these heart valve prostheses when exposed to the high-pressure circulation. We conducted a 12-month follow-up study in sheep to evaluate the in vivo functionality and neo-tissue formation of these valves in the aortic position. All valves remained free from endocarditis, thrombotic complications and macroscopic calcifications. Cell colonisation in the leaflets was mainly restricted to the hinge area, while resorption of synthetic fibers was limited. Most valves were pliable and structurally intact (10/15), however, other valves (5/15) showed cusp thickening, retraction or holes in the leaflets. Further research is needed to assess whether in-situ heart valve tissue engineering in the aortic position is possible or whether non-resorbable synthetic pliable prostheses are preferred.

Strategies to Improve Survival from Surgery for Heart Valve Implantation in Sheep.

Sheep are a commonly used and validated model for cardiovascular research and, more specifically, for heart valve research. Implanting a heart valve on the arrested heart in sheep is complex and is often complicated by difficulties in restarting the heart, causing significant on-table mortality. Therefore, optimal cardioprotective management during heart valve implantation in sheep is essential. However, little is known about successful cardioprotective management techniques in sheep. This article reports our experience in the cardioprotective management of 20 female sheep that underwent surgical aortic valve replacement with a stented tissue-engineered heart valve prosthesis. During this series of experiments, we modified our cardioprotection protocol to improve survival. We emphasize the importance of total body hypothermia and external cooling of the heart. Furthermore, we recommend repeated cardioplegia administration at 20 min intervals during surgery, with the final dosage of cardioplegia given immediately before the de-clamping of the aorta. To reduce the number of defibrillator shocks during a state of ventricular fibrillation (VF), we have learned to restart the heart by reclamping the aorta, administering cardioplegia until cardiac arrest, and de-clamping the aorta thereafter. Despite these encouraging results, more research is needed to finalize a protocol for this procedure.

Failure of decellularized porcine small intestinal submucosa as a heart valved conduit.

OBJECTIVE: Decellularized extracellular matrix made from porcine small intestinal submucosa, commercially available as CorMatrix (CorMatrix Cardiovascular, Inc, Roswell, Ga) is used off-label to reconstruct heart valves. Recently, surgeons experienced failures and words of caution were raised. The aim of this study was to evaluate decellularized porcine small intestinal submucosa as right-sided heart valved conduit in a xenogeneic animal model. METHODS: A pulmonary valve replacement was performed with custom-made valved conduits in 10 lambs and 10 sheep (1 month [3 lambs and 3 sheep], 3 months [3 lambs and 3 sheep], 6 months [4 lambs and 4 sheep]). Valve function was assessed after implantation and before the animal was put to death. Explanted conduits were inspected macroscopically and analyzed using immunohistochemistry and scanning electron microscopy. They also underwent mechanical testing and testing for biochemical composition. RESULTS: All valved conduits were successfully implanted. Five sheep and 2 lambs died due to congestive heart failure within 2 months after surgery. In the animals that died, the valve leaflets were thickened with signs of inflammation (endocarditis in 4). Five sheep and 8 lambs (1 month: 6 out of 6 animals, 3 months: 4 out of 6 animals, 6 months: 3 out of 8 animals) survived planned follow-up. At the time they were put to death, 5 lambs had significant pulmonary stenosis and 1 sheep showed severe regurgitation. A well-functioning valve was seen in 4 sheep and 3 lambs for up to 3 months. These leaflets showed limited signs of remodeling. CONCLUSIONS: Fifty percent of sheep and 20% of lambs died due to valve failure before the planned follow-up period was complete. A well-functioning valve was seen in 35% of animals, albeit with limited signs of tissue remodeling at ≤3 months after implantation. Further analysis is needed to understand the disturbing dichotomous outcome before clinical application can be advised.

Added Value of Interactive 3-D Stereo Vision Echocardiography in the Heart Valve Team: A Post Hoc Analysis for Optimal Decision Making in Patients With Mitral Valve Regurgitation.

OBJECTIVE: We assessed the added value of advanced echocardiography post hoc analysis for optimal decision-making in the Heart Valve Team (HVT) using an interactive, dynamic, live visualization system with true three-dimensional (3-D) stereo vision. METHODS: HVT scrutinized the incremental value of 3 consecutive methods of presentation of full-volume echocardiographic data sets in terms of diagnosis and possibility of repair in 11 selected patients having mitral regurgitation (MR)(Table 1). The questionnaire investigated consecutively (a) standard two-dimensional (2-D) transesophageal echocardiography, (b) single-beat 3-D zoom of the surgical view of the mitral valve, and (c) advanced 3-D volumetric rendering technology (Personal Space Station, Vesalius 3D software, PS-Medtech, Netherlands). RESULTS: In 4 of 11 reviews (36%), single-beat 3-D zoom had additional value over 2-D echocardiography in terms of mechanism/adjustments or adjustment of confirmation of diagnosis. Single-beat 3-D zoom had no additional value over 2-D echocardiography in terms of proposal/probability of repair. In 7 out of 11 (64%) reviews, true stereo 3-D visualization had additional value in terms of mechanism of pathology compared to 2-D and 3-D zoom and in 5 out of 11 (45%) reviews in confirmation of diagnosis. In 3 out of 11 (27%) reviews, true stereo 3-D visualization had additional value in terms of proposal of repair and in 4 of 11 (36%) in probability of repair over 2-D and 3-D zoom. CONCLUSIONS: Advanced easy-to-use true 3-D echocardiography limited differences in interpretation and strengthened the confidence in understanding the mechanisms and suitability for repair of mitral valve regurgitation, typically in more complex valve pathology.

In Situ Remodeling Overrules Bioinspired Scaffold Architecture of Supramolecular Elastomeric Tissue-Engineered Heart Valves.

In situ tissue engineering that uses resorbable synthetic heart valve scaffolds is an affordable and practical approach for heart valve replacement; therefore, it is attractive for clinical use. This study showed no consistent collagen organization in the predefined direction of electrospun scaffolds made from a resorbable supramolecular elastomer with random or circumferentially aligned fibers, after 12 months of implantation in sheep. These unexpected findings and the observed intervalvular variability highlight the need for a mechanistic understanding of the long-term in situ remodeling processes in large animal models to improve predictability of outcome toward robust and safe clinical application.

The early days of vascular and heart valve prostheses: a historical review.

This surgical heritage article provides a historical overview of the most important early advances of vascular- and valvular surgery, that lead to the development of currently used vascular- and valvular prostheses and materials. The first writings describing techniques in vascular surgery mainly focussed on hemorrhage control and date from around 1600 B.C. The strategy of vessel ligation was first mentioned in Western literature around 200 B.C. In the 18th century, techniques of ligation were expanded towards attempts of vessel restoration. The first artificial vascular prosthesis was made in 1894. From this time on, vascular prostheses were used in animal experiments and around 1900 for the first time in humans. More than 60 years later, in 1952, the first mechanical heart valve prosthesis was implanted. Four years later, the first successful biological heart valve implantation followed. In 2000, a transcatheter heart valve was successfully implanted in a human for the first time. Over time, procedures and techniques became more efficient and effective. This led to new developments, such as the manufacturing of a tissue engineered blood vessel in 1986. Nowadays, dozens of different valve prostheses have been devised, both mechanical and biological. Still, no ideal model of vascular and heart valve prosthesis exists.

Flexible mechanoprosthesis made from woven ultra-high-molecular-weight polyethylene fibres: proof of concept in a chronic sheep model.

OBJECTIVES: Ultra-high-molecular-weight polyethylene (UHMWPE) fibres are flexible, have high tensile strength, and platelet and bacterial adhesion is low. Therefore, UHMWPE may overcome limitations of current mechanical valves and bioprostheses. In this study, the biocompatibility and functionality of prototype handmade stented valves from woven UHMWPE (U-valve) was assessed in a chronic sheep model with acetylsalicylic acid monotherapy. METHODS: Native pulmonary valves of 23 sheep were replaced by U-valves (n = 18) or Perimount bovine bioprostheses (reference group, n = 5). Sheep received 80 mg of acetylsalicylic acid daily. Follow-up was conducted at 1 week (n = 4), 1 month (n = 5), 3 months (n = 5) and 6 months (n = 4) in the U-valve group and at 3 months (n = 2) and 6 months (n = 3) in the reference group. Epicardial echocardiography and histology were used to assess valve function and tissue deposition, respectively. RESULTS: Seventeen U-valve sheep (94%) and 3 reference sheep (60%) survived the perioperative period. One reference valve sheep was sacrificed after 4 months because of congestive heart failure. At explantation, all U-valves were intact without leaflet tearing. Up to 3 months, U-valves were flexible and free of stenosis. Regurgitation was mostly mild though gradually increasing; histology showed minimal connective tissue near the leaflet base and sparse calcification. At 6 months, connective tissue was diffusely observed on the leaflets with retraction and consecutive regurgitation and leaflet thickening. CONCLUSIONS: Valves made from UHMWPE fibres demonstrated early feasibility in the pulmonary valve position with reasonably good haemodynamics and intact valve materials up to 6 months. Gradual leaflet thickening and retraction were observed after 3 months due to connective tissue overgrowth.

The "Spacemaker", a New Device for Minimally Invasive Cardiothoracic Surgery: An Evaluation and Feasibility Study.

OBJECTIVE: Our aim was to evaluate a new inflatable lung retractor, the "Spacemaker", and its efficacy in facilitating minimally invasive cardiothoracic surgery without the need of one lung ventilation or carbon dioxide overpressure insufflation. METHODS: The device was tested in 12 anesthetized pigs (90-100 kg) placed on standard endotracheal ventilation. The device was introduced into the right or left side of the chest, depending on the intended procedure to be performed, via a 3-cm incision in the fifth intercostal space. A total of seven animals were used to evaluate hemodynamic and respiratory response to the device, whereas another five animals were used to assess the feasibility of a variety of minimally invasive cardiothoracic surgical procedures. RESULTS: Introduction was easy and unhindered. The device was inflated up to 0.6 bar, thereby pushing the lung tissue gently away cranially, posteriorly, and caudally without interfering with pulmonary function or resulting in respiratory compromise. In addition, hemodynamics remained stable throughout the experiments. Different closed-chest surgical procedures such as left atrial appendage exclusion, pulmonary vein exposure, pacemaker lead placement, and endoscopic stabilization for coronary surgery, were successfully performed. Removal was quick and complete in all cases, and lung tissue showed no remnant atelectasis. CONCLUSIONS: The "Spacemaker" may represent a reliable alternative to current conventional techniques to facilitate minimally invasive cardiothoracic surgery. Further research is warranted to confirm the effectiveness and the safety of this device and to optimize the model before its use in humans and its introduction into clinical practice.

In Vitro Hemocompatibility Testing of Dyneema Purity Fibers in Blood Contact.

OBJECTIVE: Heart valve and vascular prosthesis implantation is a common procedure for patients with heart valve stenosis or regurgitation and dilated or obstructive vascular disease. Drawbacks of conventional valve prostheses are the requirement for anticoagulant drugs, moderate durability, and suboptimal resistance to fatigue and tear. Dyneema Purity fibers are made from ultra-high-molecular-weight polyethylene filaments and are very thin, flexible, and fatigue and abrasion resistant and have high strength. Therefore, prostheses made from Dyneema Purity fibers might be attractive for use in the minimally invasive treatment of valvular- and vascular diseases. The aim of this study was to test the hemocompatibility of Dyneema Purity fibers in contact with blood. METHODS: Real-time platelet adhesion in human blood of 3 volunteers was quantified after 5 minutes of perfusion on single filaments (Ø 15 µm) of Dyneema Purity and polyester fibers. Plasma thrombin generation was measured by fluoroscopy for patches of Dyneema Purity fibers and for 5 commonly used polyester and expanded polytetrafluoroethylene cardiovascular prostheses. RESULTS: Platelet adhesion per 1 mm was 6 ± 1.4 on Dyneema Purity filaments and 15 ± 3.4 on polyester filaments (P = 0.02). Total formed thrombin and the time to peak of its maximum were noninferior for patches of Dyneema Purity fibers compared with the reference materials. CONCLUSIONS: Dyneema Purity fibers are noninferior in adhesion and coagulation activation compared with commonly used cardiovascular prostheses.

Mitral valve annuloplasty rings: review of literature and comparison of functional outcome and ventricular dimensions.

In the past decades, more than 40 mitral valve annuloplasty rings of various shapes and consistency were marketed for mitral regurgitation (MR), although the effect of ring type on clinical outcome remains unclear. Our objective was to review the literature and apply a simplification method to make rings of different shapes and rigidity more comparable. We studied relevant literature from MEDLINE and EMBASE databases related to clinical studies as well as animal and finite element models. Annuloplasty rings were clustered into 3 groups as follows: rigid (R), flexible (F), and semirigid (S). Only clinical articles regarding degenerative (DEG) or ischemic/dilated cardiomyopathy (ICM) MR were included and stratified into these groups. A total of 37 rings were clustered into R, F, and S subgroups. Clinical studies with a mean follow-up of less than 1 year and a reported mean etiology of valve incompetence of less than 60% were excluded from the analysis. Forty-one publications were included. Preimplant and postimplant end points were New York Heart Association class, left ventricular ejection fraction (LVEF), left ventricular end-systolic dimension (LVESD), and left ventricular end-diastolic dimension (LVEDD). Statistical analysis included paired-samples t test and analysis of variance with post hoc Bonferroni correction. P < 0.05 indicated statistical difference. Mean ± SD follow-up was 38.6 ± 27 and 29.7 ± 13.2 months for DEG and ICM, respectively. In DEG, LVEF remained unchanged, and LVESD decreased in all subgroups. In our analysis, LVEDD decreased only in F and R, and S did not change; however, the 4 individual studies showed a significant decline. In ICM, New York Heart Association class improved in all subgroups, and LVEF increased. Moreover, LVESD and LVEDD decreased only in F and S; R was underpowered (1 study). No statistical difference among R, F, and S in either ICM or DEG could be detected for all end points. Overall, owing to underpowered data sets derived from limited available publications, major statistical differences in clinical outcome between ring types could not be substantiated. Essential end points such as recurrent MR and survival were incomparable. In conclusion, ring morphology and consistency do not seem to play a major clinical role in mitral valve repair based on the present literature. Hence, until demonstrated otherwise, surgeons may choose their ring upon their judgment, tailored to specific patient needs.

Myocardial infarction and functional outcome assessment in pigs.

Introduction of newly discovered cardiovascular therapeutics into first-in-man trials depends on a strictly regulated ethical and legal roadmap. One important prerequisite is a good understanding of all safety and efficacy aspects obtained in a large animal model that validly reflect the human scenario of myocardial infarction (MI). Pigs are widely used in this regard since their cardiac size, hemodynamics, and coronary anatomy are close to that of humans. Here, we present an effective protocol for using the porcine MI model using a closed-chest coronary balloon occlusion of the left anterior descending artery (LAD), followed by reperfusion. This approach is based on 90 min of myocardial ischemia, inducing large left ventricle infarction of the anterior, septal and inferoseptal walls. Furthermore, we present protocols for various measures of outcome that provide a wide range of information on the heart, such as cardiac systolic and diastolic function, hemodynamics, coronary flow velocity, microvascular resistance, and infarct size. This protocol can be easily tailored to meet study specific requirements for the validation of novel cardioregenerative biologics at different stages (i.e. directly after the acute ischemic insult, in the subacute setting or even in the chronic MI once scar formation has been completed). This model therefore provides a useful translational tool to study MI, subsequent adverse remodeling, and the potential of novel cardioregenerative agents.

Surgical left ventricular radius enlargement by patch insertion on the beating heart: a new experimental aneurysm model.

We presented a novel experimental aneurysm model for studies in left ventricular (LV) reconstruction techniques and assessed LV function. In eight pigs, the LV radius and geometry were enlarged surgically on the beating heart by inserting an aortic allograft construct. Haemodynamics and LV dimensions were assessed by echocardiography at baseline and under dobutamine stress. Surgery was successfully performed without lethal blood loss or arrhythmias. LV end-diastolic and end-systolic short-axis areas increased from 13.0 ± 1.7 to 17.0 ± 4.3 cm(2) (P = 0.001) and from 4.0 ± 0.9 to 13.0 ± 2.6 cm(2) (P = 0.001), respectively. Stroke volume decreased from 56 ± 11 to 33 ± 16 ml (P = 0.001). Incremental dobutamine infusion concurred with a biphasic response on fractional area shortening. Mitral valve insufficiency ranging from grades 2 to 4 was observed. In the pig, a novel, reproducible aneurysm model for acute cardiac dysfunction was created on the beating heart. Innovative (surgical) strategies for (staged) reconfiguration of the ventricle, e.g. adjustable Dor procedures and stepwise volume restraining cardiac support devices, can be tested for efficacy using this acute model.

Myocardial lesion depth with circular electroporation ablation.

BACKGROUND: Recently, we demonstrated the feasibility and safety of circular electroporation ablation in porcine pulmonary vein ostia, but the relationship between the magnitude of the application and lesion dimensions is still unknown. METHODS AND RESULTS: An in vivo porcine study was performed on left ventricular epicardium submerged under 10 mm of blood, using devices that mimic a 20-mm-diameter 7F circular ablation catheter. Model D contained 10 separate electrodes, whereas model M consisted of 1 circular electrode. Ablations were performed at 50, 100, and 200 J with model D and at 100 J with model M. Lesion dimensions were measured after 3-week survival. All applications resulted in smooth voltage waveforms demonstrating the absence of vapor globe formation, arcing, and a pressure wave. Applications up to 100 J with model D resulted in separate lesions under the electrodes. At 200 J, continuous deep circular lesions were created despite the use of separate electrodes. There was a significant relationship between applied current and median lesion depth, with a slope of 0.17 mm/A. At 100 J, there was no difference in lesion depth or width between models D and M. The electrodes and ablation site directly after ablation showed no signs of thermal damage. CONCLUSIONS: In an epicardial porcine model with blood around the application site, continuous circular lesions, deep enough for electric pulmonary vein isolation, were created with a single circular 200-J application. Lesions were continuous despite the use of separate electrodes. Lesion depth increased with the magnitude of the application.

In-vivo validation of a new non-invasive continuous ventricular stroke volume monitoring system in an animal model.

INTRODUCTION: Recently, a non-invasive, continuous ventricular stroke volume monitoring system using skin electrodes has been developed. In contrast to impedance-based methods, the new technique (ventricular field recognition) enables measurement of changes in ventricular volume. A prototype using this new method was built (the hemologic cardiac profiler, HCP) and validated against a reference method in a pig model during variations in cardiac output. METHODS: In six Dalland pigs, cardiac output was simultaneously measured with the HCP (CO-HCP), and an invasive ultrasonic flow-probe around the ascending aorta (CO-FP). Variations in CO were achieved by change in ventricular loading conditions, cardiac pacing, and dobutamine administration. Data were analysed according to Bland-Altman analysis and Pearson's correlation. RESULTS: Pearson's correlation between the CO-HCP and the CO-FP was r = 0.978. Bland-Altman analysis showed a bias of - 0.114 L/minute, and a variability of the bias (2 standard deviations, 2SD) of 0.55 L/minute. CONCLUSIONS: The results of the present study demonstrate that CO-HCP is comparable to CO-FP in an animal model of cardiac output measurements during a wide variation of CO. Therefore, the HCP has the potential to become a clinical applicable cardiac output monitor.

Computed tomography detects tissue formation in a stented engineered heart valve.

Tissue-engineered heart valves (TEHV) are being explored as an alternative to conventional heart valve prostheses. Using the classic tissue engineering paradigm, a stented tri-leaflet valve is fabricated. Subsequently, the construct is implanted into the pulmonary position in a sheep. Follow-up by means of computed tomography, magnetic resonance imaging, and echocardiography was used to assess tissue formation. After 4 weeks, the scaffold of the TEHV has degraded and new tissue is formed. However, small areas without tissue formation were present at macroscopic inspection. This phenomenon was only visible on computed tomographic images. Therefore, computed tomography appears a promising technique for in vivo follow-up of tissue formation in tissue-engineered heart valves.

Feasibility of electroporation for the creation of pulmonary vein ostial lesions.

INTRODUCTION: There is an obvious need for a better energy source for pulmonary vein (PV) antrum isolation. OBJECTIVE: We investigated the feasibility and safety of electroporation for the creation of PV ostial lesions. METHODS: After transseptal puncture, a custom 7F decapolar 20 mm circular ablation catheter was placed in the PV ostia of 10 pigs. Ablation was performed with a nonarcing, 200 J application delivered between the catheter and an indifferent patch electrode on the lower back. A single pulse was applied for each catheter position, with a maximum of 4 per ostium. Local PV electrogram amplitude and stimulation threshold were measured at multiple locations in both ostia before and directly after ablation, and after 3 weeks survival, using a regular 4 mm mapping catheter. All PV ostia were sectioned, stained, and histologically investigated. RESULTS: The 3-week survival period was uneventful. PV ostial electrogram amplitude decreased and stimulation threshold increased significantly in most ostia. PV angiograms did not show any stenosis during this short follow-up. Histologically, up to 3.5-mm-deep lesions were found. CONCLUSION: Data suggest that electroporation can safely be used to create lesions in a sensitive environment like PV ostia.

Resizable ventricular patch plasty in the porcine left ventricle: a pilot study.

OBJECTIVE: : Endoventricular circular patch plasty is a method used to reconstruct the ventricular cavity in patients with (post) ischemic left ventricular aneurysm or global dilatation. However, late redilatation with mitral regurgitation has been reported, in which postoperative apex shape seems to play an important role. We studied the feasibility of ventricular volume downsizing with a variably shaped patch in porcine hearts. METHODS: : In five in vitro and two acute animal experiments, a dyskinetic aneurysm was simulated with a pericardial insert. Reducing patch surface by changing patch shape diminished end-diastolic volume. In vitro, static end-diastolic volume was determined for each patch shape using volumetry and echocardiography. In the acute animal experiments, preliminary observations of patch behavior in live material were made, and pressure/time relationship, dPdTmax, was registered. RESULTS: : In vitro, bringing the convex patch into a flat plane reduced LV volume from 66 ± 7 mL (aneurysm) to 49 ± 5 mL. Four of 5 patch shapes further reduced volume to a mean of 38 ± 7 mL (P = 0.03). The in vitro echocardiographic measurements correlated with volumetry findings (r = 0.81). In the acute animal experiments, dPdTmax varied with patch shape, independent of volume changes. CONCLUSIONS: : In this pilot study, in vitro shape configuration of the resizable ventricular patch resulted in a calibrated end-diastolic volume reduction. The data of the two in vivo pilot experiments clearly indicate that change in patch configuration in the situation of more or less unchanged end-diastolic volume had impact on cardiac performance. Future studies must substantiate the results of this observation.

Translating autologous heart valve tissue engineering from bench to bed.

Tissue engineering is currently being actively investigated to ascertain if it can offer an alternative to prosthetic aortic heart valves that may overcome the current limitations of prosthetic aortic heart valves while at the same time conferring the advantages of a living autologous structure, such as biocompatibility, the capacity to grow, repair, and remodel. In vitro studies have shown tissue-engineered heart valves to have adequate structural and functional properties, indicating a promising future for heart valve tissue engineering. However, criteria are required to be able to evaluate autologous heart valves and to deem them satisfactory for clinical use. Preclinical animal studies are needed, as a precursor to long-term in vivo follow-up studies, to establish such criteria. The first challenge is to find appropriate techniques to evaluate the functionality of tissue-engineered heart valves in vivo without having to kill the animal. As such, the development of such noninvasive techniques that are able to assess the functionality of tissue-engineered heart valves is the next step in translational research. This review discusses methods of evaluating the functionality of autologous heart valves when translating from in vitro to in vivo studies and determines potential assessment criteria imperative to achieve clinical applicability of tissue-engineered heart valves in aortic valve replacement.

Stability of an autologous platelet clot in the pericardial sac: An experimental and clinical study.

OBJECTIVE: Autologous platelet clots serve as slow-release delivery systems for platelet-derived growth factors and cytokines. Their application to the pericardial sac might facilitate salvage and repair of ischemically injured myocardium. However, little is known about platelet clot stability in the pericardial sac. We investigated the stability of platelet clots in vitro and after administration to the pericardial sac in pigs and patients. METHODS: In 5 Yorkshire-Landrace pigs and 10 patients, in vitro manufactured autologous platelet gel (Medtronic Magellan Platelet Separator) and platelet-rich fibrin (Vivolution Vivostat System) were administered to the pericardial sac for 30 minutes. Two antifibrinolytics (tranexamic acid and aprotinin) were tested for their capacity to stabilize autologous platelet gel. In vitro clots, incubated at 37 degrees C for 48 hours, served as controls. Clot weight was measured before and after administration. RESULTS: In vitro, autologous platelet gel clots of either formula liquefied almost entirely within 60 minutes whereas platelet-rich fibrin clots remained intact. In the pig, platelet clot weight decreased to 16.7% +/- 7.8% (P < .05) and 66.4% +/- 3.2% (P < .05) of initial clot weight for autologous platelet gel and platelet-rich fibrin, respectively. Addition of antifibrinolytics to autologous platelet gel did not reduce clot degradation significantly. In patients, autologous platelet gel and platelet-rich fibrin clot weight remained 9.0% +/- 1.5% (P < .05) and 73.7% +/- 2.6% (P < .05) of initial clot weight, respectively. CONCLUSIONS: Autologous platelet gel is unstable both in vitro and in vivo, whereas platelet-rich fibrin remains intact in vitro and, compared with autologous platelet gel, is less subject to degradation in pigs and in patients.