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Heart valve bioprosthesis durability: A challenge to the new generation of porcine valves
Abstract
Long-term experience with first generation porcine valve xenografts enabled identification of the major limitations to their durability: (1) prosthetic-ventricular mismatch due to the high profile of the stent in patients with mitral stenosis and a small left ventricle; (2) high-pressure fixation with loss of natural collagen crimping in the fibrosa, and wash-out of proteoglycans in the spongiosa; (3) xenograft tissue autolysis, due to the long interval between animal slaughter and aortic valve removal fixation; (4) muscle shelf in the right coronary cusp, which created a gradient and could undergo accelerated calcification and/or spontaneous perforation with time; (5) a flexible polypropylene stent, which could creep or even fracture with consequent inward bending of the stent; (6) progressive time-related dystrophic calcification; (7) host fibrous tissue ingrowth. An awareness of these limitations stimulated technical modifications, which frequently brought about distinct improvements: (1) the reduction of the stent profile eliminated the problem of mismatch, but resulted in a higher tendency towards cusp prolapse and earlier commissural tearing; (2) natural collagen waviness, proteoglycans and cusp extensibility were preserved by employing low or even zero pressure during the fixation process; (3) earlier valve fixation enabled preservation of cell integrity; (4) a new orifice for small valves was designed by replacing the right muscular cusp, thus achieving less gradient and avoiding muscle-shelf-related complications; (5) polypropylene was replaced by Delrin as stent material; (6) calcium-retarding agents like T6 and toluidine blue were applied during commercial processing and storage in order to mitigate tissue mineralization.(ABSTRACT TRUNCATED AT 250 WORDS)
... The primary problem of bioprostheses, whether porcine or pericardial, is structural valve deterioration at long-term, mainly due to dystrophic calcification [1,2]. However, many improvements have been implemented in order to prolong long-term durability [3,4]. ...
... In cells rendered non-viable by GA fixation, calcium pumps are impaired and calcium passively diffuses into the cytoplasm and reacts with phospholipids of cells and organelles membranes [8,14,15]. Calcification of bioprostheses at first occurs upon cell membranes and then involves collagen fibers and interfibrillar spaces [3,8,16]. ...
Anticalcification strategies of glutaraldehyde-fixed xenograft tissue aim to extract lipids or to neutralize toxic aldehyde residuals. The purpose of this study was to evaluate the efficacy of octanediol compared to standard treatments of glutaraldehyde-fixed bovine pericardium in the subdermal rat model. Octanediol treatment is an ethanolic solution (40%) containing a long chain aliphatic alcohol (5% 1,2-octanediol) that removes lipids without diminishing the stability of collagen.
Octanediol and standard glutaraldehyde fixed bovine pericardium were both implanted in 24 Sprague-Dawley rats, explanted after 30-75 days (12 animals each) and submitted to X-ray (score 0-4), histology, electron microscopy and elemental analysis by spectroscopy (Ca and P content). Unimplanted octanediol and standard glutaraldehyde fixed pericardium served as control.
At 30 days octanediol-treated pericardium showed calcium content of 0.20+/-0.1 vs 20.07+/-36.79 mg/g dry weight for standard pericardium. The difference was also evident at 75 days: calcium content of 2.36+/-7.38 mg/g dry weight for octanediol vs 165.61+/-23.35 mg/g dry weight for standard (p<0.0001). Differences were also detected at X-ray (mean score 0.7+/-0.6 octanediol vs 3.8+/-0.4 standard at 75 days). Equally, mean P content was 11.69+/-21.33 mg/g dry weight for standard vs 0.60+/-1.45 mg/g dry weight for octanediol samples at 30 days, and 90.90+/-12.61 mg/g dry weight for standard vs 1.42+/-4.34 mg/g dry weight for octanediol at 75 days (p<0.0001). At electron microscopy collagen appeared well preserved regardless of the type of treatment; in octanediol treated pericardium cell membranes almost disappeared and only few profiles of endoplasmic reticulum and rare mitochondria were visible.
Treatment with octanediol strongly prevents calcification of glutaraldehyde fixed bovine pericardium in rat subdermal model, even in the long-term. Evidence of octanediol efficacy may entail important implications for new generation bioprosthetic valves.
... Morphological and structural changes occur in BHVs, particularly in the extracellular matrix of valve leaflets, following implantation. Eventually, these modifications may lead to the demise of the implant via various mechanisms including: calcification, GAG loss, collagen bundle loosening, and degeneration [2,3,26,27]. The relationship between GAGs and calcification remains highly debated although previous work by our group as well as others have shown that the preservation of endogenous GAGs within valve leaflets may play a minor role in inhibiting calcification [4,28]. ...
Numerous crosslinking chemistries and methodologies have been investigated as alternative fixatives to glutaraldehyde (GLUT) for the stabilization of bioprosthetic heart valves (BHVs). Particular attention has been paid to valve leaflet collagen and elastin stability following fixation. However, the stability of glycosaminoglycans (GAGs), the primary component of the spongiosa layer of the BHV, has been largely overlooked despite recent evidence provided by our group illustrating their structural and functional importance. In the present study we investigate the ability of two different crosslinking chemistries: sodium metaperiodate (NaIO(4)) followed by GLUT (PG) and 1-Ethyl-3-(3 dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS) followed by GLUT (ENG) to stabilize GAGs within BHV leaflets and compare resulting leaflet characteristics with that of GLUT-treated tissue. Incubation of fixed leaflets in GAG-degrading enzymes illustrated in vitro resistance of GAGs towards degradation in PG and ENG treated tissue while GLUT fixation alone was not effective in preventing GAG loss from BHV leaflets. Following subdermal implantation, significant amounts of GAGs were retained in leaflets in the ENG group in comparison to GLUT-treated tissue, although GAG loss was evident in all groups. Utilizing GAG-targeted fixation did not alter calcification potential of the leaflets while collagen stability was maintained at levels similar to that observed in conventional GLUT-treated tissue.
In recent years, there has been a revival in the use of bioprosthetic valves as aortic substitutes. For example, during 2008 in Germany, among 12,397 patients who underwent isolated aortic valve procedures, 78% received a biological prosthesis and 21% a mechanical valve, while only 1% underwent valve repair. However, whilst this situation is the reverse of that during the 1990s, the question must be asked as to whether this is simply a fluctuating fashion, or a reflection of other factors.
Background. Although stentless bioprostheses were introduced into clinical practice in the early nineties, their durability and resistance to primary degeneration of surrounding tissue have not yet been established due to limited follow-up duration. Aim. To assess the impact of stent on the rate and characteristics of the degeneration process of bioprosthetic valves in an animal experimental model. Methods. Aortic valve bioprostheses (stentless valves Toronto SPV™ - Group T, n=6, and valves with stent - Hancock® II MO - Group H, n=6) were implanted into the pulmonary artery in sheep. Echocardiographic assessment was performed three and six months after implantation. After valve removal, bioprostheses underwent macroscopic, microscopic, histological (light microscopy as well as transmission electron microscopy) and atomic absorption spectrophotometry assessment. Results. Echocardiography revealed valve insufficiency in 50% of bioprostheses in group T and in all valves in group H. Macroscopic abnormalities of valvular leaflets were seen in all group H xenografts. Valvular leaflets in group T had well preserved structure and very infrequent limited calcium deposits, whereas the valvular leaflets in group H were thickened due to significant calcification, covered by a thick layer of connective tissue, and collagen fibres were separated and smooth. The amount of calcium, assessed six months after implantation, was significantly lower in group T than in group H bioprostheses (5.59±2.50 μg/mg vs 7.89±2.31 μg/mg; p<0.05). A progressive malfunction and degeneration of valve tissue was observed in the course of time in both groups. Conclusions. Stentless biological valves are characterised by increased durability and significantly less pronounced tissue degeneration.
Objective: To evaluate the effect of postoperative follow-up duration on prosthetic valves on real-time three-dimensional echocardiography (RT3DE). Methods: Totally 80 patients with prosthetic valves underwent RT3DE for 125 times between five days and twelve years during postoperative follow-up period. Results: From five days to two months (inclusive) during the postoperative period, the quality of three-dimensional images was high in 75 % (63/84), and poor or affected by respiratory motion in 25% (21/ 84); while from two months to twelve years, high in 97.6% (40/41), and poor or affected by respiratory motion in 2.4% (1/41) (P<0.01). From five days to one year(inclusive), the mechanical prosthetic atrioventricular annulus was clear in 86.2% (50/58), and unclear in 13.8% (8/58); while from one year to twelve years, clear in 11.1% (1/9) and unclear in 88.9% (8/9) (P<0.001). Conclusion: With the prolongation of the follow-up duration, the quality of three-dimensional images will be improved, whereas the clarity of the mechanical prosthetic atrioventricular annulus will be decreased gradually. The cause of this phenomenon is due to the endocardial growing over to the surface of the prosthetic atrioventricular annulus.
The primary mechanism and most common cause of hemolytic disease in patients with prosthetic heart valves are mechanical trauma to red blood cells and paraprosthetic valvular regurgitation, respectively. Presenting features in patients with this condition include anemia, congestive heart failure, fatigue, jaundice, dark urine, and a regurgitant murmur. Various laboratory studies can be utilized to diagnose hemolytic anemia and to assess the severity of hemolysis. Transthoracic echocardiography, transesophageal echocardiography, and Doppler studies including color Doppler are useful imaging methods to assess valve function. Treatment is usually medical (oral iron); however, in patients with paravalvular regurgitation, surgery is often required to correct the anemia.
In recent years, there has been a revival in the use of bioprosthetic valves as aortic substitutes. For example, during 2008 in Germany, among 12,397 patients who underwent isolated aortic valve procedures, 78% received a biological prosthesis and 21% a mechanical valve, while only 1% underwent valve repair. However, whilst this situation is the reverse of that during the 1990s, the question must be asked as to whether this is simply a fluctuating fashion, or a reflection of other factors.
Heart valve tissue engineering requires biocompatible and hemocompatible scaffolds that undergo remodeling and repopulation, but that also withstand harsh mechanical forces immediately following implantation. We hypothesized that reversibly stabilized acellular porcine valves, seeded with endothelial cells and conditioned in pulsatile bioreactors would pave the way for next generations of tissue engineered heart valves (TEHVs). A novel valve conditioning system was first designed, manufactured and tested to adequately assess TEHVs. The bioreactor created proper closing and opening of valves and allowed for multiple mounting methods in sterile conditions. Porcine aortic heart valve roots were decellularized by chemical extractions and treated with penta-galloyl glucose (PGG) for stabilization. Properties of the novel scaffolds were evaluated by testing resistance to collagenase and elastase, biaxial mechanical analysis, and thermal denaturation profiles. Porcine aortic endothelial cells were seeded onto the leaflets and whole aortic roots were mounted within the dynamic pulsatile heart valve bioreactor system under physiologic pulmonary valve pressures and analyzed after 17 days for cell viability, morphology, and metabolic activity. Our tissue preparation methods effectively removed cells, including the potent α-Gal antigen, while leaving a well preserved extra-cellular matrix scaffold with adequate mechanical properties. PGG enhanced stabilization of extracellular matrix components but also showed the ability to be reversible. Engineered valve scaffolds encouraged attachment and survival of endothelial cells for extended periods and showed signs of widespread cell coverage after conditioning. Our novel approach shows promise toward development of sturdy and durable TEHVs capable of remodeling and cellular repopulation.
Sixty-two explanted Liotta porcine bioprostheses were examined to review the issues related to their biocompatibility, biofunctionality, and biodurability. These bioprostheses were harvested from 56 patients with implantation times ranging from only a few hours to more than nine years of implantation. There were 10 acute and short-term (< 1 year), 20 midterm (1 < t < 5 years), and 32 long-term (> 5 years) cases. The indications for the reoperations were: hemodynamic (59), thrombosis (10), and endocarditis (3). The major indications varied according to the duration of implantation: blood infiltration, fibrin buildup, thrombosis in the short-term; endocarditis and hemodynamic insufficiency in the midterm; and mineralization and tears causing hemodynamic incompetence in the long term. Mineralization proved to be the main threat to long-term durability for porcine valves. Besides a few short-term failures, these explanted devices slowly degenerated over time and were replaced to prevent congestive heart failure. Documentation of the failure modes of these porcine valves is important since the demand for bioprostheses will increase in the future, in particular for percutaneous devices. Such bioprostheses emphasize a critical biocompatibility issue following implantation because they have the capacity to remain free of thrombus in the absence of anticoagulation. The biofunctionality appears to be adequate in the absence of subsequent pathology with restoration of normal valve function. However, the documentation of such significant long-term biodurability issues raises questions that have been addressed but not fully answered yet with the new generations of bioprostheses.
Glutaraldehyde is considered a promoter of calcification by the action of toxic aldehyde group residuals from cross-linking. Post-fixation treatment with homocysteic acid (HA), besides bonding aldehyde groups and neutralizing toxicity, should enhance biocompatibility due to the strongly electronegative sulfonic group. The aim of this investigation was to evaluate HA efficacy on tissue preservation and dystrophic calcification mitigation in glutaraldehyde cross-linked bovine pericardium (BP) using a subcutaneous rat model.
Four samples of BP, two with glutaraldehyde-HA and two with glutaraldehyde treatment, were implanted in each of 24 male Sprague-Dawley rats. Three rats were killed at 14 days, eight at 28 days, eight at 56 days and five at 84 days. Unimplanted glutaraldehyde-HA- and glutaraldehyde-treated samples served as controls. All samples were studied by gross examination, mammography, light transmission and scanning electron microscopy, and atomic absorption spectroscopy. The nature of mineralization was investigated by coupling techniques of scanning electron microscopy, electron microprobe analysis and X-ray powder diffraction.
No histological and ultrastructural differences were found between glutaraldehyde-HA- and glutaraldehyde-treated BP, whether implanted or unimplanted. In both groups, calcification progressed with time, but significantly less after glutaraldehyde-HA treatment than after glutaraldehyde alone and at all time intervals (14.63 +/- 21.34 versus 43.17 +/- 15.99 at 28 days, p = 0.003; 56.42 +/- 40.20 versus 90.59 +/- 32.90 at 56 days, p = 0.008; 91.68 +/- 67.68 versus 156.23 +/- 17.85 at 84 days, p = 0.01). Differences were evident by mammography and histology (von Kossa stain). Electron microprobe analysis in both groups showed the composition of calcified nuclei to be calcium phosphate, stoichiometrically close to apatite (Ca5(PO4)3(OH)). The occurrence of crystallized apatite was supported by X-ray powder diffraction findings, the amount of crystallized apatite being higher in glutaraldehyde-treated samples.
Post-fixation treatment with HA preserves BP structural properties and significantly mitigates mineralization of long-term subcutaneous implants.
The aim of this study was to define the clinical, echocardiographic, and pathologic correlates of commissural dehiscence of aortic wall from the stent post of the porcine bioprostheses in the mitral position. This form of valve degeneration was found in 5 of 23 explanted mitral bioprostheses. A thickened, separated aortic wall at multiple commissural sites along with other evidence of valve degeneration was identified in the three patients who had chronic congestive heart failure. A large dehiscence at a single commissural site with otherwise normal valve morphology was present in the two patients who had acute heart failure. Two dimensional/Doppler echocardiography showed a prolapsing or a flail anteriorly positioned leaflet and an eccentric posteriorly directed mitral regurgitation jet in all patients. These echocardiographic findings in patients with a porcine bioprosthetic mitral valve should suggest commissural dehiscence from the aortic wall as a possible mechanism of valve failure. Exclusive involvement of the porcine aortic bioprosthesis placed in the mitral position along with involvement of strut of the bioprosthesis facing the aortic root in all cases suggests excessive hemodynamic stress on the valve in the mitral position and in particular on the anteriorly placed strut as the potential cause of this form of valve degeneration.
Heart valve substitutes have been in use for over 30 years. Bioprosthetic heart valves have many advantages, but unfortunately suffer tissue degeneration and calcification. Many approaches, such as antimineralization treatment to prevent or delay these changes, have been tried. We present the morphologic findings from a series of Hancock II (antimineralization-treated) porcine bioprostheses.
Forty-five Hancock II porcine valve bioprostheses (16 mitral, 29 aortic) surgically explanted between March 1991 and December 1995 at the Toronto Hospital were analyzed for morphologic findings and causes of failure. The prostheses were implanted in 36 adults (mean age 55+/-14.7 years, range: 27 to 75 years) and had been in place between one month and 11 years (mean 5.1+/-3.3 years).
Structural valve deterioration (SVD) characterized by tissue degeneration, calcification, cusp tears and increased stiffness, was the single most significant finding and cause of failure, affecting 56% of valves. Some degree of calcification was seen in 55% of prostheses, with severe calcification (grade 3 or 4) in 18%. Aortic bioprostheses showed more severe and earlier calcification than mitral ones (p = 0.03). Compared with the standard Hancock valve, the low incidence of significant calcification suggests a beneficial protective effect of antimineralization treatment. Severe pannus (grade 3 or 4) was seen in 60% of these prostheses. The pattern of pannus growth differs between mitral and aortic sites; mitral prostheses showed pannus on the flow and non-flow surfaces, often associated with cusp tears, mitral regurgitation and mitral leaflet preservation. A similar degree of pannus on aortic prostheses was invariably present on the flow surface and extended onto the valve cusps, leading to changes in the orifice which may cause clinical aortic stenosis. Infective endocarditis was seen in 15 prostheses (five mitral, 10 aortic) from 11 patients, and comprised the second most important cause of prosthesis failure. The risk of recurrent endocarditis was particularly high in patients who had infective endocarditis before valve replacement, even at five and six years post implantation.
SVD is the major finding in explanted Hancock II bioprostheses and is associated with cusp tears and calcification. The incidence of tissue calcification appears lower at the mitral site. These findings suggest that the antimineralization treatment had some beneficial effect. Pannus associated with prosthesis dysfunction at the mitral sites is a prominent finding and on the non-flow surface may be related to the native mitral valve-conserving procedure.
The study investigates the mechanical properties of porcine aortic valve leaflets fixed with a naturally occurring crosslinking agent, genipin, at distinct pressure heads. Fresh and the glutaraldehyde-fixed counterparts were used as controls. Subsequent to fixation, the changes in leaflet collagen crimps and its surface morphology were investigated by light microscopy and scanning electron microscopy (SEM). Also, the crosslinking characteristics of each studied group were determined by measuring its fixation index and denaturation temperature. In the mechanical testing, tissue strips made from each studied group were examined in both the circumferential and radial directions. Histological and SEM comparisons between fresh porcine aortic valve leaflet and those fixed at medium or high pressure revealed that the following changes may occur: elimination of the natural collagen crimping, and extensive loss of the endothelial layer. The denaturation temperatures of the glutaraldehyde-fixed leaflets were significantly greater than the genipin-fixed leaflets; however, their fixation indices were comparable. Generally, fixation pressure did not affect the crosslinking characteristics of the genipin- and glutaraldehyde-fixed leaflets. It was found that fixation of porcine aortic valves in genipin or glutaraldehyde did not alter the mechanical anisotropy observed in fresh valve leaflets. This indicated that the intramolecular and intermolecular crosslinks introduced into the collagen fibrils during fixation is of secondary importance to the presence of structural and mechanical anisotropy in fresh leaflet. Tissue fixation in genipin or glutaraldehyde may produce distinct crosslinking structures. However, the difference in crosslinking structure between the genipin- and glutaraldehyde-fixed leaflets did not seem to cause any significant discrepancies in their mechanical properties when compared at the same fixation pressure. Nevertheless, regardless of the crosslinking agent used, changes in mechanical properties and ruptured patterns were observed when the valve leaflets were fixed at distinct pressures.
Performance of bioprosthetic valves is limited by tissue degeneration due to calcification with reduced performance and longevity. The Mosaic bioprosthetic valve (Medtronic Heart Valves, Inc, Minneapolis, MN) combines zero pressure fixation, antimineralization properties of alpha-amino oleic acid (AOA), and a proven stent design. We tested the hypothesis that AOA treatment of Mosaic valves improves hemodynamics, antimineralization properties, and survival in a chronic ovine model.
Mitral valves were implanted in juvenile sheep with Mosaic valves with AOA treatment (n = 8) or without AOA treatment (non-AOA, n = 8), or Hancock I (HAN, n = 4) tissue valves, and explanted at 20 postoperative weeks.
Survival was equivalent in AOA and non-AOA (140 +/- 0.4 and 129 +/- 30 days), but was significantly less in HAN (82 +/- 35). Leaflet calcium (microgCa/mg tissue) was less in AOA (9.6 +/- 13.9; p < 0.05 versus non-AOA and HAN) than non-AOA (96.3 +/- 63.8) and HAN (130.8 +/- 43.2). Explant valve orifice area (cm2) was significantly preserved in the AOA group compared with the non-AOA group (1.5 +/- 0.7 vs 0.8 +/- 0.3; p < 0.05 versus non-AOA and HAN).
We conclude that AOA treatment of Mosaic valves reduces leaflet calcification and valve gradient in juvenile sheep, and that the Mosaic design and fixation features may offer survival advantages that must be confirmed in extended trials.
The Hancock II bioprosthesis is a second-generation porcine valve xenograft treated with the detergent sodium dodecyl sulphate (T6) to retard calcification. The aim of this investigation was to study the gross and microscopic features in Hancock II explants to assess the structural changes occurring with time.
Among 1382 Hancock II bioprostheses (701 isolated aortic, 421 isolated mitral, 130 double) implanted from 1983 to 1997 in 1252 patients, 22 (16 mitral, 6 aortic) were removed at reoperation until 1999 and were available for pathological investigation: infective endocarditis occurred in 5 and structural deterioration in 8, whereas in the remaining 9 xenografts reoperation was performed for nonstructural valve deterioration (paravalvular leak in 4 and prophylactic replacement in 5). Morphological investigation consisted of gross examination and x-ray, histologic, immunohistochemistry, electron microscopic, and atomic absorption spectroscopic examination.
The cause of structural valve deterioration was dystrophic calcification in 4 cases (1 aortic, 3 mitral; range of time graft was in place, 101 to 144 months), non-calcium-related tears in 3 cases (all mitral, range 121 to 163 months), and commissural dehiscence in 1 (aortic, range 156 months). Five of the nonstructural valve deterioration explants (range 42 to 122 months) showed only pinpoint mineralization at the commissures. Mean calcium content in nonstructural deterioration explants was 14.70 +/- 22.33 versus 99.11 +/- 81.52 mg/g in explants with structural valve deterioration. Electron microscopic examination showed early nuclei of mineralization mostly consisting of calcospherulae upon cell debris. Local or diffuse lipid insudation was observed in all but 2 explants and consisted of cholesterol clefts, lipid droplets, and lipid-laden macrophages featuring foam cells. The lipid insudation was the most plausible cause of tearing in 2 explants.
These pathologic findings support the clinical results of a delayed occurrence of structural failure of Hancock II bioprostheses and a mitigation of mineralization by the anti-calcification treatment. However, other factors such as lipid insudation may come into play in the long term.
BIOSA is a single sheet, two-cuspal-shaped ("bileaflet") glutaraldehyde fixed bovine pericardial valve prosthesis (BP), designed to minimize stress and prevent mechanical failure. We tested this device in the adult sheep model.
Seven BIOSA and two Baxter Carpentier-Edwards (Perimount) pericardial BPs, 25 mm in size, were implanted in the tricuspid position of adult sheep. A mid term BIOSA explant (81 days) died of infective endocarditis. The remaining BPs were divided in three early explants (0-3 days: two BIOSA and one Perimount) and five late explants (162-189 days: four BIOSA and one Perimount). Protocol of the study included gross examination, mammography X-ray (score 0-4), histology, scanning (SEM) and transmission (TEM) electron microscopy and atomic absorption spectroscopy.
(a) Early explants: The mesothelium was detached, collagen-elastic fibers and pericardiocytes of the fibrosa showed optimal preservation. Calcium content in BIOSA BPs was 6.22 mg/g dry weight (mean) versus 7.75 mg/g of the Perimount BP. (b) Late explants: At naked eye all BPs showed regular cusp pliability and coaptation, without tears, perforations, fibrous pannus or calcific deposits. X-ray was either negative (three BPs) or exhibited score 1 (two BPs). Microscopic features were excellent both in terms of collagen-elastic fibers preservation and absence of inflammation. The calcium content was 4.95 mg/g dry weight (mean) in BIOSA BPs versus 5.29 mg/g in the Perimount BP.
Tissue characteristics of BIOSA BPs were optimal, without difference with Perimount BPs and in this animal model no case of structural valve deterioration occurred in the long term.
Calcification is still a major cause of failure of implantable biomaterials. A fast and reliable in vitro model could contribute to the study of its mechanisms and to testing different anticalcification techniques. In this work, we attempted to investigate the potential calcification of biomaterials using an in vitro model. We purposed to test the ability of this model to screening possible anticalcification efficacy of different biomaterials. Porcine heart valve (PAV) and bovine pericardial (BP) tissues, fixed with glutaraldehyde were immersed into biological mimicking solution, where the pH and the initial concentrations of calcium and phosphoric ions were kept stable by the addition of precipitated ions during calcification. Kinetics of calcification was continuously monitored. The evaluation of biomaterials was carried out by comparing the kinetic rates of formation of calcific deposits. After 24 h, the calcific deposits on PAVs were found to be developed at significant higher rates (ranged from 0.81 x 10(-4)-2.18 x 10(-4)mol/min m2) than on BP (0.19 x 10(-4)-0.52 x 10(-4)mol/min m2) (one-way ANOVA, p < 0.05) depending on the experimental conditions (supersaturation of the solution). Parallel tests for similar biomaterials implanted subcutaneously in animal (rat) model showed after 49 days that significant higher amounts of total minerals deposited on PAV (236.73+/-139.12, 9 animals mg minerals/g dry net tissue) (mean+/-standard deviation) compared with that formed on BP (104.36+/-79.21, #9 mg minerals/g dry net tissue) (ANOVA, p < 0.05). There is evidence that in vitro calcification was correlated well with that of animal model and clinical data.
Bovine pericardium (BPC) and polytetrafluoroethylene (PTFE) have been widely used to reinforce staple lines in lung resection. Since limited information regarding the calcification of these biomaterials is available, we undertook an in vitro study to evaluate their calcification potential. Commercially available BPC and PTFE biomaterials were evaluated and compared with custom-prepared BPC tissue. In vitro calcification was performed via submersion in supersaturated solution in a double-walled glass reactor at 37.0 degrees C +/- 0.1 degrees C, pH 7.4 +/- 0.1, mimicking most ion concentrations of human blood plasma. In processing of calcification, the pH decrease of the solution simulated the addition of consumed H(+), Ca(2+), and PO(4)(3-) ions from titrant solutions, the concentrations of which were based on the stoichiometry of octacalcium phosphate. The molar ion addition with time was recorded, and the initial slope of the curve was computed for each experiment. The rate of calcification developed (molar calcium phosphate ion addition rate per time and total surface area) (R) was computed after that with respect to the relative supersaturation (sigma) used in each experiment. R for custom-prepared BPC tissues was found to be in the range of 0.19 +/- 0.08 to 0.52 +/- 0.19 (n = 17) in sigma range of 0.72 to 1.42. Commercial BPC was found to be 0.016 to 0.052 (n = 4), and PTFE was 0.005 to 0.05 (n = 8) in the same sigma range. Both clinically applied biomaterials, BPC and PTFE, seemed to be calcified with rates of at least one order of magnitude lower than the custom-prepared BPC tissue. This data suggested that BPC and PTFE biomaterials showed a similar, relatively very low tendency for calcification compared with custom-prepared BPC tissue. Although further studies are necessary, staple line reinforcement by these two biomaterials should be considered safe from the calcification point of view.
