Pigs as Models to Test Cardiovascular Devices.

Pigs as laboratory animals are used in preclinical studies aimed at developing medical devices for cardiac surgery. The anatomy of the cardiovascular system of these animals has been well studied and acknowledged as suitable for use and the testing of new cardiovascular devices developed for humans. However, there are no morphometric characteristics of the aortic root and thoraco-abdominal part of porcine aorta. This can lead to difficulties in experimental surgery and even result in the death of experimental animals due to the mismatch in the size of the implantable devices. Thus, such information is essential to enhance the efficiency of surgical technologies used for eliminating aortic pathologies in their various sections. The purpose of our research is to study the anatomy of the aorta in mini pigs and to assess whether the size, age, and sex of the animals affect the size of the main structures in their aortas. In addition, we attempted to compare the results obtained by transesophageal echocardiography (TEE) and angiography. We studied 28 laboratory mini pigs, dividing them into three groups by body weight (40-70 kg, 71-90 kg, and 90 kg). We did not find any relationship between the external somatometric characteristics of the animals and the size of their aortas. Animals have individual anatomical variability in their cardiovascular systems, which means that they need to be examined in terms of preoperative planning by any available method-echocardiography, angiography, or multispiral computed tomography (CT).

Which Gelatin and Antibiotic Should Be Chosen to Seal a Woven Vascular Graft?

Among the vascular prostheses used for aortic replacement, 95% are woven or knitted grafts from polyester fibers. Such grafts require sealing, for which gelatin (Gel) is most often used. Sometimes antibiotics are added to the sealant. We used gelatin type A (GelA) or type B (GelB), containing one of the three antibiotics (Rifampicin, Ceftriaxone, or Vancomycin) in the sealant films. Our goal was to study the effect of these combinations on the mechanical and antibacterial properties and the cytocompatibility of the grafts. The mechanical characteristics were evaluated using water permeability and kinking radius. Antibacterial properties were studied using the disk diffusion method. Cytocompatibility with EA.hy926 endothelial cells was assessed via indirect cytotoxicity, cell adhesion, and viability upon direct contact with the samples (3, 7, and 14 days). Scanning electron microscopy (SEM) and energy dispersive spectrometry (EDS) were used to visualize the cells in the deep layers of the graft wall. "GelA + Vancomycin" and "GelB + vancomycin" grafts showed similar good mechanical characteristics (permeability~10 mL/min/cm2, kinking radius 21 mm) and antibacterial properties (inhibition zones for Staphilococcus aureus~15 mm, for Enterococcus faecalis~12 mm). The other samples did not exhibit any antibacterial properties. The cytocompatibility was good in all the tested groups, but endothelial cells exhibited the ability to self-organize capillary-like structures only when interacting with the "GelB + antibiotics" coatings. Based on the results obtained, we consider "GelB + vancomycin" sealant to be the most promising.

An insight into the effect of interaction with protein on antibacterial activity of chitosan derivatives.

Antimicrobial activity of chitosan in protein-rich media is of a particular interest for various protein-based drug delivery and other systems. For the first time, bacteriostatic activity of chitosan derivatives in the presence of caseinate sodium (CAS) was studied and discussed. Complexation of chitosan derivatives soluble in acidic (CH and RCH) or alkalescent (RCH) media with CAS was confirmed by fluorescent spectroscopy, turbodimetry, light scattering data and measurement of electrical potentials of CAS/chitosan derivative complexes. An addition of CH and RCH caused a static quenching of CAS. Binding constants Kb determined for CH/CAS and RCH/CAS complexes at pH 6.0 were equal to 29.8 × 106 M-1 and 8.9 × 106 M-1, respectively. Kb value of RCH/CAS complex at pH 7.4 was equal to 1.1 × 105'M-1. The poisoned food method was used for counting the number and the direct measurement of the size of bacterial colonies on the surfaces of turbid agar media containing CAS/chitosan derivative complexex. Complete suppression of E. coli cells growth and restriction of S. aureus cells growth were observed on the surface of acidic media. A high concentration of CAS reduced the activity. The activity of RCH in alkalescent media is low or absent. These results can be promising for preparation of microbiologically stable protein-based drug delivery systems.

Epoxy- versus Glutaraldehyde-Treated Bovine Jugular Vein Conduit for Pulmonary Valve Replacement: A Comparison of Morphological Changes in a Pig Model.

Valved conduits are often required to replace pulmonary arteries (PA). A widely used Contegra device is made of bovine jugular vein (BJV), preserved with glutaraldehyde (GA) and iso-propanol. However, it has several drawbacks that may be attributed to its chemical treatment. We hypothesized that the use of an alternative preservation compound may significantly improve BJV conduit performance. This study aimed to compare the macroscopic and microscopic properties of the BJV treated with diepoxide (DE) and GA in a porcine model. Twelve DE-BJVs and four Contegra conduits were used for PA replacement in minipigs. To assess the isolated influence of GA, we included an additional control group-BJV treated with 0.625% GA (n = 4). The animals were withdrawn after 6 months of follow-up and the conduits were examined. Explanted DE-BJV had a soft elastic wall with no signs of thrombosis or calcification and good conduit integration, including myofibroblast germination, an ingrowth of soft connective tissue formations and remarkable neoangiogenesis. The inner surface of DE-BJVs was covered by a thin neointimal layer with a solid endothelium. Contegra grafts had a stiffer wall with thrombosis on the leaflets. Calcified foci, chondroid metaplasia, and hyalinosis were observed within the wall. The distal anastomotic sites had hyperplastic neointima, partially covered with the endothelium. The wall of GA-BJV was stiff and rigid with degenerative changes, a substantial amount of calcium deposits and dense fibrotic formations in adventitia. An irregular neointimal layer was presented in the anastomotic sites without endothelial cover in the GA BJV wall. These results demonstrate that DE treatment improves conduit integration and the endothelialization of the inner surface while preventing the mineralization of the BJV, which may reduce the risk of early conduit dysfunction.

Remote Outcomes with Poly-ε-Caprolactone Aortic Grafts in Rats.

Poly-ε-caprolactone ((1,7)-polyoxepan-2-one; PCL) is a biodegradable polymer widely used in various fields of bioengineering, but its behavior in long-term studies appears to depend on many conditions, such as application specificity, chemical structure, in vivo test systems, and even environmental conditions in which the construction is exploited in. In this study, we offer an observation of the remote outcomes of PCL tubular grafts for abdominal aorta replacement in an in vivo experiment on a rat model. Adult Wistar rats were implanted with PCL vascular matrices and observed for 180 days. The results of ultrasound diagnostics and X-ray tomography (CBCT) show that the grafts maintained patency for the entire follow-up period without thrombosis, leakage, or interruptions, but different types of tissue reactions were found at this time point. By the day of examination, all the implants revealed a confluent endothelial monolayer covering layers of hyperplastic neointima formed on the luminal surface of the grafts. Foreign body reactions were found in several explants including those without signs of stenosis. Most of the scaffolds showed a pronounced infiltration with fibroblastic cells. All the samples revealed subintimal calcium phosphate deposits. A correlation between chondroid metaplasia in profound cells of neointima and the process of mineralization was supported by immunohistochemical (IHC) staining for S100 proteins and EDS mapping. Microscopy showed that the scaffolds with an intensive inflammatory response or formed fibrotic capsules retain their fibrillar structure even on day 180 after implantation, but matrices infiltrated with viable cells partially save the original fibrillary network. This research highlights the advantages of PCL vascular scaffolds, such as graft permeability, revitalization, and good surgical outcomes. The disadvantages are low biodegradation rates and exceptionally high risks of mineralization and intimal hyperplasia.

Influence of Single-Wall Carbon Nanotube Suspension on the Mechanical Properties of Polymeric Films and Electrospun Scaffolds.

Carbon nanotubes (CNTs) are used in applications ranging from electrical engineering to medical device manufacturing. It is well known that the addition of nanotubes can influence the mechanical properties of various industrial materials, including plastics. Electrospinning is a popular method for fabricating nanomaterials, widely suggested for polymer scaffold manufacturing. In this study, we aimed to describe the influence of single-walled carbon nanotube (SWCNT) suspensions on polymeric poured films and electrospun scaffolds and to investigate their structural and mechanical properties obtained from various compositions. To obtain films and electrospun scaffolds of 8 mm diameter, we used poly-ε-caprolactone (PCL) and poly(cyclohexene carbonate) (PCHC) solutions containing several mass fractions of SWCNT. The samples were characterized using tensile tests, atomic force and scanning electronic microscopy (AFM and SEM). All the studied SWCNT concentrations were shown to decrease the extensibility and strength of electrospun scaffolds, so SWCNT use was considered unsuitable for this technique. The 0.01% mass fraction of SWCNT in PCL films increased the polymer strength, while fractions of 0.03% and more significantly decreased the polymer strength and extensibility compared to the undoped polymer. The PHCH polymeric films showed a similar behavior with an extremum at 0.02% concentration for strength at break.

Effect of Soft Preheating of Bovine Serum Albumin on the Complexation with Oligochitosan: Structure and Conformation of BSA in the Complex.

Phase analysis, spectroscopic, and light scattering methods are applied to investigate the peculiarities of the interaction of oligochitosan (OCHI) with native and preheated bovine serum albumin (BSA) as well as the conformational and structural changes of BSA in BSA/OCHI complex. As shown, untreated BSA binds with OCHI mainly forming soluble electrostatic nanocomplexes, with the binding causing an increase in BSA helicity without a change in the local tertiary structure and thermal stability of BSA. In contrast, soft preheating at 56 °C enhances the complexation of BSA with OCHI and slightly destabilizes the secondary and local tertiary structures of BSA within the complex particles. Preheating at 64 °C (below the irreversible stage of BSA thermodenaturation) leads to further enhancement in the complexation and formation of insoluble complexes stabilized by both Coulomb forces and hydrophobic interactions. The finding can be promising for the preparation of biodegradable BSA/chitosan-based drug delivery systems.

Calcification of Various Bioprosthetic Materials in Rats: Is It Really Different?

The causes of heart valve bioprosthetic calcification are still not clear. In this paper, we compared the calcification in the porcine aorta (Ao) and the bovine jugular vein (Ve) walls, as well as the bovine pericardium (Pe). Biomaterials were crosslinked with glutaraldehyde (GA) and diepoxide (DE), after which they were implanted subcutaneously in young rats for 10, 20, and 30 days. Collagen, elastin, and fibrillin were visualized in non-implanted samples. Atomic absorption spectroscopy, histological methods, scanning electron microscopy, and Fourier-transform infrared spectroscopy were used to study the dynamics of calcification. By the 30th day, calcium accumulated most intensively in the collagen fibers of the GA-Pe. In elastin-rich materials, calcium deposits were associated with elastin fibers and localized differences in the walls of Ao and Ve. The DE-Pe did not calcify at all for 30 days. Alkaline phosphatase does not affect calcification since it was not found in the implant tissue. Fibrillin surrounds elastin fibers in the Ao and Ve, but its involvement in calcification is questionable. In the subcutaneous space of young rats, which are used to model the implants' calcification, the content of phosphorus was five times higher than in aging animals. We hypothesize that the centers of calcium phosphate nucleation are the positively charged nitrogen of the pyridinium rings, which is the main one in fresh elastin and appears in collagen as a result of GA preservation. Nucleation can be significantly accelerated at high concentrations of phosphorus in biological fluids. The hypothesis needs further experimental confirmation.

Effect of the Nanorough Surface of TiO2 Thin Films on the Compatibility with Endothelial Cells.

The cytocompatibility of titanium oxides (TiO2) and oxynitrides (N-TiO2, TiOxNy) thin films depends heavily on the surface topography. Considering that the initial relief of the substrate and the coating are summed up in the final topography of the surface, it can be expected that the same sputtering modes result in different surface topography if the substrate differs. Here, we investigated the problem by examining 16 groups of samples differing in surface topography; 8 of them were hand-abraded and 8 were machine-polished. Magnetron sputtering was performed in a reaction gas medium with various N2:O2 ratios and bias voltages. Abraded and polished uncoated samples served as controls. The surfaces were studied using atomic force microscopy (AFM). The cytocompatibility of coatings was evaluated in terms of cytotoxicity, adhesion, viability, and NO production. It has been shown that the cytocompatibility of thin films largely depends on the surface nanostructure. Both excessively low and excessively high density of peaks, high and low kurtosis of height distribution (Sku), and low rates of mean summit curvature (Ssc) have a negative effect. Optimal cytocompatibility was demonstrated by abraded surface with a TiOxNy thin film sputtered at N2:O2 = 1:1 and Ub = 0 V. The nanopeaks of this surface had a maximum height, a density of about 0.5 per 1 µm2, Sku from 4 to 5, and an Ssc greater than 0.6. We believe that the excessive sharpness of surface nanostructures formed during magnetron sputtering of TiO2 and N-TiO2 films, especially at a high density of these structures, prevents both adhesion of endothelial cells, and their further proliferation and functioning. This effect is apparently due to damage to the cell membrane. At low height, kurtosis, and peak density, the main factor affecting the cell/surface interface is inefficient cell adhesion.

Complexation of oligochitosan with sodium caseinate in alkalescent and weakly acidic media.

Сomplexation of oligochitosan (OCHI) having the degree of acetylation (DA 26 %) with sodium caseinate (SC) at pH 5.8 and 7.2 is described and compared with the complexation of OCHI (DA 2 %) at pH 5.8. In the alkalescent medium, the complexation of OCHI (DA 26 %) is weaker and dualistic depending on SC concentration in the system. In the diluted alkalescent system, the formation of only soluble complexes is observed at OCHI/SC ratio ≤0.9. In the semi diluted one, the complexation results in the formation of insoluble complexes those composition changes symbatically with the OCHI/SC ratio in the system. At pH 5.8, OCHI/SC ratio in insoluble complexes remains the same regardless of OCHI/SC ratio in the solution. At pH 5.8, the electrostatic complexation weakens with an increase in DA and is completely suppressed at a high ionic strength. These results can be promising for construction of biodegradable protein/chitosan drug delivery systems.

Integral Algorithms to Evaluate TiO2 and N-TiO2 Thin Films' Cytocompatibility.

Titanium oxide (TiO2) and oxynitride (N-TiO2) coatings can increase nitinol stents' cytocompatibility with endothelial cells. Methods of TiO2 and N-TiO2 sputtering and cytocompatibility assessments vary significantly among different research groups, making it difficult to compare results. The aim of this work was to develop an integral cytocompatibility index (ICI) and a decision tree algorithm (DTA) using the "EA.hy926 cell/TiO2 or N-TiO2 coating" model and to determine the optimal cytocompatible coating. Magnetron sputtering was performed in a reaction gas medium with various N2:O2 ratios and bias voltages. The samples' morphology was studied by scanning electron microscopy (SEM) and Raman spectroscopy. The cytocompatibility of the coatings was evaluated in terms of their cytotoxicity, adhesion, viability, and NO production. The ICI and DTA were developed to assess the cytocompatibility of the samples. Both algorithms demonstrated the best cytocompatibility for the sample sputtered at Ubias = 0 V and a gas ratio of N2:O2 = 2:1, in which the rutile phase dominated. The DTA provided more detailed information about the cytocompatibility, which depended on the sputtering mode, surface morphology, and crystalline phase. The proposed mathematical models relate the cytocompatibility and the studied physical characteristics.

In Vivo Evaluation of PCL Vascular Grafts Implanted in Rat Abdominal Aorta.

Electrospun tissue-engineered grafts made of biodegradable materials have become a perspective search field in terms of vascular replacement, and more research is required to describe their in vivo transformation. This study aimed to give a detailed observation of hemodynamic and structural properties of electrospun, monolayered poly-ε-caprolactone (PCL) grafts in an in vivo experiment using a rat aorta replacement model at 10, 30, 60 and 90 implantation days. It was shown using ultrasound diagnostic and X-ray tomography that PCL grafts maintain patency throughout the entire follow-up period, without stenosis or thrombosis. Vascular compliance, assessed by the resistance index (RI), remains at the stable level from the 10th to the 90th day. A histological study using hematoxylin-eosin (H&E), von Kossa and Russell-Movat pentachrome staining demonstrated the dynamics of tissue response to the implant. By the 10th day, an endothelial monolayer was forming on the graft luminal surface, followed by the gradual growth and compaction of the neointima up to the 90th day. The intense inflammatory cellular reaction observed on the 10th day in the thickness of the scaffold was changed by the fibroblast and myofibroblast penetration by the 30th day. The cellularity maximum was reached on the 60th day, but by the 90th day the cellularity significantly (p = 0.02) decreased. From the 60th day, in some samples, the calcium phosphate depositions were revealed at the scaffold-neointima interface. Scanning electron microscopy showed that the scaffolds retained their fibrillar structure up to the 90th day. Thus, we have shown that the advantages of PCL scaffolds are excellent endothelialization and good surgical outcome. The disadvantages include their slow biodegradation, ineffective cellularization, and risks for mineralization and intimal hyperplasia.

A feasible approach to tune the interaction of chitosan with sodium dodecyl sulfate.

Interaction of binary chitosan/nonionic surfactant (NIS) system with sodium dodecyl sulfate (SDS) in aqueous solution is described using turbodimetry, light scattering, electophoretic mobility and cryogenic electron microscopy. The formation of insoluble CHI/SDS complexes is weakened with a decrease in molecular weight of chitosan and critical micelle concentration of NIS as well as with an increase in NIS concentration. Soluble chitosan/NIS complexes absorb SDS molecules until the charge of mixed chitosan/NIS/SDS complexes reaches a critical value that depends on chitosan molecular weight followed by aggregation of primary electrostatic complexes via hydrogen bonding to complex nanoparticles. In contrast to formation of asymmetric swarm-like structures in the binary chitosan/SDS system, the aggregation of complex nanoparticles in the ternary chitosan/NIS/SDS system occurs by a head-to-tail binding mechanism with formation of elongated filamentous microstructures. The finding can be promising for preparation of microbiologically stable pharmaceutical and cosmetic compositions and drug delivery systems containing mixed surfactants.

Effects of Electrospinning Parameter Adjustment on the Mechanical Behavior of Poly-ε-caprolactone Vascular Scaffolds.

Electrospinning is a perspective method widely suggested for use in bioengineering applications, but the variability in currently available data and equipment necessitates additional research to ascertain the desirable methodology. In this study, we aimed to describe the effects of electrospinning technique alterations on the structural and mechanical properties of (1,7)-polyoxepan-2-one (poly-ε-caprolactone, PCL) scaffolds, such as circumferential and longitudinal stress/strain curves, in comparison with corresponding properties of fresh rat aorta samples. Scaffolds manufactured under different electrospinning modes were analyzed and evaluated using scanning electronic microscopy as well as uniaxial longitudinal and circumferential tensile tests. Fiber diameter was shown to be the most crucial characteristic of the scaffold, correlating with its mechanical properties.

Bovine jugular vein conduit: What affects its elastomechanical properties and thermostability?

The aim of this study was to compare the mechanical properties and thermal stability of the venous wall depending on the treatment method used, and, accordingly, on those structural changes in the tissue that this treatment causes. Bovine jugular vein walls (BJVWs) cross-linked with glutaraldehyde (GA), ethylene glycol diglycidyl ether (DE), and Contegra commercial conduit were evaluated using uniaxial stretching [with and without pre-conditioning (PreC)], differential scanning calorimetry, amino acid analysis, and attenuated total reflection infrared spectroscopy. Fresh BJVW was used as a control. It was shown that failure stress in non-PreC GA-treated and DE-treated materials was lower than that in fresh and Contegra counterparts. Contegra samples were the stiffest among the tested materials. Cyclic preloading leads to distortion of the mechanical behavior of this material, which is heterogeneous in composition and structure. The denaturation temperatures (Td ) of all cross-linked BJVWs were higher than the Td of the fresh vein. The microstructures of the tested BJVWs did not exhibit any differences, but the cross-linking density and hydration of the DE-vein were the highest. GA-cross-linking or DE-cross-linking and isopropanol exposure (Contegra) changed the protein secondary structures of the tested materials in different ways. We hypothesized that the protein secondary structure and hydration degree are the main causes of differences in the mechanical properties and thermal stability of BJVW.

Toward a model of learning innovation integration: TPACK-SAMR based analysis of the introduction of a digital learning environment in three Russian universities.

Digitalisation is penetrating higher education (HE) in Russia. The study explores how three Russian universities have been integrating the Skyes digital learning platform to transform English as a foreign language (EFL) educational practices. The article outlines the main characteristics of the platform, recounts the steps in adopting the technology and addresses the emerging changes to learning and teaching. Students' and teachers' perceptions are examined to review the implementation process. The research is designed as a case study; the methods used are semi-structured interviews and online questionnaires. The study framework combines the Technological Pedagogical Content Knowledge (TPACK) model and the Substitution, Augmentation, Modification, and Redefinition (SAMR) model for a holistic approach to investigating innovation integration in a university setting. TPACK is used to analyse the changes associated with teachers' knowledge of technology, pedagogy and content. SAMR helps to reflect on the current use of the platform showcasing technology integration along substitution, augmentation and modification levels. Based on the findings, the paper discusses the factors influencing the implementation of the Skyes platform and proposes recommendations for its successful integration; they highlight the importance of motivation, feedback both from teachers and students, clear assessment of the learning outcomes, TPACK transformation plan and alignment between the innovation and educational regulations.

Allografts and xenografts for right ventricular outflow tract reconstruction in Ross patients.

OBJECTIVES: Pulmonary allografts (AG) are the gold standard for right ventricular outflow tract (RVOT) reconstruction during the Ross procedure. However, there is limited availability of AG in some countries, and the use of alternative grafts for RVOT reconstruction remains controversial. This study aimed to compare the rates of freedom from RVOT graft dysfunction for AG and diepoxide-treated pericardial xenografts (DPXG). METHODS: Between 1998 and 2015, 793 adult patients underwent the Ross procedure in our centre. Using propensity score matching, the clinical outcomes and echocardiographic results of AG and DPXG were compared. RESULTS: Propensity score matching resulted in 2 groups (AG and DPXG) of 122 patients each. No difference was found in early mortality (2.5%) in both groups. The freedom from RVOT graft dysfunction curves were comparable between the AG and DPXG groups (P = 0.186) and the 8-year rates of freedom from graft dysfunction were 91.8% and 82.2%, respectively. The survival rates at 8 years were 90.5% and 90.1%, and the rates of freedom from RVOT reintervention at 8 years were 100% and 96.8% for the AG and DPXG groups, respectively. At discharge and follow-up, transprosthetic gradients were significantly higher in the DPXG group. The rate of the RVOT gradient progression was also higher in the DPXG group than in the AG group (1.80 ± 0.06 vs 1.39 ± 0.04 mmHg/year, P < 0.001). CONCLUSIONS: There was no difference in freedom from RVOT graft dysfunction by 8 years when using AG and DPXG in adult Ross patients, nor in survival and freedom from RVOT conduit reintervention. Long-term results need further evaluation.

Immobilized Bisphosphonates as Potential Inhibitors of Bioprosthetic Calcification: Effects on Various Xenogeneic Cardiovascular Tissues.

Calcification is the major factor limiting the clinical use of bioprostheses. It may be prevented by the immobilization of bisphosphonic compounds (BPs) on the biomaterial. In this study, we assessed the accumulation and structure of calcium phosphate deposits in collagen-rich bovine pericardium (Pe) and elastin-rich porcine aortic wall (Ao) and bovine jugular vein wall (Ve) cross-linked with glutaraldehyde (GA) or diepoxy compound (DE). These tissues were then modified with pamidronic (PAM) acid or 2-(2'-carboxyethylamino)ethylidene-1,1-bisphosphonic (CEABA) acid. Tissue transformations were studied using Fourier-transform infrared spectroscopy. After subcutaneous implantation of the biomaterials in 220 rats, calcification dynamics were examined using atomic absorption spectrophotometry, light microscopy after von Kossa staining, and scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy The calcium content in all GA-cross-linked tissues and DE-cross-linked Ao increased to 100-160 mg/g on day 60 after implantation. BPs prevented the accumulation of phosphates on the surface of all materials and most effectively inhibited calcification in GA-cross-linked Ao and DE-cross-linked Pe. PAM containing -OH in the R1 group was more effective than CEABA containing -H in R1. The calcification-inhibitory effect of BPs may be realized through their ability to block nucleation and prevent the growth of hydroxyapatite crystals.

Cross-linking method using pentaepoxide for improving bovine and porcine bioprosthetic pericardia: A multiparametric assessment study.

Bioprosthetic heart valves made from bovine pericardium (BP) and porcine pericardium (PP) preserved with glutaraldehyde (GA) are commonly used in valve surgeries but prone to calcification in many patients. In this study, we compared BP and PP preserved with GA, ethylene glycol diglycidyl ether (DE), and 1,2,3,4,6-penta-O-{1-[2-(glycidyloxy)ethoxy]ethyl}-d-glucopyranose (PE). We studied the stabilities of DE and PE in preservation media along with the amino acid (AA) compositions, Fourier-transform infrared spectra, mechanical properties, surface morphologies, thermal stability, calcification, and the cytocompatibility of BP and PP treated with 0.625% GA, 5% DE, 2% PE, and alternating 5% DE and 2% PE for 3 + 11 d and 10 + 10 d, respectively. Both epoxides were stable in the water-buffer solutions (pH 7.4). DE provided high linkage densities in BP and PP owing to reactions with Hyl, Lys, His, Arg, Ser, and Tyr. PE reacted weakly with these AAs but strongly with Met. High cross-linking density obtained using the 10 d + 10 d method provided satisfactory thermal stability of biomaterials. The epoxy preservations improved cytocompatibility and resistance to calcification. PE enhanced the stress/strain properties of the xenogeneic pericardia, perhaps by forming nanostructures that were clearly visualised in BP using scanning electron microscopy. The DE + PE combination, in an alternating cross-linking manner, thus constitutes a promising option for developing bioprosthetic pericardia.

Treatment with bisphosphonates to mitigate calcification of elastin-containing bioprosthetic materials.

This study evaluated the ability of bisphosphonates (BPAs) of different molecular structures to mitigate the calcification of porcine aortic wall (PAW) and bovine jugular vein wall (BJVW). Tissues cross-linked with glutaraldehyde (GA) or diepoxide (DE) were modified with pamidronic acid (PAM), alendronic acid (ALE), neridronic acid (NER) (type 1 BPAs); 2-(2'-carboxyethylamino)ethylidene-1,1-bisphosphonic acid (CEABA), 2-(5-carboxypentylamino)ethylidene-1,1-bisphosphonic acid (CPABA) (type 2); and zoledronic acid (ZOL) (type 3). After implanting the tissue samples subcutaneously in 100 rats, calcification was examined using atomic absorption spectrophotometry (60-day explants) and light microscopy after von Kossa staining (10- and 30-day explants). The calcium contents in GA-BJVW and GA- and DE-PAW increased up to 100-120 mg/g after 60 days, while being 3 times lower in DE-BJVW. In modified and nonmodified PAW samples, calcium phosphates appeared by day 10 and were associated with elastic fibers and devitalized cellular elements. In all groups of BJVW samples, mineralization began in elastic fibers near the subendothelial layer. In addition, calcified collagen was found in the GA-BJVW samples. Minimal calcification was found in GA-PAW treated with type 1 BPAs and CEABA. For DE-PAW and GA-BJVW, the calcium level significantly decreased with PAM and CEABA. Meanwhile, ALE and NER were effective for DE-BJVW.