Search Results (22)

The adhesion between an implant and a wound could result in over-bleeding when attempting to separate the two. To address this issue, a cell-repelling implant is preferred. In this study, a cell-repelling membrane was prepared by modifying decellularized porcine pericardium with multi-arm polyethylene glycol. With this modification technology, we switched the surface properties of the decellularized porcine pericardium from cell-adhering to cell-repelling. The result showed that this pericardium was successfully modified without any effect on the original properties of the pericardium and also maintained a low inflammatory response. The level of cell adhesion on the surface of the membrane was significantly reduced. Full article

The production of biomedical devices able to appropriately interact with the biological environment is still a great challenge. Synthetic materials are often employed, but they fail to replicate the biological and functional properties of native tissues, leading to a variety of adverse effects. Several commercial products are based on chemically treated xenogeneic tissues: their principal drawback is due to weak mechanical stability and low durability. Recently, decellularization has been proposed to bypass the drawbacks of both synthetic and biological materials. Acellular materials can integrate with host tissues avoiding/mitigating any foreign body response, but they often lack sufficient patency and impermeability. The present paper investigates an innovative approach to the realization of hybrid materials that combine decellularized bovine pericardium with polycarbonate urethanes. These hybrid materials benefit from the superior biocompatibility of the biological tissue and the mechanical properties of the synthetic polymers. They were assessed from physicochemical, structural, mechanical, and biological points of view; their ability to promote cell growth was also investigated. The decellularized pericardium and the polymer appeared to well adhere to each other, and the two sides were distinguishable. The maximum elongation of hybrid materials was mainly affected by the pericardium, which allows for lower elongation than the polymer; this latter, in turn, influenced the maximum strength achieved. The results confirmed the promising features of hybrid materials for the production of vascular grafts able to be repopulated by circulating cells, thus, improving blood compatibility. Full article

The aim of this study was to compare the viscoelastic properties of a decellularized mesh from the porcine esophagus, prepared by our group, with two commercial acellular tissues derived from porcine small intestine submucosa and bovine pericardium for use in medical devices. The tissues’ viscoelastic properties were characterized by creep tests in tension, applying the load in the direction of the fibers or the transverse direction, and also by dynamic-shear mechanical tests between parallel plates or in tension at frequencies between 0.1 and 35 Hz. All the tests were performed in triplicate at a constant temperature of 37 °C immersed in distilled water. The tissues’ surface and cross-sectional microstructure were observed by scanning electron microscopy (SEM) to characterize the orientation of the fibers. The matrices of the porcine esophagus present an elastic modulus in the order of 60 MPa when loaded in the longitudinal direction while those of the porcine intestine submucosa and bovine pericardium have an elastic modulus below 5 MPa. Nevertheless, the shear modulus of bovine pericardium nearly triplicates that of the esophageal matrix. The viscoelasticity of decellularized esophageal mucosa is characterized by a fast change in the creep compliance with time. The slope of the creep curve in the double logarithmic plot is twice that of the control samples. These results are consistent with the microstructure observed under electron microscopy regarding the orientation of the fibers that make up the matrices. Full article

Myocardial infarction (MI) is a serious cardiovascular disease as the leading cause of death globally. Hence, reconstruction of the cardiac tissue comes at the forefront of strategies adopted to restore heart functions following MI. In this investigation, we studied the capacity of rat adipose-derived mesenchymal stem cells (r-AdMSCs) and decellularized porcine pericardium (DPP) to restore heart functions in MI animals. MI was induced in four different groups, three of which were treated either using DPP (MI-DPP group), stem cells (MI-SC group), or both (MI-SC/DPP group). Cardiac functions of these groups and the Sham group were evaluated using echocardiography, the intraventricular pressure gradient (IVPG) on weeks 2 and 4, and intraventricular hemodynamics on week 4. On day 31, the animals were euthanized for histological analysis. Echocardiographic, IVPG and hemodynamic findings indicated that the three treatment strategies shared effectively in the regeneration process. However, the MI-SC/DPP group had a unique synergistic ability to restore heart functions superior to the other treatment protocols. Histology showed that the MI-SC/DPP group presented the lowest (p < 0.05) degeneration score and fibrosis % compared to the other groups. Conclusively, stem cell-seeded DPP is a promising platform for the delivery of stem cells and restoration of heart functions post-MI. Full article

Defects in the dura matter can be caused by head injury, and many cases require neurosurgeons to use artificial dura matter. Bovine pericardium is an option due to its abundant availability, adjustable size and characteristics, and because it has more collagen than porcine or equine pericardia. Nevertheless, the drawback of bovine pericardium is that it has a higher inflammatory effect than other synthetic dura matters. Chitosan has been shown to have a strong anti-inflammatory effect and has good tensile strength; thus, the idea was formulated to use chitosan as a coating for bovine pericardium. This study used decellularized bovine pericardial membranes with 0.5% sodium dodecyl sulphate and coatings containing chitosan at concentrations of 0.25%, 0.5%, 0.75%, and 1%. An FTIR test showed the presence of a C=N functional group as a bovine pericardium–chitosan bond. Morphological tests of the 0.25% and 0.5% chitosan concentrations showed standard pore sizes. The highest tensile strength percentage was shown by the membrane with a chitosan concentration of 1%. The highest degradation rate of the membrane was observed on the 7th and 14th days for 0.75% and 1% concentrations, and the lowest swelling ratio was observed for the 0.25% concentration. The highest level of cell viability was found for 0.75% chitosan. The bovine pericardium membrane with a 0.75% concentration chitosan coating was considered the optimal sample for use as artificial dura matter. Full article

Pathological aseptic calcification is the most common form of structural valvular degeneration (SVD), leading to premature failure of heart valve bioprostheses (BHVs). The processing methods used to obtain GA-fixed pericardium-based biomaterials determine the hemodynamic characteristics and durability of BHVs. This article presents a comparative study of the effects of several processing methods on the degree of damage to the ECM of GA-fixed pericardium-based biomaterials as well as on their biostability, biocompatibility, and resistance to calcification. Based on the assumption that preservation of the native ECM structure will enable the creation of calcinosis-resistant materials, this study provides a soft biomimetic approach for the manufacture of GA-fixed biomaterials using gentle decellularization and washing methods. It has been shown that the use of soft methods for preimplantation processing of materials, ensuring maximum preservation of the intactness of the pericardial ECM, radically increases the resistance of biomaterials to calcification. These obtained data are of interest for the development of new calcinosis-resistant biomaterials for the manufacture of BHVs. Full article

Animal-derived xenogeneic biomaterials utilized in different surgeries are promising for various applications in tissue engineering. However, tissue decellularization is necessary to attain a bioactive extracellular matrix (ECM) that can be safely transplanted. The main objective of the present study is to assess the structural integrity, biocompatibility, and potential use of various acellular biomaterials for tissue engineering applications. Hence, a bovine pericardium (BP), porcine pericardium (PP), and porcine tunica vaginalis (PTV) were decellularized using a Trypsin, Triton X (TX), and sodium dodecyl sulfate (SDS) (Trypsin + TX + SDS) protocol. The results reveal effective elimination of the cellular antigens with preservation of the ECM integrity confirmed via staining and electron microscopy. The elasticity of the decellularized PP (DPP) was markedly (p < 0.0001) increased. The tensile strength of DBP, and DPP was not affected after decellularization. All decellularized tissues were biocompatible with persistent growth of the adipose stem cells over 30 days. The staining confirmed cell adherence either to the peripheries of the materials or within their matrices. Moreover, the in vivo investigation confirmed the biocompatibility and degradability of the decellularized scaffolds. Conclusively, Trypsin + TX + SDS is a successful new protocol for tissue decellularization. Moreover, decellularized pericardia and tunica vaginalis are promising scaffolds for the engineering of different tissues with higher potential for the use of DPP in cardiovascular applications and DBP and DPTV in the reconstruction of higher-stress-bearing abdominal walls. Full article

The covalent functionalization of synthetic peptides allows the modification of different biomaterials (metallic, polymeric, and ceramic), which are enriched with biologically active sequences to guide cell behavior. Recently, this strategy has also been applied to decellularized biological matrices. In this study, the covalent anchorage of a synthetic peptide (REDV) to a pericardial matrix decellularized via Schiff base is realized starting from concentrated peptide solutions (10⁻⁴ M and 10⁻³ M). The use of a labeled peptide demonstrated that as the concentration of the working solution increased, the surface density of the anchored peptide increased as well. These data are essential to pinpointing the concentration window in which the peptide promotes the desired cellular activity. The matrices were extensively characterized by Water Contact Angle (WCA) analysis, Differential Scanning Calorimetry (DSC) analysis, geometric feature evaluation, biomechanical tests, and preliminary in vitro bioassays. Full article

Regenerative grafts for myocardial reconstruction are often mechanically not stable enough to withstand the left ventricle’s high blood pressure. Hence, decellularized pericardium may serve as a stabilizing structure for biological myocardium prostheses. The efficacy of detergent- and enzyme-based protocols to decellularize porcine pericardium was compared. Then, the decellularized pericardium was employed for a primary cover of a transmural left ventricular defect in minipigs (n = 9). This pericardium patch was applied to mitigate the high-pressure load on an autologous stomach tissue, which was utilized as a regenerative tissue prosthesis. Decellularization of the porcine pericardium with deoxycholic acid (DOA)- and enzyme-based protocols (trypsin/EDTA) removed 90% of the original cells (p < 0.001). The trypsin/EDTA protocol significantly altered the matrix architecture compared to the DOA protocol. There were no infections or clinical signs of graft rejection following the transplantation of the decellularized pericardium and the autologous segment of the stomach in the surviving animals (n = 7). A good left ventricular function could be detected via MRI six months following surgery. The biological integration of the graft into the host’s tissue was found histologically. The stabilization of initially fragile grafts with decellularized pericardium facilitates the application of regenerative myocardial prostheses even on the left ventricle. Full article

Implantation of scaffolds causes a local inflammatory response whereby the early recruitment of neutrophils is of great importance not only for fighting the infection, but also for facilitating effective regeneration. We used luminol-dependent chemiluminescence, flow cytometry, ELISA, and confocal microscopy to assess the responses of neutrophils after the exposure to the scaffold-decellularized bovine pericardium (collagen type I) crosslinked with genipin (DBPG). We demonstrated that DBPG activated neutrophils in whole blood causing respiratory burst, myeloperoxidase (MPO) secretion, and formation of neutrophil extracellular trap-like structures (NETs). In addition, we studied platelets, another important player of the immediate immune host response. We found that platelets triggered redox-activation of isolated neutrophils by the pericardium scaffold, and likely participate in the NETs formation. Free radicals generated by neutrophils and hypochlorous acid produced by MPO are potent oxidizing agents which can oxidatively degrade biological structures. Understanding the mechanisms and consequences of redox activation of neutrophils by pericardium scaffolds is important for the development of new approaches to increase the efficiency of tissue regeneration. Full article

The degeneration of heart valve bioprostheses due to calcification processes is caused by the intercalation of calciumhydroxyapatite in pericardium collagen bundles. Variations of the protein secondary structure of biomaterials according to preparation are relevant for this mineralization process and thus the structural characterization of innovative bioprostheses materials is of great importance. The gold standard for prostheses preparation is glutaraldehyde (GA)-fixation of bovine pericardium that adversely promotes calcification. The novel GA-free SULEEI-treatment of bovine pericardium includes decellularization, UV-crosslinking, and electron beam sterilization. The aim of this study is the structural characterization of SULEEI-treated and GA-fixed bovine pericardium. IR spectroscopic imaging combined with multivariate data and curve fit analysis was applied to investigate the amide I and amide II regions of SULEEI-treated and GA-fixed samples. The spectroscopic images of GA-fixed pericardial tissue exhibited a generally high content of amine groups and side chains providing nucleation points for calcification processes. In contrast, in SULEEI-treated tissue, the typical α-helical structure was retained and was supposed to be less prone to deterioration. Full article

Over the past decade regenerative branches of dentistry have taken on more and more importance, resulting in the development of performing scaffold materials. These should induce cell adhesion, support, and guide the tissues’ growth. Among the developed materials, we can include resorbable or non-membranes. The purpose of this study was to investigate the proliferation abilities and the attachment of human periodontal ligament fibroblasts (HPLIFs) over two bovine pericardium membranes with different thicknesses, 0.2 mm and 0.4 mm, respectively. These membranes have been decellularized by the manufacturer, preserving the three-dimensional collagen’s structure. The HPLFs were cultured in standard conditions and exposed to the tested materials. XTT was performed to assess cell proliferation, while light microscopy (LM) and scanning electron microscopy (SEM) observations assessed fibroblast morphology at different times (T1, T2, and T3). Proliferation assays have shown a statistically significant difference in growth at T1 (p < 0.05) in the cells cultured with a thicker membrane compared to the thinner one. LM analysis showed healthy fibroblasts in contact with the membranes, appearing larger and with a polygonal shape. SEM observation demonstrated thickening of the fibroblasts which continued to adhere to the membrane’s surface, with enlarged polygonal shape and developed filipodia and lamellipodia. These results showed a similar cell behavior over the two bovine pericardium membranes, demonstrating a cellular migration along and within the layers of the membrane, binding with membrane fibers by means of filopodial extensions. Knowledge of the effects of the collagen membranes derived from bovine pericardium on cellular behavior will help clinicians choose the type of scaffolds according to the required clinical situation. Full article

Recent advancements in regenerative medicine have enhanced the development of biomaterials as multi-functional dressings, capable of accelerating wound healing and addressing the challenge of chronic wounds. Hydrogels obtained from decellularized tissues have a complex composition, comparable to the native extracellular environment, showing highly interesting characteristics for wound healing applications. In this study, a bovine pericardium decellularized extracellular matrix (dECM) hydrogel was characterized in terms of macromolecules content, and its immunomodulatory, angiogenic and wound healing potential has been evaluated. The polarization profile of human monocytes-derived macrophages seeded on dECM hydrogel was assessed by RT-qPCR. Angiogenic markers expression has been evaluated by Western blot and antibody array on cell lysates derived from endothelial cells cultured on dECM hydrogel, and a murine in vivo model of hindlimb ischemia was used to evaluate the angiogenic potential. Fibroblast migration was assessed by a transwell migration assay, and an in vivo murine wound healing model treated with dECM hydrogels was also used. The results showed a complex composition, of which the major component is collagen type I. The dECM hydrogel is biocompatible, able to drive M2 phenotype polarization, stimulate the expression of angiogenic markers in vitro, and prevent loss of functionality in hindlimb ischemia model. Furthermore, it drives fibroblast migration and shows ability to facilitate wound closure in vivo, demonstrating its great potential for regenerative applications. Full article

Human and animal pericardia are among the most widely exploited materials suitable to repair damaged tissues in the cardiovascular surgery context. Autologous, xenogeneic (chemically treated) and homologous pericardia are largely utilized, but they do exhibit some crucial drawbacks. Any tissue treated with glutaraldehyde is known to be prone to calcification in vivo, lacks regeneration potential, has limited durability, and can result in cytotoxicity. Moreover, autologous tissues have limited availability. Decellularized biological tissues represent a promising alternative: decellularization removes cellular and nuclear components from native tissues and makes them suitable for repopulation by autologous cells upon implantation into the body. The present work aims to assess the effects of a new detergent, i.e., Tergitol, for decellularizing bovine and porcine pericardia. The decellularization procedure successfully removed cells, while preserving the histoarchitecture of the extracellular matrix. No cytotoxic effect was observed. Therefore, decellularized pericardia showed potential to be used as scaffold for cardiovascular tissue regeneration. Full article

Recent applications of decellularized tissues have included the ectopic use of their sheets and powders for three-dimensional (3D) tissue reconstruction. Decellularized tissues are fabricated with the desired functions to employ them to a target tissue. The aim of this study was to develop a 3D reconstruction method using a recellularized pericardium to overcome the difficulties in cell infiltration into tight and dense tissues, such as ligament and tendon tissues. Decellularized pericardial tissues were prepared using the high hydrostatic pressurization (HHP) and surfactant methods. The pericardium consisted of bundles of aligned fibers. The bundles were slightly disordered in the surfactant decellularization method compared to the HHP decellularization method. The mechanical properties of the pericardium were maintained after the HHP and surfactant decellularizations. The HHP-decellularized pericardium was rolled up into a cylindrical formation. Its mechanical behavior was similar to that of a porcine anterior cruciate ligament in tensile testing. NIH3T3, C2C12, and mesenchymal stem cells were adhered with elongation and alignment on the HHP- and surfactant-decellularized pericardia, with dependences on the cell type and decellularization method. When the recellularized pericardium was rolled up into a cylinder formation and cultured by hanging circulation for 2 days, the cylinder formation and cellular elongation and alignment were maintained on the decellularized pericardium, resulting in a layer structure of cells in a cross-section. According to these results, the 3D-reconstructed decellularized pericardium with cells has the potential to be an attractive alternative to living tissues, such as ligament and tendon tissues. Full article