Matrix metalloproteinases as a possible factor of biological prosthetic

Aim. To identify the expression and possible sources of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in bioprosthetic heart valves (BHVs).Methods. The material for the study was epoxy-treated BHVs (NeoKor Ltd, Kemerovo) obtained during valve re-prosthetics. Cellular infiltration and ECM degradation were assessed by haematoxylin and eosin staining while lipid deposition and calcification were analysed by Oil Red O and Alizarin Red S staining. Сellular typing and detection of MMP-1/-2/-8/-9/-12 and TIMP-1/-2 expression in samples were performed using immunohistochemical staining with antibodies to CD45, CD68, СD3, CD19, myeloperoxidase, and to the corresponding MMPs and TIMPs. Analysis of samples was performed by light microscopy.Results. We examined 7 xenoaortic and 7 xenopericardial BHVs which were removed during re-replacement from the aortic (n = 2) and mitral (n = 12) positions. In studied leaflets from 13 explanted BHVs, sporadic infiltrates consisting of macrophages and neutrophils were revealed. Semi-quantitative analysis showed that more aggressive cellular infiltration is characteristic of xenoaortic BHVs (p = 0.038). MMP-1/-2/-8/-12 and TIMP-1/-2 were weakly expressed and co-localised with infiltrating cells whilst MMP-9 was abundant in the loosened extracellular matrix (ECM) devoid of host cells.Conclusion. The recipient cells infiltrating BHVs are sources of MMP-1/-2/-8/-9/-12 and TIMP-1/-2. In addition, MMP-9 can diffuse into BHVs leaflets from the blood of patients.

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TExpression of matrix metalloproteinases 1, 2, 9, 12 in xenogenic tissues of epoxy-crosslinked bioprosthetic heart valves explanted due to dysfunction

Russian Journal of Cardiology ◽

10.15829/1560-4071-2020-3978 ◽

2020 ◽

Vol 25 (10) ◽

pp. 3978

Author(s):

A. E. Kostyunin ◽

T. V. Glushkova

Keyword(s):

Matrix Metalloproteinases ◽

Heart Valves ◽

Immune Cell ◽

Blood Clot ◽

Expression Patterns ◽

Bioprosthetic Valve ◽

Positive Staining ◽

Intense Staining ◽

Bioprosthetic Heart Valves ◽

Microscopic Studies

Aim. To study the expression patterns of matrix metalloproteinases (MMPs) -1, -2, -9, -12 in the leaflets of the epoxy-treated bioprostheses explanted due to dysfunction and to identify the pathways for the accumulation of these enzymes in the xenogenic tissues.Methods. 19 leaflets from seven epoxy-treated bioprostheses (Kemcor (n = 2), PeriCor (n = 2), UniLine (n = 2) and TiAra (n = 1)) explanted from the mitral or aortic positions during repeat heart valve replacements were included in a study. Sections for microscopic studies were cut on a standard rotary microtome. Cell typing and the expression of MMP-1, -2, -9, -12 were evaluated using immunohistochemical staining with the antibodies against PTPRC/CD45, CD68, neutrophil myeloperoxidase and the corresponding MMPs. Stained samples were examined by light microscopy.Results. Sporadic cell infiltrates, mainly composed of macrophages (PTPRC/CD45+, CD68+), were found in 17 leaflets from six explanted bioprostheses. Positive staining for MMP-1, -2, -9, -12 was colocalized with immune cell infiltrates. It is worth noting that MMP-9 staining was visualized even in the absence of cell infiltration, while more intense staining was found in the areas with a loose extracellular matrix. There were no signs of macrophage infiltration or MMP expression in xenotissues of pericardial bioprostheses failed due to thrombosis and explanted two days after implantation. However, a blood clot formed on its surface showed intense MMP-9 staining and included a large proportion of neutrophils positive for myeloperoxidase.Conclusion. Macrophages and other immune cells that infiltrate xenotissues of epoxy-treated bioprostheses are sources of MMP-1, -2, -9, -12. In addition, MMP-9 can diffuse into bioprosthetic valve leaflets from blood plasma of patients. Thus, MMPs deposition in xenotissues may contribute to the leaflet ruptures and calcifications leading to the development of bioprosthetic valve dysfunction.

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303 IN VITRO NEURONAL DIFFERENTIATION OF MESENCHYMAL STEM CELLS DERIVED FROM CANINE EAR SKIN

Reproduction Fertility and Development ◽

10.1071/rdv23n1ab303 ◽

2011 ◽

Vol 23 (1) ◽

pp. 248

Author(s):

J. H. Lee ◽

Y. M. Lee ◽

G. H. Maeng ◽

S. L. Lee ◽

G. J. Rho

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Immunohistochemical Staining ◽

Transcriptional Factors ◽

Surface Markers ◽

Rt Pcr ◽

Alizarin Red ◽

Oil Red O

The canine has been a useful animal model for the study of fundamental mechanisms and the testing of new therapies for several human pathologies using mesenchymal stem cells (MSC). For preclinical applications, the most commonly used source of canine MSC is bone marrow. Because the amount of autologous bone marrow that can be obtained is limited, skin tissue could supply a noninvasive alternative with large quantities available for the establishment of MSC. In this study, we isolated canine MSC (cMSC) from ear skin and evaluated the expression of transcriptional factors and in vitro differentiation into multiple mesenchymal lineages. The cMSC isolated from the ear skin of a female beagle dog (6 years old) were cultured in advanced-DMEM/F12 (1:1, v/v) supplemented with 10% serum replacement at 37°C, 5% CO2 in a humidified atmosphere. The cMSC at passage 3 were analysed for expression of surface markers (CD44, CD90, and CD105) and transcriptional factors (Oct-4, Nanog, and Sox2) using flow cytometry, immunohistochemical staining and RT-PCR, respectively. Differentiations into adipocytes and osteocytes of cMSC were carried out under controlled conditions for 2 and 4 weeks and evaluated by staining (Oil Red O, von Kossa and Alizarin Red S, respectively). The cMSC were induced to differentiate into neural cells in the controlled condition for 6 h. Neuronal differentiated cMSC were evaluated by immunohistochemical staining, RT-PCR, and Western blot of specific markers of neuron, such as Î2-tubulin, microtubule associated protein (MAP-2), neuronfilament M (NF-M), nerve growth factor (NGF), and nestin. The MSC surface markers such as CD44, 90, and 105 were highly detected, and transcriptional factors (Oct-4, Nanog, and Sox2) were expressed in cMSC. Adipocyte induced cells were positive by staining with Oil Red O, and osteocytes were stained by von Kossa and Alizarin Red S. Neuronal specific markers such as Î2-tubulin, MAP-2, NF-M, NGF, and nestin were expressed in the neuron induced cMSC. In conclusion, canine ear-skin-derived MSC have the capacity for differentiation into multiple lineages and have a confirmed great capability for neuronal differentiation. Hence, canine ear skin tissue could be considered a source for applications of MSC for neuronal regeneration therapy of canine and a preclinical research model for human. This work was supported by Grant No. 2007031034040 from Bio-organ and Grant No. 200908FHT010204005 from Biogreen21.

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Cellular Composition of Calcified Bioprosthetic Heart Valves

Annals of the Russian academy of medical sciences ◽

10.15690/vramn560 ◽

2015 ◽

Vol 70 (6) ◽

pp. 662-668 ◽

Cited By ~ 5

Author(s):

Rinat Avkhadievich Mukhamadiyarov ◽

Natal'ya Vital'evna Rutkovskaya ◽

Ol'ga Dmitrievna Sidorova ◽

Leonid Semenovich Barbarash

Keyword(s):

Smooth Muscle ◽

Heart Valves ◽

Plasma Cells ◽

Giant Cells ◽

Cellular Infiltration ◽

Cellular Composition ◽

Multinucleated Giant Cells ◽

Bioprosthetic Heart Valves ◽

Calcium Deposits ◽

Multiple Cells

With the aim to assess the mechanisms of the structural dysfunctions associated with xenograft tissue calcification, we investigated the cellular composition of the explanted xenoaortic epoxy-treated bioprosthetic heart valves. In the leaflets, we revealed multiple cells with retained internal structure. Most of them located on the leaflet surface, at the areas of collagen destruction, and near calcium deposits. Monocytes were the predominant cell fraction on the leaflet surface whilst immune (macrophages, multinucleated giant cells, plasma cells, neutrophils) and connective tissue (fibroblasts, fibrocytes, endothelial, and smooth muscle cells) cells prevailed at the areas of collagen destruction and near calcium deposits. Calcification of the leaflets was accompanied by cellular infiltration, therefore suggesting that pathological mineralization may be associated with cell-mediated processes.

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Comparative Study of the L-Hydro Process and Glutaraldehyde Preservation

Asian Cardiovascular and Thoracic Annals ◽

10.1177/021849230501300302 ◽

2005 ◽

Vol 13 (3) ◽

pp. 203-207

Author(s):

Vinicius JS Nina ◽

Pablo MA Pomerantzeff ◽

Ivan SJ Casagrande ◽

David T Cheung ◽

Carlos MA Brandão ◽

...

Keyword(s):

Endothelial Cells ◽

Heart Valves ◽

Host Cells ◽

Complete Coverage ◽

Bioprosthetic Heart Valves ◽

Porcine Heart ◽

Transmission Electron ◽

Mitral Position ◽

Von Willebrand

Commercial bioprosthetic heart valves are commonly preserved in glutaraldehyde and are cytotoxic to host cells, preventing spontaneous endothelialization. The aim of this study was to demonstrate the potential for in vivo endothelialization of bioprostheses treated by the L-Hydro process which consists of mild extraction of antigenic substances and incorporation of antiinflammatory and antithrombotic agents. Seven stented porcine heart valves treated by the L-Hydro process and 3 glutaraldehyde-fixed porcine heart valves were implanted in the mitral position in juvenile sheep. The valves were evaluated by echocardiography, angiography, histology, and histochemistry. No hemodynamic differences were observed, but scanning and transmission electron microscopy showed nearly complete coverage by endothelial cells of all leaflets in the L-Hydro-treated valves after 5 months of implantation. The endothelial cells were in direct contact with the underlying collagen and expressed von Willebrand-related antigens. The surfaces of the glutaraldehyde-treated valves were covered by fibrin, macrophages, calcium, and thrombotic material; only sparse endothelial cells were observed and contact with the underlying tissue was incomplete. These data indicate that L-Hydro-treated porcine valves are capable of inducing spontaneous endothelialization.

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Thrombosis of Surgical Bioprosthetic Heart Valves-Insights from Reports to International Medical Device Vigilance Systems

The Thoracic and Cardiovascular Surgeon ◽

10.1055/s-0037-1598875 ◽

2017 ◽

Vol 65 (S 01) ◽

pp. S1-S110

Author(s):

C. Gestrich ◽

J.E. Klein ◽

B. Toctam ◽

G.D. Dürr ◽

J.M. Sinning ◽

...

Keyword(s):

Medical Device ◽

Heart Valves ◽

International Medical ◽

Bioprosthetic Heart Valves

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Role of Matrix Metalloproteinases in Degenerative Kidney Disorders

Current Medicinal Chemistry ◽

10.2174/0929867325666171205143441 ◽

2018 ◽

Vol 25 (15) ◽

pp. 1805-1816 ◽

Cited By ~ 6

Author(s):

Shifa Narula ◽

Chanderdeep Tandon ◽

Simran Tandon

Keyword(s):

Matrix Metalloproteinases ◽

Transforming Growth Factor ◽

Kidney Diseases ◽

Transforming Growth Factor Β1 ◽

Bioactive Molecules ◽

Tissue Inhibitors Of Metalloproteinases ◽

Ecm Remodeling ◽

Surface Receptors ◽

Calcium Dependent ◽

Ecm Degradation

Matrix metalloproteinases (MMPs) are members of calcium dependent-zinc containing endopeptidases that play a pivotal role in extracellular matrix (ECM) remodeling. MMPs are also known to cleave non-matrix proteins, including cell surface receptors, TNF-α, angiotensin-II, growth factors, (especially transforming growth factor-β1, ΤGF- β1) plasminogen, endothelin and other bioactive molecules. The tissue inhibitors of metalloproteinases (TIMPs) inhibit the activity of MMPs and decrease ECM degradation. Various patho-physiological conditions have been linked with the imbalance of ECM synthesis and degradation. Numerous studies have reported the significance of MMPs and TIMPs in the progression of kidney pathologies, including glomerulonephritis, diabetic nephropathy, renal cancer, and nephrolithiasis. Although dysregulated activity of MMPs could directly or indirectly lead to pathological morbidities, their contribution in disease progression is still understated. Specifically, MMP activity in the kidneys and it's relation to kidney diseases has been the subject of a limited number of investigations. Therefore, the aim of the present review is to provide an updated insight of the involvement of MMPs and TIMPs in the pathogenesis of inflammatory and degenerative kidney disorders.

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In Situ “Humanization” of Porcine Bioprostheses: Demonstration of Tendon Bioprostheses Conversion into Human ACL and Possible Implications for Heart Valve Bioprostheses

Bioengineering ◽

10.3390/bioengineering8010010 ◽

2021 ◽

Vol 8 (1) ◽

pp. 10

Author(s):

Uri Galili ◽

Kevin R. Stone

Keyword(s):

Extracellular Matrix ◽

Heart Valves ◽

Cruciate Ligament ◽

Anticoagulation Therapy ◽

Young Patients ◽

Collagen Fibers ◽

Reconstruction Process ◽

Bioprosthetic Heart Valves ◽

Porcine Tissue ◽

Anterior Cruciate

This review describes the first studies on successful conversion of porcine soft-tissue bioprostheses into viable permanently functional tissue in humans. This process includes gradual degradation of the porcine tissue, with concomitant neo-vascularization and reconstruction of the implanted bioprosthesis with human cells and extracellular matrix. Such a reconstruction process is referred to in this review as “humanization”. Humanization was achieved with porcine bone-patellar-tendon-bone (BTB), replacing torn anterior-cruciate-ligament (ACL) in patients. In addition to its possible use in orthopedic surgery, it is suggested that this humanization method should be studied as a possible mechanism for converting implanted porcine bioprosthetic heart-valves (BHV) into viable tissue valves in young patients. Presently, these patients are only implanted with mechanical heart-valves, which require constant anticoagulation therapy. The processing of porcine bioprostheses, which enables humanization, includes elimination of α-gal epitopes and partial (incomplete) crosslinking with glutaraldehyde. Studies on implantation of porcine BTB bioprostheses indicated that enzymatic elimination of α-gal epitopes prevents subsequent accelerated destruction of implanted tissues by the natural anti-Gal antibody, whereas the partial crosslinking by glutaraldehyde molecules results in their function as “speed bumps” that slow the infiltration of macrophages. Anti-non gal antibodies produced against porcine antigens in implanted bioprostheses recruit macrophages, which infiltrate at a pace that enables slow degradation of the porcine tissue, neo-vascularization, and infiltration of fibroblasts. These fibroblasts align with the porcine collagen-fibers scaffold, secrete their collagen-fibers and other extracellular-matrix (ECM) components, and gradually replace porcine tissues degraded by macrophages with autologous functional viable tissue. Porcine BTB implanted in patients completes humanization into autologous ACL within ~2 years. The similarities in cells and ECM comprising heart-valves and tendons, raises the possibility that porcine BHV undergoing a similar processing, may also undergo humanization, resulting in formation of an autologous, viable, permanently functional, non-calcifying heart-valves.

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