114 research outputs found
Heart stem cells: hope or myth?
The search and study of endogenous heart repair remains an urgent issue in modern regenerative medicine. It is generally accepted that the human heart has a limited regenerative potential, but recent studies show that functionally significant regeneration is possible. However, the mechanisms underlying these processes remain poorly understood. In the heart, there are populations of resident mesenchymal cells that have some properties of stem cells that carry certain markers, such as c-kit+, Sca-1, etc. The ability of these cells to differentiate directly into cardiomyocytes remains controversial, but their use in clinical trials has shown improved cardiac function in patients with myocardial infarction. Currently, approaches are being developed to use, mainly, induced pluripotent stem cells as a promising regenerative therapy, but the cardioprotective role of cardiac mesenchymal cells remains the subject of active study due to their paracrine signaling
Magnetron plasma mediated immobilization of hyaluronic acid for the development of functional double-sided biodegradable vascular graft
The clinical need for vascular grafts is associated with cardiovascular
diseases frequently leading to fatal outcomes. Artificial vessels based on
bioresorbable polymers can replace the damaged vascular tissue or create a
bypass path for blood flow while stimulating regeneration of a blood vessel in
situ. However, the problem of proper conditions for the cells to grow on the
vascular graft from the adventitia while maintaining its mechanical integrity
of the luminal surface remains a challenge. In this work, we propose a
two-stage technology for processing electrospun vascular graft from
polycaprolactone, which consists of plasma treatment and subsequent
immobilization of hyaluronic acid on its surface producing thin double-sided
graft with one hydrophilic and one hydrophobic side. Plasma modification
activates the polymer surfaces and produces a thin layer for linker-free
immobilization of bioactive molecules, thereby producing materials with unique
properties. The proposed modification does not affect the morphology or
mechanical properties of the graft and improves cell adhesion. The proposed
approach can potentially be used for various biodegradable polymers such as
polylactic acid, polyglycolide, and their copolymers and blends, with a
hydrophilic inner surface and a hydrophobic outer surface
Transfer of synthetic human chromosome into human induced pluripotent stem cells for biomedical applications
Alphoid(tetO)-type human artificial chromosome (HAC) has been recently synthetized as a novel class of gene delivery vectors for induced pluripotent stem cell (iPSC)-based tissue replacement therapeutic approach. This HAC vector was designed to deliver copies of genes into patients with genetic diseases caused by the loss of a particular gene function. The alphoid(tetO)-HAC vector has been successfully transferred into murine embryonic stem cells (ESCs) and maintained stably as an independent chromosome during the proliferation and differentiation of these cells. Human ESCs and iPSCs have significant differences in culturing conditions and pluripotency state in comparison with the murine naΓ―ve-type ESCs and iPSCs. To date, transferring alphoid(tetO)-HAC vector into human iPSCs (hiPSCs) remains a challenging task. In this study, we performed the microcell-mediated chromosome transfer (MMCT) of alphoid(tetO)-HAC expressing the green fluorescent protein into newly generated hiPSCs. We used a recently modified MMCT method that employs an envelope protein of amphotropic murine leukemia virus as a targeting cell fusion agent. Our data provide evidence that a totally artificial vector, alphoid(tetO)-HAC, can be transferred and maintained in human iPSCs as an independent autonomous chromosome without affecting pluripotent properties of the cells. These data also open new perspectives for implementing alphoid(tetO)-HAC as a gene therapy tool in future biomedical applications
Protective Role of Mytilus edulis Hydrolysate in Lipopolysaccharide-Galactosamine Acute Liver Injury
Acute liver injury in its terminal phase trigger systemic inflammatory response syndrome with multiple organ failure. An uncontrolled inflammatory reaction is difficult to treat and contributes to high mortality. Therefore, to solve this problem a search for new therapeutic approaches remains urgent. This study aimed to explore the protective effects of M. edulis hydrolysate (N2-01) against Lipopolysaccharide-D-Galactosamine (LPS/D-GalN)-induced murine acute liver injure and the underlying mechanisms. N2-01 analysis, using Liquid Chromatography Mass Spectrometry (LCMS) metabolomic and proteomic platforms, confirmed composition, molecular-weight distribution, and high reproducibility between M. edulis hydrolysate manufactured batches. N2-01 efficiently protected mice against LPS/D-GalN-induced acute liver injury. The most prominent result (100% survival rate) was obtained by the constant subcutaneous administration of small doses of the drug. N2-01 decreased Vascular Cell Adhesion Molecule-1 (VCAM-1) expression from 4.648 Β± 0.445 to 1.503 Β± 0.091 Mean Fluorescence Intensity (MFI) and Interleukin-6 (IL-6) production in activated Human Umbilical Vein Endothelial Cells (HUVECs) from 7.473 Β± 0.666 to 2.980 Β± 0.130Β ng/ml in vitro. The drug increased Nitric Oxide (NO) production by HUVECs from 27.203 Β± 2.890 to 69.200 Β± 4.716 MFI but significantly decreased inducible Nitric Oxide Synthase (iNOS) expression from 24.030 Β± 2.776 to 15.300 Β± 1.290 MFI and NO production by murine peritoneal lavage cells from 6.777 Β± 0.373 Β΅m to 2.175 Β± 0.279Β Β΅m. The capability of the preparation to enhance the endothelium barrier function and to reduce vascular permeability was confirmed in Electrical Cell-substrate Impedance Sensor (ECIS) test in vitro and Miles assay in vivo. These results suggest N2-01 as a promising agent for treating a wide range of conditions associated with uncontrolled inflammation and endothelial dysfunction
A short G1 phase is an intrinsic determinant of naΓ―ve embryonic stem cell pluripotency
AbstractA short G1 phase is a characteristic feature of mouse embryonic stem cells (ESCs). To determine if there is a causal relationship between G1 phase restriction and pluripotency, we made use of the Fluorescence Ubiquitination Cell Cycle Indicator (FUCCI) reporter system to FACS-sort ESCs in the different cell cycle phases. Hence, the G1 phase cells appeared to be more susceptible to differentiation, particularly when ESCs self-renewed in the naΓ―ve state of pluripotency. Transitions from ground to naΓ―ve, then from naΓ―ve to primed states of pluripotency were associated with increased durations of the G1 phase, and cyclin E-mediated alteration of the G1/S transition altered the balance between self-renewal and differentiation. LIF withdrawal resulted in a lengthening of the G1 phase in naΓ―ve ESCs, which occurred prior to the appearance of early lineage-specific markers, and could be reversed upon LIF supplementation. We concluded that the short G1 phase observed in murine ESCs was a determinant of naΓ―ve pluripotency and was partially under the control of LIF signaling
Π£ΡΠ°ΡΡΠΈΠ΅ ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΈΠΎΠ½Π½ΠΎΠ³ΠΎ ΡΠ°ΠΊΡΠΎΡΠ° ZBTB16 Π² ΠΏΡΠΎΡΠ΅ΡΡΠ°Ρ ΡΠΈΠ·ΠΈΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΠΎΠ±ΡΠ°Π·ΠΎΠ²Π°Π½ΠΈΡ ΠΊΠΎΡΡΠ½ΠΎΠΉ ΡΠΊΠ°Π½ΠΈ ΠΈ ΠΏΡΠΈ ΠΏΠ°ΡΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠΉ ΠΊΠ°Π»ΡΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ Π°ΠΎΡΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»Π°ΠΏΠ°Π½Π°
Degenerative calcific aortic valve stenosis is the most common type of heart valve disease in the Western world. Patients with severe stenosis are associated with 50 percent chance of mortality within two years in the absence of intervention. Surgical interventions are the only treatment method for severe calcific aortic valve stenosis to date. Pharmacological approaches have so far failed to affect the course of the disease. Thus, there is an urgent need to develop novel treatment strategies that could slow down the progression of the stenosis. ZBTB16 is a zinc finger protein with N-term BTB/POZ domain (protein-protein interaction motif) and 9 zinc finger domains (DNA binding motif) in C-term. There is growing evidence proving the participation of ZBTB16 in skeletal development. ZBTB16 has been shown to play a role in the specification of limb and axial skeleton patterning. Moreover, the expression of ZBTB16 is increased in patients with ectopic bone formation. Nowadays, the evidence supports that the mechanisms that play key roles in the formation of bone tissue are similar to the processes occurring during the development of ectopic ossification of the aortic valve. Thus, it can be assumed that ZBTB16 is heavily involved in osteogenic transformation in the aortic valve. Understanding similarities and differences in the mechanisms that mediate osteogenic differentiation of stem cells during bone formation and pathological ossification of tissues can help to find the ways to control the osteogenic differentiation in the human body. The aim of this review is to summarize data on the role of ZBTB16 and its products in the regulation of differentiation and proliferation of cells involved in osteogenesis and in the development of ectopic calcification of the aortic valve. The study of the dynamic changes of ZBTB16 expression in aortic valve calcification is a new and relevant study field.ΠΠ΅Π³Π΅Π½Π΅ΡΠ°ΡΠΈΠ²Π½ΡΠΉ ΠΊΠ°Π»ΡΡΠΈΠ½ΠΈΡΠΎΠ²Π°Π½Π½ΡΠΉ ΡΡΠ΅Π½ΠΎΠ· Π°ΠΎΡΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»Π°ΠΏΠ°Π½Π° β Π½Π°ΠΈΠ±ΠΎΠ»Π΅Π΅ ΡΠ°ΡΠΏΡΠΎΡΡΡΠ°Π½Π΅Π½Π½Π°Ρ Π² ΠΌΠΈΡΠ΅ ΠΏΠ°ΡΠΎΠ»ΠΎΠ³ΠΈΡ ΠΊΠ»Π°ΠΏΠ°Π½ΠΎΠ² ΡΠ΅ΡΠ΄ΡΠ°. ΠΡΠΈ ΡΠ°Π·Π²ΠΈΡΠΈΠΈ ΠΊΠ°Π»ΡΡΠΈΠ½ΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠ³ΠΎ ΡΡΠ΅Π½ΠΎΠ·Π° Π°ΠΎΡΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»Π°ΠΏΠ°Π½Π° ΠΏΡΠΎΠ³Π½ΠΎΠ· Π΄Π²ΡΡ
Π»Π΅ΡΠ½Π΅ΠΉ Π²ΡΠΆΠΈΠ²Π°Π΅ΠΌΠΎΡΡΠΈ Π±Π΅Π· Ρ
ΠΈΡΡΡΠ³ΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ Π²ΠΌΠ΅ΡΠ°ΡΠ΅Π»ΡΡΡΠ²Π° ΡΠΎΡΡΠ°Π²Π»ΡΠ΅Ρ 50%. ΠΠ΄ΠΈΠ½ΡΡΠ²Π΅Π½Π½ΡΠΌ Π½Π° ΡΠ΅Π³ΠΎΠ΄Π½ΡΡΠ½ΠΈΠΉ Π΄Π΅Π½Ρ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ Π»Π΅ΡΠ΅Π½ΠΈΡ ΡΡΠΆΠ΅Π»ΠΎΠΉ ΠΊΠ°Π»ΡΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ ΠΊΠ»Π°ΠΏΠ°Π½Π° Π²ΡΡΡΡΠΏΠ°Π΅Ρ Ρ
ΠΈΡΡΡΠ³ΠΈΡΠ΅ΡΠΊΠΎΠ΅ Π²ΠΌΠ΅ΡΠ°ΡΠ΅Π»ΡΡΡΠ²ΠΎ. Π€Π°ΡΠΌΠ°ΠΊΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈΠ΅ ΠΏΠΎΠ΄Ρ
ΠΎΠ΄Ρ Π΄ΠΎ ΡΠΈΡ
ΠΏΠΎΡ Π½Π΅ ΡΠΌΠΎΠ³Π»ΠΈ ΠΈΠ·ΠΌΠ΅Π½ΠΈΡΡ ΡΠ΅ΡΠ΅Π½ΠΈΠ΅ Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΡ, ΠΏΠΎΡΡΠΎΠΌΡ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠΊΠ° Π½ΠΎΠ²ΡΡ
ΡΡΡΠ°ΡΠ΅Π³ΠΈΠΉ Π»Π΅ΡΠ΅Π½ΠΈΡ, Π·Π°ΠΌΠ΅Π΄Π»ΡΡΡΠΈΡ
ΡΠ°Π·Π²ΠΈΡΠΈΠ΅ ΡΡΠ΅Π½ΠΎΠ·Π° Π°ΠΎΡΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»Π°ΠΏΠ°Π½Π°, ΠΏΡΠ΅Π΄ΡΡΠ°Π²Π»ΡΠ΅Ρ Π°ΠΊΡΡΠ°Π»ΡΠ½ΡΡ ΠΊΠ»ΠΈΠ½ΠΈΡΠ΅ΡΠΊΡΡ ΠΏΠΎΡΡΠ΅Π±Π½ΠΎΡΡΡ. ΠΠ΅Π»ΠΎΠΊ ZBTB16 ΡΠ²Π»ΡΠ΅ΡΡΡ ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΈΠΎΠ½Π½ΡΠΌ ΡΠ°ΠΊΡΠΎΡΠΎΠΌ Ρ N-ΠΊΠΎΠ½ΡΠ΅Π²ΡΠΌ BTB/POZ-Π΄ΠΎΠΌΠ΅Π½ΠΎΠΌ Π΄Π»Ρ Π±Π΅Π»ΠΎΠΊ-Π±Π΅Π»ΠΊΠΎΠ²ΠΎΠ³ΠΎ Π²Π·Π°ΠΈΠΌΠΎΠ΄Π΅ΠΉΡΡΠ²ΠΈΡ ΠΈ Π΄Π΅Π²ΡΡΡΡ C-ΠΊΠΎΠ½ΡΠ΅Π²ΡΠΌΠΈ Π΄ΠΎΠΌΠ΅Π½Π°ΠΌΠΈ ΡΠΈΠΏΠ° ΡΠΈΠ½ΠΊΠΎΠ²ΡΠΉ ΠΏΠ°Π»Π΅Ρ Π΄Π»Ρ ΡΠ²ΡΠ·ΡΠ²Π°Π½ΠΈΡ ΠΠΠ. Π Π»ΠΈΡΠ΅ΡΠ°ΡΡΡΠ΅ ΠΏΡΠ΅Π΄ΡΡΠ°Π²Π»Π΅Π½Ρ Π΄Π°Π½Π½ΡΠ΅ ΠΎΠ± ΡΡΠ°ΡΡΠΈΠΈ ZBTB16 Π² ΡΠ°Π·Π²ΠΈΡΠΈΠΈ ΡΠΊΠ΅Π»Π΅ΡΠ°. ΠΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ ZBTB16 ΠΈΠ³ΡΠ°Π΅Ρ ΡΠΎΠ»Ρ Π² ΡΠΏΠ΅ΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ ΠΏΠ°ΡΡΠ΅ΡΠ½ΠΎΠ² Π°ΠΊΡΠΈΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΡΠΊΠ΅Π»Π΅ΡΠ° ΠΈ ΠΊΠΎΠ½Π΅ΡΠ½ΠΎΡΡΠ΅ΠΉ. ΠΡΠΎΠΌΠ΅ ΡΠΎΠ³ΠΎ, ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΡ ZBTB16 ΠΏΠΎΠ²ΡΡΠ΅Π½Π° Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ
ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ², ΡΡΡΠ°Π΄Π°ΡΡΠΈΡ
ΡΠΊΡΠΎΠΏΠΈΡΠ΅ΡΠΊΠΈΠΌ ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΠΊΠΎΡΡΠ½ΠΎΠΉ ΡΠΊΠ°Π½ΠΈ. ΠΠ° ΡΠ΅Π³ΠΎΠ΄Π½ΡΡΠ½ΠΈΠΉ Π΄Π΅Π½Ρ ΠΌΡ ΠΈΠΌΠ΅Π΅ΠΌ ΠΌΠ½ΠΎΠΆΠ΅ΡΡΠ²ΠΎ ΠΏΠΎΠ΄ΡΠ²Π΅ΡΠΆΠ΄Π΅Π½ΠΈΠΉ ΡΠΎΠΌΡ, ΡΡΠΎ ΠΊΠ»ΡΡΠ΅Π²ΡΠ΅ ΠΌΠ΅Ρ
Π°Π½ΠΈΠ·ΠΌΡ ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΊΠΎΡΡΠ½ΠΎΠΉ ΡΠΊΠ°Π½ΠΈ Π² Π½ΠΎΡΠΌΠ΅, ΡΡ
ΠΎΠ΄Π½Ρ Ρ ΠΏΡΠΎΡΠ΅ΡΡΠ°ΠΌΠΈ ΠΏΡΠΈ ΡΠΊΡΠΎΠΏΠΈΡΠ΅ΡΠΊΠΎΠΉ ΠΎΡΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ ΡΠΊΠ°Π½Π΅ΠΉ Π°ΠΎΡΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»Π°ΠΏΠ°Π½Π°. Π’Π°ΠΊΠΈΠΌ ΠΎΠ±ΡΠ°Π·ΠΎΠΌ, ΠΌΠΎΠΆΠ½ΠΎ ΡΠ΄Π΅Π»Π°ΡΡ ΠΏΡΠ΅Π΄ΠΏΠΎΠ»ΠΎΠΆΠ΅Π½ΠΈΠ΅ ΠΎ Π·Π½Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎΠΌ ΡΡΠ°ΡΡΠΈΠΈ ZBTB16 ΠΈ Π² ΠΎΡΡΠ΅ΠΎΠ³Π΅Π½Π½ΠΎΠΉ ΡΡΠ°Π½ΡΡΠΎΡΠΌΠ°ΡΠΈΠΈ ΠΊΠ»Π΅ΡΠΎΠΊ ΠΊΠ»Π°ΠΏΠ°Π½Π° Π°ΠΎΡΡΡ. ΠΠΎΠ½ΠΈΠΌΠ°Π½ΠΈΠ΅ ΡΡ
ΠΎΠ΄ΡΡΠ² ΠΈ ΡΠ°Π·Π»ΠΈΡΠΈΠΉ ΠΌΠ΅Ρ
Π°Π½ΠΈΠ·ΠΌΠΎΠ², ΠΎΠΏΠΎΡΡΠ΅Π΄ΡΡΡΠΈΡ
ΠΎΡΡΠ΅ΠΎΠ³Π΅Π½Π½ΡΡ Π΄ΠΈΡΡΠ΅ΡΠ΅Π½ΡΠΈΡΠΎΠ²ΠΊΡ ΠΊΠ»Π΅ΡΠΎΠΊ Π²ΠΎ Π²ΡΠ΅ΠΌΡ ΡΠΈΠ·ΠΈΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΊΠΎΡΡΠΈ ΠΈ ΠΏΠ°ΡΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠΉ ΠΎΡΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ ΡΠΊΠ°Π½Π΅ΠΉ, ΠΌΠΎΠΆΠ΅Ρ Π΄Π°ΡΡ ΠΏΡΠ΅Π΄ΠΏΠΎΡΡΠ»ΠΊΠΈ Π΄Π»Ρ Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎΡΡΠΈ ΡΠΏΡΠ°Π²Π»Π΅Π½ΠΈΡ ΠΏΡΠΎΡΠ΅ΡΡΠ°ΠΌΠΈ ΠΎΡΡΠ΅ΠΎΠ³Π΅Π½Π½ΠΎΠΉ Π΄ΠΈΡΡΠ΅ΡΠ΅Π½ΡΠΈΡΠΎΠ²ΠΊΠΈ Π² ΠΎΡΠ³Π°Π½ΠΈΠ·ΠΌΠ΅ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ°. Π¦Π΅Π»ΡΡ Π΄Π°Π½Π½ΠΎΠ³ΠΎ ΠΎΠ±Π·ΠΎΡΠ° ΡΡΠ°Π»ΠΎ ΠΎΠ±ΠΎΠ±ΡΠ΅Π½ΠΈΠ΅ ΡΠ²Π΅Π΄Π΅Π½ΠΈΠΉ ΠΎ ΡΠΎΠ»ΠΈ ZBTB16 ΠΈ Π΅Π³ΠΎ ΠΏΡΠΎΠ΄ΡΠΊΡΠΎΠ² Π² ΡΠ΅Π³ΡΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΠΈ Π΄ΠΈΡΡΠ΅ΡΠ΅Π½ΡΠΈΡΠΎΠ²ΠΊΠΈ ΠΈ ΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠΈ ΠΊΠ»Π΅ΡΠΎΠΊ, ΡΡΠ°ΡΡΠ²ΡΡΡΠΈΡ
Π² ΠΏΡΠΎΡΠ΅ΡΡΠ°Ρ
ΡΠΈΠ·ΠΈΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΠΎΡΡΠ΅ΠΎΠ³Π΅Π½Π΅Π·Π° ΠΈ ΠΏΡΠΈ ΡΠΊΡΠΎΠΏΠΈΡΠ΅ΡΠΊΠΎΠΉ ΠΊΠ°Π»ΡΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ ΡΠΊΠ°Π½Π΅ΠΉ, Π² ΡΠΎΠΌ ΡΠΈΡΠ»Π΅ Π°ΠΎΡΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»Π°ΠΏΠ°Π½Π°. ΠΠ·ΡΡΠ΅Π½ΠΈΠ΅ Π΄ΠΈΠ½Π°ΠΌΠΈΡΠ΅ΡΠΊΠΎΠΉ Π²Π°ΡΠΈΠ°Π±Π΅Π»ΡΠ½ΠΎΡΡΠΈ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ ZBTB16 Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ
Π°ΠΎΡΡΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»Π°ΠΏΠ°Π½Π° ΠΏΡΠΈ ΠΊΠ°Π»ΡΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ ΡΠΊΠ°Π½ΠΈ ΡΠ²Π»ΡΠ΅ΡΡΡ Π½ΠΎΠ²ΡΠΌ ΠΈ Π°ΠΊΡΡΠ°Π»ΡΠ½ΡΠΌ Π½Π°ΠΏΡΠ°Π²Π»Π΅Π½ΠΈΠ΅ΠΌ
Inflammation and Mechanical Stress Stimulate Osteogenic Differentiation of Human Aortic Valve Interstitial Cells
Background: Aortic valve calcification is an active proliferative process, where interstitial cells of the valve transform into either myofibroblasts or osteoblast-like cells causing valve deformation, thickening of cusps and finally stenosis. This process may be triggered by several factors including inflammation, mechanical stress or interaction of cells with certain components of extracellular matrix. The matrix is different on the two sides of the valve leaflets. We hypothesize that inflammation and mechanical stress stimulate osteogenic differentiation of human aortic valve interstitial cells (VICs) and this may depend on the side of the leaflet.Methods: Interstitial cells isolated from healthy and calcified human aortic valves were cultured on collagen or elastin coated plates with flexible bottoms, simulating the matrix on the aortic and ventricular side of the valve leaflets, respectively. The cells were subjected to 10% stretch at 1 Hz (FlexCell bioreactor) or treated with 0.1 ΞΌg/ml lipopolysaccharide, or both during 24 h. Gene expression of myofibroblast- and osteoblast-specific genes was analyzed by qPCR. VICs cultured in presence of osteogenic medium together with lipopolysaccharide, 10% stretch or both for 14 days were stained for calcification using Alizarin Red.Results: Treatment with lipopolysaccharide increased expression of osteogenic gene bone morphogenetic protein 2 (BMP2) (5-fold increase from control; p = 0.02) and decreased expression of mRNA of myofibroblastic markers: Ξ±-smooth muscle actin (ACTA2) (50% reduction from control; p = 0.0006) and calponin (CNN1) (80% reduction from control; p = 0.0001) when cells from calcified valves were cultured on collagen, but not on elastin. Mechanical stretch of VICs cultured on collagen augmented the effect of lipopolysaccharide. Expression of periostin (POSTN) was inhibited in cells from calcified donors after treatment with lipopolysaccharide on collagen (70% reduction from control, p = 0.001), but not on elastin. Lipopolysaccharide and stretch both enhanced the pro-calcific effect of osteogenic medium, further increasing the effect when combined for cells cultured on collagen, but not on elastin.Conclusion: Inflammation and mechanical stress trigger expression of osteogenic genes in VICs in a side-specific manner, while inhibiting the myofibroblastic pathway. Stretch and lipopolysaccharide synergistically increase calcification
Generation of two iPSC lines (FAMRCi007-A and FAMRCi007-B) from patient with Emery-Dreifuss muscular dystrophy and heart rhythm abnormalities carrying genetic variant LMNA p.Arg249Gln.
Human iPSC lines were generated from peripheral blood mononuclear cells of patient carrying LMNA mutation associated with EmeryβDreifuss muscular dystrophy accompanied by atrioventricular block and paroxysmal atrial fibrillation. Reprogramming factors OCT4, KLF4, SOX2, CMYC were delivered using Sendai virus transduction. iPSCs were characterized in order to prove the pluripotency markers expression, normal karyotype, ability to differentiate into three embryonic germ layers. Generated iPSC lines would be useful model to investigate disease development associated with genetic variants in LMNA gene
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