ABSTRACT
Adverse cardiac remodeling after myocardial infarction (MI) causes impaired ventricular function and heart failure. Histopathological characterization is commonly used to detect the location, size and shape of MI sites. However, the information about chemical composition, physical structure and molecular mobility of peri- and infarct zones post-MI is rather limited. The main objective of this work was to explore the spatiotemporal biochemical and biophysical alterations of key cardiac components post-MI. The FTIR spectra of healthy and remote myocardial tissue shows amides A, I, II and III associated with proteins in freeze-died tissue as major absorptions bands. In infarcted myocardium, the spectrum of these main absorptions was deeply altered. FITR evidenced an increase of the amide A band and the distinct feature of the collagen specific absorption band at 1338cm-1 in the infarct area at 21days post-MI. At 21days post-MI, it also appears an important shift of amide I from 1646cm-1 to 1637cm-1 that suggests the predominance of the triple helical conformation in the proteins. The new spectra bands also indicate an increase in proteoglycans, residues of carbohydrates in proteins and polysaccharides in ischemic areas. Thermal analysis indicates a deep increase of unfreezable water/freezable water in peri- and infarcted tissues. In infarcted tissue is evidenced the impairment of myofibrillar proteins thermal profile and the emergence of a new structure. In conclusion, our results indicate a profound evolution of protein secondary structures in association with collagen deposition and reorganization of water involved in the scar maturation of peri- and infarct zones post-MI.
Subject(s)
Muscle Proteins/metabolism , Myocardial Infarction/metabolism , Myocardium/metabolism , Ventricular Remodeling , Animals , Male , Mice , Myocardial Infarction/pathology , Myocardial Infarction/physiopathology , Myocardium/pathology , Protein Structure, Secondary , Spectroscopy, Fourier Transform Infrared/methodsABSTRACT
A major challenge of cardiac tissue engineering is directing cells to establish the physiological structure and function of the myocardium being replaced. Our aim was to examine the effect of electrical stimulation on the cardiodifferentiation potential of cardiac adipose tissue-derived progenitor cells (cardiac ATDPCs). Three different electrical stimulation protocols were tested; the selected protocol consisted of 2 ms monophasic square-wave pulses of 50 mV/cm at 1 Hz over 14 days. Cardiac and subcutaneous ATDPCs were grown on biocompatible patterned surfaces. Cardiomyogenic differentiation was examined by real-time PCR and immunocytofluorescence. In cardiac ATDPCs, MEF2A and GATA-4 were significantly upregulated at day 14 after stimulation, while subcutaneous ATDPCs only exhibited increased Cx43 expression. In response to electrical stimulation, cardiac ATDPCs elongated, and both cardiac and subcutaneous ATDPCs became aligned following the linear surface pattern of the construct. Cardiac ATDPC length increased by 11.3%, while subcutaneous ATDPC length diminished by 11.2% (p = 0.013 and p = 0.030 vs unstimulated controls, respectively). Compared to controls, electrostimulated cells became aligned better to the patterned surfaces when the pattern was perpendicular to the electric field (89.71 ± 28.47º for cardiac ATDPCs and 92.15 ± 15.21º for subcutaneous ATDPCs). Electrical stimulation of cardiac ATDPCs caused changes in cell phenotype and genetic machinery, making them more suitable for cardiac regeneration approaches. Thus, it seems advisable to use electrical cell training before delivery as a cell suspension or within engineered tissue.
Subject(s)
Adipose Tissue/cytology , Myocardium/metabolism , Stem Cells/metabolism , Tissue Engineering/methods , Biocompatible Materials/chemistry , Cell Differentiation , Cells, Cultured , Electric Stimulation Therapy , Humans , Ions/chemistry , Microscopy, Fluorescence , Myocardium/pathology , Myocytes, Cardiac/cytology , Phalloidine/chemistry , Phenotype , Real-Time Polymerase Chain Reaction , Regeneration , Signal Transduction , Stem Cells/cytology , Up-RegulationABSTRACT
La regeneración del corazón dañado ha sido objeto de intensa investigación durante la década pasada. Las diferentes estrategias que han sido desarrrolladas (como por ejemplo la terapia celular basada en células madre (ES)), esclarecen la información acerca de las moléculas y los factores de transcripción involucrados en las cardiomiogénesis. Sin embargo, todavía no es posible programar eficientemente las ES para que se desarrollen a cardiomiocitos. Los mecanismos celulares y moleculares inherentes en el desarrollo embrionario del corazón, así como las interconexiones entre ellos, pueden aportar datos acerca de las rutas bioquímicas necesarias para la diferenciación de las ES embrionarias a células cardíacas. Nosotros proponemos que un modelo cuantitativo que puede servir para descifrar las elaboradas rutas involucradas en la cardiomiogénesis. Esta aproximación podría revelar la etiología de los defectos cardíacos y permitiría producir cardiomiocitos con propósitos clínicos en la regeneracíon y la toxicología entre otros (AU)
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Subject(s)
Humans , Heart/embryology , Cell- and Tissue-Based Therapy/methods , Myocytes, Cardiac , Calcium Signaling/physiology , Regeneration/physiology , Embryonic Stem Cells , Gene Expression , Cell Differentiation/physiologyABSTRACT
BACKGROUND: Recent reports refute the classic paradigm by which human heart is unable to repair itself following disease or injury. Cardiac and noncardiac stem cells with cardiac regeneration potential have been documented. We studied whether untreated mesenchymal stem cells express markers of cardiomyogenic lineage in vitro. METHODS: Mesenchymal stem cells were obtained from human iliac crest marrow aspirates. Cells were isolated and characterized using flow cytometry by surface expression of CD105, CD166, CD29, CD44, CD14, and CD34. To evaluate their cardiomyogenic potential, presence of cardiac proteins (cardiac troponin I, sarcomeric alpha-actinin, beta myosin heavy chain (beta-MyHC), connexin-43, and SERCA-2), and transcription factors (GATA-4) were assessed. RESULTS: Mesenchymal stem cells expressed CD105 (4.25 +/- 0.35), CD166 (27.83 +/- 1.89), and CD29 (9.4 +/- 0.57) and were negative for CD34, CD14, and CD45. In absence of additional stimuli in the culture media, these cells expressed connexin-43, alpha-actinin, and GATA-4, and were negative for SERCA-2, cardiac troponin I, and beta-MyHC. CONCLUSIONS: Human adult mesenchymal stem cells spontaneously exhibit markers of cardiac phenotype in vitro. In the appropiate myocardial environment, these cells may transdifferentiate into mature cardiomyocytes.
