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Spatial and Temporal Analysis of Extracellular Matrix Proteins in the Developing Murine Heart: A Blueprint for Regeneration.
MedLine Citation:
PMID:  23273220     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
The extracellular matrix (ECM) of the embryonic heart guides assembly and maturation of cardiac cell types and thus may serve as a useful template, or blueprint, for fabrication of scaffolds for cardiac tissue engineering. Surprisingly, characterization of the ECM with cardiac development is scattered and fails to comprehensively reflect spatiotemporal dynamics making it difficult to apply to tissue engineering efforts. The objective of this work was to define a blueprint of the spatiotemporal organization, localization, and relative amount of four essential ECM proteins, collagen types I and IV (COLI, COLIV), elastin (ELN), and fibronectin (FN) in the left ventricle of the murine heart at embryonic stages E12.5, E14.5, and E16.5 and 2 days postnatal (P2). Second harmonic generation identified fibrillar collagens at E14.5, with increasing density over time. Subsequently, immunohistochemistry was used to compare the spatial distribution, organization and relative amounts of each ECM protein. COLIV was found throughout the developing heart, progressing in amount and organization from E12.5 to P2. The amount of COLI was greatest at E12.5 particularly within the epicardium. For all stages FN was present in the epicardium (except E16.5), myocardium (except E12.5), and endocardium. ELN remained relatively constant in appearance and amount throughout the developmental stages except for a transient increase at E16.5. Expression of ECM mRNA was determined using quantitative polymerase chain reaction (qPCR) and allowed for comparison of amounts of ECM molecules at each time point. Generally, COLI and COLIII mRNA expression levels were comparatively high, while COLIV, LN and FN were expressed at intermediate levels throughout the time period studied. Interestingly, levels of ELN mRNA were relatively low at early time points (E12.5), but increased significantly by P2. Thus, we identified changes in the spatial and temporal localization of primary ECM of the developing ventricle. This characterization can serve as a blueprint for fabrication techniques, which we illustrate by using multiphoton excitation photochemistry to create a synthetic scaffold based on COLIV organization at P2. Similarly fabricated scaffolds, generated using ECM components, could be utilized for ventricular repair.
Authors:
Kevin Hanson; Jangwook Philip Jung; Quyen Tran; Shao-Pu Hsu; Rioko Iida; Visar Ajeti; Paul Campagnola; Kevin W Eliceiri; Jayne M Squirrell; Gary Lyons; Brenda Ogle
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-12-31
Journal Detail:
Title:  Tissue engineering. Part A     Volume:  -     ISSN:  1937-335X     ISO Abbreviation:  Tissue Eng Part A     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-31     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101466659     Medline TA:  Tissue Eng Part A     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
University of Wisconsin-Madison, Biomedical Engineering, Madison, Wisconsin, United States; kphanson@wisc.edu.
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