Fluorescence microscopic distinction between elastin and collagen

Puchtler, Holde; Waldrop, Faye Sweat; Valentine, Linda S.

doi:10.1007/bf00303661

Cited by 28 publications

(5 citation statements)

References 34 publications

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“…Puchtler et al (1967) first demonstrated that collagen could be distinguished using fluorescence microscopy and picro-sirius red staining, later using their technique to distinguish between collagen and elastin (Puchtler et al 1973;Rodgers et al 1967). Dolber and Spach (1987) introduced an improved method in which tissue sections are first treated with 0.2% phosphomolybidic acid (PMA) for 1-5 min, followed by the standard 90 min staining in 0.1% Sirius red F3BA in saturated aqueous picric acid.…”

Section: Transmural Collagen Concentration Distribution Determinationmentioning

confidence: 97%

Prediction of extracellular matrix stiffness in engineered heart valve tissues based on nonwoven scaffolds

Engelmayr

Sacks

2007

Biomech Model Mechanobiol

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The in vitro development of tissue engineered heart valves (TEHV) exhibiting appropriate structural and mechanical characteristics remains a significant challenge. An important step yet to be addressed is establishing the relationship between scaffold and extracellular matrix (ECM) mechanical properties. In the present study, a composite beam model accounting for nonwoven scaffold-ECM coupling and the transmural collagen concentration distribution was developed, and utilized to retrospectively estimate the ECM effective stiffness in TEHV specimens incubated under static and cyclic flexure conditions (Engelmayr Jr et~al. in Biomaterials 26(2):175-187 2005). The ECM effective stiffness was expressed as the product of the local collagen concentration and the collagen specific stiffness (i.e., stiffness/concentration), and was related to the overall TEHV effective stiffness via an empirically determined scaffold-ECM coupling parameter and measured transmural collagen concentration distributions. The scaffold-ECM coupling parameter was determined by flexural mechanical testing of polyacrylamide gels (i.e., ECM analogs) of variable stiffness and associated scaffold-polyacrylamide gel composites (i.e., engineered tissue analogs). The transmural collagen concentration distributions were quantified from fluorescence micrographs of picro-sirius red stained TEHV sections. As suggested by a previous structural model of the nonwoven scaffold (Engelmayr Jr and Sacks in J Biomech Eng 128(4):610-622, 2006), nonwoven scaffold-ECM composites did not follow a traditional rule of mixtures. The present study provided further evidence that the primary mode of reinforcement in nonwoven scaffold-ECM composites is an increase in the number fiber-fiber bonds with a concomitant increase in the effective stiffness of the spring-like fiber segments. Simulations of potential ECM deposition scenarios using the current model indicated that the present approach is sensitive to the specific time course of tissue deposition, and is thus very suitable for studies of ECM formation in engineered heart valve tissues.

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Section: Transmural Collagen Concentration Distribution Determinationmentioning

confidence: 97%

Prediction of extracellular matrix stiffness in engineered heart valve tissues based on nonwoven scaffolds

Engelmayr

Sacks

2007

Biomech Model Mechanobiol

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“…Insoluble elastin particles tend to autofluorescence under the DAPI (4',6-diamidino-2-phenylindole) filter set [26]. Therefore, autofluorescence imaging of Col-IE fibers was performed to confirm the incorporation and distribution of insoluble elastin within the ELAC fibers.…”

Section: Confirmation Of Elastin Incorporation Into Elac Fibersmentioning

confidence: 99%

Impact of elastin incorporation into electrochemically aligned collagen fibers on mechanical properties and smooth muscle cell phenotype

2016

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Application of tissue-engineered vascular grafts (TEVGs) for the replacement of small-diameter arteries is limited due to thrombosis and intimal hyperplasia. Previous studies have attempted to address the limitations of TEVGs by developing scaffolds that mimic the composition (collagen and elastin) of native arteries to better match the mechanical properties of the graft with the native tissue. However, most existing scaffolds do not recapitulate the aligned topography of the collagen fibers found in native vessels. In the current study, based on the principles of isoelectric focusing, two different types of elastin (soluble and insoluble) were incorporated into highly oriented electrochemically aligned collagen (ELAC) fibers and the effect of elastin incorporation on the mechanical properties of the ELAC fibers and smooth muscle cell (SMC) phenotype was investigated. The results indicate that elastin incorporation significantly decreased the modulus of ELAC fibers to converge upon that of native vessels. Further, a significant increase in yield strain and decrease in Young's modulus was observed on all fibers post SMC culture compared with before the culture. Real-time polymerase chain reaction results showed a significant increase in the expression of α-smooth muscle actin and calponin on ELAC fibers with insoluble elastin, suggesting that incorporation of insoluble elastin induces a contractile phenotype in SMCs after two weeks of culture on ELAC fibers. Immunofluorescence results showed that calponin expression increased with time on all fibers. In conclusion, insoluble elastin incorporated ELAC fibers have the potential to be used for the development of functional TEVGs for the repair and replacement of small-diameter arteries.

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“…Furthermore, in TEM studies, fibrils without the periodic cross-striation of collagen could have been interpreted as evidence of elastin fibers. However, collagen fibrils withqut striation have long been known to collagen chemists (20). The non-striated fibrils described in pulpal arterioles (24) may thus have been pseudoelastica consisting of aperiodic type III collagen or "elastic fiber microfibrils".…”

Section: Discussionmentioning

confidence: 99%

Elastic (pseudoelastic) tissue in arterioles of the human and dog dental pulp

Halsm

Tønder

1981

European J Oral Sciences

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– Formalin‐fixed pulps from seven human and five dog teeth were investigated for the occurrence of vascular elastic tissue with special reference to the difference between elastin and pseudoelasuca. The latter term denotes structures which show the histochemical and fluorescence microscopic. properties of collage, but also bind the so‐called elastica stains. Hanuman aorta, and aorta, lips, tongues and gingiva from dogs served as control material. Whereas conventional elastica stains, in this stydy represented by orcinol‐new fuchsin, only differentiate elastic tissue and cikkage; a thiazine red R fluorescence method also permits differentiation between elastin memberances and pseudoelastica. Other methods for light and fluorescence microscopy verified this conclusion. Pseudoelastic fibers/memberances occurred in arterioles of both human and dog pulps, but were more frequent in the latter. The distribution of these elements in the arteiolar walls was different in the human and dog pulp. No elastin elements were observed.

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Fluorescence microscopic distinction between elastin and collagen

Cited by 28 publications

References 34 publications

Prediction of extracellular matrix stiffness in engineered heart valve tissues based on nonwoven scaffolds

Prediction of extracellular matrix stiffness in engineered heart valve tissues based on nonwoven scaffolds

Impact of elastin incorporation into electrochemically aligned collagen fibers on mechanical properties and smooth muscle cell phenotype

Elastic (pseudoelastic) tissue in arterioles of the human and dog dental pulp

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