Inhibition of aortic wall calcification in bioprosthetic heart valves by ethanol pretreatment: Biochemical and biophysical mechanisms

Lee, Chi-Hyun; Vyavahare, Narendra R.; Zand, Ramin; Kruth, Howard S.; Schoen, Frederick J.; Bianco, Richard W.; Levy, Robert J.

doi:10.1002/(sici)1097-4636(199810)42:1<30::aid-jbm5>3.0.co;2-p

Cited by 57 publications

(55 citation statements)

References 30 publications

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“…A second approach consisted of treating isolated calcospherulites on electron microscopy grids with ice-cold 70% ethanol in order to selectively remove phospholipids ( Fig. 4B; ethanoltreated TEM) [27,28], and submitting them to EDS. Such analysis revealed the presence of ethanol-extractable phosphorus from isolated calcospherulites ( Fig.…”

Section: Isolated Calcospherulites Likely Contain Phospholipidsmentioning

confidence: 87%

“…Our studies revealed the presence of ethanol-extractable phosphorus in isolated calcospherulites, and future studies will identify and quantify the nature of this extractable phosphorus. Since nucleic acids, proteins, and complex carbohydrates are insoluble in cold ethanol solutions, then the most likely molecular candidate to explain this ethanolextractable phosphorus would be phospholipids [27,28]. If true, then an apparent association of phospholipids with calcospherulites would be consistent with the concept that some type of matrix vesicle aggregation phenomenon could permit the transition of some of their contents to become calcospherulites [35].…”

Section: Discussionmentioning

confidence: 99%

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Calcospherulites11Calcospherulites (calco: calcium salt+spherulite: spherical crystalline body); calcium-containing, spherical bodies have also been referred to as calcified microspheres, mineral clusters, crystal ghost aggregates, calcification nodules, calcospherites, and small calcified spheres. isolated from the mineralization front of bone induce the mineralization of type I collagen

Midura

Vasanji

et al. 2007

Bone

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Previous work has suggested that "calcospherulites" actively participate in the mineralization of developing and healing bone. This study sought to directly test this hypothesis by developing a method to isolate calcospherulites and analyzing their capacity to seed mineralization of fibrillar collagen. The periosteal surface of juvenile rat tibial diaphysis was enriched in spherulites of ~0.5-micron diameter exhibiting a Ca/P ratio of 1.3. Their identity as calcospherulites was confirmed by their uptake of calcein at the tibial mineralization front 24 h following in vivo injection. Periosteum was dissected and unmineralized osteoid removed by collagenase in order to expose calcospherulites. Calcein-labeled calcospherulites were then released from the mineralization front by dispase digestion and isolated via fluorescence flow sorting. X-ray diffraction analysis revealed they contained apatite crystals (c-axis length of 17.5 ± 0.2 nm), though their Ca/P ratio of 1.3 is lower than that of hydroxyapatite. Much of their non-mineral phosphorous content was removed by icecold ethanol, elevating their Ca/P ratio to 1.6, suggesting the presence of phospholipids. Western * Calcospherulites (calco: calcium salt + spherulite: spherical crystalline body); calcium-containing, spherical bodies have also been referred to as calcified microspheres, mineral clusters, crystal ghost aggregates, calcification nodules, calcospherites, and small calcified spheres.Address correspondence to: Ronald J. Midura, Dept. of Biomedical Engineering-ND20, Cleveland Clinic, 9500 Euclid Ave., Cleveland, Ohio, 44195; Tel. 216 445-3212 (phone); Fax. 216 444-9198; E-mail: midurar@ccf.org. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access Author ManuscriptBone. Author manuscript; available in PMC 2008 December 1. Published in final edited form as:Bone. 2007 December ; 41(6): 1005-1016. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript blot analyses showed the presence of bone matrix proteins and type I collagen in these preparations. Incubating isolated calcospherulites in collagen hydrogels demonstrated that they could seed a mineralization reaction on type I collagen fibers in vitro. Ultrastructural analyses revealed crystals on the collagen fibers that were distributed rather uniformly along the fiber lengths. Furthermore, crystals were observed at distances well away from the observed calcospherulites. Our results directly support an active role for calcospherulites in inducing the mineralization of type I collagen fibers at the mineralization front of bone.

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Section: Isolated Calcospherulites Likely Contain Phospholipidsmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Calcospherulites11Calcospherulites (calco: calcium salt+spherulite: spherical crystalline body); calcium-containing, spherical bodies have also been referred to as calcified microspheres, mineral clusters, crystal ghost aggregates, calcification nodules, calcospherites, and small calcified spheres. isolated from the mineralization front of bone induce the mineralization of type I collagen

Midura

Vasanji

et al. 2007

Bone

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“…Gramicidin D, BA, and radioactive [carboxy- 14 C] BA (specific activity, 250 mCi/mmol) were purchased from Sigma (St Louis, MO). HEPES-buffered (25 mM ) Hank's balanced salt solution (HHBSS) was used as a receptor solution.…”

Section: Methodsmentioning

confidence: 99%

“…Following 12-hour permeation studies using radioactive [carboxy- 14 C] BA (100 µ g), skin samples were carefully rinsed 5 times with 1 ml of water. The skin was placed epidermis side up on a board lined with a parafilm.…”

Section: Evaluation Of Drug Distribution In the Membranementioning

confidence: 99%

Effect of gramicidin on percutaneous permeation of a model drug

Lee

Choi

2000

AAPS PharmSciTech

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This study investigated the enhancement effect of gramicidin, a cationic ionophore, on percutaneous absorption of a model drug, benzoic acid (BA), through rat abdominal skin. The mechanisms by which gramicidin increased skin permeability to BA were also investigated. Degree of hydration measured by the Karl Fisher method, the concentration gradient measured by cryostat analysis, and lipid concentration measured by the Fiske-Subbarow method were evaluated and compared. The results showed that BA permeation profiles through rat abdominal skin followed dose-and volume-dependent patterns. The pretreatment of gramicidin increased the permeation rate of BA through rat abdominal skin compared with the untreated control (18.89 vs. 10.86 µ g/cm 2 /hour). Change in skin permeation rate of BA after gramicidin pretreatment was closely correlated with the remaining skin water content. There were no significant differences in the amounts of phospholipid phosphorous between gramicidin pretreated and untreated skin. The enhancing effect of gramicidin on percutaneous absorption of a model drug is mainly attributed to increasing the diffusivity in the hydration domain of the skin and rearranging the lipid bilayer in the stratum corneum.

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“…O etanol apresentou efeito anticalcificante no pericárdio bovino de implantes subcutâneos em ratos e de próteses mitrais em ovelhas (18)(19)(20) . Vários estudos têm demonstrado a eficácia do etanol como inibidor da calcificação de biopróteses porcinas, sendo seu principal efeito a extração lipídica (1,18,20,21) .…”

Section: Introductionunclassified

Estudo comparativo da eficácia do etanol e do ácido L-glutâmico na prevenção da calcificação das cúspides e parede aórtica porcina: estudo experimental em ratos

Rosa

Costa

Mesquita

et al. 2002

Rev Bras Cir Cardiovasc

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Estudo comparativo da eficácia do etanol e do ácido L-glutâmico na prevenção da calcificação das cúspides e parede aórtica porcina: estudo experimental em ratos Rev Bras Cir Cardiovasc 2002; 17(2): 40-51.RBCCV George Ronald SONCINI DA ROSA*, Francisco Diniz Affonso da COSTA* **, Lismari MESQUITA*, Iseu de Santo Elias Affonso da COSTA* RESUMO: Introdução: As cúspides porcinas tratadas com glutaraldeído (GDA) são um dos tecidos biológicos mais utilizados em biopróteses, porém a calcificação tardia pós-implante constitui a causa predominante de sua falência.Objetivo: Analisar comparativamente dois métodos de prevenção da calcificação (etanol 80% e ácido L-glutâmico 0,8%) em cúspide e parede aórtica porcina implantadas no subcutâneo em ratos, tendo como grupo controle as cúspides e os segmentos de parede aórtica fixadas em glutaraldeído (GDA), num período de 15, 30 e 60 dias após implante.Material e Métodos: Foram utilizados 45 ratos jovens, distribuídos em 3 grupos contendo 15 animais cada, que por sua vez foram subdivididos em 3 subgrupos de 5 animais, nos quais foram implantados, em 2 bolsas no subcutâneo, uma cúspide e um segmento de parede aórtica em cada. Em cada grupo assim nominados: GDA (grupo controle), E80% (grupo cujas estruturas foram pré-tratadas com etanol 80%) e o AG 0,8% (grupo cujas estruturas foram pré-tratadas com ácido L-glutâmico 0,8%) foram realizadas a mensuração do cálcio e análise microscópica quanto à presença de calcificação (localização e intensidade da mesma) e de Infiltrado inflamatório (localização e tipo), no período de 15, 30 e 60 dias após implante.Resultados: Na mensuração do cálcio na cúspide aórtica encontrou-se no grupo E80% 15 dias (1,30±0,21 mg cálcio/mg tecido), E80% 30 dias (1,05±0,22 mg cálcio/mg tecido) e E80% 60 dias (0,53±0,42 mg cálcio/mg tecido); no grupo AG 0,8% 15 dias (12,17±0,66 mg cálcio/mg tecido), AG 0,8% 30 dias (15,31±2,82 mg cálcio/mg tecido) e AG 0,8% 60 dias (34,24±16,28 mg cálcio/mg tecido) com o grupo controle GDA 15 dias (12,44±2,26 mg cálcio/mg tecido), GDA 30 dias (13,44±3,34 mg cálcio/mg tecido) e GDA 60 dias (50,85±8,71 mg cálcio/mg tecido). Em relação à mensuração do cálcio na parede aórtica, encontrou-se no grupo de E80% 15 dias (4,62±0,68 mg cálcio/ mg tecido), E80% 30 dias (9,47 ± 2,59mg cálcio/mg tecido) e E80% 60 dias (23,56±7,75 mg cálcio/mg tecido) no grupo de AG 0,8% 15 dias (4,31±0,85 mg cálcio/mg tecido), AG 0,8% 30 dias (7,69±1,48 mg cálcio/mg tecido) e AG 0,8% 60 dias (20,50± 1,22 mg cálcio/mg tecido) com o grupo controle GDA 15 dias (7,34±1,32 mg cálcio/mg tecido), GDA 30 dias (9,28±0,76 mg cálcio/mg tecido) e GDA 60 dias (27,60±1,08 mg cálcio/mg tecido). Na avaliação microscópica da cúspide aórtica houve uma progressiva calcificação naquelas subme-

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Inhibition of aortic wall calcification in bioprosthetic heart valves by ethanol pretreatment: Biochemical and biophysical mechanisms

Cited by 57 publications

References 30 publications

Effect of gramicidin on percutaneous permeation of a model drug

Estudo comparativo da eficácia do etanol e do ácido L-glutâmico na prevenção da calcificação das cúspides e parede aórtica porcina: estudo experimental em ratos

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