Long-Term Cryopreservation of Human Donor Corneas

Canals, M.; Costa, José Miguel; Potau, Josep Maria; Merindano, M D; Pita, Demetrio; Ruano, D.

doi:10.1177/112067219600600302

Cited by 15 publications

(9 citation statements)

References 10 publications

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“…It is important to stress that the interpretation of our study is based upon the use of primary aortic endothelial cells from multiple donors. Previous reports have shown interindividual variability (Canals et al ., 1996; Rice et al ., 1996) and heterogeneity within the vasculature of the same organ (Antonov et al ., 1986; Ghitescu and Robert, 2002) in experiments conducted utilizing primary endothelial cells. We observed some variability in the amounts of chemokine and cytokine produced in response to the various stimuli employed in this study; however, similar trends were observed in the results from all donors.…”

Section: Discussionmentioning

confidence: 99%

Fimbria-dependent activation of pro-inflammatory molecules in Porphyromonas gingivalis infected human aortic endothelial cells

et al. 2006

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SummaryEpidemiological studies support that chronic periodontal infections are associated with an increased risk of cardiovascular disease. Previously, we reported that the periodontal pathogen Porphyromonas gingivalis accelerated atherosclerotic plaque formation in hyperlipidemic apoE -/-mice, while an isogenic fimbria-deficient (FimA-) mutant did not. In this study, we utilized 41 kDa (major) and 67 kDa (minor) fimbria mutants to demonstrate that major fimbria are required for efficient P. gingivalis invasion of human aortic endothelial cells (HAEC). Enzymelinked immunosorbent assay (ELISA) revealed that only invasive P. gingivalis strains induced HAEC production of pro-inflammatory molecules interleukin (IL)-1 β β β β , IL-8, monocyte chemoattractant protein (MCP)-1, intracellular adhesion molecule (ICAM)-1, vascular cellular adhesion molecule (VCAM)-1 and Eselectin. The purified native forms of major and minor fimbria induced chemokine and adhesion molecule expression similar to invasive P. gingivalis , but failed to elicit IL-1 β β β β production. In addition, the major and minor fimbria-mediated production of MCP-1 and IL-8 was inhibited in a dose-dependent manner by P. gingivalis lipopolysaccharide (LPS). Both P. gingivalis LPS and heat-killed organisms failed to stimulate HAEC. Treatment of endothelial cells with cytochalasin D abolished the observed pro-inflammatory MCP-1 and IL-8 response to invasive P. gingivalis and both purified fimbria, but did not affect P. gingivalis induction of IL-1 β β β β . These results suggest that major and minor fimbria elicit chemokine production in HAEC through actin cytoskeletal rearrangements; however, induction of IL-1 β β β β appears to occur via a separate mechanism. Collectively, these data support that invasive P. gingivalis and fimbria stimulate endothelial cell activation, a necessary initial event in the development of atherogenesis.

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Section: Discussionmentioning

confidence: 99%

Fimbria-dependent activation of pro-inflammatory molecules in Porphyromonas gingivalis infected human aortic endothelial cells

et al. 2006

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“…More recently, Delbosc et al [1984] and Böhnke et al [1986] describe endothelial cell necrosis phenomena and enlargement of intercellular spaces in human cryopreserved corneas. Canals et al [1996] calculate 12.2% dead endothelial cells in human corneas cryopreserved at 1¤°C/min in 7% dimethylsulfoxide.…”

Section: Discussionmentioning

confidence: 99%

“…A maximum postthawing endothelial cell survival of 90% was achieved when using 7% dimethylsulphoxide, a 1¤°C/min cooling rate and thawing in a water bath at 37¤°C. The cryopreservation process used in the present study has also been tested in human corneas at different cooling rates, showing optimum results when using a 1¤°C/min cooling speed [Canals et al, 1996].…”

Section: Cryopreservationmentioning

confidence: 99%

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Morphological Study of Cryopreserved Human Corneal Endothelium

Canals

Costa-Vila

Potau

et al. 1999

Cells Tissues Organs

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The aim of the present paper is to describe the morphological changes that occur in human corneal endothelium as an immediate consequence of corneal cryopreservation. Therefore, 16 human donor corneas were cryopreserved with an original procedure at a 1 °C/min cooling rate in a freezing solution cryoprotected with 7% dimethylsulphoxide until a final temperature of –100 °C was reached. After storage of the corneas in liquid nitrogen for periods ranging from 1 to 96 days (mean: 34.31 days), the corneas were thawed in a water bath at +37 °C. Eight additional control corneas were processed without cryopreservation. Morphological assessment of the endothelial layer was performed by scanning electron microscopy and trypan blue and alizarin red S vital staining. Results showed cryoinduced damage at variable degrees in all cryopreserved corneas. They were classified into three groups according to the intensity and extension of the cryoinduced damage: group I (n = 10): corneas with minor endothelial alterations consisting in the presence of microholes in the posterior cell membrane; group II (n = 1): corneas with generalized disruption of endothelial intercellular junctions and intact cell membranes; group III (n = 5): corneas with severe endothelial damage consisting of massive cell necrosis and complete alteration of the morphological pattern of the endothelium. All control corneas had intact endothelial layers. Cryoinduced damage cannot be completely avoided with the cryopreservation protocol tested. The high interindividual variability of the results observed is not related to the storage time of the cornea in liquid nitrogen.

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“…Studies have been reported on cryoprotection of porcine cornea in dextran and Me 2 SO of concentrations higher than those used in our experimental system (Canals et al, 1996;Wusterman et al, 1999). The human cornea has been shown to successfully recover from freezing cryoprotected in Me 2 SO (Canals et al, 1999), Me 2 SO, formamide and 1,2-propanediol , Me 2 SO, glycerol and 2,3-butanediol .…”

Section: Discussionmentioning

confidence: 99%

Cryoprotection of porcine cornea: a scanning electron microscopy study

Neronov¹,

Giurov²,

Cholakova³

et al. 2005

Vet. Med. - Czech

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Porcine corneas were frozen with Me 2 SO, glycerol, 1,2-propanediol and PEG-400. The effects of the range of concentrations (5% and 10%) and temperature regimen (1ºC/min and 5ºC/min) were investigated. The integrity of corneal endothelial cells was evaluated by scanning electron microscopy and trypan blue staining. The presence of 5-10% PEG-400 in the protective medium was the most effective in minimizing changes in the integrity of the corneal endothelium during freezing-thawing procedures.

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Long-Term Cryopreservation of Human Donor Corneas

Cited by 15 publications

References 10 publications

Fimbria-dependent activation of pro-inflammatory molecules in Porphyromonas gingivalis infected human aortic endothelial cells

Fimbria-dependent activation of pro-inflammatory molecules in Porphyromonas gingivalis infected human aortic endothelial cells

Morphological Study of Cryopreserved Human Corneal Endothelium

Cryoprotection of porcine cornea: a scanning electron microscopy study

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