Functional assessment of cryopreserved human aortas for pharmaceutical research

Vázquez, M.; Santos, Mauricio; Mallo, Rosa; Ibáñez, Jacinto Sánchez; Iglesias, R Segura; Capó, G. Matheu; Fernández, P. Filgueira; Díaz, Sonia Pértega; González, T Bermúdez; Núñez, C. Andión

doi:10.1023/b:catb.0000034087.26468.5a

Cited by 7 publications

(2 citation statements)

References 25 publications

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“…smooth muscle cells) and extra‐cellular (i.e. elastin and collagen fibres) constituents of the arterial wall have been described [11,12,32,33]. The damage has been related to many factors such as the cooling and thawing rate, temperature range and cryoprotectant toxicity.…”

Section: Discussionmentioning

confidence: 99%

“…The damage has been related to many factors such as the cooling and thawing rate, temperature range and cryoprotectant toxicity. The deleterious effects of the different factors have been ascribed to mechanic, osmotic, biochemical and thermal stresses that build up during cryopreservation [18–21,32,33]. These factors were considered, and our technique of cryopreservation is in agreement with the techniques, which have shown the best results [19–21,32,34].…”

Section: Discussionmentioning

confidence: 99%

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Femoral arteries energy dissipation and filtering function remain unchanged after cryopreservation procedure

et al. 2005

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Summary The aim was to evaluate our cryopreservation method effects on the mechanical properties and filtering function of human superficial femoral arteries (SFA). SFA segments from 10 multiorgan donors were divided into two groups: fresh, tested 24–48 h after harvesting, and cryopreserved/defrosted, tested after 1 month of cryopreservation. The cooling process was carried out in three steps: 2 °C/min until −40 °C; 5 °C/min until −90 °C and finally a rapid cooling by transferring the bag to vapour phase of liquid nitrogen (−142 °C). Thawing was made in two steps, a slow warming time by exposing the bag to 20 °C during 20 min, followed by a rapid warming by immersion in a 40 °C warm bath until defrost. In a circulation mock, arterial pressure [Pressure signal (P)] and diameter [Diameter (D)] were registered at similar stretch‐frequency, P and flow levels. A compliance transfer function (D/P) was used for the on‐line assessment of the arterial wall elastic (E), viscous (η), and inertial (M) properties. To evaluate the arterial wall filter function, the arterial wall D/P frequency response was characterized, the cut‐off frequency (fc) was quantified, and the viscous energy dissipation (W) was calculated. After cryopreservation, there were not significant changes in E, η, M, W, and fc.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Femoral arteries energy dissipation and filtering function remain unchanged after cryopreservation procedure

et al. 2005

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Cryopreservation procedure does not modify human carotid homografts mechanical properties: an isobaric and dynamic analysis

Bia

Pessana

Armentano

et al. 2006

Cell Tissue Banking

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The viscoelastic and inertial properties of the arterial wall are responsible for the arterial functional role in the cardiovascular system. Cryopreservation is widely used to preserve blood vessels for vascular reconstruction but it is controversially suspected to affect the dynamic behaviour of these allografts. The aim of this work was to assess the cryopreservation's effects on human arteries mechanical properties. Common carotid artery (CCA) segments harvested from donors were divided into two groups: Fresh (n = 18), tested for 24-48 h after harvesting, and Cryopreserved (n = 18) for an average time of 30 days in gas-nitrogen phase, and finally defrosted. Each segment was tested in a circulation mock, and its pressure and diameter were registered at similar pump frequency, pulse and mean pressure levels, including those of normotensive and hypertensive conditions. A compliance transfer function (diameter/pressure) derived from a mathematical adaptive modelling was designed for the on line assessment of the arterial wall dynamics and its frequency response. Assessment of arterial wall dynamics was made by measuring its viscous (eta), inertial (M) and elastic (E) properties, and creep and stress relaxation time constant (tauC and tauSR, respectively). The frequency response characterization allowed to evaluate the arterial wall filter or buffer function. Results showed that non-significant differences exist between wall dynamics and buffer function of fresh and cryopreserved segments of human CCA. In conclusion, our cryopreservation method maintains arterial wall functional properties, close to their fresh values.

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The “in situ” expression of Human Leukocyte Antigen Class I antigens is not altered by cryopreservation in human arterial allografts

Pasquinelli

Pistillo²,

Ricci

et al. 2006

Cell Tissue Banking

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This study was aimed to establish whether the cryopreservation procedure we currently use in clinics can modify arterial homograft antigenicity. To this purpose, we performed an immunohistochemical study on fresh and cryopreserved human arterial homografts to visualize the expression of HLA class I heavy and light chains "in situ" by using the HC-10 and Namb-1 monoclonal antibodies. Human femoral arteries and thoracic aortas were harvested from 18 heart-beating donors and sampled before and after cryopreservation. Arterial segments were frozen in liquid nitrogen vapors in a controlled rate freezing system. After thawing, samples were processed for routine immunohistochemistry. To standardize immunostaining, flow-cytometry indirect immunofluorescence analysis was performed on HUVEC; immunohistochemistry of human ovarian cortical vessels was performed as an additional positive control. Negative controls were performed by omitting tissue incubation with primary antibodies. HLA-class I antigens were markedly expressed by endothelial cells lining surface intima and adventitial vasa vasorum; a moderate expression was found in medial smooth muscle cells. Except for the surface unreactivity caused by loss of endothelium, results from cryopreserved arterial allografts were strictly comparable to those observed in fresh, unfrozen tissues. These results support the view that cryopreserved arterial allografts are immunogenic as their fresh counterparts; apart from smooth muscle cells which retained a moderate expression of HLA class I antigens following cryopreservation, our study suggests that the highly HC-10 positive endothelial cells we found to line the rich adventitial network of vasa vasorum are expected to be one of the major targets of the serological response in the recipient.

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Functional assessment of cryopreserved human aortas for pharmaceutical research

Cited by 7 publications

References 25 publications

Femoral arteries energy dissipation and filtering function remain unchanged after cryopreservation procedure

Femoral arteries energy dissipation and filtering function remain unchanged after cryopreservation procedure

Cryopreservation procedure does not modify human carotid homografts mechanical properties: an isobaric and dynamic analysis

The “in situ” expression of Human Leukocyte Antigen Class I antigens is not altered by cryopreservation in human arterial allografts

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