Biomedical subjects

P Petronini

Publications and source records attributed to P Petronini.

6 recordsLinked to original sources

PubMed1998

The effect of dextran for hypothermic (10 degrees C) preservation of human fetal lung fibroblasts.

BACKGROUND: Low-potassium (LP) solution with Dextran (Dx) improves lung preservation. Nevertheless, the role of Dx in simple cold storage is not established yet. This study was designed to investigate the relationship between molecular weight and concentration of Dx in LP solutions and its effects on cell viability after prolonged hypothermic preservation. METHODS: Human fetal lung fibroblasts (WI-38) were preserved at 10 degrees C for 16 hrs in five solutions containing respectively Dx11, Dx17, Dx39.2, Dx71, Dx178 at 2% and at 5% concentrations and in LP solution without Dx. Cell viability was assessed by means of both the analysis of the total protein content (cytotoxicity index) and the rate of protein synthesis (index of cellular functioning). RESULTS: No differences were recorded in total protein content among the solutions tested. By contrast, the index of cellular functioning was significantly higher using LPDx178 at both concentrations. However, LPDx178 exerted a more significant cytotoxic effect than did LP alone. CONCLUSIONS: These effects were not mediated by the variation of osmolarity; two factors probably influenced this protection: the low oncotic pressure of the LPDx178 solution and an effect chemically specific due to the increased molecular weight of Dx still unknown. Nevertheless, during 10 degrees C preservation, WI-38 cells were better preserved with LP solution without Dx confirming, thus, that during simple cold storage the presence of an oncotic pressure might be harmful.

Cell Survival↗

PubMed1996

An effective solution for prolonged preservation of cultured human pulmonary artery endothelial cells.

Pulmonary endothelium is considered the compartment most susceptible to preservation damage. This investigation was designed to analyze the efficacy of an original, University of Parma low-potassium-albumin solution (SPAL UP) on cultured human pulmonary artery endothelial cells (HPAEC) and to compare its effects with those of University of Wisconsin solution (UW) and Euro-Collins solution (EC). Cryopreserved HPAEC tertiary cultures were inoculated at the density of 5000 cells/cm2 in 9-cm2 well-plates; subcultures were then incubated at 10 degrees C for 6 hr and 16 hr in 2 ml/well of SPAL UP, UW, and EC. The HPAEC viability after incubation was assessed by evaluating the total protein content and the expression of cytotoxicity, and by analyzing the rate of protein synthesis and expression of cellular functionality after stress. Results after 6 hr of preservation showed that SPAL UP had a less significant cytotoxic effect than EC, exerted a less depressing effect on cellular metabolism, and enhanced functional recovery of endothelial cells compared with UW. At the second time interval (16 hr), SPAL UP provided a less cytotoxic effect than UW; besides, SPAL UP-induced cytotoxicity was similar to that of warm control. In conclusion, in vitro preliminary data regarding the use of SPAL UP in HPAEC preservation suggest its suitability as solution for prolonged lung protection.

Cells, Cultured↗

PubMed1995

Methodology for the assessment of lung protection. Human pulmonary artery endothelial cell preservation using haemaccel.

This investigation was designed to show an original methodology for the assessment of lung preservation and to analyze the efficacy of a low potassium polygelin solution (haemaccel [HM]) on isolated human pulmonary artery endothelial cells. The effects of HM were compared with those of low potassium dextran (LPD), Belzer (University of Wisconsin [UWS]), and Euro-Collins solutions. The viability of the endothelial cultures was assessed by means of both total protein content and recovery of metabolic cellular function expressed as the protein synthesis rate after 6 hr and 16 hr of incubation at 10 degrees C. Our results failed to show any significant difference in the total protein content for HM, LPD, and UWS, both after 6 and 16 hr of incubation; however, the Euro-Collins-preserved sample revealed a significant drop in this parameter as early as 6 hr after the start. This finding was regarded as a clear indication of cellular cytotoxicity. In contrast, the metabolism recovery capacity of the cells varied significantly between HM and UWS at 6 hr and among HM, LPD, and UWS at 16 hr; at 6 hr, however, no significant difference was observed between HM and LPD. In conclusion, HM appears to exert a more significant effect on human pulmonary artery endothelial cell metabolism recovery than do the other fluids, thus suggesting its suitability as a long-term pulmonary perfusate.

Cell Division↗

PubMed1995

A new extracellular type solution for lung preservation: "in vitro" comparison with Beltzer, low potassium Dextran and Euro-Collins solutions by means of human lung fibroblasts.

UNLABELLED: The attempt to synthesise efficacious solution to prolong lung preservation is, at present one of the most interesting challenges in transplant research. Recently, several issues emphasise the central role of ionic composition of lung-flush solutions, underlining, however, that colloid-free solutions are clearly detrimental. We have been studying a complex extracellular type solution (SPAL UP) synthesised to minimise the pathological events that occur during both preservation and reperfusion period. We report the results of toxicity of SPAL UP on normal human fibroblasts obtained from foetal lung (WI-38). WI-38 cells were seeded at 1.4 x 10(4)/cm2 in disposable plastic 12-well plates. After 3 days, cells were incubated in SPAL UP, Beltzer (UWS), Low Potassium Dextran (LPD) and Eurocollins (ECS) solutions for 6 hours at 10 degrees C. Cellular viability was evaluated by the rate of protein synthesis exploiting the incorporation of 35S-Methionine (2 microCi/ml) in growth medium with 10 mM unlabelled Methionine during 30 minutes incubation at 37 degrees C. The results were expressed as nmol. 35S-Methionine/mg of proteins/minute, and presented as means +/- SD of data of three (n = 3) well for each solution studied. RESULTS: the viability at time 0 before incubation (considered as control) was 1.65 +/- 0.1; after hypothermic preservation the data were respectively as follow: SPAL UP 0.51 +/- 0.09; UW 0.24 +/- 0.02; ECS 0.19 +/- 0.01; LDP 0.19 +/- 0.05. CONCLUSIONS: in this "in vitro" model SPAL UP solution provides a significantly (p < 0.05) better cell preservation than regular UW, ECS and LPD solutions.

Adenosine↗

PubMed1994

Haemmaccel, a key for lung preservation?

Several and different solutions have been used for lung preservation but, at present, fluids and solutions are quite alike. In the last years extracellular type solutions have been progressively tested in experimental researches and have shown a better protection vs intracellular one. In this research we have studied the effect of a low-potassium solution normally used as plasma expander (Haemmaccel, HM) on isolated foetal human fibroblasts (WI-38). HM has been compared with Belzer solution (UWS) after 16 hrs incubation at 10 degrees C and low-potassium solutions with dextran 2% and 5% after 6 hrs and 16 hrs incubation at 10 degrees C. Wi-38 cells have been seeded at the density of 9x10(4)/cm2 onto plastic well plates. Cellular viability was measuring using the rate of protein synthesis through the incorporation of 3H leucine in growth medium during a 1 hr incubation at 37 degrees C. The results were expressed as nmol 3 H leu/mg of proteins/minute and presented as means +/- SD; the comparison has been performed by the one way variance analysis test. After 16 hrs incubation at 10 degrees C HM preserves WI-38 cells significantly better than UWS. Comparing HM with LPD 2% and 5% a significant difference both at 6 hrs and at 16 hrs was observed. Our preliminary in vitro results confirm that low-potassium solutions are less toxic on isolated lung cells than intracellular one. Polygelin contained in HM could be considered a suitable colloidal substance that determine a better preservation if compared with Low-Potassium Dextran solutions.

Culture Techniques↗

PubMed1994

Verapamil used in lung transplantation: could it have a cytoprotective effect during the ischemic phase of pulmonary preservation? Study on alveolar type II cells.

It has been shown that Verapamil (V) improves lung preservation, but it is not clear whether the effect is due to vasodilation, with a more homogeneous cooling effect during hypothermic preservation, or a direct cytoprotection. Attempting to verify this, we studied the cellular viability of rats' pneumocytes type II primary cultures separately incubated in the first experiment for 6 hours in Ringer Lactate (RL) +/- V (10 mg/l) at 25 degrees C and at 4 degrees C, and in the second experiment for the same period in RL, EuroCollins (EC) and Collins (CL) solution +/- V (10 mg/l) at 4 degrees C. We analyzed the influence of the drug using two methods: the total protein content in each culture and the metabolic function of the cells using the rate of protein synthesis by means of 35 S methionine uptake assay. All data showed that there were no significant statistical difference (Student t-test, p > 0.05) in the use of verapamil during alveolar type II cells either in normothermic or hypothermic preservation. Thus we conclude that the protection of this calcium channel blocker during lung preservation is probably due to the effect on pulmonary vessels rather than to a direct cytoprotective effect on the lung cells.

Animals↗