Oxygen consumption and carbon dioxide production during liquid ventilation

Liquid ventilation with perfluorocarbon (PFCV) has advantages over conventional gas ventilation (GV) in premature and lung-injured newborn animals. Indirect calorimetric measurement of both oxygen consumption (VO2) and carbon dioxide production (VCO2) during PFCV has not been previously performed. In addition, comparison to indirect calorimetric measurement of VO2 and VCO2 during GV has not been evaluated. Ten fasted normal cats weighing 2.6 to 3.9 kg were anesthetized with pentobarbital and pancuronium. Tracheostomy was performed. Gas exchange was measured across the native lung during GV and across the membrane lung of the liquid ventilator during PFCV. VO2 was measured using a modification of a previously described, indirect, closed-circuit, volumetric technique. VCO2 was analyzed by capnographic assay of the mixed-expired closed-circuit air. The VCO2/VO2 ratio (RQ) was calculated. There was no change in VO2, VCO2, or RQ during PFCV when compared with GV (VO2: GV = 5.7 +/- 0.3 mL/kg/min, PFCV = 5.6 +/- 0.5 mL/kg/min [P = NS]; VCO2 : GV = 4.9 +/- 1.1 mL/kg/min, PFCV = 4.8 +/- 0.9 mL/kg/min [P = NS]; RQ: GV = 0.85 +/- 0.21, PFCV = 0.86 +/- 0.21 [P = NS]). During GV the PaO2 was higher than during PFCV (PaO2: GV = 335 +/- 70 mm Hg, PFCV = 267 +/- 83 mm Hg [P = .04]), as is expected because of the relative reduction in the inspiratory PiO2 of the perfluorocarbon during liquid ventilation. There was no significant change in the PaCO2 (PaCO2: GV = 37.3 +/- 2.2 mm Hg, PFCV = 40.4 +/- 5.3 mm Hg [P = NS] or the pH (pH: GV = 7.34 +/- 0.04, PFCV = 7.35 +/- 0.06 [P = NS]). This study demonstrates the efficacy of measuring VO2 and VCO2 during gas and liquid ventilation using an indirect calorimetric technique. The data demonstrate that VO2 and VCO2 do not change during liquid ventilation and that excellent gas exchange can be accomplished through PFCV.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/30885/1/0000553.pd

Perfluorochemical Liquid-Adenovirus Suspensions Enhance Gene Delivery to the Distal Lung

We compared lung delivery methods of recombinant adenovirus (rAd): (1) rAd suspended in saline, (2) rAd suspended in saline followed by a pulse-chase of a perfluorochemical (PFC) liquid mixture, and (3) a PFC-rAd suspension. Cell uptake, distribution, and temporal expression of rAd were examined using A549 cells, a murine model using luciferase bioluminescence, and histological analyses. Relative to saline, a 4X increase in transduction efficiency was observed in A549 cells exposed to PFC-rAd for 2–4 h. rAd transgene expression was improved in alveolar epithelial cells, and the level and distribution of luciferase expression when delivered in PFC-rAd suspensions consistently peaked at 24 h. These results demonstrate that PFC-rAd suspensions improve distribution and enhance rAd-mediated gene expression which has important implications in improving lung function by gene therapy

The effects of lung protective ventilation or hypercapnic acidosis on gas exchange and lung injury in surfactant deficient rabbits

Background: Permissive hypercapnia has been shown to reduce lung injury in subjects with surfactant deficiency. Experimental studies suggest that hypercapnic acidosis by itself rather than decreased tidal volume may be a key protective factor. Objectives: To study the differential effects of a lung protective ventilatory strategy or hypercapnic acidosis on gas exchange, hemodynamics and lung injury in an animal model of surfactant deficiency. Methods: 30 anesthetized, surfactant-depleted rabbits were mechanically ventilated (FiO2 = 0.8, PEEP = 7cmH2O) and randomized into three groups: Normoventilation-Normocapnia (NN)- group: tidal volume (Vt) = 7.5 ml/kg, target PaCO2 = 40 mmHg; Normoventilation-Hypercapnia (NH)-group: Vt = 7.5 ml/kg, target PaCO2 = 80 mmHg by increasing FiCO2; and a Hypoventilation- Hypercapnia (HH)-group: Vt = 4.5 ml/kg, target PaCO2 = 80 mmHg. Plasma lactate and interleukin (IL)-8 were measured every 2 h. Animals were sacrificed after 6 h to perform bronchoalveolar lavage (BAL), to measure lung wet-to-dry weight, lung tissue IL-8, and to obtain lung histology. Results: PaO2 was significantly higher in the HH-group compared to the NN-group (p<0.05), with values of the NH-group between the HH- and NN-groups. Other markers of lung injury (wetdry- weight, BAL-Protein, histology-score, plasma-IL-8 and lung tissue IL-8) resulted in significantly lower values for the HH-group compared to the NN-group and trends for the NHgroup towards lower values compared to the NN-group. Lactate was significantly lower in both hypercapnia groups compared to the NN-group. Conclusion: Whereas hypercapnic acidosis may have some beneficial effects, a significant effect on lung injury and systemic inflammatory response is dependent upon a lower tidal volume rather than resultant arterial CO2 tensions and pH alone

A Novel Approach for Ovine Primary Alveolar Epithelial Type II Cell Isolation and Culture from Fresh and Cryopreserved Tissue Obtained from Premature and Juvenile Animals.

The in vivo ovine model provides a clinically relevant platform to study cardiopulmonary mechanisms and treatments of disease; however, a robust ovine primary alveolar epithelial type II (ATII) cell culture model is lacking. The objective of this study was to develop and optimize ovine lung tissue cryopreservation and primary ATII cell culture methodologies for the purposes of dissecting mechanisms at the cellular level to elucidate responses observed in vivo. To address this, we established in vitro submerged and air-liquid interface cultures of primary ovine ATII cells isolated from fresh or cryopreserved lung tissues obtained from mechanically ventilated sheep (128 days gestation-6 months of age). Presence, abundance, and mRNA expression of surfactant proteins was assessed by immunocytochemistry, Western Blot, and quantitative PCR respectively on the day of isolation, and throughout the 7 day cell culture study period. All biomarkers were significantly greater from cells isolated from fresh than cryopreserved tissue, and those cultured in air-liquid interface as compared to submerged culture conditions at all time points. Surfactant protein expression remained in the air-liquid interface culture system while that of cells cultured in the submerged system dissipated over time. Despite differences in biomarker magnitude between cells isolated from fresh and cryopreserved tissue, cells isolated from cryopreserved tissue remained metabolically active and demonstrated a similar response as cells from fresh tissue through 72 hr period of hyperoxia. These data demonstrate a cell culture methodology using fresh or cryopreserved tissue to support study of ovine primary ATII cell function and responses, to support expanded use of biobanked tissues, and to further understanding of mechanisms that contribute to in vivo function of the lung

Physicochemical properties of perfluorochemical liquids influence ventilatory requirements, pulmonary mechanics, and microvascular permeability during partial liquid ventilation following intestinal ischemia/reperfusion injury.

OBJECTIVE: To test the hypothesis that the physicochemical properties of perfluorochemical liquid used in partial liquid ventilation can influence ventilatory requirements, pulmonary mechanics, microvascular permeability, and vasoactive mediator release in the abnormal lung. DESIGN: Prospective, controlled animal study. SETTING: Research laboratory in a university setting. SUBJECTS Male Sprague-Dawley rats: sham and intestinal ischemia/reperfusion injury. INTERVENTIONS: Treatment with perfluorochemical partial liquid ventilation (PLV: PP-5 or H-130) or conventional mechanical ventilation (CMV) over 60 mins of superior mesenteric artery occlusion and 60 mins of reperfusion. MEASUREMENTS AND MAIN RESULTS: Gas exchange, ventilatory requirements, and pulmonary mechanics were measured in vivo. Subsequently, pulmonary vascular resistance, microvascular permeability, and thromboxane were measured by using the isolated perfused lung preparation. PLV with PP-5 required significantly (p CONCLUSION: We conclude that PLV with perfluorochemical liquids attenuates pulmonary sequelae resulting from remote organ injury and that the extent of lung protection depends on the physicochemical properties of the perfluorochemical liquids

ATII cell identification.

<p>Freshly isolated cells were stained according to modified Papanicolau protocol. Type II cells are identified by the presence of dark purple inclusions (arrows) representing lamellar bodies. Purity of ATII cell preparations as assessed using this method was comparable for fresh (85.4 ± 2.2% SD; n = 10 isolations; shown in figure) as compared to cryopreserved tissue (85.1 ± 1% SD; n = 10 isolations); bar = 10μm.</p