Morphometric characterisation of the fine structure of human type II pneumocytes

Fehrenbach, Heinz; Schmiedl, Andreas; Wahlers, T; Hirt, Stefan; Brasch, Frank; Riemann, D.; Richter, Joachim

doi:10.1002/ar.1092430107

Cited by 19 publications

(16 citation statements)

References 64 publications

(50 reference statements)

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“…While one donor lung was transplanted according to the techniques initially described by Cooper et al [5], the contralateral donor lung was fixed at the time of transplantation as soon as the clinical procedure allowed. Fixation was performed by instillation via the airways to ensure rapid and uniform fixation of the whole organ for subsequent ultrastructural examination [3,12,13,31]. Donor lungs were only used for investigation of lung structure on the understanding that they could not be made available for another suitable recipient by The Eurotransplant Foundation Centre, Leiden or they were excluded from transplantation by the explanting surgeon for clinical reasons.…”

Section: Methodsmentioning

confidence: 99%

“…After separation of the double lung bloc, the contralateral donor lung was fixed as described in detail elsewhere [13,31]. Briefly, a 0.1 M cacodylate-buffered mixture of 1.5% glutaraldehyde and 1.5% paraformaldehyde was instilled via the airways at a pressure of 25 cmH 2 O until the flow of the fixative ceased automatically.…”

Section: Methodsmentioning

confidence: 99%

“…For SEM, about 5 additional 5 × 5 × 2 mm blocks of tissue were collected according to systematic random sampling. SEM samples were processed according to a modified OTOTO (osmium-thiocarbohydrazide) method [21] as detailed recently [13,31]. After critical-point drying, specimens were directly observed by means of a Zeiss DSM 960 with the omission of metalsputtering.…”

Section: Methodsmentioning

confidence: 99%

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Ultrastructural pathology of the alveolar type II pneumocytes of human donor lungs

et al. 1998

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Alveolar type II pneumocytes (PII) were studied in 12 human donor lungs perfused with modified Euro-Collins solution during single-lung transplantation (SLTx). While one lung was transplanted, the contralateral donor lung (cDL) was fixed at the time of SLTx for examination by electron microscopy, stereology, and microanalysis. Three groups were then formed: group A (n = 7), cDL without contusions, uneventful early postoperative course; group B (n = 3), cDL with conclusions, uneventful early postoperative course; group C (n = 2), cDL without contusions, early postoperative respiratory dysfunction. The major findings were that the presence of contusions had no effect on PII ultrastructure and that intracellular surfactant-storing lamellar bodies of cDL in group C were characterized by a higher volume-to-surface ratio (VsR) and larger area per cell profile than group A. Correlation analysis based on pooled data (groups A and C) showed that ischaemic time had little effect on PII ultrastructure and bore no relationship to postoperative clinical variables. The duration of preoperative donor intubation had a pronounced influence on ultrastructure and postoperative clinical variables. The stereologically estimated amount of intracellular surfactant and mitochondrial VsR were the only ultrastructural parameters that were significantly associated with early postoperative oxygenation. Lamellar bodies were the only ultrastructural components found to have a significant relationship to postoperative intubation time. The ultrastructural integrity of type II pneumocytes of human donor lungs is an important determinant of early respiratory function following clinical lung transplantation.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Ultrastructural pathology of the alveolar type II pneumocytes of human donor lungs

et al. 1998

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“…surface of the apical membrane related to a unit volume, and the mean apical surface area per unit cell volume in the AMRC, IMRC and pavement cell and (iii) the volume density and outer surface/volume ratio of mitochondria in the AMRC. In addition, the mean volume weighted volume of mitochondria in AMRC and the volume density (V V ) of the AMRC related to a defined reference space independent from the cell size were determined to exclude cellular and mitochondrial swelling, respectively [22][23][24]. Since these two parameters did not differ between the two experimental groups (P > 0.05), it was concluded that the experimental design induced neither cellular nor mitochondrial swelling.…”

Section: Stereologymentioning

confidence: 99%

Exposure of the gill epithelial cells of larval lampreys to an ion-deficient environment: a stereological study

Bartels

Schmiedl

Rosenbruch

et al. 2009

Journal of Electron Microscopy

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Three kinds of epithelial cells comprise the surfaces of the gill filaments and lamellae of larval lampreys (ammocoetes): ammocoete mitochondria-rich cells (AMRCs), intercalated mitochondria-rich cells (IMRCs) and pavement cells. Selected characteristics of these cell types in ammocoetes of Geotria australis held in distilled water and in 10% sea water were compared using an ultrastructural stereological approach to determine which of those cell type(s) respond to exposure to an ion-deficient environment in a manner that indicates that they are involved in ion uptake. Particular focus was placed on the enigmatic AMRC, which comprises ca 60% of the cells and contains numerous mitochondria. The mean percentage contributions of both AMRCs and pavement cells to the total number of the three cell types in the two experimental groups were not significantly different, whereas that of IMRCs was >7% in distilled water and <1% in 10% sea water (P < 0.001). Furthermore, the mean apical surface areas of neither AMRCs nor pavement cells differed significantly between the two experimental groups, whereas that of IMRCs was nearly 3-fold greater in distilled water than in 10% sea water. The volume densities and size of mitochondria in AMRCs did not differ between the two exposure regimes. The above comparisons provide no indications that the uptake of Na + and Cl − in the gill epithelium of ammocoetes involves either the AMRC or pavement cell but, when considered in conjunction with data on ion-transporting cells in other vertebrates, they are consistent with the conclusion that the IMRC plays a crucial role in this process.

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“…Sections of 1 Ìm were cut from each block (10-12/lung) and stained with methylene blue/Azur II (1:2). The relative volume fraction of intra-alveolar edema formation was determined by established stereological techniques [11,12] using a Cast-Grid-System 2.0 (Olympus, Denmark).…”

Section: Stereological Analysismentioning

confidence: 99%

Lung Retrieval from Non-Heart-Beating Donors: First Experience with an Innovative Preservation Strategy in a Pig Lung Transplantation Model

et al. 2004

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Objective: Lung transplantation is limited by the scarcity of donor organs. Lung retrieval from non-heart-beating donors (NHBD) might extend the donor pool and has been reported recently. However, no studies in NHBD exist using the novel approach of retrograde preservation with Perfadex solution. Methods: Heparinized asystolic pigs (n = 5, 30–35 kg) were ventilated for 90 min. The lungs were retrogradely preserved with Perfadex solution and stored inflated at 4°C for 3 h. Left lung transplantation in the recipient was followed by exclusion of the right lung. Results were compared to sham-operated animals. Oxygenation, hemodynamics and dynamic compliance were monitored for 4 h. Infiltration of polymorphonuclear cells (PMNs) and stereological quantification of alveolar edema was performed. Statistical analysis comprised Kruskal-Wallis and Mann-Whitney tests and ANOVA analysis with repeated measures. Results: No mortality was observed. During preservation, continuous elimination of blood clots via the pulmonary artery venting site was observed. Oxygenation and compliance were similar between groups, but sham controls showed significantly lower pulmonary vascular resistance. Stereological quantification revealed higher volume fractions of intra-alveolar edema in NHBD grafts, while PMN infiltration was comparable to sham controls. Conclusions: Use of NHBD lungs results in excellent outcome after 90 min of warm ischemia followed by retrograde preservation with Perfadex solution. This novel approach can optimize lung preservation by eliminating clots from the pulmonary circulation and might clinically be considered in brain-dead organ donors who become hemodynamically unstable prior to organ harvest. Further trials with longer warm and cold ischemic periods are necessary to further elucidate this promising approach to donor pool expansion.

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Morphometric characterisation of the fine structure of human type II pneumocytes

Abstract: Two to 7.5 hours of cold ischemia following ECS preservation do not deteriorate the fine structure of type II pneumocytes of human donor lungs. For reliable assessment of fine structural variations, morphometric parameters are required that are independent of variations in cell size.

Cited by 19 publications

References 64 publications

Ultrastructural pathology of the alveolar type II pneumocytes of human donor lungs

Ultrastructural pathology of the alveolar type II pneumocytes of human donor lungs

Exposure of the gill epithelial cells of larval lampreys to an ion-deficient environment: a stereological study

Lung Retrieval from Non-Heart-Beating Donors: First Experience with an Innovative Preservation Strategy in a Pig Lung Transplantation Model

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