PMID- 36436720 OWN - NLM STAT- MEDLINE DCOM- 20230206 LR - 20230206 IS - 1618-0402 (Electronic) IS - 0940-9602 (Linking) VI - 246 DP - 2023 Feb TI - Preterm rabbit-derived Precision Cut Lung Slices as alternative model of bronchopulmonary dysplasia in preclinical study: a morphological fine-tuning approach. PG - 152039 LID - S0940-9602(22)00154-6 [pii] LID - 10.1016/j.aanat.2022.152039 [doi] AB - Bronchopulmonary dysplasia (BPD) is the most common complication of preterm delivery, with significant morbidity and mortality in a neonatal intensive care setting. Research in this field aims to identify the mechanisms of late lung development with possible therapeutic targets and the improvement of medical management. Rabbits represent a suitable lab preclinical tool for mimicking the clinical BPD phenotype. Rabbits are born at term in the alveolar phase as occurs in large animals and humans and in addition, they can be delivered prematurely in contrast to mice and rats. Continuous exposure to high oxygen concentration (95% O(2)) for 7 days induces functional and morphological lung changes in preterm rabbits that resemble those observed in BPD-affected babies. The preclinical research pays great attention to optimize the experimental procedures, reduce the number of animals used in experiments and, where possible, replace animal models with alternative assays, following the principle of the 3 Rs (Replace, Reduce and Refine). The use of in vitro assays based on the ex vivo culture of Precision Cut Lung Slices (PCLS) goes in this direction, representing a good compromise between controlled and flexible in vitro models and the more physiologically relevant in vivo ones. This work aims to set up morphological analyses to be applied in preclinical tests using preterm rabbits derived PCLS, cultured up to 7 days in different oxygen conditions, as a model. After a preliminary optimization of both lung preparation and histological processing methods of the lung slices of 300 microm, the morphological analysis was conducted evaluating a series of histomorphometric parameters derived from those widely used to follow the phases of lung development and its alterations in vivo. Our histomorphometric results demonstrated that the greatest differences from pseudo-normoxia and hyperoxia exposed samples at day 0, used as starting points to compare changes due to treatments and time, are detectable after 4 days of in vitro culture, representing the most suitable time point for analysis in preclinical screening. The combination of parameters suitable for evaluating PCLS morphology in vitro resulted to be Tissue Density and Septal Thickness. Shape Factor and Roughness, evaluated to highlight the increasing complexity of the airspaces, due to the formation of septal crests, gave useful information, however, without significant differences up to day 4. Other parameters like Mean Linear Intercept and Septal Density did not allow to highlight significant differences between different oxygen conditions and time points. Instead, Radial Alveolar Count, could not be applied to PCLS, due to the tissue changes following agar infusion and culture conditions. CI - Copyright (c) 2022 Elsevier GmbH. All rights reserved. FAU - Ragionieri, Luisa AU - Ragionieri L AD - Dept. of Veterinary Science, University of Parma, Via del Taglio 10, Parma 43126, Italy. FAU - Scalera, Enrica AU - Scalera E AD - Corporate R&D Preclinical Department, Chiesi Farmaceutici S.p.A, Largo Belloli, 11/A, 43122 Parma, Italy. FAU - Zoboli, Matteo AU - Zoboli M AD - Dept. of Veterinary Science, University of Parma, Via del Taglio 10, Parma 43126, Italy. FAU - Ciccimarra, Roberta AU - Ciccimarra R AD - Dept. of Veterinary Science, University of Parma, Via del Taglio 10, Parma 43126, Italy. FAU - Petracco, Giulia AU - Petracco G AD - Corporate R&D Preclinical Department, Chiesi Farmaceutici S.p.A, Largo Belloli, 11/A, 43122 Parma, Italy. FAU - Gazza, Ferdinando AU - Gazza F AD - Dept. of Veterinary Science, University of Parma, Via del Taglio 10, Parma 43126, Italy. Electronic address: ferdinando.gazza@unipr.it. FAU - Cacchioli, Antonio AU - Cacchioli A AD - Dept. of Veterinary Science, University of Parma, Via del Taglio 10, Parma 43126, Italy. FAU - Storti, Matteo AU - Storti M AD - Corporate R&D Preclinical Department, Chiesi Farmaceutici S.p.A, Largo Belloli, 11/A, 43122 Parma, Italy. FAU - Catozzi, Chiara AU - Catozzi C AD - Corporate R&D Preclinical Department, Chiesi Farmaceutici S.p.A, Largo Belloli, 11/A, 43122 Parma, Italy. FAU - Ricci, Francesca AU - Ricci F AD - Corporate R&D Preclinical Department, Chiesi Farmaceutici S.p.A, Largo Belloli, 11/A, 43122 Parma, Italy. FAU - Ravanetti, Francesca AU - Ravanetti F AD - Dept. of Veterinary Science, University of Parma, Via del Taglio 10, Parma 43126, Italy. LA - eng PT - Journal Article DEP - 20221124 PL - Germany TA - Ann Anat JT - Annals of anatomy = Anatomischer Anzeiger : official organ of the Anatomische Gesellschaft JID - 100963897 RN - S88TT14065 (Oxygen) SB - IM MH - Infant, Newborn MH - Humans MH - Rabbits MH - Animals MH - Mice MH - Rats MH - *Bronchopulmonary Dysplasia/etiology MH - Animals, Newborn MH - Lung/pathology MH - *Lung Injury/etiology MH - *Hyperoxia/complications/genetics MH - Oxygen MH - Disease Models, Animal OTO - NOTNLM OT - Bronchopulmonary dysplasia OT - Ex vivo model OT - Histomorphometry OT - Precision cut lung slices OT - Preterm rabbit COIS- Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. EDAT- 2022/11/28 06:00 MHDA- 2023/02/07 06:00 CRDT- 2022/11/27 19:25 PHST- 2022/10/21 00:00 [received] PHST- 2022/11/10 00:00 [revised] PHST- 2022/11/16 00:00 [accepted] PHST- 2022/11/28 06:00 [pubmed] PHST- 2023/02/07 06:00 [medline] PHST- 2022/11/27 19:25 [entrez] AID - S0940-9602(22)00154-6 [pii] AID - 10.1016/j.aanat.2022.152039 [doi] PST - ppublish SO - Ann Anat. 2023 Feb;246:152039. doi: 10.1016/j.aanat.2022.152039. Epub 2022 Nov 24.