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Medicine

A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes

Published: October 28th, 2021

DOI:

10.3791/62445

1Department of Surgery Division of Cardiac Surgery, The Ohio State University Wexner Medical Center, 2The Collaboration for Organ Perfusion, Protection, Engineering and Regeneration (COPPER) Laboratory, The Ohio State University College of Medicine, 3William G. Lowrie Department of Chemical and Biomolecular Engineering, College of Engineering, The Ohio State University, 4Comprehensive Transplant Center, The Ohio State University Wexner Medical Center

Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection.

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods for choosing appropriate cuff sizes for the pulmonary vein (PV), pulmonary artery (PA), or bronchus (Br) are varied, thus making the determination of proper cuff size during rat lung transplantation an exercise of trial and error. By standardizing the cuffing technique to use the smallest effective cuff appropriate for the size of the vessel or bronchus, one can make the transplantation procedure safer, faster, and more successful. Since diameters of the PV, PA, and Br are related to the body weight of the rat, we present a strategy to choosing an appropriate size using a weight-based guide. Since lung volume is also related to body weight, we recommend that this relationship should also be considered when choosing the proper volume of air for donor lung inflation during warm ischemia as well as for the proper volume of PBS to be instilled during bronchoalveolar lavage (BAL) fluid collection. We also describe methods for 4th intercostal space dissection, wound closure, and sample collection from both the native and transplanted lobes.

For over three decades, researchers have been modifying and improving rat lung transplantation models so that the data generated are more consistent and more reflective of the actual clinical condition. In our laboratory's time performing this model, we have determined four areas of improvement: cuffing techniques for anastomoses, identification of the recipient's 4th intercostal space, lung inflation and wound closure during the recipient's procedure, and the harvesting of samples for analysis.

Cuffing technique modifications for anastomoses can improve the entire transplantation procedure by shortening handling time....

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Male Sprague-Dawley rats (180-270 g body weight) were purchased commercially (e.g., Envigo) and were housed under pathogen-free conditions at The Ohio State University Animal Facility. All procedures were humanely performed according to the NIH and the National Research Council's Guide for the Humane Care and Use of Laboratory Animals and with the approval of The Ohio State University Institutional Animal Care and Use Committee (IACUC Protocol # 2012A00000135-R2).

1. Initial setup

    .......

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In order to measure pulmonary edema, the wet-to-dry weight ratio was calculated. The donor's native lobe, the transplanted lobe, and the recipient's native lobe were collected as described in the protocol and weighed immediately for wet weight, dried at 60 °C for 48 h, and then weighed again for the dry weight. An increased wet-to-dry weight ratio would be indicative of pulmonary edema. Our results indicate that the transplanted lobe did have a significant increase in wet-to-dry .......

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In this report, we have intervened at several critical steps in a rat lung transplantation protocol to optimize the procedure. While various cuffing techniques for rat lung transplantation has been reported1,2,3,4,5,6,7,8,9.......

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None.

....

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Name Company Catalog Number Comments
12 Gauge angio-catheter BD 382277
14 Gauge angio-catheter B. Braun 4251717-02
16 Gauge angio-catheter B. Braun 4252586-02
18 Gauge angio-catheter B. Braun 4251679-02
20 Gauge angio-catheter B. Braun 4252527-02
4-0 silk suture Surgical Specialties Corp. SP116
6-0 nylon suture AD Surgical S-N618R13
7-0 nylon suture AD Surgical S-N718SP13
8-0 nylon suture AD Surgical XXS-N807T6
Betadine Spray Avrio Health L.P UPC 367618160039
Clippers VWR MSPP-023326
Castroviejo micro dissecting spring scissors Roboz Surgical Instrument Co RS-5668
Dumont #5 - Fine Forceps Fine Science Tools 11254-20
Electrocautery Macan MV-7A
Endotracheal intubation kit Kent Scientific ETI-MSE
Forceps Fine Science Tools 11027-12
Halsted-mosquito hemostat Roboz Surgical Instrument Co RS-7112
Heparin Fresnius Medical Care C504701
Insulin syringe Life Technologies B328446
Isoflurane Piramal Critical Care NDC 66794-017-25
Isopropyl Alcohol Swabs BD 326895
Ketamine Hikma Pharmaceuticals PLC NDC 0413-9505-10
Dieffenbach Bulldog Clamp World Precision Instruments WPI14117
Needle holder/Forceps, Curved Micrins MI1542
Needle holder/Forceps, Straight Micrins MI1540
Perfadex Plus (Organ Preservation Solution) XVIVO Perfusion AB REF# 19950
PhysioSuite Kent Scientific PS-MSTAT-RT Used to check SpO2 and heartbeat
Retractor Roboz Surgical Instrument Co RS-6560
Saline PP Pharmaceuticals LLC NDC 63323-186-10
Scissors Fine Science Tools 14090-11
SomnoSuite Small Animal Anesthesia System Kent Scientific SS-MVG-Module
Sterile  Cotton Gauze Pad Fisherbrand 22-415-469
Surgical Microscope Leica M500-N w/ OHS
Syringe 5mL BD 309646
Vannas-Tubingen Spring Scissors Fine Science Tools 15008-08
Xylazine Korn Pharmaceuticals Corp NDC 59399-110-20
Yasargil Clamp Aesculap, Inc FT351T Used to clamp bronchus
Yasargil Clamp Aesculap, Inc FT261T Used to clamp hilum
Yasargil Clamp Applicator Aesculap, Inc FT484T

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