scholarly journals The efficacy and safety of pinocembrin in a sheep model of bleomycin-induced pulmonary fibrosis

PLoS ONE ◽  
2021 ◽  
Vol 16 (12) ◽  
pp. e0260719
Author(s):  
Habtamu B. Derseh ◽  
Jason Q. D. Goodger ◽  
Jean-Pierre Y. Scheerlinck ◽  
Chrishan S. Samuel ◽  
Ian E. Woodrow ◽  
...  
Keyword(s):  
Animal Model ◽  
Pulmonary Fibrosis ◽  
Inflammatory Cells ◽  
Lung Parenchyma ◽  
Lung Compliance ◽  
Large Animal Model ◽  
Large Animal ◽  
Lung Pathology ◽  
Sheep Model ◽  
Anti Inflammatory

The primary flavonoid, pinocembrin, is thought to have a variety of medical uses which relate to its reported anti-oxidant, anti-inflammatory, anti-microbial and anti-cancer properties. Some studies have reported that this flavonoid has anti-fibrotic activities. In this study, we investigated whether pinocembrin would impede fibrosis, dampen inflammation and improve lung function in a large animal model of pulmonary fibrosis. Fibrosis was induced in two localized lung segments in each of the 10 sheep participating in the study. This was achieved via two infusions of bleomycin delivered bronchoscopically at a two-week interval. Another lung segment in the same sheep was left untreated, and was used as a healthy control. The animals were kept for a little over 5 weeks after the final infusion of bleomycin. Pinocembrin, isolated from Eucalyptus leaves, was administered to one of the two bleomycin damaged lung segments at a dose of 7 mg. This dose was given once-weekly over 4-weeks, starting one week after the final bleomycin infusion. Lung compliance (as a measure of stiffness) was significantly improved after four weekly administrations of pinocembrin to bleomycin-damaged lung segments. There were significantly lower numbers of neutrophils and inflammatory cells in the bronchoalveolar lavage of bleomycin-infused lung segments that were treated with pinocembrin. Compared to bleomycin damaged lung segments without drug treatment, pinocembrin administration was associated with significantly lower numbers of immuno-positive CD8+ and CD4+ T cells in the lung parenchyma. Histopathology scoring data showed that pinocembrin treatment was associated with significant improvement in inflammation and overall pathology scores. Hydroxy proline analysis showed that the administration of pinocembrin did not reduce the increased collagen content that was induced by bleomycin in this model. Analyses of Masson’s Trichrome stained sections showed that pinocembrin treatment significantly reduced the connective tissue content in lung segments exposed to bleomycin when compared to bleomycin-infused lungs that did not receive pinocembrin. The striking anti-inflammatory and modest anti-fibrotic remodelling effects of pinocembrin administration were likely linked to the compound’s ability to improve lung pathology and functional compliance in this animal model of pulmonary fibrosis.

scholarly journals Tetrathiomolybdate Treatment Attenuates Bleomycin-Induced Angiogenesis and Lung Pathology in a Sheep Model of Pulmonary Fibrosis

2021 ◽  
Vol 12 ◽  
Author(s):  
Habtamu B. Derseh ◽  
Kopiyawaththage U. E. Perera ◽  
Sasika N. Vithana Dewage ◽  
Andrew Stent ◽  
Emmanuel Koumoundouros ◽  
...  
Keyword(s):  
Pulmonary Fibrosis ◽  
Lung Compliance ◽  
Large Animal Model ◽  
Saline Control ◽  
Collagen Type Iv ◽  
Vascular Density ◽  
Large Animal ◽  
Lung Pathology ◽  
Sheep Model ◽  
Sterile Saline

Idiopathic pulmonary fibrosis (IPF) is a progressive chronic lung disease characterized by excessive extracellular matrix (ECM) deposition in the parenchyma of the lung. Accompanying the fibrotic remodeling, dysregulated angiogenesis has been observed and implicated in the development and progression of pulmonary fibrosis. Copper is known to be required for key processes involved in fibrosis and angiogenesis. We therefore hypothesized that lowering bioavailable serum copper with tetrathiomolybdate could be of therapeutic value for treating pulmonary fibrosis. This study aimed to investigate the effect of tetrathiomolybdate on angiogenesis and fibrosis induced in sheep lung segments infused with bleomycin. Twenty sheep received two fortnightly infusions of either bleomycin (3U), or saline (control) into two spatially separate lung segments. A week after the final bleomycin/saline infusions, sheep were randomly assigned into two groups (n = 10 per group) and received twice-weekly intravenous administrations of either 50 mg tetrathiomolybdate, or sterile saline (vehicle control), for 6 weeks. Vascular density, expressed as the percentage of capillary area to the total area of parenchyma, was determined in lung tissue sections immuno-stained with antibodies against CD34 and collagen type IV. The degree of fibrosis was assessed by histopathology scoring of H&E stained sections and collagen content using Masson’s trichrome staining. Lung compliance was measured via a wedged bronchoscope procedure prior to and 7 weeks following final bleomycin infusion. In this large animal model, we show that copper lowering by tetrathiomolybdate chelation attenuates both bleomycin-induced angiogenesis and pulmonary fibrosis. Moreover, tetrathiomolybdate treatment downregulates vascular endothelial growth factor (VEGF) expression, and improved lung function in bleomycin-induced pulmonary fibrosis. Tetrathiomolybdate also suppressed the accumulation of inflammatory cells in bronchoalveolar lavage fluid 2 weeks after bleomycin injury. The molecular mechanism(s) underpinning copper modulation of fibrotic pathways is an important area for future investigation, and it represents a potential therapeutic target for pulmonary fibrosis.

Tetrathiomolybdate Treatment Attenuates Bleomycin-Induced Pulmonary Fibrosis in a Large Animal Model

2019 ◽  
Author(s):  
H.B. Derseh ◽  
K.U.E. Perera ◽  
V.D.S. Nimanthi ◽  
E. Koumoundouros ◽  
A. Stent ◽  
...  
Keyword(s):  
Animal Model ◽  
Pulmonary Fibrosis ◽  
Large Animal Model ◽  
Large Animal

scholarly journals Inhibition of the KCa3.1 Channel Alleviates Established Pulmonary Fibrosis in a Large Animal Model

2017 ◽  
Vol 56 (4) ◽  
pp. 539-550 ◽  
Author(s):  
Louise Organ ◽  
Barbara Bacci ◽  
Emmanuel Koumoundouros ◽  
Wayne G. Kimpton ◽  
Chrishan S. Samuel ◽  
...  
Keyword(s):  
Animal Model ◽  
Pulmonary Fibrosis ◽  
Large Animal Model ◽  
Large Animal

scholarly journals Structural and functional correlations in a large animal model of bleomycin-induced pulmonary fibrosis

2015 ◽  
Vol 15 (1) ◽  
Author(s):  
Louise Organ ◽  
Barbara Bacci ◽  
Emmanuel Koumoundouros ◽  
Garry Barcham ◽  
Marjorie Milne ◽  
...  
Keyword(s):  
Animal Model ◽  
Pulmonary Fibrosis ◽  
Large Animal Model ◽  
Large Animal

The effect of mast cell stabilisation in a large animal model of pulmonary fibrosis

2019 ◽  
Author(s):  
S Nimanthi Vithana Dewage ◽  
Andrew Stent ◽  
Habtamu Biyazen Derseh ◽  
K Udari Eshani Perera ◽  
Louise Organ ◽  
...  
Keyword(s):  
Animal Model ◽  
Mast Cell ◽  
Pulmonary Fibrosis ◽  
Large Animal Model ◽  
Large Animal

scholarly journals Naturally-occurring myopia and loss of cone function in a sheep model of achromatopsia

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Maya Ross ◽  
Ron Ofri ◽  
Itzhak Aizenberg ◽  
Mazen Abu–Siam ◽  
Oren Pe’er ◽  
...  
Keyword(s):  
Animal Model ◽  
Axial Length ◽  
Statistical Tests ◽  
Red Light ◽  
Large Animal Model ◽  
Large Animal ◽  
Sheep Model ◽  
Awassi Sheep ◽  
Naturally Occurring ◽  
Cone Function

Abstract Achromatopsia is an inherited retinal disease characterized by loss of cone photoreceptor function. Day blind CNGA3 mutant Improved Awassi sheep provide a large animal model for achromatopsia. This study measured refractive error and axial length parameters of the eye in this model and evaluated chromatic pupillary light reflex (cPLR) testing as a potential screening test for loss of cone function. Twenty-one CNGA3 mutant, Improved Awassi, 12 control Afec-Assaf and 12 control breed-matched wild-type (WT) Awassi sheep were examined using streak retinoscopy and B-mode ocular ultrasonography. Four CNGA3 mutant and four Afec-Assaf control sheep underwent cPLR testing. Statistical tests showed that day-blind sheep are significantly more myopic than both Afec-Assaf and WT Awassi controls. Day-blind sheep had significantly longer vitreous axial length compared to WT Awassi (1.43 ± 0.13 and 1.23 ± 0.06 cm, respectively, p < 0.0002) and no response to bright red light compared to both controls. Lack of response to bright red light is consistent with cone dysfunction, demonstrating that cPLR can be used to diagnose day blindness in sheep. Day-blind sheep were found to exhibit myopia and increased vitreous chamber depth, providing a naturally occurring large animal model of myopia.

Development and Characterization of a Novel CD34 Monoclonal Antibody That Identifies Sheep Hematopoietic Stem/Progenitor Cells.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4909-4909 ◽  
Author(s):  
Christopher D. Porada ◽  
Dee Harrison-Findik ◽  
Chad Sanada ◽  
Vincent Valiente ◽  
Paul J. Simmons ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Animal Model ◽  
Monoclonal Antibodies ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Large Animal Model ◽  
Large Animal ◽  
Sheep Model ◽  
Cd34 Cells ◽  
Hsc Transplantation

Abstract To date, the cell surface marker CD34 has been the antigen most widely used for identification and isolation of HSC in humans and numerous other species. CD34 is an integral membrane sialomucin present on roughly 1–3% of BM mononuclear cells (MNC) in a normal adult human, yet its precise function remains largely unknown. The CD34+ population is now known to produce durable reconstitution of all blood lineages in both autologous and allogeneic transplantations, providing evidence that CD34 is expressed on some of the most primitive long-term engrafting HSC. We and others have used the fetal sheep model extensively to study the potential and behavior of human HSC. Unfortunately, no reagents exist that allow sheep HSC to be identified or purified, impeding the development of experimental HSC transplantation strategies in this clinically relevant large animal model. We therefore developed monoclonal antibodies (MoAbs) to the ovine homologue of CD34. We PCR cloned and sequenced an 858bp cDNA corresponding to the extracellular domain of the sheep CD34 antigen, which was found to share 92% homology with the bovine CD34 homologue. When aligned to human CD34, the sheep homologue shared 90% homology from nucleotides 1–84, 78% homology from nucleotides 453–858, and exhibited minimal homology between nucleotides 85 and 452. This cDNA was cloned into an expression vector and used to genetically immunize mice and create monoclonal antibodies. One antibody (8D11) was selected for all subsequent studies. Using flow cytometry, 8D11 identified a small, discrete population of CD45+ cells within the peripheral blood, cord blood, and BM of sheep. This population comprised 0.5–2.5% of the total sheep BM MNC, a proportion in close accord with the previously reported incidence of CD34+ cells in adult human BM. The ability of 8D11 to enrich for sheep hematopoietic progenitors was tested by magnetically sorting 8D11+cells and assessing colony-forming potential (CFU) and the CAFC frequency. Ovine CD34+ cells were 35- to 150-fold enriched for CFU and CAFC as compared with BM mononuclear cells, whereas CD34-negative cells were correspondingly depleted of progenitors. G-CSF mobilization of sheep resulted in a 14-fold increase in the levels of circulating CD34+ cells on day 4, providing further evidence of the utility of 8D11 as a marker of primitive hematopoietic cells in the sheep model. Antibody 8D11 also robustly labeled the lining of blood vessels in both frozen and paraffin-embedded sheep tissues, further extending the utility of this antibody to include analysis of tissue sections. In conclusion, this first successful generation of a monoclonal antibody to sheep CD34 will greatly facilitate the use of the sheep as a clinically relevant large animal model system to study allogeneic HSC transplantation both in utero and in post-natal recipients using bone marrow, mobilized peripheral blood, and cord blood as cell sources.

scholarly journals Porcine model of sepsis-induced systemic inflammation and acute lung injury in donor lungs

2020 ◽  
Author(s):  
Basil Saad Nasir ◽  
Ahmed Menaouar ◽  
Caroline Landry ◽  
Jean-Francois Germain ◽  
Patrick Laplante ◽  
...  
Keyword(s):  
Animal Model ◽  
Acute Lung Injury ◽  
Lung Injury ◽  
Systemic Inflammation ◽  
Ex Vivo ◽  
Large Animal Model ◽  
Large Animal ◽  
Edema Formation ◽  
Marginal Donor ◽  
Anti Inflammatory

Abstract Background: The shortage of organ donors is a major challenge in lung transplantation. To expand the lung donor pool, ex-vivo lung perfusion (EVLP) has emerged as a platform for assessment and reconditioning marginal donor lungs. In this study a stable and reproducible large animal model of lipopolysaccharide (LPS) induced systemic inflammation and acute lung injury (ALI) was developed.Methods: Pigs (n=6) were anesthetized and monitored. After infusion of LPS (20 μg/kg) for 1 hour, followed by a 90-minute response period, lungs were procured and kept on ice for 2 hours, followed by 4 hours of EVLP. Pulmonary function, inflammatory biomarkers and edema formation were measured in vivo before procurement and during EVLP. Pro and anti-inflammatory cytokines were assayed in blood and in EVLP perfusate, which were collected before and every 30 minutes after LPS administration and EVLP.Results: LPS infusion resulted in significant hemodynamic instability, characterized by marked pulmonary hypertension, decreased systemic blood pressure and increased heart rate. This was associated with increased levels of TNFα, IL-10, IL-6, but no change in IL-1β. Ex vivo assessment of injured lungs showed graft dysfunction characterized by impaired gas exchange and edema formation. The inflammatory profile showed stable but elevated TNFα levels, and continuous production of interleukins during EVLP.Conclusion: We describe a reproducible large animal model of LPS-induced systemic inflammation and ALI. EVLP alone was unable to recondition severely injured lungs. These findings suggest that the EVLP platform requires adjuncts such as targeted anti-inflammatory agents to allow reconditioning of marginal donor lungs.

scholarly journals DXA reference values of the humanoid sheep model in preclinical studies

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11183
Author(s):  
Christoph Biehl ◽  
Jakob Schmitt ◽  
Sabine Stoetzel ◽  
Deeksha Malhan ◽  
Fathi Hassan ◽  
...  
Keyword(s):  
Animal Model ◽  
Linear Regression ◽  
Reference Group ◽  
Regression Line ◽  
Large Animal Model ◽  
Large Animal ◽  
Sheep Model ◽  
Linear Regression Line ◽  
T Score ◽  
Reference Parameters

Background Merino land sheep are a popular pre-clinical large animal model in research on systemic skeletal diseases such as osteoporosis. Interpretation of studies is difficult because many reference parameters are missing or not established. This study aims to determine the reference parameters of the skeletal system (peak bone mass = PBM, T-Score). A defined standard allows an easier comparison of the study data of the animal model with human studies (T-Score). Materials and methods A total of 116 Dual Energy X-ray Absorptiometry DXA measurements were performed on 74 untreated sheep. The average age of the animals was 57 months. The BMD, BMC, and fat content of the sheep were determined by the relevant human region of interest (ROI). From this, the PBM and from this the T-score for each of the animals were calculated. Results Using 682 DXA measurements BMD and BMC were determined to provide an indication to PBM. For BMD a significant correlation to the age of the animals was observed (p = 0.043). A significant correlation was also seen for BMC (B) (p ≤ 0.001). In the age-dependent analysis, a widespread of values above the linear regression line was measured for both BMD and BMC between the 50th and 90th months of life. From an age of about 90 months, a wider spread of values below the linear regression line was found, although the average values continued to rise. Discussion The evaluation of the 116 DXA measurements allowed the determination of the PBM for merino land sheep. With the help of the PBM, a T-score was calculated for each animal. The statistical analysis shows significant differences in BMD values between the different animal groups in each of the four ROIs investigated. Individual control or sham groups per study are therefore not sufficient. To improve comparability, an independent reference group should be established. Conclusion An independent reference group for PBM and a T-score was established from four to six-year-old animals. The bone density increases with the age of the animals. Around the fourth year of life, a first peak could be observed. Also, after the seventh year of life, a further peak with the beginning plateau phase was observed. When compiling a group of animals for an osteoporosis model, animals from the age of seven years should, therefore, be used.

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