Lung accumulation of niosome-entrapped gentamicin sulfate follows intravenous and intratracheal administration in rats

Abstract Purpose Idiopathic pulmonary fibrosis (IPF) is a progressive disease with poor outcome and limited therapeutic options. Imaging of IPF is limited to high-resolution computed tomography (HRCT) which is often not sufficient for a definite diagnosis and has a limited impact on therapeutic decision and patient management. Hypoxia of the lung is a significant feature of IPF but its role on disease progression remains elusive. Thus, the aim of our study was to evaluate hypoxia imaging with [18F]FMISO as a predictive biomarker of disease progression and therapy efficacy in preclinical models of lung fibrosis in comparison with [18F]FDG. Methods Eight-week-old C57/BL6 mice received an intratracheal administration of bleomycin (BLM) at day (D) 0 to initiate lung fibrosis. Mice received pirfenidone (300 mg/kg) or nintedanib (60 mg/kg) by daily gavage from D9 to D23. Mice underwent successive PET/CT imaging at several stages of the disease (baseline, D8/D9, D15/D16, D22/D23) with [18F]FDG and [18F]FMISO. Histological determination of the lung expression of HIF-1α and GLUT-1 was performed at D23. Results We demonstrate that mean lung density on CT as well as [18F]FDG and [18F]FMISO uptakes are upregulated in established lung fibrosis (1.4-, 2.6- and 3.2-fold increase respectively). At early stages, lung areas with [18F]FMISO uptake are still appearing normal on CT scans and correspond to areas which will deteriorate towards fibrotic lesions at later timepoints. Nintedanib and pirfenidone dramatically and rapidly decreased mean lung density on CT as well as [18F]FDG and [18F]FMISO lung uptakes (pirfenidone: 1.2-, 2.9- and 2.6-fold decrease; nintedanib: 1.2-, 2.3- and 2.5-fold decrease respectively). Early [18F]FMISO lung uptake was correlated with aggressive disease progression and better nintedanib efficacy. Conclusion [18F]FMISO PET imaging is a promising tool to early detect and monitor lung fibrosis progression and therapy efficacy.

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POS0331 LUNG TARGETED DELIVERY OF EVEROLIMUS AS A NEW TREATMENT OF SCLERODERMA-RELATED INTERSTITIAL LUNG DISEASE (SSc-ILD) DEVELOPED BY PSGL-1 KO MICE

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2021-eular.2498 ◽

2021 ◽

Vol 80 (Suppl 1) ◽

pp. 393.1-393

Author(s):

E. González Sánchez ◽

J. Gómez-Román ◽

A. Muñoz-Callejas ◽

A. Marengo ◽

N. Tsapis ◽

...

Keyword(s):

T Cells ◽

Hyaluronic Acid ◽

Interstitial Lung Disease ◽

Observational Study ◽

Lung Disease ◽

High Dose ◽

Lung Pathology ◽

Intratracheal Administration ◽

Interstitial Inflammation ◽

Autoimmune Syndrome

Background:Interstitial lung disease (ILD), the main cause of mortality in scleroderma (SSc) patients (1), has no treatment (2). P-selectin glycoprotein ligand 1 (PSGL-1), the main ligand for P-Selectin, is expressed on leukocytes and responsible for the initial steps of extravasation (3). The absence of PSGL-1 in mice spontaneously develops an autoimmune syndrome similar to human SSc with fibrosis, vascular damage, autoantibodies and pulmonary arterial hypertension in females, and almost 60% of animals older than 12 months develop ILD with aging (4). In this work, the therapeutic action of everolimus-loaded nanomedicine given by local administration as a treatment for ILD was evaluated. The intratracheal administration of everolimus loaded into in liposomes decorated with hyaluronic acid (HA) is studied as an administration strategy to reach the inflammatory and fibrotic cells, targeting these cells and avoiding systemic effects and possible toxicity on epithelial cellsObjectives:1) To study the effect of everolimus on bronchoalveolar lavage (BAL) cell populations and in lung pathology in SSc-ILD PSGL-1 KO mice2) To analyze the intratracheal application of everolimus included in empty liposomes (Lip+Ev) vs. liposomes decorated with hyaluronic acid (Lip-HA+Ev) as an administration strategy to decrease drug toxicity and increase drug effectivityMethods:In an observational study, PSGL-1−/− C57BL/6 males older than 12 months (n=4) were treated intratracheally with 4 doses of Lip or Lip-HA (with or without everolimus included), once a week (Lip+Ev 295.67µg/mL; Lip+Ev 82.73µg/mL; Lip-HA+Ev 82.73µg/mL). Then, animals were euthanatized and BAL and lungs were obtained. BAL cells were stained for flow cytometry analysis. Lungs were embedded in paraffin blocks for blind histological analysis by a pathologist and evaluated for interstitial inflammation and fibrosis degree. Lip-HA was selected as the treatment of choice for a second experiment (n=8) following the same experimental design (86.22µg/mL)Results:The observational study showed an increase in CD45+, alveolar macrophages (AM), eosinophils (Eos), granulocytes (Gr1+) and T cells in the BAL of untreated PSGL-1-/- mice compared with WT mice. Everolimus reduced these populations to WT levels in all casesLip-HA+Ev administration was chosen for further experiments because a lower dose of the drug gave a better result than the high dose in undecorated liposomes. Reduction of CD45+, AM, eosinophils, and CD45- cells populations by Lip-HA+Ev was confirmed. Lip-HA treatment increased the number of neutrophils and T cells, but this effect is controlled by the everolimus administrationHistological lung analysis showed an increase in interstitial inflammation and fibrosis in untreated PSGL-1-/- and empty Lip-HA experimental groups. Treatment with everolimus included in Lip-HA reduced the fibrotic and inflammatory interstitial lung lesions, reaching values similar to those observed in WT miceConclusion:PSGL-1 KO mice present ILD associated with scleroderma (SSc-ILD) with an increase of CD45+, Gr1+, Eos, T cells and AM populations in the BAL. Intratracheal treatment with everolimus included in liposomes decorated with hyaluronic acid reduces immune cell infiltration and fibrosis once SSc-ILD is establishedReferences:[1]Solomon JJ, Olson AL, Fischer A, Bull T, Brown KK, Raghu G (2013). Scleroderma lung disease[2]Singh D, Parihar AK, Patel S, Srivastava S, Diwan P, Singh MR (2019). Scleroderma: An insight into causes, pathogenesis and treatment strategies. Pathophysiology, 26(2)[3]Zarbock A, McEver RP, Hidalgo A (2011). Leukocyte Ligands for Endothelial Selectins: Specialized Glycoconjugates That Mediate Rolling and Signaling Under Flow. BLOOD[4]Pérez-Frías A, Núñez-Andrade N, et al. (2014). Development of an autoimmune syndrome affecting the skin and internal organs in P-selectin glycoprotein ligand 1 leukocyte receptor-deficient mice. Arthritis RheumatolDisclosure of Interests:None declared

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Consecutive daily administration of intratracheal surfactant and human umbilical cord-derived mesenchymal stem cells attenuates hyperoxia-induced lung injury in neonatal rats

Stem Cell Research & Therapy ◽

10.1186/s13287-021-02335-4 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Hsiu-Chu Chou ◽

Chien-Hsiang Chang ◽

Chien-Han Chen ◽

Willie Lin ◽

Chung-Ming Chen

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Lung Injury ◽

Umbilical Cord ◽

Daily Administration ◽

Vascular Density ◽

Human Umbilical Cord ◽

Neonatal Rats ◽

Vegf Expression ◽

Intratracheal Administration

Abstract Background Surfactant therapy is a standard of care for preterm infants with respiratory distress and reduces the incidence of death and bronchopulmonary dysplasia in these patients. Our previous study found that mesenchymal stem cells (MSCs) attenuated hyperoxia-induced lung injury and the combination therapy of surfactant and human umbilical cord-derived MSCs (hUC-MSCs) did not have additive effects on hyperoxia-induced lung injury in neonatal rats. The aim is to evaluate the effects of 2 consecutive days of intratracheal administration of surfactant and hUC-MSCs on hyperoxia-induced lung injury. Methods Neonatal Sprague Dawley rats were reared in either room air (RA) or hyperoxia (85% O2) from postnatal days 1 to 14. On postnatal day 4, the rats received intratracheal injections of either 20 μL of normal saline (NS) or 20 μL of surfactant. On postnatal day 5, the rats reared in RA received intratracheal NS, and the rats reared in O2 received intratracheal NS or hUC-MSCs (3 × 104 or 3 × 105 cells). Six study groups were examined: RA + NS + NS, RA + surfactant + NS, O2 + NS + NS, O2 + surfactant + NS, O2 + surfactant + hUC-MSCs (3 × 104 cells), and O2 + surfactant + hUC-MSCs (3 × 105 cells). The lungs were excised for histological, western blot, and cytokine analyses. Results The rats reared in hyperoxia and treated with NS yielded significantly higher mean linear intercepts (MLIs) and interleukin (IL)-1β and IL-6 levels and significantly lower vascular endothelial growth factors (VEGFs), platelet-derived growth factor protein expression, and vascular density than did those reared in RA and treated with NS or surfactant. The lowered MLIs and cytokines and the increased VEGF expression and vascular density indicated that the surfactant and surfactant + hUC-MSCs (3 × 104 cells) treatment attenuated hyperoxia-induced lung injury. The surfactant + hUC-MSCs (3 × 105 cells) group exhibited a significantly lower MLI and significantly higher VEGF expression and vascular density than the surfactant + hUC-MSCs (3 × 104 cells) group did. Conclusions Consecutive daily administration of intratracheal surfactant and hUC-MSCs can be an effective regimen for treating hyperoxia-induced lung injury in neonates.

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TLR4-dependent GM-CSF protects against lung injury in Gram-negative bacterial pneumonia

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00415.2010 ◽

2012 ◽

Vol 302 (5) ◽

pp. L447-L454 ◽

Cited By ~ 41

Author(s):

Louis R. Standiford ◽

Theodore J. Standiford ◽

Michael J. Newstead ◽

Xianying Zeng ◽

Megan N. Ballinger ◽

...

Keyword(s):

Lung Injury ◽

Bacterial Pneumonia ◽

Lavage Fluid ◽

Alveolar Epithelial Cells ◽

Intratracheal Administration ◽

Bacterial Colony ◽

Gram Negative ◽

Gm Csf

Toll-like receptors (TLRs) are required for protective host defense against bacterial pathogens. However, the role of TLRs in regulating lung injury during Gram-negative bacterial pneumonia has not been thoroughly investigated. In this study, experiments were performed to evaluate the role of TLR4 in pulmonary responses against Klebsiella pneumoniae (Kp). Compared with wild-type (WT) (Balb/c) mice, mice with defective TLR4 signaling (TLR4lps-d mice) had substantially higher lung bacterial colony-forming units after intratracheal challenge with Kp, which was associated with considerably greater lung permeability and lung cell death. Reduced expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA and protein was noted in lungs and bronchoalveolar lavage fluid of TLR4 mutant mice postintratracheal Kp compared with WT mice, and primary alveolar epithelial cells (AEC) harvested from TLR4lps-d mice produced significantly less GM-CSF in vitro in response to heat-killed Kp compared with WT AEC. TLR4lps-d AEC underwent significantly more apoptosis in response to heat-killed Kp in vitro, and treatment with GM-CSF protected these cells from apoptosis in response to Kp. Finally, intratracheal administration of GM-CSF in TLR4lps-d mice significantly decreased albumin leak, lung cell apoptosis, and bacteremia in Kp-infected mice. Based on these observations, we conclude that TLR4 plays a protective role on lung epithelium during Gram-negative bacterial pneumonia, an effect that is partially mediated by GM-CSF.

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