Porcine Lung Research Articles

Tolerable Duration of Warm Ischaemia After Circulatory Death Is Safe For At Least One Hour in Porcine Lungs: Functional Assessment with Ex Vivo Lung Perfusion.

Lungs from donation after circulatory death (DCD) may be an underused resource for transplantation. The aim was to investigate, with a DCD pig model, if it was possible to recondition lungs exposed for up to 2 h of warm ischaemia with ex vivo lung perfusion (EVLP). Danish domestic pigs (N = 17) were randomized into three groups. In the two study groups, lungs were exposed to either 1 or 2 h of warm ischaemia. All lungs were reconditioned and evaluated after 83 ± 38 minutes of perfusion at FiO2 1.0 and 0.21 with EVLP. Outcome measures were gas exchange, pulmonary physiology, inflammatory markers, and histopathologic assessment score. Lungs exposed for 2 h of warm ischaemia did not meet the criteria: PaO2 > 13 kPa required for donation compared with lungs subjected to 0 and 1 h of warm ischaemia (11.0 kPa vs. 14.2 kPa, P < 0.001). These lungs also developed an increased amount of foam and fluid in the airways. No differences in PaCO2, compliance, or pulmonary vascular resistance were observed. Results show that while lungs subjected to 0 or 1 h of warm ischaemia meet the criteria for transplantation based on EVLP evaluation, lungs subjected to 2 h of warm ischaemia did not.

Viral co-infections of the porcine respiratory tract: Insight into the local cytokine response

Infections of the porcine respiratory tract are frequently multifactorial, with more than one pathogen involved. They have a significant impact on the efficiency of pig production. One example of such a mixed infection is the porcine respiratory disease complex (PRDC). PRDC can be caused by various viral or bacterial agents. The main viral agents associated with PRDC and considered the primary pathogens are porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), and porcine circovirus type 2 (PCV2). PRRSV, SIV, and PCV2 are known as inducers of inflammatory cytokines. Cytokines play an important role in all aspects of immune responses, but their uncontrolled release in virus-induced diseases may aggravate the course of the disease and the severity of pathological lesions. Although data regarding the kinetics of the local cytokine response in porcine lungs during mono-infection with these pathogens are abundant, their impact on each other during simultaneous infection in different combinations is not thoroughly understood. This paper aims to present the available data on interactions between SIV, PRRSV, and PCV2 in mixed infections of the porcine respiratory tract and the influence of co-infections on local cytokine profiles in the lungs.

Decellularized organ biomatrices facilitate quantifiable in vitro 3D cancer metastasis models.

Metastatic cancers are chemoresistant, involving complex interplay between disseminated cancer cell aggregates and the distant organ microenvironment (extracellular matrix and stromal cells). Conventional metastasis surrogates (scratch/wound healing, Transwell migration assays) lack 3D architecture and ECM presence. Metastasis studies can therefore significantly benefit from biomimetic 3D in vitro models recapitulating the complex cascade of distant organ invasion and colonization by collective clusters of cells. We aimed to engineer reproducible and quantifiable 3D models of highly therapy-resistant cancer processes: (i) colorectal cancer liver metastasis; and (ii) breast cancer lung metastasis. Metastatic seeds are engineered using 3D tumor spheroids to recapitulate the 3D aggregation of cancer cells both in the tumor and in circulation throughout the metastatic cascade of many cancers. Metastatic soil was engineered by decellularizing porcine livers and lungs to generate biomatrix scaffolds, followed by extensive materials characterization. HCT116 colorectal and MDA-MB-231 breast cancer spheroids were generated on hanging drop arrays to initiate clustered metastatic seeding into liver and lung biomatrix scaffolds, respectively. Between days 3-7, biomatrix cellular colonization was apparent with increased metabolic activity and the presence of cellular nests evaluated via multiphoton microscopy. HCT116 and MDA-MB-231 cells colonized liver and lung biomatrices, and at least 15% of the cells invaded more than 20 μm from the surface. Engineered metastases also expressed increased signatures of genes associated with the metastatic epithelial to mesenchymal transition (EMT). Importantly, inhibition of matrix metalloproteinase-9 inhibited metastatic invasion into the biomatrix. Furthermore, metastatic nests were significantly more chemoresistant (>3 times) to the anti-cancer drug oxaliplatin, compared to 3D spheroids. Together, our data indicated that HCT116 and MDA-MB-231 spheroids invade, colonize, and proliferate in livers and lungs establishing metastatic nests in 3D settings in vitro. The metastatic nature of these cells was confirmed with functional readouts regarding EMT and chemoresistance. Modeling the dynamic metastatic cascade in vitro has potential to identify therapeutic targets to treat or prevent metastatic progression in chemoresistant metastatic cancers.

Primary Macrophage Culture from Porcine Blood and Lungs.

Primary cultures represent the most reliable method to isolate and propagate field isolates of African swine fever virus (ASFV ). Within the pig ASFV predominantly targets the reticuloendothelial system for replication; therefore, primary macrophage cell cultures are commonly used to isolate, propagate, and study the virus life cycle in the laboratory. In this chapter we will describe methods for the direct isolation of pulmonary alveolar macrophages by lung lavage and the culture of monocyte-derived macrophages from pig blood. We also include a method for the positive selection of CD14+ monocytes as a source for monocyte-derived macrophages from pig blood using microbeads.

Genomic and metabolic features of the Lactobacillus sakei JD10 revealed potential probiotic traits

Lactic acid bacteria that inhabit in the lung play important roles in maintaining the microbiome balance by interacting with the host immune system. Numerous metabolites (e.g., short chain fatty acids, bacteriocins, and hydrogen peroxide) produced by Lactobacillus sakei possess a special inhibitory spectrum against invading pathogens. In this research, the whole genome of L. sakei JD10 strain isolated from the porcine lung was sequenced and investigated. The whole size of the L. sakei JD10 chromosome was 1,989,921 bp, which encoded a total of 1951 predicted genes. Genome analyses revealed that many genes encoded carbohydrate-active enzymes (CAZymes) were predicted, which were responsible for the carbohydrate degradation and short chain fatty acids production. The metabolic profiles of short chain fatty acids in the L. sakei JD10 culture medium were measured by GC/TOFMS, and their regulatory effects on bacterial phagocytosis of RAW264.7 cells were also determined. The bacteriocin-producing genes of the L. sakei JD10 genome were also predicted, and a bacteriocin gene encoding carnocin was characterized and its molecular structure was analyzed. Two CRISPR-Cas system related genes were identified from the L. sakei JD10 genome, revealed that precise and efficient genome editing technologies could be applied for genetic engineering-manipulation. In all, investigation on the genomic features and metabolic features of L. sakei JD10 showed the potential probiotic traits to fight against pathogenic infection and regulate the host immune function.

Potential health risks of the interaction of microplastics and lung surfactant

Microplastics (MPs), as pollutants of environmental concern, are correlated with increased risk of various respiratory diseases. Nevertheless, whether or not MPs have adverse influences on the interfacial properties of lung surfactant (LS), and its effect on the generation of reactive oxygen species are poorly understood. In the present study, natural LS extracted from porcine lungs was used to investigate the interaction with polystyrene as a representative MPs. The results showed that the phase behavior, surface tension, and membrane structure of the LS were altered in the presence of polystyrene. Adsorption experiments demonstrated that in the mixed system of polystyrene and LS (the main active ingredients are phospholipids and proteins), adsorption of phospholipid components by polystyrene was notably higher than that of proteins. Moreover, polystyrene can accelerate the conversion between ascorbic acid and deoxyascorbic acid, thereby producing hydrogen peroxide (HOOH) in simulated lung fluid (containing LS) and further giving rise to an increase in the content of hydroxyl radicals (•OH). This work provides new insight into the potential hazard of MPs in human respiratory system, which is helpful for deeply understanding the unfavorable physicochemical effects of MPs exposure and the role of inhaled MPs on lung health.

Experimental determination of modulation power of lung tissue for particle therapy

In particle therapy of lung tumors, modulating effects on the particle beam may occur due to the microscopic structure of the lung tissue. These effects are caused by the heterogeneous nature of the lung tissue and cannot be completely taken into account during treatment planning, because these micro structures are too small to be fully resolved in the planning CT. In several publications, a new material parameter called modulation power (P mod ) was introduced to characterize the effect. For various artificial lung surrogates, this parameter was measured and published by other groups and ranges up to approximately 1000 μm. Studies investigating the influence of the modulation power on the dose distribution during irradiation are using this parameter in the rang of 100–800 μm. More precise measurements for P mod on real lung tissue have not yet been published. In this work, the modulation power of real lung tissue was measured using porcine lungs in order to produce more reliable data of P mod for real lung tissue. For this purpose, ex-vivo porcine lungs were frozen in a ventilated state and measurements in a carbon ion-beam were performed. Due to the way the lungs were prepared and transferred to a solid state, the lung structures that modulate the beam could also be examined in detail using micro CT imaging. An optimization of the established methods of measuring the modulation power, which takes better account of the typical structures within lung tissue, was developed as well.

Optical oxygen saturation imaging in cellular ex vivo lung perfusion to assess lobular pulmonary function.

Ex vivo lung perfusion (EVLP) is an emerging tool to evaluate marginal lungs in lung transplantation. However, there is no objective metric to monitor lobular regional oxygenation during EVLP. In this study, we developed oxygen saturation (SaO2) imaging to quantitatively assess the regional gas exchange potential of the lower lobes. Ten porcine lungs were randomly divided into control and donation after circulatory death (DCD) groups (n = 5, each). Lungs were perfused in cellular EVLP for 2 h, and multispectral images were continuously collected from the dorsal sides of the lower lobes. We examined whether lower lobe SaO2 correlated with PaO2/FiO2 (P/F) ratios in lower pulmonary veins (PV). The wet/dry ratio in lower lobes was measured and Monte Carlo simulations were performed to investigate the method's feasibility. There was a significant correlation between lower lobe SaO2 and the P/F ratio in lower PV (r = 0.855, P < 0.001). The DCD group was associated with lower SaO2 and higher wet/dry ratio than the control group (P < 0.001). The error of estimated SaO2 was limited according to Monte Carlo simulations. The developed technology provides a noninvasive and regional evaluative tool of quantitative lobular function in EVLP.

Efficacy of Lung-Tuned Monopole Antenna for Microwave Ablation: Analytical Solution and Validation in a Ventilator-Controlled ex Vivo Porcine Lung Model.

The goal of this study was to optimize a lung-tuned monopole antenna to deliver microwave energy at 2.45 GHz into a novel ventilator-controlled ex vivo lung model. An analytic and parametric approach was utilized to create an optimized monopole antenna that was impedance-matched to aerated lung tissue. This lung-tuned antenna was then fabricated using a copper 0.085" semi-rigid copper coaxial cable. For validation, the lung-tuned antenna was inserted centrally into lobes of a ex vivo porcine lung that was fully inflated to physiologically appropriate volumes. Microwave ablations were then created at 50 and 100 W for 1 minute and 5 minutes. Reflected power, cross sectional ablation sizes and spherical shape of the lung-tuned antenna were compared against a liver-tuned antenna in the ventilator-controlled ex vivo lung tissue. The study showed that the lung-tuned antennas delivered energy significantly more efficiently, with less reflected power, compared to the conventionally-used liver-tuned antennas at 50 W at 1 minute (11.8±3.0 vs 16.3±3.1 W; p value=0.03) and 5 minutes (16.2±2.8 vs 19.4±2.9 W; p value=0.04), although this was only true using 100 W at the 1 minute time point (29.0±3.5 vs 38.0±5.3 W; p value=0.02). While overall ablation zone sizes were comparable between the two types of antenna, the lung-tuned antenna did create a significantly more spherical ablation zone compared to the liver-tuned antenna at the 1 minute, 50 W setting (aspect ratio: 0.43±0.07 vs 0.38±0.04; p value=0.04). In both antenna groups, there was a significant rise in the ablation zone aspect ratio between 1 and 5 minutes, indicating that higher power and time settings can increase the spherical shape of ablation zones when using tuned antennas. Adapting this combined analytic and parametric approach to antenna design can be implemented in adaptive tissue-tuning for real-time microwave ablation optimization in lung tissue.

Developing a Lung Model in the Age of COVID-19: A Digital Image Correlation and Inverse Finite Element Analysis Framework.

Pulmonary diseases, driven by pollution, industrial farming, vaping, and the infamous COVID-19 pandemic, lead morbidity and mortality rates worldwide. Computational biomechanical models can enhance predictive capabilities to understand fundamental lung physiology; however, such investigations are hindered by the lung’s complex and hierarchical structure, and the lack of mechanical experiments linking the load-bearing organ-level response to local behaviors. In this study we address these impedances by introducing a novel reduced-order surface model of the lung, combining the response of the intricate bronchial network, parenchymal tissue, and visceral pleura. The inverse finite element analysis (IFEA) framework is developed using 3-D digital image correlation (DIC) from experimentally measured non-contact strains and displacements from an ex-vivo porcine lung specimen for the first time. A custom-designed inflation device is employed to uniquely correlate the multiscale classical pressure-volume bulk breathing measures to local-level deformation topologies and principal expansion directions. Optimal material parameters are found by minimizing the error between experimental and simulation-based lung surface displacement values, using both classes of gradient-based and gradient-free optimization algorithms and by developing an adjoint formulation for efficiency. The heterogeneous and anisotropic characteristics of pulmonary breathing are represented using various hyperelastic continuum formulations to divulge compound material parameters and evaluate the best performing model. While accounting for tissue anisotropy with fibers assumed along medial-lateral direction did not benefit model calibration, allowing for regional material heterogeneity enabled accurate reconstruction of lung deformations when compared to the homogeneous model. The proof-of-concept framework established here can be readily applied to investigate the impact of assorted organ-level ventilation strategies on local pulmonary force and strain distributions, and to further explore how diseased states may alter the load-bearing material behavior of the lung. In the age of a respiratory pandemic, advancing our understanding of lung biomechanics is more pressing than ever before.

Compositional, structural and functional properties of discrete coexisting complexes within bronchoalveolar pulmonary surfactant

Lung surfactant (LS) stabilizes the respiratory surface by forming a film at the alveolar air-liquid interface that reduces surface tension and minimizes the work of breathing. Typically, this surface-active agent has been isolated from animal lungs both for research and biomedical applications. However, these materials are constituted by complex membranous architectures including surface-active and inactive lipid/protein assemblies. In this work, we describe the composition, structure and surface activity of discrete membranous entities that are part of a LS preparation isolated from bronchoalveolar lavages of porcine lungs. Seven different fractions could be resolved from whole surfactant subjected to sucrose density gradient centrifugation. Detailed compositional characterization revealed differences in protein and cholesterol content but no distinct saturated:unsaturated phosphatidylcholine ratios. Moreover, no significant differences were detected regarding apparent hydration at the headgroup region of membranes, as reported by the probe Laurdan, and lipid chain mobility analysed by electron spin resonance (ESR) in spite of the variety of membranous assemblies observed by transmission electron microscopy. In addition, six of the seven separated LS subfractions formed similar, essentially disordered-like, interfacial films and performed efficient surface activity, under physiologically relevant conditions. Altogether, our work show that a LS isolated from porcine lungs is comprised by a heterogenous population of membranous assemblies lacking freshly secreted unused LS complexes sustaining highly dehydrated and ordered membranous assemblies as previously reported. We propose that surfactant subfractions may illustrate intermediates in sequential structural steps within the structural transformations occurring along the respiratory compression-expansion cycles.

Insulin-Degrading Enzyme Regulates the Proliferation and Apoptosis of Porcine Skeletal Muscle Stem Cells via Myostatin/MYOD Pathway.

Identifying the genes relevant for muscle development is pivotal to improve meat production and quality in pigs. Insulin-degrading enzyme (IDE), a thiol zinc-metalloendopeptidase, has been known to regulate the myogenic process of mouse and rat myoblast cell lines, while its myogenic role in pigs remained elusive. Therefore, the current study aimed to identify the effects of IDE on the proliferation and apoptosis of porcine skeletal muscle stem cells (PSMSCs) and underlying molecular mechanism. We found that IDE was widely expressed in porcine tissues, including kidney, lung, spleen, liver, heart, and skeletal muscle. Then, to explore the effects of IDE on the proliferation and apoptosis of PSMSCs, we subjected the cells to siRNA-mediated knockdown of IDE expression, which resulted in promoted cell proliferation and reduced apoptosis. As one of key transcription factors in myogenesis, MYOD, its expression was also decreased with IDE knockdown. To further elucidate the underlying molecular mechanism, RNA sequencing was performed. Among transcripts perturbed by the IDE knockdown after, a downregulated gene myostatin (MSTN) which is known as a negative regulator for muscle growth attracted our interest. Indeed, MSTN knockdown led to similar results as those of the IDE knockdown, with upregulation of cell cycle-related genes, downregulation of MYOD as well as apoptosis-related genes, and enhanced cell proliferation. Taken together, our findings suggest that IDE regulates the proliferation and apoptosis of PSMSCs via MSTN/MYOD pathway. Thus, we recruit IDE to the gene family of regulators for porcine skeletal muscle development and propose IDE as an example of gene to prioritize in order to improve pork production.

Transbronchial lung parenchyma cryoablation with a novel flexible cryoprobe in an in vivo porcine model

PurposeThe purpose of this study was to evaluate the feasibility and safety of transbronchial cryoablation with a novel flexible cryoprobe using nitrogen as the refrigerant in an in vivo porcine model of lung parenchyma. Materials and methodsA novel flexible cryoprobe using nitrogen as the refrigerant was used for transbronchial cryoablation of lung parenchyma in six normal female pigs. The cryoprobe was delivered to the distal bronchus in the bilateral porcine lungs via the bronchoscopic working channel under virtual bronchoscopy guidance. The position was confirmed with real-time computed tomography (CT). The whole procedure included two freeze-thaw cycles (15 min and 2 min, respectively). CT images were obtained during cryoablation and at 24 h, one week, two weeks and four weeks after the treatment to assess the effectiveness and safety of the procedure. Ablation zone tissue samples were obtained at 24 h and four weeks after the cryoablation for further histopathological analysis. ResultsAll ablation procedures (12/12; 100%) were performed successfully. No major complications occurred during the procedure or the observation period. The ablation zones were clearly depicted on CT with a maximal ablation zone volume at 24 h (21.88 ± 12.61 [SD] cm3) compared to 3.64 ± 2.06 (SD) cm3 and 10.73 ± 3.84 (SD) cm3 at the end of the 1st and 2nd freeze-thaw cycles, respectively (P < 0.001). Histopathological analysis revealed that a coagulative necrotic zone was formed along the target bronchus, with obvious vascular occlusion and hemorrhage 24 h after treatment. The lesions gradually formed fibrosis after four weeks. ConclusionThe novel flexible bronchoscopy-guided cryoablation is a feasible, safe and effective modality in an in vivo porcine model of peripheral normal lung parenchyma, suggesting potential capabilities for the treatment of peripheral lung cancer in humans.

Quantitative proteomics reveals tissue-specific toxic mechanisms for acute hydrogen sulfide-induced injury of diverse organs in pig

Hydrogen sulfide (H2S) is a highly toxic gas in many environmental and occupational places. It can induce multiple organ injuries particularly in lung, trachea and liver, but the relevant mechanisms remain poorly understood. In this study, we used a TMT-based discovery proteomics to identify key proteins and correlated molecular pathways involved in the pathogenesis of acute H2S-induced toxicity in porcine lung, trachea and liver tissues. Pigs were subjected to acute inhalation exposure of up to 250 ppm of H2S for 5 h for the first time. Changes in hematology and biochemical indexes, serum inflammatory cytokines and histopathology demonstrated that acute H2S exposure induced organs inflammatory injury and dysfunction in the porcine lung, trachea and liver. The proteomic data showed 51, 99 and 84 proteins that were significantly altered in lung, trachea and liver, respectively. Gene ontology (GO) annotation, KEGG pathway and protein-protein interaction (PPI) network analysis revealed that acute H2S exposure affected the three organs via different mechanisms that were relatively similar between lung and trachea. Further analysis showed that acute H2S exposure caused inflammatory damages in the porcine lung and trachea through activating complement and coagulation cascades, and regulating the hyaluronan metabolic process. Whereas antigen presentation was found in the lung but oxidative stress and cell apoptosis was observed exclusively in the trachea. In the liver, an induced dysfunction was associated with protein processing in the endoplasmic reticulum and lipid metabolism. Further validation of some H2S responsive proteins using western blotting indicated that our proteomics data were highly reliable. Collectively, these findings provide insight into toxic molecular mechanisms that could potentially be targeted for therapeutic intervention for acute H2S intoxication.

Validation of proton dose calculation on scatter corrected 4D cone beam computed tomography using a porcine lung phantom

Proton therapy treatment for lungs remains challenging as images enabling the detection of inter- and intra-fractional motion, which could be used for proton dose adaptation, are not readily available. 4D computed tomography (4DCT) provides high image quality but is rarely available in-room, while in-room 4D cone beam computed tomography (4DCBCT) suffers from image quality limitations stemming mostly from scatter detection. This study investigated the feasibility of using virtual 4D computed tomography (4DvCT) as a prior for a phase-per-phase scatter correction algorithm yielding a 4D scatter corrected cone beam computed tomography image (4DCBCTcor), which can be used for proton dose calculation. 4DCT and 4DCBCT scans of a porcine lung phantom, which generated reproducible ventilation, were acquired with matching breathing patterns. Diffeomorphic Morphons, a deformable image registration algorithm, was used to register the mid-position 4DCT to the mid-position 4DCBCT and yield a 4DvCT. The 4DCBCT was reconstructed using motion-aware reconstruction based on spatial and temporal regularization (MA-ROOSTER). Successively for each phase, digitally reconstructed radiographs of the 4DvCT, simulated without scatter, were exploited to correct scatter in the corresponding CBCT projections. The 4DCBCTcor was then reconstructed with MA-ROOSTER using the corrected CBCT projections and the same settings and deformation vector fields as those already used for reconstructing the 4DCBCT. The 4DCBCTcor and the 4DvCT were evaluated phase-by-phase, performing proton dose calculations and comparison to those of a ground truth 4DCT by means of dose-volume-histograms (DVH) and gamma pass-rates (PR). For accumulated doses, DVH parameters deviated by at most 1.7% in the 4DvCT and 2.0% in the 4DCBCTcor case. The gamma PR for a (2%, 2 mm) criterion with 10% threshold were at least 93.2% (4DvCT) and 94.2% (4DCBCTcor), respectively. The 4DCBCTcor technique enabled accurate proton dose calculation, which indicates the potential for applicability to clinical 4DCBCT scans.

Rapid Detection and Typing of Actinobacillus pleuropneumoniae Serovars Directly From Clinical Samples: Combining FTA® Card Technology With Multiplex PCR.

Actinobacillus pleuropneumoniae (APP), the causative agent of porcine pleuropneumonia, is highly contagious and responsible for high morbidity, mortality, and economic losses in the swine industry worldwide, but quick serotyping and diagnosis are still not widely available. In this study, we sought to validate the use of Whatman FTA® cards for collection and processing of A. pleuropneumoniae isolates, or porcine lung tissue samples, for direct use in diagnostic multiplex PCRs. We have optimized the processing of 3-mm discs punched from FTA® cards loaded with cultured A. pleuropneumoniae, or imprinted on lesioned regions of lung tissue, with only three distilled water washes before addition into our APP-multiplex PCR (mPCR) assay for rapid, low-cost identification and serotyping. DNA captured on FTA® cards generated the same diagnostic PCR results as DNA extracted using commercial kits for 85 A. pleuropneumoniae clinical isolate cultures and 22 lung samples. Additionally, bacterial DNA bound to FTA® cards was detectable by PCR after 6 months of storage at 37°C. This study provides simple, efficient, rapid, and practical sample processing for detection and molecular serotyping of A. pleuropneumoniae.

Rapid self-healing and self-adhesive chitosan-based hydrogels by host-guest interaction and dynamic covalent bond as flexible sensor

A sensor used to monitor tissue deformation requires good flexibility, stretchability, self-adhesion, cyto-compatibility, and antibacterial property. Here, we prepared hydrogel sensor based on O-carboxymethyl chitosan (O-CMCS) and poly(vinyl alcohol) (PVA) for monitoring human and organ motions. Based on the host-guest complexing of poly(β-cyclodextrin) with diamantane, a cross-linker containing multiple aldehyde groups was prepared for cross-linking with O-CMCS through Schiff base linkages. Borax was used as the second cross-linker to cross-link PVA through dynamic borate ester bonds. Carbon nanotubes (CNTs) were added into the hydrogels to improve their electrical conductivity and mechanical properties. The obtained hydrogel exhibited rapid self-healing ability with healing efficiency as high as 97%–103% (in 15 s), good adhesion to human skin and wet organ, good antibacterial property, cyto-compatibility, and stretchability. Furthermore, the hydrogel sensor can monitor the respiratory movement of porcine lungs and the beating of rat hearts.

Engineered mucoadhesive microparticles of formoterol/budesonide for pulmonary administration

In the present study, a multi-component system comprised of dipalmitylphospatidylcholine (DPPC), Chitosan, Lactose, and L-Leucine was developed for pulmonary delivery. Microparticles were engineered by the spray drying process and the selection of the critical parameters was performed by applying experimental design. The microcarriers with the appropriate size and yield were co-formulated with two active pharmaceutical ingredients (APIs), namely, Formoterol fumarate and Budesonide, and they were further investigated. All formulations exhibited spherical shape, appropriate aerodynamic performance, satisfying entrapment efficiency, and drug load. Their physicochemical properties were evaluated using Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FT-IR), and Differential Scanning Calorimetry (DSC). The aerodynamic particle size characterization was determined using an eight-stage Andersen cascade impactor, whereas the release of the actives was monitored in vitro in simulated lung fluid. Additional evaluation of the microparticles’ mucoadhesive properties was performed by ζ-potential measurements and ex vivo mucoadhesion study applying a falling liquid film method using porcine lung tissue. Cytotoxicity and cellular uptake studies in Calu-3 lung epithelial cell line were conducted to further investigate the safety and efficacy of the developed formulations.

Bioprintable Lung Extracellular Matrix Hydrogel Scaffolds for 3D Culture of Mesenchymal Stromal Cells

Mesenchymal stromal cell (MSC)-based cell therapy in acute respiratory diseases is based on MSC secretion of paracrine factors. Several strategies have proposed to improve this are being explored including pre-conditioning the MSCs prior to administration. We here propose a strategy for improving the therapeutic efficacy of MSCs based on cell preconditioning by growing them in native extracellular matrix (ECM) derived from the lung. To this end, a bioink with tunable stiffness based on decellularized porcine lung ECM hydrogels was developed and characterized. The bioink was suitable for 3D culturing of lung-resident MSCs without the need for additional chemical or physical crosslinking. MSCs showed good viability, and contraction assays showed the existence of cell–matrix interactions in the bioprinted scaffolds. Adhesion capacity and length of the focal adhesions formed were increased for the cells cultured within the lung hydrogel scaffolds. Also, there was more than a 20-fold increase of the expression of the CXCR4 receptor in the 3D-cultured cells compared to the cells cultured in plastic. Secretion of cytokines when cultured in an in vitro model of lung injury showed a decreased secretion of pro-inflammatory mediators for the cells cultured in the 3D scaffolds. Moreover, the morphology of the harvested cells was markedly different with respect to conventionally (2D) cultured MSCs. In conclusion, the developed bioink can be used to bioprint structures aimed to improve preconditioning MSCs for therapeutic purposes.

Interfacial interaction between benzo[a]pyrene and pulmonary surfactant: Adverse effects on lung health

Inhaled polycyclic aromatic hydrocarbons (PAHs) can directly interact with the lung surfactant (PS) lining of alveoli, thereby affecting the normal physiological functions of PS, which is a serious threat to lung health. In spite of the extensive study of benzo[a]pyrene (BaP, a representative of PAHs), its potential biophysical influence on the natural PS is still largely unknown. In this study, the interfacial interaction between PS (extracted from porcine lungs) and BaP is investigated in vitro. The results showed that the surface tension, phase behavior, and interfacial structure of the PS monolayers were obviously altered in the presence of BaP. A solubilization test manifested that PS and its major components (dipalmitoyl phosphatidylcholine, DPPC; bovine serum albumin, BSA) could in turn accelerate the dissolution of BaP, which followed the order: PS > DPPC > BSA, and mixed phospholipids were significantly responsible for the solubilization of BaP by PS. In addition, solubilization of BaP also enhanced the consumption of hydroxyl radicals (·OH) in the simulated lung fluid, which could disturb the balance between oxidation and antioxidation.