Idiopathic pulmonary fibrosis (IPF) is characterized by maladaptive epithelial–mesenchymal crosstalk and progressive extracellular matrix accumulation, whereas currently available antifibrotic agents merely decelerate functional decline.This study investigated whether exosomes derived from human mesenchymal stem cells derived from embryonic stem cells (ESC-MSCs) restore epithelial stress responses and attenuate fibrotic remodeling. Human IPF lung transcriptomes were integrated with a bleomycin-induced murine model analyzed by RNA sequencing and protein signaling, together with cigarette smoke extract-induced injury in A549 epithelial cells. ESC-MSCs-derived exosomes exhibited typical morphology and size distribution, enrichment of tetraspanins, and absence of endoplasmic reticulum contamination, consistent with high-purity preparations. Across human IPF and bleomycin-injured lungs, transcriptomic profiling revealed prominent enrichment of extracellular matrix and cytoskeletal gene programs, whereas mitogen-activated protein kinase (MAPK) and Smad families displayed only modest alterations at the mRNA level. In vivo administration of exosomes during the fibrotic remodeling phase, via either intravenous or intratracheal delivery, resulted in improved body weight, reduced lung weight-to-body weight ratios, and decreased collagen deposition and Ashcroft scores. These structural and functional improvements were accompanied by suppression of profibrotic and mesenchymal markers and selective attenuation of activator protein-1 (AP-1) activity. In epithelial injury models, ESC-MSCs-derived exosomes enhanced cell viability, restored redox homeostasis, and constrained stress-induced mesenchymal gene expression and MAPK phosphorylation in both co-treatment and post-treatment settings. Collectively, these data support an epithelial-centered mechanism in which ESC-MSCs-derived exosomes re-establish oxidative balance and selectively restrict AP-1-driven stress signaling, thereby secondarily limiting extracellular matrix accumulation and fibrotic remodeling.

Background and Objectives: O-cyclic phytosphingosine-1-phosphate (cP1P) is a synthetic chemical and has a structure like sphingosine-1-phosphate (S1P). S1P is known to promote cell migration, invasion, proliferation, and anti-apoptosis through hippocampal signals. However, S1P mediated cellular-, molecular mechanism is still remained in the lung.Acute lung injury (ALI) and its severe form acute respiratory distress syndrome (ARDS) are characterized by excessive immune response, increased vascular permeability, alveolar-peritoneal barrier collapse, and edema. In this study, we determined whether cP1P primed human dermal derived mesenchymal stem cells (hdMSCs) ameliorate lung injury and its therapeutic pathway in ALI mice. Methods: and Results: cP1P treatment significantly stimulated MSC migration and invasion ability. In cytokine array, secretion of vascular-related factors was increased in cP1P primed hdMSCs (hdMSCcP1P ), and cP1P treatment induced inhibition of Lats while increased phosphorylation of Yap. We next determined whether hdMSCcP1P reduce inflammatory response in LPS exposed mice. hdMSCcP1P further decreased infiltration of macrophage and neutrophil, and release of TNF-α, IL-1β, and IL-6 were reduced rather than naïve hdMSC treatment. In addition, phosphorylation of STAT1 and expression of iNOS were significantly decreased in the lungs of MSCcP1P treated mice. Conclusions Taken together, these data suggest that cP1P treatment enhances hdMSC migration in regulation of Hippo signaling and MSCcP1P provide a therapeutic potential for ALI/ARDS treatment.

Mesenchymal stromal/stem cells (MSCs) possess immunoregulatory properties and their regulatory functions represent a potential therapy for acute lung injury (ALI). However, uncertainties remain with respect to defining MSCs-derived immunomodulatory pathways.Therefore, this study aimed to investigate the mechanism underlying the enhanced effect of human recombinant bone morphogenic protein-2 (rhBMP-2) primed ES-MSCs (MSC BMP2 ) in promoting Tregs in ALI mice. MSC were preconditioned with 100 ng/ml rhBMP-2 for 24 h, and then administrated to mice by intravenous injection after intratracheal injection of 1 mg/kg LPS. Treating MSCs with rhBMP-2 significantly increased cellular proliferation and migration, and cytokines array reveled that cytokines release by MSC BMP2 were associated with migration and growth. MSC BMP2 ameliorated LPS induced lung injury and reduced myeloperoxidase activity and permeability in mice exposed to LPS. Levels of inducible nitric oxide synthase were decreased while levels of total glutathione and superoxide dismutase activity were further increased via inhibition of phosphorylated STAT1 in ALI mice treated with MSC BMP2 . MSC BMP2 treatment increased the protein level of IDO1, indicating an increase in Treg cells, and Foxp3 + CD25 + Treg of CD4 + cells were further increased in ALI mice treated with MSC BMP2 . In co-culture assays with MSCs and RAW264.7 cells, the protein level of IDO1 was further induced in MSC BMP2 . Additionally, cytokine release of IL-10 was enhanced while both IL-6 and TNF-α were further inhibited. In conclusion, these findings suggest that MSC BMP2 has therapeutic potential to reduce massive inflammation of respiratory diseases by promoting Treg cells.

Background: Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive interstitial lung disease. Despite alveolar epithelial cells is crucial role in lung, its contribution and the associated biomarker remain unknown in the pathogenesis of IPF. Recently, environmental factors including stone dust, silica and cigarette smoking were found as risk factors involved in IPF. Receptor for advanced glycation end products (RAGE) is a member of the immunoglobulin super family of cell surface receptors. It has been shown that interaction between RAGE and its ligands on immune cells mediates cellular migration and regulation of pro-inflammation. RAGE is highly expressed in the lung, in particular, alveolar epithelial cells. Therefore, we determined whether RAGE expression is associated with fibrosis-associated genes in patients with IPF and mice. Results: When bleomycin (BLM) was intratracheally administered to C57BL/6 mice for 1, 2 weeks, macrophage and neutrophils were significantly increased. The fibrotic nodule formed and accumulation of collagen was determined after BLM injection in H&E- and Masson’s trichrome staining. Levels of elastin, Col1a1 and fibronectin were increased in quantitative real-time PCR and protein levels of α-SMA was increased in western blot analysis. In the lung tissues of 1 mg/kg BLM-induced mice, RAGE expression was gradually decreased in 1- and 2 weeks in immunohistochemistry and western blot analysis, and 3 mg/kg of BLM-induced mice exhibited decreased RAGE levels while α-SMA expression was increased. We next determined RAGE expression in the lungs of IPF patients using immunohistochemistry.As a result, RAGE expression was decreased, while α-SMA expression was increased compared with non-IPF subjects. Conclusions Our findings suggest that reduced RAGE was associated with increased fibrotic genes in BLM-induced mice and patients with IPF. Therefore, RAGE could be applied with a biomarker for prognosis and diagnosis in the pathogenesis of IPF.

BACKGROUND: Idiopathic pulmonary fibrosis involves irreversible alveolar destruction. Although alveolar epithelial type II cells are key functional participants within the lung parenchyma, how epithelial cells are affected upon bleomycin (BLM) exposure remains unknown. In this study, we determined whether BLM could induce cell cycle arrest via regulation of Schlafen (SLFN) family genes, a group of cell cycle regulators known to mediate growth-inhibitory responses and apoptosis in alveolar epithelial type II cells. METHODS: Mouse AE II cell line MLE-12 were exposed to 1–10 µg/mL BLM and 0.01–100 µM baicalein (Bai), a G1/G2 cell cycle inhibitor, for 24 hours. Cell viability and levels of pro-inflammatory cytokines were analyzed by MTT and enzyme-linked immunosorbent assay, respectively. Apoptosis-related gene expression was evaluated by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Cellular morphology was determined after DAPI and Hoechst 33258 staining. To verify cell cycle arrest, propidium iodide (PI) staining was performed for MLE-12 after exposure to BLM. RESULTS: BLM decreased the proliferation of MLE-12 cells. However, it significantly increased expression levels of interleukin 6, tumor necrosis factor α, and transforming growth factor β1. Based on Hoechst 33258 staining, BLM induced condensation of nuclear and fragmentation. Based on DAPI and PI staining, BLM significantly increased the size of nuclei and induced G2/M phase cell cycle arrest. Results of qRT-PCR analysis revealed that BLM increased mRNA levels of BAX but decreased those of Bcl2. In addition, BLM/Bai increased mRNA levels of p53, p21, SLFN1, 2, 4 of Schlafen family. CONCLUSION BLM exposure affects pulmonary epithelial type II cells, resulting in decreased proliferation possibly through apoptotic and cell cycle arrest associated signaling.

BACKGROUND: Idiopathic pulmonary fibrosis involves irreversible alveolar destruction. Although alveolar epithelial type II cells are key functional participants within the lung parenchyma, how epithelial cells are affected upon bleomycin (BLM) exposure remains unknown. In this study, we determined whether BLM could induce cell cycle arrest via regulation of Schlafen (SLFN) family genes, a group of cell cycle regulators known to mediate growth-inhibitory responses and apoptosis in alveolar epithelial type II cells. METHODS: Mouse AE II cell line MLE-12 were exposed to 1–10 µg/mL BLM and 0.01–100 µM baicalein (Bai), a G1/G2 cell cycle inhibitor, for 24 hours. Cell viability and levels of pro-inflammatory cytokines were analyzed by MTT and enzyme-linked immunosorbent assay, respectively. Apoptosis-related gene expression was evaluated by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Cellular morphology was determined after DAPI and Hoechst 33258 staining. To verify cell cycle arrest, propidium iodide (PI) staining was performed for MLE-12 after exposure to BLM. RESULTS: BLM decreased the proliferation of MLE-12 cells. However, it significantly increased expression levels of interleukin 6, tumor necrosis factor α, and transforming growth factor β1. Based on Hoechst 33258 staining, BLM induced condensation of nuclear and fragmentation. Based on DAPI and PI staining, BLM significantly increased the size of nuclei and induced G2/M phase cell cycle arrest. Results of qRT-PCR analysis revealed that BLM increased mRNA levels of BAX but decreased those of Bcl2. In addition, BLM/Bai increased mRNA levels of p53, p21, SLFN1, 2, 4 of Schlafen family. CONCLUSION: BLM exposure affects pulmonary epithelial type II cells, resulting in decreased proliferation possibly through apoptotic and cell cycle arrest associated signaling.
Animals ; Apoptosis ; Bisbenzimidazole ; Bleomycin ; Cell Cycle Checkpoints ; Cell Cycle ; Cell Line ; Cell Survival ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells ; Gene Expression ; Genes, vif ; Humans ; Idiopathic Pulmonary Fibrosis ; Interleukin-6 ; Lung ; Mice ; Propidium ; RNA, Messenger ; Transforming Growth Factors ; Tumor Necrosis Factor-alpha

PURPOSE: In this study we compared the surface wettability of ocular prosthesis and depositions depending on different types of artificial tear eye drops. METHODS: The artificial tear eye drops contain sodium hyaluronate (HA) 0.1%, 0.18%, 0.3%, carboxylmethylcellulose sodium (CMC), hydroxymethylcelluose + dextran (HMC), propylene glycol + polyethylene glycol (PG), polysorbate 80 (PS) povidone (Pov) were evaluated. Flat rectangular parallelepiped blocks consisting of polymethylmethacrylate (PMMA) or silicone materials were made. One artificial tear eye drop was applied on the surface of two different blocks of artificial eyes using a 23-gauge needle. Then, the static method contact angle was measured by using a contact angle goniometer. To measure the deposits, a petri dish was covered with 3 mL of artificial tear eye drops and dried for 48 hours at room temperature. Then, the light transmittance at the center of the petri dish was measured to investigate the amount of the residue. RESULTS: The contact angles of HA 0.1%, 0.18%, 0.3%, CMC, HMC, PG, PS and Pov on PMMA were 78.69degrees, 84.29degrees, 75.46degrees, 80.93degrees, 66.29degrees, 71.26degrees, 58.40degrees and 70.24degrees, respectively. The contact angles on silicone were 53.68degrees, 60.87degrees, 64.46degrees, 62.78degrees, 38.89degrees, 63.58degrees, 30.68degrees and 51.41degrees, respectively. The largest decrease in transparency was observed in the artificial tear eye drops containing HMC. CONCLUSIONS: The wettability and deposits on the surface of ocular prosthesis can vary based on the components and concentration of artificial tear eye drops. The results from this study should be considered when choosing the right artificial tear eye drops for improving dry eye symptoms in patients wearing ocular prostheses.
Anophthalmos ; Dextrans ; Dry Eye Syndromes ; Eye, Artificial* ; Humans ; Hyaluronic Acid ; Needles ; Ophthalmic Solutions* ; Polyethylene Glycols ; Polymethyl Methacrylate ; Polysorbates ; Povidone ; Propylene Glycol ; Silicones ; Sodium ; Tears* ; Wettability*

PURPOSE: In this study we compared the surface wettability of ocular prosthesis and depositions depending on different types of artificial tear eye drops. METHODS: The artificial tear eye drops contain sodium hyaluronate (HA) 0.1%, 0.18%, 0.3%, carboxylmethylcellulose sodium (CMC), hydroxymethylcelluose + dextran (HMC), propylene glycol + polyethylene glycol (PG), polysorbate 80 (PS) povidone (Pov) were evaluated. Flat rectangular parallelepiped blocks consisting of polymethylmethacrylate (PMMA) or silicone materials were made. One artificial tear eye drop was applied on the surface of two different blocks of artificial eyes using a 23-gauge needle. Then, the static method contact angle was measured by using a contact angle goniometer. To measure the deposits, a petri dish was covered with 3 mL of artificial tear eye drops and dried for 48 hours at room temperature. Then, the light transmittance at the center of the petri dish was measured to investigate the amount of the residue. RESULTS: The contact angles of HA 0.1%, 0.18%, 0.3%, CMC, HMC, PG, PS and Pov on PMMA were 78.69degrees, 84.29degrees, 75.46degrees, 80.93degrees, 66.29degrees, 71.26degrees, 58.40degrees and 70.24degrees, respectively. The contact angles on silicone were 53.68degrees, 60.87degrees, 64.46degrees, 62.78degrees, 38.89degrees, 63.58degrees, 30.68degrees and 51.41degrees, respectively. The largest decrease in transparency was observed in the artificial tear eye drops containing HMC. CONCLUSIONS: The wettability and deposits on the surface of ocular prosthesis can vary based on the components and concentration of artificial tear eye drops. The results from this study should be considered when choosing the right artificial tear eye drops for improving dry eye symptoms in patients wearing ocular prostheses.
Anophthalmos ; Dextrans ; Dry Eye Syndromes ; Eye, Artificial* ; Humans ; Hyaluronic Acid ; Needles ; Ophthalmic Solutions* ; Polyethylene Glycols ; Polymethyl Methacrylate ; Polysorbates ; Povidone ; Propylene Glycol ; Silicones ; Sodium ; Tears* ; Wettability*

PURPOSE: To compare dorzolamide-timolol fixed combination (DTFC) and latanoprost with regard to their effects on intraocular pressure (IOP) and ocular pulse amplitude (OPA). METHODS: Sixty eyes of 60 patients with open angle glaucoma or glaucoma suspect were included in the present study. Patients were divided into 2 groups, DTFC-treated (n = 30) and latanoprost-treated (n = 30). IOP and OPA were measured with dynamic contour tonometer (DCT) and Goldmann applanation tonometer (GAT), before and at least 1 month after treatment. RESULTS: GAT IOP, DCT IOP and OPA decreased by 2.25 +/- 2.23 mm Hg, 1.97 +/- 2.06 mm Hg, and 0.14 +/- 0.88 mm Hg, respectively in the DTFC-treated group. In the latanoprost-treated group, GAT IOP, DCT IOP and OPA was reduced by 2.74 +/- 2.96 mm Hg, 2.06 +/- 3.50 mm Hg, and 0.69 +/- 1.07 mm Hg, respectively. There was no significant difference (p = 0.311) in the decline of IOP between the 2 groups, but OPA of the DTFC-treated group decreased less than the latanoprost-treated group (p = 0.032). CONCLUSIONS: No significant differences were observed in the short-term decline of IOP between the 2 medications. However, the influence of DTFC on OPA appeared negligible in the latanoprost-treated group.
Glaucoma ; Glaucoma, Open-Angle ; Humans ; Intraocular Pressure*