In vitro antioxidant and wound healing properties of baru nut extract (Dipteryx alata Vog.) in pulmonary epithelial cells for therapeutic application in chronic pulmonary obstructive disease (COPD)

The effects of the surfactant proteins (SP)-A, SP-B, and SP-C on binding and endocytosis of fluorescently labeled lipid vesicles were studied in rat type II epithelial cells and in MLE-12 cells, a pulmonary adenocarcinoma cell line with alveolar cell characteristics. Incorporation of SP-C in lipid vesicles markedly stimulated binding to the cell membrane at 4 degrees C and endocytosis of lipids at 37 degrees C. SP-C enhanced lipid uptake in MLE-12 cells, type II cells, and NIH 3T3 cells. SP-B stimulated lipid uptake in MLE-12 cells, but to a lesser degree. SP-B decreased the amount of lipid uptake stimulated by SP-C, SP-A did not increase endocytosis of lipids by MLE-12 cells or type II cells, but aggregates of lipid were observed associated with the cell surface in the presence of SP-A. Maintenance of active surfactant in the lung may be achieved through the selective uptake and degradation of surfactant subfractions depleted in SP-A and SP-B.

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Sodium Nitrite Enhances Pulmonary Epithelial Cells Wound Healing Under Normoxia via Cyclic GMPindependent Signaling Pathways

Free Radical Biology and Medicine ◽

10.1016/j.freeradbiomed.2010.10.276 ◽

2010 ◽

Vol 49 ◽

pp. S104

Author(s):

Ling Wang ◽

Mark T Gladwin

Keyword(s):

Wound Healing ◽

Epithelial Cells ◽

Signaling Pathways ◽

Sodium Nitrite ◽

Pulmonary Epithelial Cells

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PTH Derivative Promotes Wound Healing via Synergistic Multicellular Stimulating and Exosomal Activities

10.21203/rs.2.18433/v3 ◽

2020 ◽

Author(s):

Yi-Fan Shen ◽

Jing-Huan Huang ◽

Kai-Yang Wang ◽

Jin Zheng ◽

Lin Cai ◽

...

Keyword(s):

Wound Healing ◽

Epithelial Cells ◽

Intercellular Communication ◽

Indirect Effects ◽

Clinical Problem ◽

Therapeutic Effects ◽

And Migration ◽

Proliferation And Migration

Abstract Background: Diabetic wounds are a disturbing and rapidly growing clinical problem. A novel peptide, parathyroid hormone related peptide (PTHrP-2), is assumed as multifunctional factor in angiogenesis, fibrogenesis and re-epithelization. This study aims to test PTHrP-2 efficiency and mechanism in wound healing. Methods: Through repair phenomenon in vivo some problems were detected, and further research on their mechanisms was made. In vivo therapeutic effects of PTHrP-2 were determined by HE, Masson, microfil and immunohistochemical staining. In vitro direct effects of PTHrP-2 were determined by proliferation, migration, Vascular Endothelial Grown Factor and collagen I secretion of cells and Akt/ Erk1/2 pathway change. In vitro indirect effects of PTHrP-2 was study via exosomes. Exosomes from PTHrP-2 untreated and treated HUVECs and HFF-1 cells were insolated and identified. Exosomes were co-cultured with original cells, HUVECs or HFF-1 cells, and epithelial cells. Proliferation and migration and pathway change were observed. PTHrP-2-HUVEC-Exos were added into in vivo wound to testify its hub role in PTHrP-2 indirect effects in wound healing. Results: In vivo, PTHrP-2 exerted multifunctional pro-angiogenesis, pro-firbogenesis and re-epithelization effects. In vitro, PTHrP-2 promoted proliferation and migration of endothelial and fibroblast cells, but had no effect on epithelial cells. Therefore, we tested PTHrP-2 indirect effects via exosomes. PTHrP-2 intensified intercellular communication between endothelial cells and fibroblasts and initiated endothelial-epithelial intercellular communication. PTHrP-2-HUVEC-Exos played a hub role in PTHrP-2 indirect effects in wound healing. Conclusion: These findings of this study indicated that PTHrP-2, a multifunctional factor, could promote wound healing via synergistic multicellular stimulating and exosomal activities. Key words PTH, multifunctional factor, diabetic wound, exosomes, synergistic effect

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New Gene Markers Involved in Molecular Processes of Tissue Repair, Response to Wounding and Regeneration Are Differently Expressed in Fibroblasts from Porcine Oral Mucosa during Long-Term Primary Culture

Animals ◽

10.3390/ani10111938 ◽

2020 ◽

Vol 10 (11) ◽

pp. 1938

Author(s):

Artur Bryja ◽

Patrycja Sujka-Kordowska ◽

Aneta Konwerska ◽

Sylwia Ciesiółka ◽

Maria Wieczorkiewicz ◽

...

Keyword(s):

Wound Healing ◽

Epithelial Cells ◽

Expression Profile ◽

Transforming Growth Factor Beta ◽

Transforming Growth Factor ◽

Primary Cultures ◽

Gene Markers ◽

Morphological Process

The mechanisms of wound healing and vascularization are crucial steps of the complex morphological process of tissue reconstruction. In addition to epithelial cells, fibroblasts play an important role in this process. They are characterized by dynamic proliferation and they form the stroma for epithelial cells. In this study, we have used primary cultures of oral fibroblasts, obtained from porcine buccal mucosa. Cells were maintained long-term in in vitro conditions, in order to investigate the expression profile of the molecular markers involved in wound healing and vascularization. Based on the Affymetrix assays, we have observed three ontological groups of markers as wound healing group, response to wounding group and vascularization group, represented by different genes characterized by their expression profile during long-term primary in vitro culture (IVC) of porcine oral fibroblasts. Following the analysis of gene expression in three previously identified groups of genes, we have identified that transforming growth factor beta 1 (TGFB1), ITGB3, PDPN, and ETS1 are involved in all three processes, suggesting that these genes could be recognized as markers of repair specific for oral fibroblasts within the porcine mucosal tissue.

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Exosomes derived from human amniotic epithelial cells accelerate diabetic wound healing via PI3K-AKT-mTOR-mediated promotion in angiogenesis and fibroblast function

Burns & Trauma ◽

10.1093/burnst/tkaa020 ◽

2020 ◽

Vol 8 ◽

Cited By ~ 2

Author(s):

Pei Wei ◽

Chenjian Zhong ◽

Xiaolan Yang ◽

Futing Shu ◽

Shichu Xiao ◽

...

Keyword(s):

Wound Healing ◽

Epithelial Cells ◽

High Throughput ◽

High Throughput Sequencing ◽

Biological Effects ◽

Mtor Pathway ◽

Diabetic Wound ◽

Diabetic Wound Healing ◽

Human Amniotic Epithelial Cells

Abstract Background Diabetic wounds are one of the most common and serious complications of diabetes mellitus, characterized by the dysfunction of wound-healing-related cells in quantity and quality. Our previous studies revealed that human amniotic epithelial cells (hAECs) could promote diabetic wound healing by paracrine action. Interestingly, numerous studies demonstrated that exosomes derived from stem cells are the critical paracrine vehicles for stem cell therapy. However, whether exosomes derived from hAECs (hAECs-Exos) mediate the effects of hAECs on diabetic wound healing remains unclear. This study aimed to investigate the biological effects of hAECs-Exos on diabetic wound healing and preliminarily elucidate the underlying mechanism. Methods hAECs-Exos were isolated by ultracentrifugation and identified by transmission electron microscopy, dynamic light scattering and flow cytometry. A series of in vitro functional analyses were performed to assess the regulatory effects of hAECs-Exos on human fibroblasts (HFBs) and human umbilical vein endothelial cells (HUVECs) in a high-glycemic microenvironment. High-throughput sequencing and bioinformatics analyses were conducted to speculate the related mechanisms of actions of hAECs-Exos on HFBs and HUVECs. Subsequently, the role of the candidate signaling pathway of hAECs-Exos in regulating the function of HUVECs and HFBs, as well as in diabetic wound healing, was assessed. Results hAECs-Exos presented a cup- or sphere-shaped morphology with a mean diameter of 105.89 ± 10.36 nm, were positive for CD63 and TSG101 and could be internalized by HFBs and HUVECs. After that, hAECs-Exos not only significantly promoted the proliferation and migration of HFBs, but also facilitated the angiogenic activity of HUVECs in vitro. High-throughput sequencing revealed enriched miRNAs of hAECs-Exos involved in wound healing. Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analyses have shown that the target genes of the top 15 miRNAs were highly enriched in the PI3K-AKT pathway. Further functional studies demonstrated that the PI3K-AKT-mTOR pathway was necessary for the induced biological effects of hAECs-Exos on HFBs and HUVECs, as well as on wound healing, in diabetic mice. Conclusions Our findings demonstrated that hAECs-Exos represent a promising, novel strategy for diabetic wound healing by promoting angiogenesis and fibroblast function via activation of the PI3K-AKT-mTOR pathway.

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Mechanical strain-induced proliferation and signaling in pulmonary epithelial H441 cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.2000.279.1.l43 ◽

2000 ◽

Vol 279 (1) ◽

pp. L43-L51 ◽

Cited By ~ 48

Author(s):

Patricia R. Chess ◽

Liana Toia ◽

Jacob N. Finkelstein

Keyword(s):

Epithelial Cells ◽

Tyrosine Kinase ◽

Cellular Proliferation ◽

Kinase Inhibitor ◽

Mechanical Strain ◽

Mobility Shift ◽

Pulmonary Epithelial Cells ◽

Mitogen Activated Protein ◽

Mobility Shift Assay

Pulmonary epithelial cells are exposed to mechanical strain during physiological breathing and mechanical ventilation. Strain regulates pulmonary growth and development and is implicated in volutrauma-induced fibrosis. The mechanisms of strain-induced effects are not well understood. It was hypothesized that mechanical strain induces proliferation of pulmonary epithelial cells and that this is mediated by signals initiated within seconds of strain. To test this hypothesis, human pulmonary adenocarcinoma H441 cells were strained in vitro. Cyclic as well as tonic strain resulted in increased cellular proliferation. Western blot analysis of strained cells demonstrated three newly phosphorylated tyrosine residues within 30 s of strain. Phosphorylation of mitogen-activated protein kinases p42/44 increased, electrophoretic mobility shift assay demonstrated activation of transcription factor activating protein-1, and immunohistochemistry demonstrated increased phosphorylation of c- jun in response to strain. The tyrosine kinase inhibitor genistein blocked the strain-induced proliferation. We conclude that strain induces proliferation in pulmonary epithelial cells and that tyrosine kinase activity is necessary to signal the proliferative response to mechanical strain.

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Surface relocation of alpha 6 beta 4 integrins and assembly of hemidesmosomes in an in vitro model of wound healing.

The Journal of Cell Biology ◽

10.1083/jcb.115.6.1737 ◽

1991 ◽

Vol 115 (6) ◽

pp. 1737-1750 ◽

Cited By ~ 109

Author(s):

M A Kurpakus ◽

V Quaranta ◽

J C Jones

Keyword(s):

Wound Healing ◽

Epithelial Cell ◽

Epithelial Cells ◽

Connective Tissue ◽

Collagen Type ◽

Polyclonal Antibodies ◽

Vitro Model ◽

Tissue Attachment ◽

Collagen Type Vii

A transmembrane extracellular matrix receptor of the integrin family, alpha 6 beta 4, is a component of the hemidesmosome, an adhesion complex of importance in epithelial cell-connective tissue attachment (Stepp, M. A., S. Spurr-Michaud, A. Tisdale, J. Elwell, and I. K. Gipson. 1990. Proc. Natl. Acad. Sci. USA. 87:8970-8974; Jones, J. C. R., M. A. Kurpakus, H. M. Cooper, and V. Quaranta. 1991. Cell Regulation. 2:427-438). Cytosolic components of hemidesmosomes include bullous pemphigoid (BP) antigens while extracellular components include a 125-kD component of anchoring filaments (CAF) and collagen type VII-containing anchoring fibrils. We have monitored the incorporation of the alpha 6 beta 4 integrins into forming hemidesmosomes in an in vitro wound-healing explant model. In epithelial cells recently migrated from the edges of unwounded sites over bare connective tissue, alpha 6 beta 4 first appears along the entire cell surface. At this stage, these cells contain little or no cytosolic hemidesmosomal components, at least as detectable by immunofluorescence using BP autoantibodies, whereas they are already positive for laminin and CAF. At a later stage, as cells become positive for cytosolic hemidesmosome components such as BP antigens as well as collagen type VII, alpha 6 beta 4 becomes concentrated along the basal pole of the epithelial cell where it abuts the connective tissue of the explant. Polyclonal antibodies to beta 4 do not interfere with the migration of epithelial cells in the explant. However, they prevent assembly of hemidesmosomal complexes and inhibit expression of collagen type VII in cells that have migrated over wound areas. In addition, they induce disruption of established hemidesmosomes in nonmigrating cells of the unwounded area of the explant. Monoclonal antibodies to alpha 6 have a more dramatic effect, since they completely detach epithelial cells in the unwounded area of the explant. Antibodies to CAF also detach epithelial cells in unwounded areas, apparently by inducing separation between epithelium and connective tissue at the lamina lucida of the basement membrane zone. These results suggest a model whereby polarization of alpha 6 beta 4 to the basal surface of the cells, perhaps induced by a putative anchoring filament-associated ligand, triggers assembly of hemidesmosome plaques.

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Tumor Necrosis Factor-α and Interleukin (IL)-1β, IL-6, and IL-8 Impair In Vitro Migration and Induce Apoptosis of Gingival Fibroblasts and Epithelial Cells, Delaying Wound Healing

Journal of Periodontology ◽

10.1902/jop.2016.150713 ◽

2016 ◽

Vol 87 (8) ◽

pp. 990-996 ◽

Cited By ~ 22

Author(s):

Fernanda G. Basso ◽

Taisa N. Pansani ◽

Ana Paula S. Turrioni ◽

Diana G. Soares ◽

Carlos Alberto de Souza Costa ◽

...

Keyword(s):

Wound Healing ◽

Tumor Necrosis Factor ◽

Epithelial Cells ◽

Tumor Necrosis ◽

Tumor Necrosis Factor Α ◽

Gingival Fibroblasts ◽

Factor Α ◽

Necrosis Factor

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In vitro potential of human mesenchymal stem cells for corneal epithelial regeneration

Regenerative Medicine ◽

10.2217/rme-2019-0067 ◽

2020 ◽

Vol 15 (3) ◽

pp. 1409-1426 ◽

Cited By ~ 2

Author(s):

Núria Nieto-Nicolau ◽

Beatriz Martín-Antonio ◽

Claudia Müller-Sánchez ◽

Ricardo P Casaroli-Marano

Keyword(s):

Stem Cells ◽

Wound Healing ◽

Mesenchymal Stem Cells ◽

Epithelial Cells ◽

Electric Resistance ◽

Corneal Epithelial Cells ◽

Corneal Epithelial ◽

Epithelial Regeneration ◽

Human Corneal Epithelial Cells

Aim: To determine the potential of mesenchymal stem cells (MSC) for corneal epithelial regeneration in vitro. Materials & methods: Bone marrow MSC (BM-MSC) and adipose tissue MSC were analyzed for corneal epithelial and mesenchymal markers, using limbal stem cells and corneal cells as controls. MSC with better potential were cultured with specific mediums for epithelial induction. Transepithelial electric resistance and wound healing assay with human corneal epithelial cells were performed. Results: BM-MSC showed better potential, increased corneal markers, and higher transepithelial electric resistance values when induced with limbal epithelial culture medium. Induced BM-MSC promoted better wound healing of human corneal epithelial cells by paracrine secretion. Conclusion: BM-MSC has potential for corneal epithelial induction in a protocol compatible with human application.

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