Interaction Mechanism of Arc, Keyhole, and Weld Pool in Keyhole Plasma Arc Welding: A Review.

The Keyhole Plasma Arc Welding (KPAW) process utilizes arc plasma highly constricted by a water-cooled cupper nozzle to produce great arc pressure for opening a keyhole in the weld pool, achieving full penetration to the thick plate. However, advanced control of welding is known to still be difficult due to the complexity of the process mechanism, in which thermal and dynamic interactions among the arc, keyhole, and weld pool are critically important. In KPAW, two large eddies are generally formed in the weld pool behind the keyhole by plasma shear force as the dominant driving force. These govern the heat transport process in the weld pool and have a strong influence on the weld pool formation process. The weld pool flow velocity is much faster than those of other welding processes such as Tungsten Inert Gas (TIG) welding and Gas Metal Arc (GMA) welding, enhancing the heat transport to lower the weld pool surface temperature. Since the strength and direction of this shear force strongly depend on the keyhole shape, it is possible to control the weld pool formation process by changing the keyhole shape by adjusting the torch design and operating parameters. If the lower eddy is relatively stronger, the heat transport to the bottom side increases and the penetration increases. However, burn-through is more likely to occur, and heat transport to the top side decreases, causing undercut. In order to realize further sophistication of KPAW, a deep theoretical understanding of the process mechanism is essential. In this article, the recent progress in studies regarding the interaction mechanism of arc, keyhole, and weld pool in KPAW is reviewed.

Individual Effects of Alkali Element and Wire Structure on Metal Transfer Process in Argon Metal-Cored Arc Welding.

This study aimed to clarify the effect of wire structure and alkaline elements in wire composition on metal transfer behavior in metal-cored arc welding (MCAW). A comparison of metal transfer in pure argon gas was carried out using a solid wire (wire 1), a metal-cored wire without an alkaline element (wire 2), and another metal-cored wire with 0.084 mass% of sodium (wire 3). The experiments were conducted under 280 and 320 A welding currents, observed by high-speed imaging techniques equipped with laser assistance and bandpass filters. At 280 A, wire 1 showed a streaming transfer mode, while the others showed a projected one. When the current was 320 A, the metal transfer of wire 2 changed to streaming, while wire 3 remained projected. As sodium has a lower ionization energy than iron, the mixing of sodium vapor into the iron plasma increases its electrical conductivity, raising the proportion of current flowing through metal vapor plasma. As a result, the current flows to the upper region of the molten metal on the wire tip, with the resulting electromagnetic force causing droplet detachment. Consequently, the metal transfer mode in wire 3 remained projected. Furthermore, weld bead formation is the best for wire 3.

Numerical Analysis of Physical Characteristics and Heat Transfer Decoupling Behavior in Bypass Coupling Variable Polarity Plasma Arc.

A novel bypass coupling variable polarity plasma arc was proposed to achieve the accurate adjusting of heat and mass transfer in the welding and additive manufacturing of aluminum alloy. However, the physical characteristics and decoupled transfer behavior remain unclear, restricting its application and development. A three-dimensional model of the bypass coupling variable polarity plasma arc was built based on Kirchhoff's law, the main arc and the bypass arc are coupled by an electromagnetic field. The model of current attachment on the tungsten electrode surface is included for simulating different heating processes of the EP and EN phases in the coupling arc. The distribution of temperature field, flow field, and current density of the bypass coupling variable polarity plasma arc was studied by the three-dimensional numerical model. The heat input on the base metal under different current conditions is quantified. To verify the model, the arc voltages are compared and the results in simulation and experiment agree with each other well. The results show that the radius of the bypass coupling arc with or without bypass current action on the base metal is different, and the flow vector of the bypass coupling arc plasma with bypass current is larger than the arc without bypass current. By comparing the heat transfer on the electrodes' boundary under different current conditions, it is found that increasing the bypass current results in the rise in heat input on the base metal. Therefore, it is concluded that using bypass current is unable to completely decouple the wire melting and the heat input of the base metal. The decoupled degree of heat transfer is one of the important factors for accurate control in the manufacturing process with this coupling arc.

3D Numerical Study of External Axial Magnetic Field-Controlled High-Current GMAW Metal Transfer Behavior.

For gas metal arc welding (GMAW), increasing the welding current is the most effective way to improve welding efficiency. However, much higher current decreases the welding quality as a result of metal rotating-spray transfer phenomena in the high-current GMAW process. In this work, the external axial magnetic field (EAMF) was applied to the high-current GMAW process to control the metal transfer and decrease the welding spatters. A unified arc-droplet coupled model for high-current GMAW using EAMFs was built to investigate the metal rotating-spray transfer behavior. The temperature fields, flow fields in the arc, and droplet were revealed. Considering all the heat transferred to the molten metal, the Joule heat was found to be the dominant factor affecting the droplet temperature rise, followed by the anode heat. The conductive heat from the arc contributed less than half the value of the other two. Considering the EAMFs of different alternating frequencies, the arc constricting effects and controlled metal transfer behaviors are discussed. The calculated results agree well with the experimental high-speed camera observations.

Numerical Simulation of the Behavior of Hydrogen Source in a Novel Welding Process to Reduce Diffusible Hydrogen.

This study aims to reduce the diffusible hydrogen content in deposited metal during gas metal arc welding (GMAW) and flux-cored arc welding (FCAW) which induces cold cracking. To achieve this, a novel welding torch with a dual gas nozzle has been developed. This special welding torch decreases the hydrogen source gas evaporated from a welding wire by the suction from the inner gas nozzle. In order to improve the suction efficiency of this evaporated gas, precise control of the suction gas flow is indispensable. In this paper, a simplified numerical simulation model of this process has been described. This model can take account of the evaporation of the hydrogen source gas from the wire while simulating the behavior of the shielding gas and the arc. Using this model, the effect of suction nozzle structure and torch operating conditions on suction gas flow pattern and suction efficiency was also investigated to understand the process mechanism. Furthermore, the diffusible hydrogen content in deposited metal was measured by chromatography as a validation step. Results show that some of the shielding gas introduced from a shielding nozzle was drawn inward and also branched into an upward flow that was sucked into the suction nozzle and a downward flow to a base metal. This branching height was defined as the suction limit height, which decisively governed the suction efficiency. As a result, in order to reduce the diffusible hydrogen, it was suggested that the suction limit height should be controlled towards below the wire position, where the evaporation rate of the hydrogen source gas peaks through optimization of the suction nozzle design and the torch operating conditions.

Study on the Decoupled Transfer of Heat and Mass in Wire Variable Polarity Plasma Arc Welding.

A hybrid arc-wire welding method based on the variable polarity plasma arc (VPPA) and variable polarity pulse metal inert-gas (VP-PMIG) was proposed for manufacturing aluminum alloys. This paper aims to clarify the decoupling control process of heat and mass transfer in the hybrid welding process. To understand the arc physics and analyze the mass transfer behavior, the hybrid arc shape and droplet cross-sectional area with different parameters were obtained by high speed video photography. Further, the melting area of the base metal was analyzed by macro-metallography of the weld bead cross-section to study the heat transfer. It is found that the hybrid arc shape changes with time. The VPPA main arc is deflected to one side by the VP-PMIG, making the temperature distribution asymmetric, and during the VP-PMIG pulse necking occurs. The cross-sectional area of the droplet is more obviously affected by the VP-PMIG current than the VPPA current. Meanwhile, the VPPA current dominates the melting area of the base metal. Therefore, we conclude that heat transfer to the base metal is from the VPPA, while droplet transfer is mainly controlled by the VP-PMIG arc. These conclusions are confirmed by analyzing the decoupling degree of heat and mass transfer of the base metal by the VPPA and VP-PMIG arc.

Influence of Current Feeding Position of Duplex Current Feeding MIG Welding on Droplet Heat Quantity.

Pure argon metal inert gas (MIG) welding is expected to offer the possibility to obtain high toughness weld joints. However, due to its arc instability and low wettability, it is difficult to apply pure argon MIG to a practical welding structure. In order to solve these problems, an improved MIG welding process with a duplex current feeding (DCF-MIG) mechanism was developed. In the DCF-MIG process, the welding current and the wire feeding speed are independently controlled by an additionally feeding secondary current from a secondary power source. Thereby, DCF-MIG can supply a large current compared to conventional MIG under the same deposition rate. In this study, to consider the influence of the secondary current feeding position of DCF-MIG on droplet heat quantity, droplet heat quantity was measured by calorimetry. As a result, the droplet heat quantity was found to be increased significantly with the increase of the distance between the primary current feeding point and secondary current feeding point. The increase of the droplet heat quantity in the DCF-MIG process had a strong effect on improving bead shape and penetration. The droplet heat quantity with the effective current value of DCF-MIG was derived from the simplified calculation and the results roughly agreed with the experimental data.

Effect of Arc Pressure on the Digging Process in Variable Polarity Plasma Arc Welding of A5052P Aluminum Alloy.

The keyhole digging process associated with variable polarity plasma arc (VPPA) welding remains unclear, resulting in poor control of welding stability. The VPPA pressure directly determines the dynamics of the keyhole and weld pool in the digging process. Here, through a high speed camera, high frequency pulsed diode laser light source and X-ray transmission imaging system, we reveal the potential physical phenomenon of a keyhole weld pool. The keyhole depth changes periodically corresponding to the polarity conversion period if the current is same in the electrode negative (EN) phase and electrode positive (EP) phase. There exist three distinct regimes of keyhole and weld pool behavior in the whole digging process, due to the arc pressure attenuation and energy accumulation effect. The pressure in the EP phase is smaller than that of the EN phase, causing the fluctuation of the weld pool free surface. Based on the influence mechanism of energy and momentum transaction, the arc pressure output is balanced by separately adjusting the current in each polarity. Finally, the keyhole fluctuation during the digging process is successfully reduced and welding stability is well controlled.

Silibinin's regulation of proliferation and collagen gene expressions of rat pancreatic ß-cells cultured on types I and V collagen involves ß-catenin nuclear translocation.

Extracellular matrix (ECM) molecules have multiple functions; prevention of cytotoxicity, provision of mechanical support, cell adhesive substrates and structural integrity in addition to mediation of cellular signaling. In this study, we report that the proliferation of INS-1 cells cultured on collagen I-coated dishes is enhanced, but it is inhibited on collagen V-coated dishes. Inhibitory proliferation on collagen V-coated is not due to apoptosis induction. Silibinin decreases hepatic glucose production and protects pancreatic ß-cells, as a potential medicine for type II diabetes. Silibinin up-regulates the proliferation of cells cultured on both collagen I- and V-coated dishes. Collagen-coating regulates gene expression of collagen in a collagen type-related manner. Silibinin increases mRNA expression of collagen I in the cells on collagen I- and V-coated dishes; however, silibinin decreases collagen V mRNA expression on collagen I- and V-coated dishes. Collagen I-coating significantly enhances nuclear translocation of ß-catenin, while collagen V-coating reduces it. Differential effects of silibinin on collagen I mRNA and collagen V mRNA can be accounted for by the finding that silibinin enhances nuclear translocation of ß-catenin on both collagen I- and V-coated dishes, since phenomenologically nuclear translocation of ß-catenin enhances collagen I mRNA but represses collagen V mRNA. These results demonstrate that nuclear translocation of ß-catenin up-regulates proliferation and collagen I gene expression, whereas it down-regulates collagen V gene expression of INS-1 cells. Differential gene expressions of collagen I and V by nuclear ß-catenin could be important for understanding fibrosis where collagen I and V may have differential effects.

Silibinin protects rat pancreatic ß-cell through up-regulation of estrogen receptors' signaling against amylin- or Aß1-42 -induced reactive oxygen species/reactive nitrogen species generation.

Amylin and amyloid-ß (Aß) were found to induce reactive oxygen species (ROS) and reactive nitrogen species (RNS) in rat pancreatic ß-cell line, INS-1 cells, leading to cell death. In this study, we report on reciprocal relationship between the expression of estrogen receptors (ERs) α and ß (ERα and ERß) and generation of ROS/RNS in amylin/Aß1-42 -treated INS-1 cells. That is, pharmacological activation of ERs in INS-1 cells significantly decreases ROS/RNS generation, but blockage of ERs increases ROS/RNS generation. Silibinin is a natural polyphenolic flavonoid isolated from milk thistle with phytoestrogen activities, also known as silybin. Treatment with silibinin down-regulated ROS/RNS production induced by treatment with amylin/Aß1-42 in the cells. Silencing ERs expression with siRNAs targeting ERs showed that the protective effect of silibinin was markedly weakened, indicating that silibinin protection is largely attributed to activation of ERs' signaling. The binding of silibinin to ERs implies that the protective effect of silibinin on amylin/Aß1-42 -treated INS-1 cells owes to down-regulation of ROS/RNS through the activation of ERs phosphorylation. Amylin and Aß1-42 cotreatment enhanced furthermore ROS/RNS generation and cytotoxicity through further down-regulation of ERs phosphorylation, and this was reversed by silibinin. Silibinin also protects INS-1 cells from amylin and Aß1-42 cotreatment. These results indicate that protective effect of silibinin is mediated by enhancement of ERs phosphorylation that depresses ROS/RNS generation in amylin/Aß1-42 -treated INS-1 cells.

Silibinin-induced autophagy mediated by PPARα-sirt1-AMPK pathway participated in the regulation of type I collagen-enhanced migration in murine 3T3-L1 preadipocytes.

Preadipocyte migration is a fundamental and important process for the development of tissue organization, especially in the development of primitive adipose tissue and adipocyte tissue wound healing. However, excessive migration may result in abnormal development and fibrosis-related diseases such as hypertrophic scar. We previously reported that type I collagen (collagen I) enhanced migration of 3T3-L1 preadipocytes via phosphorylation and/or acetylation of NF-κB p65, and the enhanced cell migration is repressed by silibinin treatment through sirt1. It is known that sirt1 has an ability to deacetylate acetylated NF-κB p65, but little is known about the effect of sirt1 on phosphorylated NF-κB p65. This study aims to examine the potential effect of sirt1 on the regulation of phosphorylated NF-κB p65 and the underlying mechanism. Autophagy is involved in many physiological and pathological processes, including regulation of cell migration as well as in cellular homeostasis. The present study demonstrates that silibinin induces autophagy in a dose-dependent manner in 3T3-L1 cells. Autophagy is under the regulation of sirt1/AMPK pathway, and inhibits collagen I-enhanced migration of 3T3-L1 cells through negative regulation of NF-κB p65 phosphorylation but not acetylation. The expression of peroxisome proliferator-activated receptor α (PPARα) is up-regulated with silibinin accompanying up-regulation of autophagy through activating sirt1 in 3T3-L1 cells. Taken together, these findings indicate that silibinin-induced autophagy is mediated by up-regulation of PPARα-sirt1-AMPK, contributing to repression of type I collagen-enhanced migration in murine 3T3-L1 preadipocytes through down-regulation of phosphorylated NF-κB p65.

Type I collagen-induced YAP nuclear expression promotes primary cilia growth and contributes to cell migration in confluent mouse embryo fibroblast 3T3-L1 cells.

The extracellular matrix (ECM) is a major biomechanical environment for all cells in vivo, and tightly controls wound healing and cancer progression. Type I collagen (Col I) is the most abundant component in ECM and plays an essential role for cell motility control and migration beyond structural support. Our previous results showed that Col I increased the length of primary cilia and the expression of primary cilia-associated proteins in 3T3-L1 cells. The Hippo/YAP pathway serves as a major integrator of cell surface-mediated signals and regulates key processes for the development and maintenance of tissue functions. In this study, we investigated the role of Hippo/YAP signaling in primary cilia growth of cells cultured on Col I-coated plate, as well as the potential link between primary cilia and migration. At 2-day post-confluence, YAP localization in the nucleus was dramatically increased when the cells were cultured on Col I-coated plate, accompanied by cilia growth. YAP inhibitor verteporfin repressed the growth of primary cilia as well as the expressions of ciliogenesis-associated proteins in confluent 3T3-L1 cells cultured on Col I-coated plate. Moreover, knockdown of either YAP or IFT88, one of the ciliogenesis-associated proteins, reversed the migration of confluent 3T3-L1 cells promoted by Col I-coating. In conclusion, activation of YAP pathway by Col I-coating of culture plate for confluent 3T3-L1 cells is positively associated with the primary cilia growth, which eventually results in promoted migration.

Activated toll-like receptor 4 is involved in oridonin-induced phagocytosis via promotion of migration and autophagy-lysosome pathway in RAW264.7 macrophages.

In our previous study, we demonstrated that oridonin enhances phagocytosis of apoptotic bodies by macrophage-like cells by inducing autophagy. However, the direct sensor of autophagy and the key event controlling phagocytosis remains unknown. Herein, we showed that Toll-like receptor 4 (TLR4), known to mediate immune responses, was activated by oridonin. Activated TLR4 contributes to phagocytosis of apoptotic cells by RAW264.7 macrophages. Indeed, inhibition or small interfering RNA (siRNA) silencing of TLR4 significantly attenuated oridonin-induced phagocytosis. Inhibition of TLR4 also decreased the level of autophagy and its associated proteins, Beclin-1 and light chain 3 (LC3), suggesting that activated TLR4 is involved in activation of autophagy. LPS-induced activation of TLR4 promoted phagocytosis and autophagy progression. Activation of TLR4 accompany increase in activities of lysosome acid phosphatase and cathepsin B as well as in up-regulation of lysosomal-associated membrane protein (LAMP 1 and 2) levels. Furthermore, TLR4 in association with translocation to cytoplasm leads to macrophage motility or migration through increased plasticity of skeleton and/or membrane structure. These results suggest that oridonin-induced phagocytosis of apoptotic bodies by macrophages is TLR4 signal pathway-mediated, via activation of the autophagy-lysosome pathway as well as increase of cell migration.

Reactive oxygen species are responsible for the cell aggregation and production of pro-inflammatory mediators in phorbol ester (PMA)-treated U937 cells on gelatin-coated dishes through upregulation of autophagy.

Purpose: Our previous studies indicate that phorbol 12-myristate 13-acetate (PMA)-treated U937 cells cultured on collagen I-coated dishes express lowered production of pro-inflammatory mediators in parallel through reduced reactive oxygen species (ROS) levels. By contrast, PMA-treated U937 cells on gelatin, the denatured collagen, show enhanced production of pro-inflammatory mediators, mediated by up-regulating autophagy levels. The present study is aimed to investigate the effect of ROS levels in PMA-treated U937 cells cultured on gelatin-coated surface. Material and methods: MTT assay, flow cytometric analysis of ROS and autophagy, biochemical detection of antioxidant levels, enzyme-linked immunosorbent assay, and western blot were used. Results: Gelatin-coating increased ROS levels in PMA-treated U937 cells. Increased ROS levels are involved in the regulation of cell aggregation and the release of pro-inflammatory mediators in gelatin-coated culture. These results lead to the query about the crosstalk between the two positive regulators, the autophagy and ROS. Autophagy induction is attenuated by N-acetyl-L-cysteine treatment, but the treatment with autophagy inhibitor, 3-methyladenine, does not affect ROS levels, suggesting ROS are upstream of autophagy in the regulation axis of differentiated U937 cells on gelatin-coated surface. Further study confirmed that upregulation of autophagy was responsible for ROS-induced cell aggregation and production of pro-inflammatory mediators. Conclusion: The results suggest that gelatin-coating promotes the aggregation of PMA-treated U937 cells and the production of pro-inflammatory mediators by ROS-autophagy signaling pathway.

Silibinin ameliorates amylin-induced pancreatic ß-cell apoptosis partly via upregulation of GLP-1R/PKA pathway.

The objective was to investigate the mechanism of the protective effect of silibinin on amylin/Aß1-42-induced INS-1 cell apoptosis, with special reference to the roles of glucagon-like peptide-1 receptor (GLP-1R) and protein kinase A (PKA). The effects of silibinin on apoptosis, insulin secretion, GLP-1R, and PKA expression in the INS-1 cells treated with amylin/Aß1-42 were examined. INS-1 cells exposed to amylin showed increased TUNEL-positive ratio, reduced expression of GLP-1R and PKA. GLP-1R antagonists or PKA inhibitor enhanced the expression of apoptosis-associated proteins and TUNEL-positive ratio. Silibinin exerted antiapoptotic effect on and upregulation of GLP-1R and PKA. However, Aß1-42-induced INS-1 cell apoptosis, GLP-1R, and PKA expressions were not changed. Our results indicate that down-regulation of GLP-1R and PKA contributes to INS-1 cell apoptosis induced with amylin. Silibinin protects INS-1 cells from amylin-induced apoptosis through activation of GLP-1R/PKA signaling. Silibinin's inhibition of the toxic effects of Aß1-42 is independent of GLP-1R/PKA pathway.

Persistent IKKα phosphorylation induced apoptosis in UVB and Poly I:C co-treated HaCaT cells plausibly through pro-apoptotic p73 and abrogation of IκBα.

Toll-like receptor 3 (TLR3), a member of pattern recognition receptors, is reported to initiate skin inflammation by recognizing double-strand RNA (dsRNA) released from UVB-irradiated cells. Recently, we have discovered the NF-κB pathway activated by TLR3 is involved in apoptosis of UVB-Poly I:C-treated HaCaT cells. The real culprit for apoptosis has not been precisely identified since the system of NF-κB pathway is complex. In this study, we silenced main transcriptional factors in NF-κB family, RelA, RelB and c-Rel, but to our surprise the results show that none of them participate in apoptosis induction in UVB-Poly I:C-treated HaCaT cells. Therefore, we moved to investigate the apoptosis-associated molecules in the upstream of NF-κB pathway. We firstly checked the expression of IκBα, an NF-κB inhibitor. UVB (4.8 mJ/cm2) and Poly I:C (0.3 µg/mL) co-treatment decreased IκBα expression level in a time-dependent manner. Silencing IκBα with siRNA further enhanced UVB-Poly I:C-induced cell death. We then investigated IκB kinase (IKK) complex that contributes to the degradation of IκBα. IKK is composed of IKKα, IKKß and NEMO. Treatment with IKK-16, an IKKα/ß inhibitor, significantly diminished UVB-Poly I:C-induced IκBα degradation and thus apoptosis. Silencing either IKKα or NEMO but not IKKß with corresponding siRNA inhibited apoptosis. Tumor repressor p73, a homologue of p53, is reported to mediate IKKα-induced apoptosis in DNA damage response. Silencing p73 reduced cell apoptosis in UVB-Poly I:C-treated HaCaT cells. In summary, UVB and Poly I:C co-treatment activates IKKα and NEMO, which diminishes anti-apoptotic IκBα, resulting in enhancement of apoptosis through p73. The findings partially clarify the possible molecular mechanism of pro-apoptotic NF-κB pathway activated by TLR3 in the fate of UVB-irradiated epidermis.

L-A03, a dihydroartemisinin derivative, promotes apoptotic cell death of human breast cancer MCF-7 cells by targeting c-Jun N-terminal kinase.

L-A03 is a dihydroartemisinin derivative and exerts distinct anti-tumor activity in vitro. Previous studies showed that induction of autophagy and deficiency in nitric oxide (NO) generation contributed to apoptotic cell death in L-A03-treated MCF-7 cells. However, the detailed mechanism is still unclear. In this study, the role of mitogen-activated protein kinases (MAPKs) in this apoptotic process was investigated. L-A03 (7.5-30 µM) selectively inhibited the activation of c-Jun N-terminal protein kinase (JNK) with no significant effect on extracellular signal related kinase (ERK) and p38. In addition, the possible mechanism of interaction between JNK and L-A03 was also investigated by molecular docking. In the presence of SP600125, a specific JNK inhibitor, induction of autophagy and apoptosis with L-A03 at 15 µM were elevated, but NO generation was attenuated, indicating that JNK inactivation is essential for apoptotic cell death. Interestingly, autophagy induction and NO generation did not affect the activation of JNK, demonstrating that JNK is upstream to autophagy and NO. Taken together, L-A03-induced JNK inactivation enhances autophagic and apoptotic cell death, but represses the generation of NO. This study provides a new insight on the mechanism of L-A03-induced cell death by targeting JNK.

Prostaglandin E2 attenuates synergistic bactericidal effects between COX inhibitors and antibiotics on Staphylococcus aureus.

PGE2 is found to attenuate the bactericidal effects of kanamycin or ampicillin in Staphylococcus aureus, as well as the methicillin-resistant S. aureus (MRSA). Co-treatment with cyclooxygenase (COX) inhibitors (celecoxib, aspirin or naproxen) synergistically enhances kanamycin or ampicillin-induced cell death of S. aureus and MRSA. COX inhibitors repressed bacterial multidrug resistance through down-regulating efflux pump activity in antibiotics-treated S. aureus and MRSA. However, this synergistic bactericidal effects are reduced by the treatment with PGE2. PGE2 restores the efflux pump activity as well as increases biofilm formation in S. aureus and MRSA. Collectively, the enhancement of efflux pump activity and biofilm formation with PGE2 might partially explain the resistance to synergistic bactericidal effects between COX inhibitors and antibiotics in PGE2-treated S. aureus.

Type I collagen induces mesenchymal cell differentiation into myofibroblasts through YAP-induced TGF-ß1 activation.

In organ fibrosis, mechanical stress and transforming growth factor beta-1 (TGF-ß1) promote differentiation into myofibroblast from mesenchymal cells, leading to extracellular matrix (ECM) remodeling or active synthesis, deposition or degradation of ECM components. A major component of ECM, type I collagen (col I) triple helical molecules assemble into fibrils or are denatured to gelatin without triple-helicity in remodeling. However, whether changes of ECM components in remodeling have influence on mesenchymal cell differentiation remains elusive. This study adopted three states of collagen I existing in ECM remodeling: molecular collagen, fibrillar collagen and gelatin to see what are characteristics in the effects on two cell lines of mesenchymal origin, murine 3T3-L1 embryonic fibroblast and murine C2C12 myoblasts. The results showed that all three forms of collagen I were capable of inducing these two cells to differentiate into myofibroblasts characterized by increased expression of alpha-smooth muscle actin (α-SMA) mRNA. The expression of α-SMA is positively regulated by TGF-ß1. Nuclear translocation of Yes-associated protein (YAP) is involved in this process. Focal adhesion kinase (FAK) is activated in the cells cultured on molecular collagen-coated plates, contributing to YAP activation. On the other hand, in the cells cultured on fibrillar collagen gel or gelatin-coated plates, oxidative stress but not FAK induce YAP activation. In conclusion, the three physicochemically distinct forms of col I induce the differentiation of mesenchymal cells into myofibroblasts through different pathways.

Differential levels of reactive oxygen species in murine preadipocyte 3T3-L1 cells cultured on type I collagen molecule-coated and gel-covered dishes exert opposite effects on NF-κB-mediated proliferation and migration.

Reactive oxygen species (ROS) participate in various cell responses in association with cell proliferation, migration, differentiation, and death. Extracellular matrix (ECM) serves as cellular microenvironments for many kinds of cells, affecting cell activities. However, whether or not ECM influences cellular ROS levels has not been well studied. In this study, cells are cultured on collagen I molecule-coated (mol. coated) dishes and collagen I fibrous gel-covered (gel) dishes to explore their influence on cell behaviours. We found that the levels of ROS in murine 3T3-L1 preadipocytes increased both in cells on mol. coated and those on the gel. Much higher ROS levels were found in the cells cultured on the gel. Cell proliferation and migration were stimulated to opposite directions between the cells on mol. coated and the cells on gel. ROS in a moderate level were positive regulators in the proliferation and migration of cells on mol. coated; however, ROS in a high level served as negative regulators in the cells on gel. These opposite effects on cell proliferation and migration affected by different ROS levels are in parallel with opposite levels of NF-κB p65 activation.