Progress of polysaccharide-based tissue adhesives.

Recently, polymer-based tissue adhesives (TAs) have gained the attention of scientists and industries as alternatives to sutures for sealing and closing wounds or incisions because of their ease of use, low cost, minimal tissue damage, and short application time. However, poor mechanical properties and weak adhesion strength limit the application of TAs, although numerous studies have attempted to develop new TAs with enhanced performance. Therefore, next-generation TAs with improved multifunctional properties are required. In this review, we address the requirements of polymeric TAs, adhesive characteristics, adhesion strength assessment methods, adhesion mechanisms, applications, advantages and disadvantages, and commercial products of polysaccharide (PS)-based TAs, including chitosan (CS), alginate (AL), dextran (DE), and hyaluronic acid (HA). Additionally, future perspectives are discussed.

Highly Resilient Noncovalently Associated Hydrogel Adhesives for Wound Sealing Patch.

The development of hydrogel adhesives with high mechanical resilience and toughness remains a challenging task. Hydrogels must exhibit high mechanical resilience to withstand the inevitable movement of the human body while simultaneously demonstrating strong wet tissue adhesion and appropriate toughness to hold and seal damaged tissues; However, tissue adhesion, toughness, and mechanical resilience are typically negatively correlated. Therefore, this paper proposes a highly resilient double-network (DN) hydrogel wound-sealing patch that exhibits a well-balanced combination of tissue adhesion, toughness, and mechanical resilience. The DN structure is formed by introducing covalently and non-covalently crosslinkable dopamine-modified crosslinkers and physically interactable linear poly(vinyl imidazole) (PVI). The resulting hydrogel adhesive exhibits high toughness and mechanical resilience due to the presence of a DN involving reversible physical intermolecular interactions such as hydrogen bonds, hydrophobic associations, cation-π interactions, π-π interactions, and chain entanglements. Moreover, the hydrogel adhesive achieves strong wet tissue adhesion through the polar hydroxyl groups of dopamine and the amine group of PVI. These mechanical attributes allow the proposed adhesive to effectively seal damaged tissues and promote wound healing by maintaining a moist environment.

Progress of tissue adhesives based on proteins and synthetic polymers.

In recent years, polymer-based tissue adhesives (TAs) have been developed as an alternative to sutures to close and seal incisions or wounds owing to their ease of use, rapid application time, low cost, and minimal tissue damage. Although significant research is being conducted to develop new TAs with improved performances using different strategies, the applications of TAs are limited by several factors, such as weak adhesion strength and poor mechanical properties. Therefore, the next-generation advanced TAs with biomimetic and multifunctional properties should be developed. Herein, we review the requirements, adhesive performances, characteristics, adhesive mechanisms, applications, commercial products, and advantages and disadvantages of proteins- and synthetic polymer-based TAs. Furthermore, future perspectives in the field of TA-based research have been discussed.

Highly Resilient Dual-Crosslinked Hydrogel Adhesives Based on a Dopamine-Modified Crosslinker.

Hydrogels are promising material for wound dressing and tissue engineering. However, owing to their low tissue adhesion in a moist environment and lack of flexibility, hydrogels are still not widely applied in movable parts, such as joints. Herein, we report a dual-crosslinked hydrogel adhesive using a dopamine-modified and acrylate-terminated crosslinker, tri(ethylene glycol) diacrylate-dopamine crosslinker (TDC). The covalent crosslinking was formed by photopolymerization between acrylic acid (AA) and TDC, and the noncovalent crosslinking was formed by intermolecular dopamine-dopamine and dopamine-AA interactions. Our resultant hydrogel demonstrated strong tissue adhesion in a moist environment (approximately 71 kPa) and high mechanical resilience (approximately 94%) with immediate recovery at a 200% strain rate. Moreover, it accelerated wound healing upon dressing the wound site properly. Our study provides the potential for advanced polymer synthesis by introducing a functional crosslinking agent.

Latent, Cross-Linkable Triazole Platform on a Carbon Fiber Surface for Enhancing Interfacial Cross-Linking within Carbon Fiber/Epoxy Composites.

A long-running need in carbon fiber composite production is to ameliorate interfacial adhesion between the polymer and carbon fibers. Here, we present a convenient and feasible strategy for controlling the carbon fiber's surface in a continuous process: syntheses of click-modified silanes via copper(I)-catalyzed azide-alkyne cycloaddition reaction and grafting them onto fiber surfaces which prepare a latent curable platform under mild processes without postmodification. As 1,2,3-triazole moieties from the click reaction were added to the epoxy/dicyandiamide system, they triggered additional reactions in the later conversion stage; approximately, a 20% increase in the total reaction enthalpy compared to the system with no additives was obtained. We expected the enhanced cross-linking between the surface and matrix to expand the interfacial area, leading to reinforcements on interfacial adhesion and stress-transfer abilities within composites. The merit of the approach is well-demonstrated by conductive atomic force microscopy, showing that the interphase can be extended up to 6-fold when the triazole platform acts as curatives and serve as bridges after the epoxy cure. Consequently, the composite's interfacial shear strength and interlaminar shear strength were increased up to 78 and 72%, respectively. This work affords a reactive platform where a custom-tailored fiber/matrix interface can be designed by virtue of versatility in clickable reactants.

Rapid Photoresponsive Switchable Pressure-Sensitive Adhesive Containing Azobenzene for the Mini-Light Emitting Diode Transfer Process.

Materials that can switch adhesive properties based on external stimuli are required in several industries for temporary bonding or transfer processes. Previously studied materials achieve this under restricted conditions (hydration, heat, and long switching times), and some materials have limitations related to reuse because of irreversible reactions or residue formation on substrates. Herein, a rapid photoresponsive switchable pressure-sensitive adhesive (PSA) fabricated using an acrylic polymer and an aliphatic monomer containing azobenzene is reported. The adhesion force of the proposed PSA can be switched by photoisomerizing the azobenzene moiety. The process induces the transition of surface energy and modulus of the PSA. Ultraviolet and visible light irradiation can switch the probe tack force from 200 to 4 kPa within 15-30 s. Adhesion switching is possible in a state wherein the PSA remains adhered to a substrate. Mini-LEDs are selectively transferred from the carrier PSA to a polydimethylsiloxane substrate following the process of partial adhesion switching of the PSA. The novel and switchable PSA, which exhibits a selective and repeatable adhesion switching property and high switching ratio when stimulated by light stimuli, may be potentially used to realize the mini-LED or micro-LED transfer processes.

Photoresponsive, switchable, pressure-sensitive adhesives: influence of UV intensity and hydrocarbon chain length of low molecular weight azobenzene compounds.

Unlike traditional adhesives with a fixed adhesive force, switchable adhesives, which have an adhesive force that can be adjusted by external stimuli, are specifically designed to be released according to user demand, or to enable the transfer of fine electronic devices. Previously developed switchable adhesives have limitations such as a slow switching rate, narrow adhesion modulation range, or the lack of reusability. Thus, we fabricated switchable pressure-sensitive adhesives (PSAs) that can overcome these limitations. The adhesive force of each switchable PSA, which comprises an azobenzene-containing acrylic polymer and low molecular weight compounds, was designed to be activated/deactivated via ultraviolet (UV) and visible light irradiation. The adhesive force and UV intensity required for the switch were found to be dependent on the aliphatic chain length of the compound. The adhesive force of the SP-C10, i.e., a switchable PSA containing a azobenzene compound with an aliphatic chain of 10 hydrocarbons, increased to 3.5 N from nearly zero in response to only 30 s of low-level (25 mW cm-2) UV irradiation. Additionally, SP-C10 did not lose its adhesive force even after 30 cycles of repeated adhesion switching. The mechanism of adhesion switching influenced by UV intensity and the structure of low molecular weight azobenzene compounds are also reported.

Phellodendron amurense Extract Protects Human Keratinocytes from PM2.5-Induced Inflammation via PAR-2 Signaling.

Dietary supplement and personal care products aiming to provide protection from air pollution have been of great interest for decades. Epidemiology demonstrated that PM10 and PM2.5 particulate matter (PM) are an actual threat to public health worldwide, but the detailed processes of how these particles attack the cells are not fully understood. Here, we report that the measurement of intracellular calcium concentration ([Ca2+]i) using human respiratory or skin cells can illustrate pollutant challenges by triggering Ca2+ influx in these cells. This signal was generated by proteinase-activated receptor-2 (PAR-2), confirmed by competition analyses, and Phellodendron amurense bark extract (PAE), a traditional medicine, was able to control the response and expression of PAR-2. Increase in proinflammatory cytokines and decrease in cell adhesion components could suggest a severe damage status by air pollutants and protection by PAE. Finally, we identified 4-O-feruloylquinic acid (FQA), an active compound of PAE, showing the same effects on Ca2+ influx and PAR-2 regulation. The results presented here should help understand the underlying mechanism of PM insults and the beneficial effect of standardized PAE as dietary supplement or cosmetical ingredient.

Dual-Blocking of PI3K and mTOR Improves Chemotherapeutic Effects on SW620 Human Colorectal Cancer Stem Cells by Inducing Differentiation.

Cancer stem cells (CSCs) have tumor initiation, self-renewal, metastasis and chemo-resistance properties in various tumors including colorectal cancer. Targeting of CSCs may be essential to prevent relapse of tumors after chemotherapy. Phosphatidylinositol-3-kinase (PI3K) and mammalian target of rapamycin (mTOR) signals are central regulators of cell growth, proliferation, differentiation, and apoptosis. These pathways are related to colorectal tumorigenesis. This study focused on PI3K and mTOR pathways by inhibition which initiate differentiation of SW620 derived CSCs and investigated its effect on tumor progression. By using rapamycin, LY294002, and NVP-BEZ235, respectively, PI3K and mTOR signals were blocked independently or dually in colorectal CSCs. Colorectal CSCs gained their differentiation property and lost their stemness properties most significantly in dual-blocked CSCs. After treated with anti-cancer drug (paclitaxel) on the differentiated CSCs cell viability, self-renewal ability and differentiation status were analyzed. As a result dual-blocking group has most enhanced sensitivity for anti-cancer drug. Xenograft tumorigenesis assay by using immunodeficiency mice also shows that dual-inhibited group more effectively increased drug sensitivity and suppressed tumor growth compared to single-inhibited groups. Therefore it could have potent anti-cancer effects that dual-blocking of PI3K and mTOR induces differentiation and improves chemotherapeutic effects on SW620 human colorectal CSCs.

Targeting ROR1 inhibits the self-renewal and invasive ability of glioblastoma stem cells.

Glioblastoma is the most malignant of brain tumours and is difficult to cure because of interruption of drug delivery by the blood-brain barrier system, its high metastatic capacity and the existence of cancer stem cells (CSCs). Although CSCs are present as a small population in malignant tumours, CSCs have been studied as they are responsible for causing recurrence, metastasis and resistance to chemotherapy and radiotherapy for cancer. CSCs have self-renewal characteristics like normal stem cells. The aim of this study was to investigate whether receptor tyrosine kinase-like orphan receptor 1 (ROR1) is involved in stem cell maintenance and malignant properties in human glioblastoma. Knockdown of ROR1 caused reduction of stemness and sphere formation capacity. Moreover, down-regulation of ROR1 suppressed the expression of epithelial-mesenchymal transition-related genes and the tumour migratory and invasive abilities. The results of this study indicate that targeting ROR1 can induce differentiation of CSCs and inhibit metastasis in glioblastoma. In addition, ROR1 may be used as a potential marker for glioblastoma stem cells as well as a potential target for glioblastoma stem cell therapy.

Inhibition of c-Yes induces differentiation of HT-29 human colon cancer stem cells through midbody elongation.

Recent research suggests that a small group of cells, named cancer stem cells (CSCs), is responsible for initiating tumor formation, recurrence, and metastasis. c-Yes, a proto-oncogene that is a subfamily of Src family kinase, is often activated in human colon cancer; this implicates c-Yes in the onset and progression of the disease. The objective of this study was to investigate the correlation between c-Yes and CSCs. We performed a sphere formation assay and reverse transcription-polymerase chain reaction for studying the differentiation of HT-29 human colon CSCs. To demonstrate the specific role of c-Yes in CSCs, we performed live cell microscopy and a cell cycle assay. These study shows, for the first time, that c-Yes is enriched in CD133+ CSCs, compared to their CD133- counterparts, and that c-Yes depletion in CD133+ cells induces cell differentiation. Moreover, c-Yes depletion was found to elongate the midbody and increase the proliferation doubling time. This also suggested that the misregulation of microtubules during chromosomal separation causes aneuploidy. Our results suggest that c-Yes may play a crucial role in initiating, maintaining, and driving the tumorigenic property of colon cancer.

Profilin 2 promotes migration, invasion, and stemness of HT29 human colorectal cancer stem cells.

We investigated the role of profilin 2 in the stemness, migration, and invasion of HT29 cancer stem cells (CSCs). Increased and decreased levels of profilin 2 significantly enhanced and suppressed the self-renewal, migration, and invasion ability of HT29 CSCs, respectively. Moreover, profilin 2 directly regulated the expression of stemness markers (CD133, SOX2, and ß-catenin) and epithelial mesenchymal transition (EMT) markers (E-cadherin and snail). CD133 and ß-catenin were up-regulated by overexpression of profilin 2 and down-regulated by depletion of profilin 2. SOX2 was decreased by profilin 2 depletion. E-cadherin was not influenced by profilin 2- overexpression but increased by profilin 2- knockdown. The expression of snail was suppressed by profilin 2- knockdown. We speculated that stemness and the EMT are closely linked through profilin 2-related pathways. Therefore, this study indicates that profilin 2 affects the metastatic potential and stemness of colorectal CSCs by regulating EMT- and stemness-related proteins.

Exogenous rhTRX reduces lipid accumulation under LPS-induced inflammation.

Redox-regulating molecule, recombinant human thioredoxin (rhTRX) which shows anti-inflammatory, and anti-oxidative effects against lipopolysaccharide (LPS)-stimulated inflammation and regulate protein expression levels. LPS-induced reactive oxygen intermediates (ROI) and NO production were inhibited by exogenous rhTRX. We identified up/downregulated intracellular proteins under the LPS-treated condition in exogenous rhTRX-treated A375 cells compared with non-LPS-treated cells via 2-DE proteomic analysis. Also, we quantitatively measured cytokines of in vivo mouse inflammation models using cytometry bead array. Exogenous rhTRX inhibited LPS-stimulated production of ROI and NO levels. TIP47 and ATP synthase may influence the inflammation-related lipid accumulation by affecting lipid metabolism. The modulation of skin redox environments during inflammation is most likely to prevent alterations in lipid metabolism through upregulation of TIP47 and ATP synthase and downregulation of inflammatory cytokines. Our results demonstrate that exogenous rhTRX has anti-inflammatory properties and intracellular regulatory activity in vivo and in vitro. Monitoring of LPS-stimulated pro-inflammatory conditions treated with rhTRX in A375 cells could be useful for diagnosis and follow-up of inflammation reduction related with candidate proteins. These results have a therapeutic role in skin inflammation therapy.

Hyperglycemia decreases the expression of ATP synthase ß subunit and enolase 2 in glomerular epithelial cells.

Glomerular epithelial cells (GECs) are known to play a key role in maintaining the structure and function of the glomerulus. GEC injury induced by hyperglycemia is present in early-stage diabetic nephropathy (DN), which is the most common cause of renal failure. In an attempt to identify target proteins involved in the pathogenesis of GEC injury at early DN, we performed the proteomic analysis using primary cultures of GECs, prepared from the dissected rat glomeruli. The protein expression profiles in the two-dimensional electrophoresis gels were compared between GECs treated for three days with normal glucose (5 mM) and those with high glucose (30 mM) concentrations. These concentrations correspond to blood glucose concentrations under normoglycemia and hyperglycemia, respectively. Proteins with differential expression levels were identified using ESI-Q-TOF tandem mass spectrometry. The primary GECs cultured in hyperglycemic conditions showed cellular hypertrophy and increased production of reactive oxygen species, both of which reflect the GEC injury. Our proteomic analysis identified eight proteins with differential expression profiles, depending on glucose concentrations. Among them, we selected ATP synthase ß subunit and enolase 2 that are related to energy metabolism and are down-regulated under hyperglycemia, and confirmed that hyperglycemia decreased the expression levels of ATP synthase ß subunit and enolase 2 proteins by western blotting analysis. Hyperglycemia may impair mitochondrial function and alter glycolysis in GECs by down-regulating the expression of ATP synthase ß subunit and enolase 2. The present study may provide a better understanding of the pathogenic mechanisms of GEC injury in early DN.

Proteomic profiling of brain cortex tissues in a Tau transgenic mouse model of Alzheimer's disease.

Alzheimer's disease (AD) involves regionalized neuronal death, synaptic loss, and an accumulation of intracellular neurofibrillary tangles and extracellular senile plaques. Although there have been numerous studies on tau proteins and AD in various stages of neurodegenerative disease pathology, the relationship between tau and AD is not yet fully understood. A transgenic mouse model expressing neuron-specific enolase (NSE)-controlled human wild-type tau (NSE-htau23), which displays some of the typical Alzheimer-associated pathological features, was used to analyze the brain proteome associated with tau tangle deposition. Two-dimensional electrophoresis was performed to compare the cortex proteins of transgenic mice (6- and 12-month-old) with those of control mice. Differentially expressed spots in different stages of AD were identified with ESI-Q-TOF (electrospray ionization quadruple time-of-flight) mass spectrometry and liquid chromatography/tandem mass spectrometry. Among the identified proteins, glutathione S-transferase P 1 (GSTP1) and carbonic anhydrase II (CAII) were down-regulated with the progression of AD, and secerin-1 (SCRN1) and V-type proton ATPase subunit E 1 (ATP6VE1) were up-regulated only in the early stages, and down-regulated in the later stages of AD. The proteins, which were further confirmed by RT-PCR at the mRNA level and with western blotting at the protein level, are expected to be good candidates as drug targets for AD. The study of up- and down-regulation of proteins during the progression of AD helps to explain the mechanisms associated with neuronal degeneration in AD.

The reno-protective effect of a phosphoinositide 3-kinase inhibitor wortmannin on streptozotocin-induced proteinuric renal disease rats.

Diabetic nephropathy (DN) is a progressive kidney disease that is caused by injury to kidney glomeruli. Podocytes are glomerular epithelial cells and play critical roles in the glomerular filtration barrier. Recent studies have shown the importance of regulating the podocyte actin cytoskeleton in early DN. The phosphoinositide 3-kinase (PI3K) inhibitor, wortmannin, simultaneously regulates Rac1 and Cdc42, which destabilize the podocyte actin cytoskeleton during early DN. In this study, in order to evaluate the reno-protective effects of wortmannin in early DN by regulating Rac1 and Cdc42, streptozotocin (STZ)-induced proteinuric renal disease (SPRD) rats were treated with wortmannin. The albuminuria value of the SPRD group was 3.55 ± 0.56 mg/day, whereas wortmannin group was 1.77 ± 0.48 mg/day. Also, the albumin to creatinine ratio (ACR) value of the SPRD group was 53.08 ± 10.82 mg/g, whereas wortmannin group was 20.27 ± 6.41 mg/g. Changes in the expression level of nephrin, podocin and Rac1/Cdc42, which is related to actin cytoskeleton in podocytes, by wortmannin administration were confirmed by Western blotting. The expression levels of nephrin (79.66 ± 0.02), podocin (87.81 ± 0.03) and Rac1/Cdc42 (86.12 ± 0.02) in the wortmannin group were higher than the expression levels of nephrin (55.32 ± 0.03), podocin (53.40 ± 0.06) and Rac1/Cdc42 (54.05 ± 0.04) in the SPRD group. In addition, expression and localization of nephrin, podocin and desmin were confirmed by immunofluorescence. In summary, we found for the first time that wortmannin has a reno-protective effect on SPRD rats during the early DN. The beneficial effects of wortmannin in SPRD rats indicate that this compound could be used to delay the progression of the disease during the early DN stage.

Effects of vibration on the proteome expression of anterior cruciate ligament cells.

Recent reports have suggested that vibration has beneficial effects on knee healing response; however, the biomechanism of these beneficial effects still need to be determined on the anterior cruciate ligament (ACL) cell level. In this study, we applied a 20 Hz vibration to ACL cells, which produced a 20% increase (P < 0.001) in cell activity and 17% increase (P < 0.001) in intracellular sulfated glycosaminoglycan levels. In the 20 Hz vibration-stimulated ACL cell group, eight up-regulated (100 ∼ 300%) protein spots were identified compared with the control group by proteomics analysis. Among these proteins, Annexin A2 and Prolyl 4 hydroxylase (PH4B) were shown to have a 71% and 16% higher expression, respectively, in the 20 Hz vibration-stimulated ACL cell group by Western blotting (P < 0.001). These results indicate that vibration produces a positive cellular environment, and Annexin A2 and prolyl 4 hydroxylase are expected to help ligament repair and ACL cell proliferation by controlling cell membrane and extracellular matrix formation.

Transgelin promotes migration and invasion of cancer stem cells.

Recent studies have suggested the existence of a small subset of cancer cells called cancer stem cells (CSCs), which possess the ability to initiate malignancies, promote tumor formation, drive metastasis, and evade conventional chemotherapies. Elucidation of the specific signaling pathway and mechanism underlying the action of CSCs might improve the efficacy of cancer treatments. In this study, we analyzed differentially expressed proteins between tumerigenic and nontumorigenic cells isolated from the human hepatocellular carcinoma (HCC) cell line, Huh7, via proteomic analysis to identify proteins correlated with specific features of CSCs. The expression level of Transgelin was 25-fold higher in tumorigenic cells than nontumorigenic cells. Similar results were also observed in tumorigenic cells derived from colorectal adenocarcinoma and prostate carcinoma. More importantly, the elevated levels of Transgelin significantly increased the invasiveness of tumorigenic cells, whereas reduced levels decreased the invasive potential. Moreover, in tumors derived from Huh7-induced xenografts, Transgelin was also co-expressed with CXCR4, which is responsible for tumor invasion. Taken together, these results indicate that the metastatic potential of CSCs arises from highly expressed Transgelin.

Sample preparation method for plasma membrane proteome analysis.

The preparation of plasma membrane (PM) proteome samples is seriously difficult and time-consuming, owing to their profound hydrophobicity and low abundance. We have developed an efficient PM sample preparation method using Ultracentrifugation with Percoll and an aqueous two-phase extraction. The developed method was rapid (3 h) and provided high purities (26-fold of cell lysate) with a high yield (2.6% of whole cell lysate proteins). This method is especially useful for PM proteome studies using 2D gel electrophoresis.

Proteome analysis of Halobacterium salinarum and characterization of proteins related to the degradation of isopropyl alcohol.

We reported in a previous study that proteomic approach, coupled with genomic techniques, could be used to screen and develop multiple candidates for halophilic enzymes from Halobacterium salinarum. In order to evaluate the biodegradation of isopropyl alcohol (IPA) by H. salinarum, the amounts of residual IPA and acetone generated in the growth media were determined using a gas chromatography-flame ionization detector (GC-FID). The protein expression profiles of cells which had been cultured with IPA were obtained with the two-dimensional gel electrophoresis. Proteins evidencing different expression levels in the presence of 0.5% IPA were identified by electrospray ionization-quadruple-time of flight (ESI-Q-TOF) mass spectrometry. We found 12 proteins which were down-regulated, and another 12 proteins which were up-regulated, in the presence of 0.5% IPA and we further identified 17 proteins among them using ESI-TOF MS/MS. Among these identified proteins, we selected glyceraldehyde 3-phosphate dehydrogenase (GAPDH) for further characterization as a halophilic enzyme. We have demonstrated for the first time that H. salinarum possesses the ability to degrade IPA and GAPDH was both stable and active at high salt concentrations, with maximum activity occurring at 1 M NaCl, although the optimal salt concentration with regard to the growth of H. salinarum is 4.3 M.