Injectable composite hydrogels embedded with gallium-based liquid metal particles for solid breast cancer treatment via chemo-photothermal combination.

Photothermal therapy (PTT) holds great promise as a cancer treatment modality by generating localized heat at the tumor site. Among various photothermal agents, gallium-based liquid metal (LM) has been widely used as a new photothermal-inducible metallic compound due to its structural transformability. To overcome limitations of random aggregation and dissipation of administrated LM particles into a human body, we developed LM-containing injectable composite hydrogel platforms capable of achieving spatiotemporal PTT and chemotherapy. Eutectic gallium-indium LM particles were first stabilized with 1,2-Distearoyl-sn­glycero-3-phosphoethanolamine (DSPE) lipids. They were then incorporated into an interpenetrating hydrogel network composed of thiolated gelatin conjugated with 6-mercaptopurine (MP) chemodrug and poly(ethylene glycol)-diacrylate. The resulted composite hydrogel exhibited sufficient capability to induce MDA-MB-231 breast cancer cell death through a multi-step mechanism: (1) hyperthermic cancer cell death due to temperature elevation by near-infrared laser irradiation via LM particles, (2) leakage of glutathione (GSH) and cleavage of disulfide bonds due to destruction of cancer cells. As a consequence, additional chemotherapy was facilitated by GSH, leading to accelerated release of MP within the tumor microenvironment. The effectiveness of our composite hydrogel system was evaluated both in vitro and in vivo, demonstrating significant tumor suppression and killing. These results demonstrate the potential of this injectable composite hydrogel for spatiotemporal cancer treatment. In conclusion, integration of PTT and chemotherapy within our hydrogel platform offers enhanced therapeutic efficacy, suggesting promising prospects for future clinical applications. STATEMENT OF SIGNIFICANCE: Our research pioneers a breakthrough in cancer treatments by developing an injectable hydrogel platform incorporating liquid metal (LM) particle-mediated photothermal therapy and 6-mercaptopurine (MP)-based chemotherapy. The combination of gallium-based LM and MP achieves synergistic anticancer effects, and our injectable composite hydrogel acts as a localized reservoir for specific delivery of both therapeutic agents. This platform induces a multi-step anticancer mechanism, combining NIR-mediated hyperthermic tumor death and drug release triggered by released glutathione from damaged cancer populations. The synergistic efficacy validated in vitro and in vivo studies highlights significant tumor suppression. This injectable composite hydrogel with synergistic therapeutic efficacy holds immense promise for biomaterial-mediated spatiotemporal treatment of solid tumors, offering a potent targeted therapy for triple negative breast cancers.

Optimized Design of Hyaluronic Acid-Lipid Conjugate Biomaterial for Augmenting CD44 Recognition of Surface-Engineered NK Cells.

Triple-negative breast cancer (TNBC) presents treatment challenges due to a lack of detectable surface receptors. Natural killer (NK) cell-based adaptive immunotherapy is a promising treatment because of the characteristic anticancer effects of killing malignant cells directly by secreting cytokines and lytic granules. To maximize the cancer recognition ability of NK cells, biomaterial-mediated ex vivo cell surface engineering has been developed for sufficient cell membrane immobilization of tumor-targeting ligands via hydrophobic anchoring. In this study, we optimized amphiphilic balances of NK cell coating materials composed of CD44-targeting hyaluronic acid (HA)-poly(ethylene glycol) (PEG)-lipid to improve TNBC recognition and the anticancer effect. Changes in the modular design of our material by differentiating hydrophilic PEG length and incorporating lipid amount into HA backbones precisely regulated the amphiphilic nature of HA-PEG-lipid conjugates. The optimized biomaterial demonstrated improved anchoring into NK cell membranes and facilitating the surface presentation level of HA onto NK cell surfaces. This led to enhanced cancer targeting via increasing the formation of immune synapse, thereby augmenting the anticancer capability of NK cells specifically toward CD44-positive TNBC cells. Our approach addresses targeting ability of NK cell to solid tumors with a deficiency of surface tumor-specific antigens while offering a valuable material design strategy using amphiphilic balance in immune cell surface engineering techniques.

Tailoring tumor-recognizable hyaluronic acid-lipid conjugates to enhance anticancer efficacies of surface-engineered natural killer cells.

Natural killer (NK) cells have clinical advantages in adoptive cell therapy owing to their inherent anticancer efficacy and their ability to identify and eliminate malignant tumors. However, insufficient cancer-targeting ligands on NK cell surfaces often inhibit their immunotherapeutic performance, especially in immunosuppressive tumor microenvironment. To facilitate tumor recognition and subsequent anticancer function of NK cells, we developed hyaluronic acid (HA, ligands to target CD44 overexpressed onto cancer cells)-poly(ethylene glycol) (PEG, cytoplasmic penetration blocker)-Lipid (molecular anchor for NK cell membrane decoration through hydrophobic interaction) conjugates for biomaterial-mediated ex vivo NK cell surface engineering. Among these major compartments (i.e., Lipid, PEG and HA), optimization of lipid anchors (in terms of chemical structure and intrinsic amphiphilicity) is the most important design parameter to modulate hydrophobic interaction with dynamic NK cell membranes. Here, three different lipid types including 1,2-dimyristoyl-sn-glycero-3-phosphati-dylethanolamine (C14:0), 1,2-distearoyl-sn-glycero-3-phosphatidylethanolamine (DSPE, C18:0), and cholesterol were evaluated to maximize membrane coating efficacy and associated anticancer performance of surface-engineered NK cells (HALipid-NK cells). Our results demonstrated that NK cells coated with HA-PEG-DSPE conjugates exhibited significantly enhanced anticancer efficacies toward MDA-MB-231 breast cancer cells without an off-target effect on human fibroblasts specifically via increased NK cell membrane coating efficacy and prolonged surface duration of HA onto NK cell surfaces, thereby improving HA-CD44 recognition. These results suggest that our HALipid-NK cells with tumor-recognizable HA-PEG-DSPE conjugates could be further utilized in various cancer immunotherapies.

Tumor Microenvironment-Responsive 6-Mercaptopurine-Releasing Injectable Hydrogel for Colon Cancer Treatment.

Colon cancer is a significant health concern. The development of effective drug delivery systems is critical for improving treatment outcomes. In this study, we developed a drug delivery system for colon cancer treatment by embedding 6-mercaptopurine (6-MP), an anticancer drug, in a thiolated gelatin/polyethylene glycol diacrylate hydrogel (6MP-GPGel). The 6MP-GPGel continuously released 6-MP, the anticancer drug. The release rate of 6-MP was further accelerated in an acidic or glutathione environment that mimicked a tumor microenvironment. In addition, when pure 6-MP was used for treatment, cancer cells proliferated again from day 5, whereas a continuous supply of 6-MP from the 6MP-GPGel continuously suppressed the survival rate of cancer cells. In conclusion, our study demonstrates that embedding 6-MP in a hydrogel formulation can improve the efficacy of colon cancer treatment and may serve as a promising minimally invasive and localized drug delivery system for future development.

Current progress in gallium-based liquid metals for combinatory phototherapeutic anticancer applications.

A variety of therapeutic approaches using liquid metal (LM) have been intensively investigated, due to its unique physico-chemical properties that include high surface tension, fluidity, shape deformability, thermal conductivity, and electrical conductivity. Among a series of LMs, the relatively lower toxicity and minimal volatility of gallium (Ga)-based LMs (GaLMs) enables their usage in a series of potential biomedical applications, especially implantable platforms, to treat multiple diseases. In addition, the highly efficient conversion of light energy into thermal or chemical energy via GaLMs has led to recent developments in photothermal and photodynamic applications for anticancer treatments. As attractive photothermal agents or photosensitizers, a systematic interpretation of the structural characteristics and photo-responsive behaviors of GaLMs is necessary to develop effective anticancer engineering applications. Therefore, the aim of this review is to provide a comprehensive summary of currently suggested GaLM-mediated photo-therapeutic cancer treatments. In particular, the review summarizes (1) surface coating techniques to form stable and multifunctional GaLM particulates, (2) currently investigated GaLM-mediated photothermal and photodynamic anticancer therapies, (3) synergistic efficacies with the aid of additional interventions, and (4) 3D composite gels embedded with GaLMs particles, to convey the potential technological advances of LM in this field.

Fabrication Parameter-Dependent Physico-Chemical Properties of Thiolated Gelatin/PEGDA Interpenetrating Network Hydrogels.

BACKGROUND: The development of three-dimensional hydrogels using polymeric biomaterials is a key technology for tissue engineering and regenerative medicine. Successful tissue engineering requires the control and identification of the physicochemical properties of hydrogels. METHODS: Interpenetrating network (IPN) hydrogel was developed using thiolated gelatin (GSH) and poly(ethylene glycol) diacrylate (PEGDA), with the aid of ammonium persulfate (APS) and N,N,N,N'-tetramethylethylenediamine (TEMED) as radical initiators. Each component was prepared in the following concentrations, respectively: 2.5 and 5% GSH (LG and HG), 12.5 and 25% PEGDA (LP and HP), 3% APS/1.5% TEMED (LI), and 4% APS/2% TEMED (HI). IPN hydrogel was fabricated by the mixing of GSH, PEGDA, and initiators in 5:4:1 volume ratios, and incubated at 37 °C for 30 min in the following 6 experimental formulations: (1) HG-LP-LI, (2) HG-LP-HI, (3) LG-HP-LI, (4) LG-HP-HI, (5) HG-HP-HI, and (6) HG-HP-LI. Herein, the physico-chemical characteristics of IPN hydrogels, including their morphological structures, hydrolytic degradation properties, mechanical properties, embedded protein release kinetics, and biocompatibility, were investigated. RESULTS: The characteristics of the hydrogel were significantly manipulated by the concentration of the polymer, especially the conversion between HP and LP, rather than the concentration of the initiator, and no hydrogel formulation exhibited any toxicity to fibroblast and HaCaT cells. CONCLUSION: We provide structural-physical relationships of the hydrogels by which means their physical properties could be conveniently controlled through component control, which could be versatilely utilized for various organizational engineering strategies.

Relation of Pulmonary Function Impairment and Coronary Artery Calcification by Multi-detector Computed Tomography in Group Exposed to Inorganic Dusts.

BACKGROUND: The purpose of this study was to evaluate the relationship of pulmonary function impairment (PFI) and coronary artery calcification (CAC) by multi-detector computed tomography (MDCT), and the effect of pneumoconiosis on CAC or PFI. METHODS: Seventy-six subjects exposed to inorganic dusts underwent coronary artery calcium scoring by MDCT, spirometry, laboratory tests, and a standardized questionnaire. CAC was quantified using a commercial software (Rapidia ver. 2.8), and all the subjects were divided into two categories according to total calcium scores (TCSs), either the non-calcified (<1) or the calcified (≥1) group. Obstructive pulmonary function impairment (OPFI) was defined as forced expiratory volume in one second/forced vital capacity (FEV1/FVC, %)<70, and as FEV1/FVC (%)≥70 and FVC<80 for restrictive pulmonary function impairment (RPFI) by spirometry. All subjects were classified as either the case (profusion≥1/0) or the control (profusion≤0/1) group by pneumoconiosis findings on simple digital radiograph. RESULTS: Of the 76 subjects, 35 subjects (46.1%) had a CAC. Age and hypertension were different significantly between the non-calcified and the calcified group (p<0.05). Subjects with pneumoconiosis were more frequent in the calcified group than those in the non-calcified group (p=0.099). FEV1/FVC (%) was significantly correlated with TCSs (r=-0.316, p=0.005). Subjects with OPFI tended to increase significantly with increasing of TCS (4.82, p=0.028), but not significantly in RPFI (2.18, p=0.140). Subjects with OPFI were significantly increased in the case group compared to those in the control group. CONCLUSION: CAC is significantly correlated with OPFI, and CAC and OPFI may be affected by pneumoconiosis findings.

Reliability and validity of soft copy images based on flat-panel detector in pneumoconiosis classification: comparison with the analog radiographs.

RATIONALE AND OBJECTIVES: The aim of this study was to evaluate the reliability and validity of soft copy images based on flat-panel detector of digital radiography (DR-FPD soft copy images) compared to analog radiographs (ARs) in pneumoconiosis classification and diagnosis. MATERIALS AND METHODS: DR-FPD soft copy images and ARs from 349 subjects were independently read by four-experienced readers according to the International Labor Organization 2000 guidelines. DR-FPD soft copy images were used to obtain consensus reading (CR) by all readers as the gold standard. Reliability and validity were evaluated by a κ and receiver operating characteristic analysis, respectively. RESULTS: In small opacity, overall interreader agreement of DR-FPD soft copy images was significantly higher than that of ARs, but it was not significantly different in large opacity and costophrenic angle obliteration. In small opacity, agreement of DR-FPD soft copy images with CR was significantly higher than that of ARs with CR. It was also higher than that of ARs with CR in pleural plaque and thickening. Receiver operating characteristic areas were not different significantly between DR-FPD soft copy images and ARs. CONCLUSIONS: DR-FPD soft copy images showed accurate and reliable results in pneumoconiosis classification and diagnosis compared to ARs.

Utility of digital radiography for the screening of pneumoconiosis as compared to analog radiography: radiation dose, image quality, and pneumoconiosis classification.

The purpose of this study was to compare digital radiography (DR) and analog radiography (AR) for the screening of pneumoconiosis with respect to radiation dose, image quality, and pneumoconiosis classification. DR was performed on 50 subjects who were enrolled for an examination of pneumoconiosis (Digital Diagnost™, Philips, Netherlands), and AR (MXO-15B, Toshiba, Japan) was performed the same day after the study was approved by the Institutional Review Board and written informed consent was obtained from all subjects. Entrance surface doses (ESDs) of DR and AR were measured using a glass dosimeter attached to a Rando human phantom (Alderson Co., U.S.) under exposure conditions commonly used in clinical practice in Korea. Visibilities on all images were evaluated using a 5-point scale by four chest radiologists using a modified form of the European Chest Guidelines (EUR 16260). All the images were classified using the ILO's guidelines by referencing standard analog radiographs. ESDs of DR were significantly lower than those of AR (0.15 mGy vs. 0.21 mGy, p < 0.05). All anatomic structures were significantly more visible by DR images (p < 0.0001), especially the left main bronchus, ribs, and thoracic spine. Body mass index did not correlate with anatomic structure visibility by DR (r = -0.029, p = 0.842) or AR images (r = -0.076, p = 0.602). Overall intra- and inter-reader agreements for DR images were significantly higher than for AR images. DR offers improved image quality with a significant reduction of up to 23.6% in radiation dose and more accurate pneumoconiosis classification than AR.

Development of standard digital images for pneumoconiosis.

We developed the standard digital images (SDIs) to be used in the classification and recognition of pneumoconiosis. From July 3, 2006 through August 31, 2007, 531 retired male workers exposed to inorganic dust were examined by digital (DR) and analog radiography (AR) on the same day, after being approved by our institutional review board and obtaining informed consent from all participants. All images were twice classified according to the International Labour Office (ILO) 2000 guidelines with reference to ILO standard analog radiographs (SARs) by four chest radiologists. After consensus reading on 349 digital images matched with the first selected analog images, 120 digital images were selected as the SDIs that considered the distribution of pneumoconiosis findings. Images with profusion category 0/1, 1, 2, and 3 were 12, 50, 40, and 15, respectively, and a large opacity were in 43 images (A = 20, B = 22, C = 1). Among pleural abnormality, costophrenic angle obliteration, pleural plaque and thickening were in 11 (9.2%), 31 (25.8%), and 9 (7.5%) images, respectively. Twenty-one of 29 symbols were present except cp, ef, ho, id, me, pa, ra, and rp. A set of 120 SDIs had more various pneumoconiosis findings than ILO SARs that were developed from adequate methods. It can be used as digital reference images for the recognition and classification of pneumoconiosis.

Blood Levels of IL-Iß, IL-6, IL-8, TNF-α, and MCP-1 in Pneumoconiosis Patients Exposed to Inorganic Dusts.

Inhaled inorganic dusts such as coal can cause inflammation and fibrosis in the lung called pneumoconiosis. Chronic inflammatory process in the lung is associated with various cytokines and reactive oxygen species (ROS) formation. Expression of some cytokines mediates inflammation and leads to tissue damage or fibrosis. The aim of the present study was to compare the levels of blood cytokines interleukin (IL)-1ß, IL-6, IL-8, tumor necrosis factor (TNF)-α and monocyte chemoattractant protein (MCP)-1 among 124 subjects (control 38 and pneumoconiosis patient 86) with category of chest x-ray according to International Labor Organization (ILO) classification. The levels of serum IL-8 (p = 0.003), TNF-α (p = 0.026), and MCP-1 (p = 0.010) of pneumoconiosis patients were higher than those of subjects with the control. The level of serum IL-8 in the severe group with the small opacity (ILO category II or III) was higher than that of the control (p = 0.035). There was significant correlation between the profusion of radiological findings with small opacity and serum levels of IL-1ß (rho = 0.218, p < 0.05), IL-8 (rho = 0.224, p < 0.05), TNF-α (rho = 0.306, p < 0.01), and MCP-1 (rho = 0.213, p < 0.01). The serum levels of IL-6 and IL-8, however, did not show significant difference between pneumoconiosis patients and the control. There was no significant correlation between serum levels of measured cytokines and other associated variables such as lung function, age, BMI, and exposure period of dusts. Future studies will be required to investigate the cytokine profile that is present in pneumoconiosis patient using lung specific specimens such as bronchoalveolar lavage fluid (BALF), exhaled breath condensate, and lung tissue.

The PI3 kinase-Akt pathway mediates Wnt3a-induced proliferation.

Wnt3a activates proliferation of fibroblasts cells via activation of both extracellular signal-regulated kinase (ERK) and Wnt/beta-catenin signaling pathways. In this study, we show that the phosphatidyl inositol 3 kinases (PI3K)-Akt pathway is also involved in the Wnt3a-induced proliferation. Akt was activated within 30 min by Wnt3a in NIH3T3 cells. By Wnt3a treatment, activated Akt was transiently accumulated in nucleus although beta-catenin was accumulated in the nucleus of cells in a prolonged manner. The Wnt3a-induced Akt activation was not affected by siRNA-mediated reduction of beta-catenin, indicating that Wnt3a-induced Akt activation may occur independently of beta-catenin. The Wnt3a-induced Akt activation was abolished by pre-treatment with PI3K inhibitor, LY294002 and Wortmanin, but not by MEK inhibitor, U0126, indicating that Wnt3a activates Akt via PI3K. The growth and proliferation induced by Wnt3a were blocked by treatments of the PI3K inhibitors. Furthermore, Wnt3a-induced proliferation was blocked by Akt siRNA. These results reveal that the PI3K-Akt pathway mediates the Wnt3a-induced growth and proliferation of NIH3T3 cells.

Identification of proteins interacting with the catalytic subunit of PP2A by proteomics.

The protein phosphatase 2A (PP2A) is a serine/threonine phosphatase involved in the regulation of multiple signaling pathways including the Wnt/beta-catenin and the ERK pathways. To understand the complex signaling networking associated with PP2A, we searched proteins interacting with the catalytic subunit of protein phosphatase 2A (PP2Ac) by a pull-down analysis followed by 2-D gel electrophoresis and proteomic analyses. The probability of identification of the proteins interacting with PP2Ac was increased by searching proteins differently interacting with PP2Ac according to stimulation of Wnt3a, which regulates both the Wnt/beta-catenin and the ERK pathways. Around 100 proteins, pulled-down by His-tagged PP2Ac, were identified in 2-D gels stained with CBB. By MALDI-TOF-MS analyses of 45 protein spots, we identified several proteins that were previously known to interact with PP2A, such as Axin and CaMK IV. In addition, we also identified many proteins that potentially interact with PP2Ac. The interactions of several candidate proteins, such as tuberous sclerosis complex 2, RhoB, R-Ras, and Nm23H2, with PP2Ac, were confirmed by in vitro binding analyses and/or coimmunoprecipitation experiments.