Correction: Investigation of craquelure patterns in oil paintings using precise 3D morphological analysis for art authentication.

[This corrects the article DOI: 10.1371/journal.pone.0272078.].

Spontaneous Osteogenic Differentiation of Human Mesenchymal Stem Cells by Tuna-Bone-Derived Hydroxyapatite Composites with Green Tea Polyphenol-Reduced Graphene Oxide.

In recent years, bone tissue engineering (BTE) has made significant progress in promoting the direct and functional connection between bone and graft, including osseointegration and osteoconduction, to facilitate the healing of damaged bone tissues. Herein, we introduce a new, environmentally friendly, and cost-effective method for synthesizing reduced graphene oxide (rGO) and hydroxyapatite (HAp). The method uses epigallocatechin-3-O-gallate (EGCG) as a reducing agent to synthesize rGO (E-rGO), and HAp powder is obtained from Atlantic bluefin tuna (Thunnus thynnus). The physicochemical analysis indicated that the E-rGO/HAp composites had exceptional properties for use as BTE scaffolds, as well as high purity. Moreover, we discovered that E-rGO/HAp composites facilitated not only the proliferation, but also early and late osteogenic differentiation of human mesenchymal stem cells (hMSCs). Our work suggests that E-rGO/HAp composites may play a significant role in promoting the spontaneous osteogenic differentiation of hMSCs, and we envision that E-rGO/HAp composites could serve as promising candidates for BTE scaffolds, stem-cell differentiation stimulators, and implantable device components because of their biocompatible and bioactive properties. Overall, we suggest a new approach for developing cost-effective and environmentally friendly E-rGO/HAp composite materials for BTE application.

Efficient label-free in vivo photoacoustic imaging of melanoma cells using a condensed NIR-I spectral window.

In this paper, we propose an efficient label-free in vivo photoacoustic (PA) imaging of melanoma using a condensed near infrared-I (NIR-I) supercontinuum light source. Although NIR-II spectral window is advantageous such as longer penetration depth compared to the NIR-I region, supercontinuum light sources emitting both NIR-I and NIR-II region could lower the efficiency to target melanoma because of low optical power density in the melanoma's absorption spectra. To exploit efficient in vivo PA imaging of melanoma, we demonstrated the light source emitting from visible (532-600 nm) to NIR-I (600-1000 nm) by optimizing stimulated Raman scattering induced supercontinuum generation. The melanoma's structure is successfully differentiated from blood vessels at a high pulse energy of 2.5 µJ and a flexible pulse repetition rate (PRR) of 5-50 kHz. The proposed light source with the microjoules energies and tens of kHz of PRR can potentially accelerate clinical trials such as early diagnosis of melanoma.

Computational analysis of drug free silver triangular nanoprism theranostic probe plasmonic behavior for in-situ tumor imaging and photothermal therapy.

INTRODUCTION: The advanced features of plasmonic nanomaterials enable initial high accuracy detection with different therapeutic intervention. Computational simulations could estimate the plasmonic heat generation with a high accuracy and could be reliably compared to experimental results. This proposed combined theoretical-experimental strategy may help researchers to better understand other nanoparticles in terms of plasmonic efficiency and usability for future nano-theranostic research. OBJECTIVES: To develop innovative computationally-driven approach to quantify any plasmonic nanoparticles photothermal efficiency and effects before their use as therapeutic agents. METHODS: This report introduces drug free plasmonic silver triangular nanoprisms coated with polyvinyl alcohol biopolymer (PVA-SNT), for in vivo photoacoustic imaging (PAI) guided photothermal treatment (PTT) of triple-negative breast cancer mouse models. The synthesized PVA-SNT nanoparticles were characterized and a computational electrodynamic analysis was performed to evaluate and predict the optical and plasmonic photothermal properties. The in vitro biocompatibility and in vivo tumor abalation study was performed with MDA-MB-231 human breast cancer cell line and in nude mice model. RESULTS: The drug free 140 µg∙mL-1 PVA-SNT nanoparticles with 1.0 W∙cm-2 laser irradiation for 7 min proved to be an effective and optimized theranostic approach in terms of PAI guided triple negative breast cancer treatment. The PVA-SNT nanoparticles exhibits excellent biosafety, photostability, and strong efficiency as PAI contrast agent to visualize tumors. Histological analysis and fluorescence-assisted cell shorter assay results post-treatment apoptotic cells, more importantly, it shows substantial damage to in vivo tumor tissues, killing almost all affected cells, with no recurrence. CONCLUSION: This is a first complete study on computational simulations to estimate the plasmonic heat generation followed by drug free plasmonic PAI guided PTT for cancer treatment. This computationally-driven theranostic approach demonstrates an innovative thought regarding the nanoparticles shape, size, concentration, and composition which could be useful for the prediction of photothermal heat generation in precise nanomedicine applications.

Investigation of craquelure patterns in oil paintings using precise 3D morphological analysis for art authentication.

The development of scientific technology for art authentication has elicited multidimensional evidence to distinguish forgeries from original artwork. Here, we analyzed the three-dimensional morphology of cracks that contain information, such as the painting features of artworks, using optical coherence tomography. The forgeries were produced by an expert from original oil paintings with cracks that occur owing to paint drying, canvas aging, and physical damage. Parameters, such as shape, width, and depth, were compared based on the cross-sectional images of the original and fake cracks. The original cracks were rectangular and inverted, but the fake cracks were relatively simple inverted triangles. The original cracks were as deep as the thickness of the upper layer and mostly were "thin/deep" or "wide/shallow". The fake cracks were observed to be "'thin/shallow" or "wide/deep". This study aims to improve the understanding of crack characteristics and promote the development of techniques for determining art authenticity.

Multiphoton excitation imaging via an actively mode-locked tunable fiber-cavity SOA laser around 800 nm.

In this study, an active mode-locked tunable pulsed laser (AML-TPL) is proposed to excite picosecond pulsed light with a rapid wavelength tunability of approximately 800 nm for multiphoton microscopy. The AML-TPL is schematically based on a fiber-cavity semiconductor optical amplifier (SOA) configuration to implement a robust and align-free pulsed light source with a duration of 1.6 ps, a repetition rate of 27.9271 MHz, and average output power of over 600 mW. A custom-built multiphoton imaging system was also built to demonstrate the imaging performance of the proposed AML-TPL by comparing with the commercial Ti:Sapphire femtosecond laser. Two-photon excited fluorescence images were successfully acquired using a human breast cancer cell line (MDA-MB-231) stained with acridine orange.

Ultra-widefield photoacoustic microscopy with a dual-channel slider-crank laser-scanning apparatus for in vivo biomedical study.

Photoacoustic microscopy (PAM) is an important imaging tool that can noninvasively visualize the anatomical structure of living animals. However, the limited scanning area restricts traditional PAM systems for scanning a large animal. Here, we firstly report a dual-channel PAM system based on a custom-made slider-crank scanner. This novel scanner allows us to stably capture an ultra-widefield scanning area of 24 mm at a high B-scan speed of 32 Hz while maintaining a high signal-to-noise ratio. Our system's spatial resolution is measured at ∼3.4 µm and ∼37 µm for lateral and axial resolution, respectively. Without any contrast agent, a dragonfly wing, a nude mouse ear, an entire rat ear, and a portion of mouse sagittal are successfully imaged. Furthermore, for hemodynamic monitoring, the mimicking circulating tumor cells using magnetic contrast agent is rapidly captured in vitro. The experimental results demonstrated that our device is a promising tool for biological applications.

Coadministration of endothelial and smooth muscle cells derived from human induced pluripotent stem cells as a therapy for critical limb ischemia.

Critical limb ischemia is a condition in which tissue necrosis occurs due to arterial occlusion, resulting in limb amputation in severe cases. Both endothelial cells (ECs) and vascular smooth muscle cells (SMCs) are needed for the regeneration of peripheral arteries in ischemic tissues. However, it is difficult to isolate and cultivate primary EC and SMC from patients for therapeutic angiogenesis. Induced pluripotent stem cells (iPSCs) are regarded as useful stem cells due to their pluripotent differentiation potential. In this study, we explored the therapeutic efficacy of human iPSC-derived EC and iPSC-derived SMC in peripheral artery disease model. After the induction of mesodermal differentiation of iPSC, CD34+ progenitor cells were isolated by magnetic-activated cell sorting. Cultivation of the CD34+ progenitor cells in endothelial culture medium induced the expression of endothelial markers and phenotypes. Moreover, the CD34+ cells could be differentiated into SMC by cultivation in SMC culture medium. In a murine hindlimb ischemia model, cotransplantation of EC with SMC improved blood perfusion and increased the limb salvage rate in ischemic limbs compared to transplantation of either EC or SMC alone. Moreover, cotransplantation of EC and SMC stimulated angiogenesis and led to the formation of capillaries and arteries/arterioles in vivo. Conditioned medium derived from SMC stimulated the migration, proliferation, and tubulation of EC in vitro, and these effects were recapitulated by exosomes isolated from the SMC-conditioned medium. Together, these results suggest that iPSC-derived SMC enhance the therapeutic efficacy of iPSC-derived EC in peripheral artery disease via an exosome-mediated paracrine mechanism.

Folic acid-conjugated chitosan-functionalized graphene oxide for highly efficient photoacoustic imaging-guided tumor-targeted photothermal therapy.

Multifunctional theranostic agents have recently attracted a great deal of attention in field of biomedicine. In the present work, folic acid-conjugated chitosan-functionalized graphene oxide (FA-CS-GO) has been developed as a new type of multifunctional nanomaterial for near-infrared fluorescence (FL)/photoacoustic imaging-(PAI) guided photothermal therapy (PTT) of cancer. In vitro results showed that the FA-CS-GO was able to completely destroy cancer cells under laser irradiation. More importantly, in vivo experiments showed that in the presence of targeted FA-CS-GO with laser irradiation, the tumors were completely inhibited, with no recurrence within 20 days. A high photoacoustic signal was detected in the tumor area of mice 24 h after the injection of FA-CS-GO, demonstrating the ability of FA-CS-GO to function as a new PAI contrast agent. Altogether, FA-CS-GO showed a high tumor-targeting efficiency, powerful photothermal effect, and outstanding PAI. This study is considered the first where multifunctional nanomaterials were used for highly efficient FL/PAI-guided tumor-targeted PTT, which is a promising avenue for theranostic nanomedicine.

Anti-EGFR antibody conjugated thiol chitosan-layered gold nanoshells for dual-modal imaging-guided cancer combination therapy.

Developing a novel multifunctional theranostic agent for cancer combination therapy has attracted tremendous attention in recent years. In this report, we designed and developed a new multifunctional nanocarrier based on anti-epidermal growth factor receptor antibody-conjugated and paclitaxel loaded-thiol chitosan-layered gold nanoshells (anti-EGFR-PTX-TCS-GNSs) as a theranostic agent for the first time used for fluorescence/photoacoustic dual-modal imaging-guided chemophotothermal synergistic therapy. The resulting anti-EGFR-PTX-TCS-GNSs showed excellent biosafety, biocompatibility, broad near-infrared (NIR) absorbance, photostability, fast and laser irradiation-controllable drug release, and higher targeting efficiency for efficient chemophotothermal combination therapy of cancer under the guidance of photoacoustic imaging (PAI). The combination therapy was investigated in vitro and in vivo, displaying a powerful anticancer efficiency. More importantly, an in vivo experiment of anti-EGFR-PTX-TCS-GNSs with laser irradiation showed heavy damage to the tumor tissue, killing the tumor cells almost completely. Anti-EGFR-PTX-TCS-GNSs also showed a powerful capacity to visualize tumors, and therefore it is considered a new PAI contrast agent for subsequent therapy. Histological analysis and TUNEL assay further showed much more apoptotic cells, confirming the value of anti-EGFR-PTX-TCS-GNSs. Our results provide a new concept and a promising strategy to develop a novel multifunctional nanotheranostic agent for future clinical applications in diagnosis and therapy.

Akinetic swept-source optical coherence tomography based on a pulse-modulated active mode locking fiber laser for human retinal imaging.

Optical coherence tomography (OCT) is a noninvasive imaging modality that can provide high-resolution, cross-sectional images of tissues. Especially in retinal imaging, OCT has become one of the most valuable imaging tools for diagnosing eye diseases. Considering the scattering and absorption properties of the eye, the 1000-nm OCT system is preferred for retinal imaging. In this study, we describe the use of an akinetic swept-source OCT system based on a pulse-modulated active mode locking (AML) fiber laser at a 1080-nm wavelength for in-vivo human retinal imaging. The akinetic AML wavelength-swept fiber laser was constructed with polarization-maintaining fiber that has an average linewidth of 0.625 nm, a spectral bandwidth of 81.15 nm, and duty ratio of 90% without the buffering method. We successfully obtained in-vivo human retinal images using the proposed OCT system without the additional k-clock and the frequency shifter that provides a wide field of view of 43.1°. The main retina layers, such as the retinal pigment epithelium, can be distinguished from the OCT image with an axial resolution of 6.3 µm with this OCT system.

Quasi-Distributed Active-Mode-Locking Laser Interrogation with Multiple Partially Reflecting Segment Sensors.

A new type of quasi-distributed sensor system is implemented using an active mode locking (AML) laser cavity with multiple partially reflecting segments. The mode locking frequency of the AML laser is linearly proportional to the overall lasing cavity length. To implement multiple resonators having multiple reflection points installed in a sensing fiber, two types of partial reflectors (PRs) are implemented for an in-line configuration, one with fiber Bragg grating and the other with a fiber Fabry⁻Perot interferometer. Since the laser has oscillated only when the modulation frequencies for the mode locking frequency match with the corresponding resonator lengths, it is possible to read the multiple partially reflecting segments along the sensing fiber. The difference between two corresponding mode locking frequencies is changing proportionally with the segment length variation between two PRs upon strain application. The segment length change caused by the applied strain can be successfully measured with a linear sensitivity between mode locking frequency and displacement, linearity over 0.99, and spatial position resolution below meter order.

A Potent Tyrosinase Inhibitor, (E)-3-(2,4-Dihydroxyphenyl)-1-(thiophen-2-yl)prop-2-en-1-one, with Anti-Melanogenesis Properties in α-MSH and IBMX-Induced B16F10 Melanoma Cells.

In this study, we designed and synthesized eight thiophene chalcone derivatives (1a⁻h) as tyrosinase inhibitors and evaluated their mushroom tyrosinase inhibitory activities. Of these eight compounds, (E)-3-(2,4-dihydroxyphenyl)-1-(thiophen-2-yl)prop-2-en-1-one (1c) showed strong competitive inhibition activity against mushroom tyrosinase with IC50 values of 0.013 µM for tyrosine hydroxylase and 0.93 µM for dopa oxidase. In addition, we used enzyme kinetics study and docking program to further evaluate the inhibitory mechanism of 1c toward tyrosinase. As an underlying mechanism of 1c mediated anti-melanogenic effect, we investigated the inhibitory activity against melanin contents and cellular tyrosinase in B16F10 melanoma cells. As the results, the enzyme kinetics and docking results supports that 1c highly interacts with tyrosinase residues in the tyrosinase active site and it can directly inhibit tyrosinase as competitive inhibitor. In addition, 1c exhibited dose-dependent inhibitory effects in melanin contents and intracellular tyrosinase on α-MSH and IBMX-induced B16F10 cells. Overall, our results suggested that 1c might be considered potent tyrosinase inhibitor for use in the development of therapeutic agents for diseases associated with hyperpigment disorders.

Photoacoustic Imaging-Guided Photothermal Therapy with Tumor-Targeting HA-FeOOH@PPy Nanorods.

Cancer theragnosis agents with both cancer diagnosis and therapy abilities would be the next generation of cancer treatment. Recently, nanomaterials with strong absorption in near-infrared (NIR) region have been explored as promising cancer theragnosis agents for bio-imaging and photothermal therapy (PTT). Herein, we reported the synthesis and application of a novel multifunctional theranostic nanoagent based on hyaluronan (HA)-coated FeOOH@polypyrrole (FeOOH@PPy) nanorods (HA-FeOOH@PPy NRs) for photoacoustic imaging (PAI)-guided PTT. The nanoparticles were intentionally designed with rod-like shape and conjugated with tumor-targeting ligands to enhance the accumulation and achieve the entire tumor distribution of nanoparticles. The prepared HA-FeOOH@PPy NRs showed excellent biocompatible and physiological stabilities in different media. Importantly, HA-FeOOH@PPy NRs exhibited strong NIR absorbance, remarkable photothermal conversion capability, and conversion stability. Furthermore, HA-FeOOH@PPy NRs could act as strong contrast agents to enhance PAI, conducting accurate locating of cancerous tissue, as well as precise guidance for PTT. The in vitro and in vivo photothermal anticancer activity results of the designed nanoparticles evidenced their promising potential in cancer treatment. The tumor-bearing mice completely recovered after 17 days of PTT treatment without obvious side effects. Thus, our work highlights the great potential of using HA-FeOOH@PPy NRs as a theranostic nanoplatform for cancer imaging-guided therapy.

Modality switching between therapy and imaging based on the excitation wavelength dependence of dual-function agents in folic acid-conjugated graphene oxides.

Owing to its near infrared (NIR) absorption, graphene oxide (GO) is promising for both photothermal (PT) therapy and multiphoton (MP) imaging. Novel therapy/imaging modality switching is proposed here based on the selected excitation wavelength of femtosecond (FS) laser. GO-based destruction of cancer cells is demonstrated when the laser power of 800-nm-wavelength FS laser is increased above 7 mW. However, GO-based imaging is mainly monitored without damaging the sample when using 1200-nm wavelength FS laser in the same laser power range. Folic acid (FA) conjugated graphene oxide (FA-GO) was synthesized for selective cancer cell targeting. Dual-function FA-GO-based cancer cell targeting agents were experimentally optimized to enable therapy/imaging modality switching.

In vivo photoacoustic monitoring using 700-nm region Raman source for targeting Prussian blue nanoparticles in mouse tumor model.

Photoacoustic imaging (PAI) is a noninvasive imaging tool to visualize optical absorbing contrast agents. Due to high ultrasonic resolution and superior optical sensitivity, PAI can be used to monitor nanoparticle-mediated cancer therapy. The current study synthesized Food and Drug Administration-approved Prussian blue (PB) in the form of nanoparticles (NPs) with the peak absorption at 712 nm for photoacoustically imaging tumor-bearing mouse models. To monitor PB NPs from the background tissue in vivo, we also developed a new 700-nm-region stimulated Raman scattering (SRS) source (pulse energy up to 200 nJ and repetition rate up to 50 kHz) and implemented optical-resolution photoacoustic microscopy (OR-PAM). The SRS-assisted OR-PAM system was able to monitor PB NPs in the tumor model with micrometer resolution. Due to strong light absorption at 712 nm, the developed SRS light yielded a two-fold higher contrast from PB NPs, in comparison with a 532-nm pumping source. The proposed laser source involved cost-effective and simple system implementation along with high compatibility with the fiber-based OR-PAM system. The study highlights the OR-PAM system in conjunction with the tunable-color SRS light source as a feasible tool to assist NP-mediated cancer therapy.

Effect of Shot Noise on Simultaneous Sensing in Frequency Division Multiplexed Diffuse Optical Tomographic Imaging Process.

Diffuse optical tomography (DOT) has been studied for use in the detection of breast cancer, cerebral oxygenation, and cognitive brain signals. As optical imaging studies have increased significantly, acquiring imaging data in real time has become increasingly important. We have developed frequency-division multiplexing (FDM) DOT systems to analyze their performance with respect to acquisition time and imaging quality, in comparison with the conventional time-division multiplexing (TDM) DOT. A large tomographic area of a cylindrical phantom 60 mm in diameter could be successfully reconstructed using both TDM DOT and FDM DOT systems. In our experiment with 6 source-detector (S-D) pairs, the TDM DOT and FDM DOT systems required 6.18 and 1 s, respectively, to obtain a single tomographic data set. While the absorption coefficient of the reconstruction image was underestimated in the case of the FDM DOT, we experimentally confirmed that the abnormal region can be clearly distinguished from the background phantom using both methods.

Increased EGFR expression induced by a novel oncogene, CUG2, confers resistance to doxorubicin through Stat1-HDAC4 signaling.

BACKGROUND: Previously, it has been found that the cancer upregulated gene 2 (CUG2) and the epidermal growth factor receptor (EGFR) both contribute to drug resistance of cancer cells. Here, we explored whether CUG2 may exert its anticancer drug resistance by increasing the expression of EGFR. METHODS: EGFR expression was assessed using Western blotting, immunofluorescence and capacitance assays in A549 lung cancer and immortalized bronchial BEAS-2B cells, respectively, stably transfected with a CUG2 expression vector (A549-CUG2; BEAS-CUG2) or an empty control vector (A549-Vec; BEAS-Vec). After siRNA-mediated EGFR, Stat1 and HDAC4 silencing, antioxidant and multidrug resistance protein and mRNA levels were assessed using Western blotting and RT-PCR. In addition, the respective cells were treated with doxorubicin after which apoptosis and reactive oxygen species (ROS) levels were measured. Stat1 acetylation was assessed by immunoprecipitation. RESULTS: We found that exogenous CUG2 overexpression induced EGFR upregulation in A549 and BEAS-2B cells, whereas EGFR silencing sensitized these cells to doxorubicin-induced apoptosis. In addition, we found that exogenous CUG2 overexpression reduced the formation of ROS during doxorubicin treatment by enhancing the expression of antioxidant and multidrug resistant proteins such as MnSOD, Foxo1, Foxo4, MRP2 and BCRP, whereas EGFR silencing congruently increased the levels of ROS by decreasing the expression of these proteins. We also found that EGFR silencing and its concomitant Akt, ERK, JNK and p38 MAPK inhibition resulted in a decreased Stat1 phosphorylation and, thus, a decreased activation. Since also acetylation can affect Stat1 activation via a phospho-acetyl switch, HDAC inhibition may sensitize cells to doxorubicin-induced apoptosis. Interestingly, we found that exogenous CUG2 overexpression upregulated HDAC4, but not HDAC2 or HDAC3. Conversely, we found that HDAC4 silencing sensitized the cells to doxorubicin resistance by decreasing Stat1 phosphorylation and EGFR expression, thus indicating an interplay between HDAC4, Stat1 and EGFR. CONCLUSION: Taken together, we conclude that CUG2-induced EGFR upregulation confers doxorubicin resistance to lung (cancer) cells through Stat1-HDAC4 signaling.

Complete genome sequence of bellflower vein chlorosis virus, a novel putative member of the genus Waikavirus.

The complete genome sequence of a new virus isolated from a bellflower (Campanula takesimana) plant was determined. The genome of this virus is composed of monopartite single-stranded RNA of 11,649 nucleotides in length. BLAST searches of protein databases showed that the encoded polyprotein has a maximum amino acid sequence identity of 42% (with 99% coverage) to the polyprotein of the isolate Orissa of rice tungro spherical virus (RTSV; genus Waikavirus). Phylogenetic analysis strongly supports that the identified virus is a member of a new species of the genus Waikavirus. The name bellflower vein chlorosis virus (BVCV) is proposed for this new virus.

High-speed broadband frequency sweep of continuous-wave terahertz radiation.

We present a new technical implementation of a high-speed broadband frequency sweep of continuous-wave terahertz (THz) radiation. THz frequency sweeping with a kHz sweep rate and a THz sweep range is implemented using THz photomixing in which an optical beat source consists of a wavelength-swept laser and a distributed feedback laser diode. During the frequency sweep, frequency-domain THz interferograms are measured using the coherent homodyne detection employing signal averaging for noise reduction, which can give time-of-flight information via fast Fourier transform. Multiple reflections in a Si wafer and the thickness of the wafer are measured to demonstrate the potential of this method for fast THz tomography and thickness measurement.