Development of an earthworm-based soft robot for colon sampling.

Colorectal cancer as a major disease that poses a serious threat to human health continues to rise in incidence. And the timely colon examinations are crucial for the prevention, diagnosis, and treatment of this disease. Clinically, gastroscopy is used as a universal means of examination, prevention and diagnosis of this disease, but this detection method is not patient-friendly and can easily cause damage to the intestinal mucosa. Soft robots as an emerging technology offer a promising approach to examining, diagnosing, and treating intestinal diseases due to their high flexibility and patient-friendly interaction. However, existing research on intestinal soft robots mainly focuses on controlled movement and observation within the colon or colon-like environments, lacking additional functionalities such as sample collection from the intestine. Here, we designed and developed an earthworm-like soft robot specifically for colon sampling. It consists of a robot body with an earthworm-like structure for movement in the narrow and soft pipe-environments, and a sampling part with a flexible arm structure resembling an elephant trunk for bidirectional bending sampling. This soft robot is capable of flexible movement and sample collection within an colon-like environment. By successfully demonstrating the feasibility of utilizing soft robots for colon sampling, this work introduces a novel method for non-destructive inspection and sampling in the colon. It represents a significant advancement in the field of medical robotics, offering a potential solution for more efficient and accurate examination and diagnosis of intestinal diseases, specifically for colorectal cancer.

An AFM-Based Model-Fitting-Free Viscoelasticity Characterization Method for Accurate Grading of Primary Prostate Tumor.

Viscoelasticity is a crucial property of cells, which plays an important role in label-free cell characterization. This paper reports a model-fitting-free viscoelasticity calculation method, correcting the effects of frequency, surface adhesion and liquid resistance on AFM force-distance (FD) curves. As demonstrated by quantifying the viscosity and elastic modulus of PC-3 cells, this method shows high self-consistency and little dependence on experimental parameters such as loading frequency, and loading mode (Force-volume vs. PeakForce Tapping). The rapid calculating speed of less than 1ms per curve without the need for a model fitting process is another advantage. Furthermore, this method was utilized to characterize the viscoelastic properties of primary clinical prostate cells from 38 patients. The results demonstrate that the reported characterization method a comparable performance with the Gleason Score system in grading prostate cancer cells, This method achieves a high average accuracy of 97.6% in distinguishing low-risk prostate tumors (BPH and GS6) from higher-risk (GS7-GS10) prostate tumors and a high average accuracy of 93.3% in distinguishing BPH from prostate cancer.

Development of Microrobot with Optical Magnetic Dual Control for Regulation of Gut Microbiota.

Microrobots have emerged as a promising precision therapy approach that has been widely used in minimally invasive treatments, targeted drug delivery, and wound cleansing, and they also offer a potential new method for actively modulating gut microbiota. Here, a double-faced microrobot was designed to carry gut bacteria via covalently immobilizing the antibodies, and a corresponding integrated optical and magnetic dual-driving control system was also developed for precise control of the microrobot. The microrobot utilizes magnetic microsphere as its core, with one side coated in gold, which serves as the optical receptor surface and the interface for bacterial attachment. The specific gut bacterium, S. cerevisiae, was immobilized on the gold-coated side using the corresponding antibodies. The dual-driving control system enables the precise modulation of gut bacteria by synergistically manipulating the microrobots' movement via the optical field and magnetic field. The feasibility of independent and coordinated control using optical fields and magnetic fields was validated through experimental and numerical simulation approaches. This work introduces a novel method for the precise modulation of gut microbiota, providing a new avenue for disease treatments based on gut bacteria.

Upconversion optogenetics-driven biohybrid sensor for infrared sensing and imaging.

Living organisms are far superior to state-of-the-art devices in visual perception as they have evolved a wide number of capabilities that encompass our most advanced technologies. By leveraging the performance of living organisms and directly interfacing them with artificial components, it can use the intricacy and metabolic efficiency of biological visual sensing within artificial machines. Inspired by the molecular basis (transient receptor potential, TRP) for infrared detection of pit-bearing organisms, we propose a TRP-like biohybrid sensor by integrating upconversion nanoparticles (UCNP) and optogenetically engineered cells on a graphene transistor for infrared sensing and imaging. The UCNP converts infrared light irradiation into blue light, the blue light activates the cells expressed with channelrhodopsin-2 (ChR2) and induces transmembrane photocurrent, and the photocurrent is detected by a biocompatible graphene transistor. Stepwise and overall experimental results show that, upon infrared light irradiation, the UCNP can rapidly mediate cellular photocurrents, which further translates into the extra output current of the graphene transistor. More notably, the response speed of the biohybrid sensor is 1∼3 orders of magnitude faster than those of TRPs heterologously expressed in cell lines in the literature, which confirms the response time advantage of the combination of UCNP and ChR2 within the sensor in place of TRPs. The biohybrid sensor can successfully image infrared targets, proving the feasibility of developing bionic infrared sensing devices by biohybrid integration of nonliving nanomaterials and biological components. This work opens up an avenue for biohybrid sensors to develop the bionic infrared vision that promisingly reproduces the functional superiority of natural organisms. STATEMENT OF SIGNIFICANCE: Infrared sensing and imaging have a wide range of military and civilian applications. Organisms have evolved excellent infrared vision with the molecular basis, transient receptor potential (TRP), and the performance is superior to existing state-of-the-art infrared devices. Inspired by this, a TRP-like biohybrid sensor based on upconversion optogenetics and a 2D material-based device is developed for infrared sensing and imaging. The biohybrid sensor has a relatively fast response speed that is 1∼3 orders of magnitude faster than that of the heterologously expressed TRPs, which enables its capability of infrared imaging with a single pixel-based method. This work broadens the spectrum of biohybrid sensing based on engineered cells to infrared, advancing the process of reproducing the excellent infrared detection of organisms.

Accurate Micromanipulation of Optically Induced Dielectrophoresis Based on a Data-Driven Kinematic Model.

The precise control method plays a crucial role in improving the accuracy and efficiency of the micromanipulation of optically induced dielectrophoresis (ODEP). However, the unmeasurable nature of the ODEP force is a great challenge for the precise automatic manipulation of ODEP. Here, we propose a data-driven kinematic model to build an automatic control system for the precise manipulation of ODEP. The kinematic model is established by collecting the input displacement of the optical pattern and the output displacements of the manipulated object. Then, the control system based on the model was designed, and its feasibility and control precise were validated by numerical simulations and actual experiments on microsphere manipulation. In addition, the applications of ODEP manipulation in two typical scenarios further demonstrated the feasibility of the designed control system. This work proposes a new method to realize the precise manipulation of ODEP technology by establishing a kinematic model and a control system for micromanipulation, and it also provides a general approach for the improvement of the manipulation accuracy of other optoelectronic tweezers.

Isolation method of Saccharomyces cerevisiae from red blood cells based on the optically induced dielectrophoresis technique for the rapid detection of fungal infections.

Saccharomyces cerevisiae (S. cerevisiae) has been classically used to treat diarrhea and diarrhea-related diseases. However, in the past two decades, fungal infections caused by S. cerevisiae have been increasing among immunocompromised patients, and it takes too long to isolate S. cerevisiae from blood to diagnose it in time. In this paper, a new method for the isolation and selection of S. cerevisiae from red blood cells (RBC) is proposed by designing a microfluidic chip with an optically-induced dielectrophoresis (ODEP) system. It was verified by theory and experiments that the magnitude and direction of the dielectrophoresis force applied on RBCs and S. cerevisiae are different, which determine that the S. cerevisiae can be isolated from RBCs by the ODEP system. By designing the specific light images and the dynamic separation mode, the optimal operating conditions were experimentally achieved for acquiring higher purity of S. cerevisiae. The purity ranges were up to 95.9%-97.3%. This work demonstrates a promising tool for efficient and effective purification of S. cerevisiae from RBCs and provides a novel method of S. cerevisiae isolation for the timely diagnosis of fungal infections.

Label-free rapid isolation of saccharomyces cerevisiae with optically induced dielectrophoresis-based automatic micromanipulation.

Saccharomyces cerevisiae is well-known in the baking and brewing industries and always used for the preparation of probiotics, especially its subtype, Saccharomyces boulardii, to prevent and treat various diarrhea and intestinal diseases. However, case reports on the side effects of a wide range of serious infections for the elderly, immunocompromised and critically ill patients after treatment with the S. cerevisiae have been increasing in recent years. The existing diagnose methods of the invasive S. cerevisiae infections in clinical, especially, the key step of the method-cell isolation, is time-consuming that always miss timey diagnose and early prevention. Here, we propose a new automatic micromanipulation method to label-free rapid isolation of S. cerevisiae based on the optically-induced dielectrophoresis (ODEP) technology, combining with image processing and recognition. S. cerevisiae is firstly identified by the image recognition method and then, automatically captured and moved to the target location by designing optical patterns. The results indicate the method can flexibly and automatically manipulate multiple S. cerevisiae cells simultaneously, such as, arranging S. cerevisiae cells, moving an array of the cells at any directions, aggregating the cells, and separating S. cerevisiae from the solution mixed with impurities. This work represents a step toward the use of automatic micromanipulation of ODEP technology to automatically and rapidly isolate S. cerevisiae for the detection of the invasive S. cerevisiae infections.

Rapid isolation method of Saccharomyces cerevisiae based on optically induced dielectrophoresis technique for fungal infection diagnosis.

Saccharomyces cerevisiae(S. cerevisiae) has been classically used as a treatment for diarrhea and diarrhea-related diseases. However, cases of the fungal infections caused by S. cerevisiae have been increasing in the last two decades among immunocompromised patients, while a long time was spent on S. cerevisiae isolation clinically so it was difficult to achieve timely diagnosis the diseases. Here, a novel approach for isolation and selection of S. cerevisiae is proposed by designing a microfluidic chip with an optically induced dielectrophoresis (ODEP) system. S. cerevisiae was isolated from the surroundings by ODEP due to different dielectrophoretic forces. Two special light images were designed and used to block and separate S. cerevisiae, respectively, and several manipulation parameters of ODEP were experimentally optimized to acquire the maximum isolation efficiency of S. cerevisiae. The results on the S. cerevisiae isolation declared that the purity of the S. cerevisiae selected by the method was up to 99.5%±0.05, and the capture efficiency was up to 65.0%±2.5 within 10 min. This work provides a general method to isolate S. cerevisiae as well as other microbial cells with high accuracy and efficiency and paves a road for biological research in which the isolation of high-purity cells is required.

Design, Synthesis, and Activity Evaluation of Novel Acyclic Nucleosides as Potential Anticancer Agents In Vitro and In Vivo.

In the present work, 103 novel acyclic nucleosides were designed, synthesized, and evaluated for their anticancer activities in vitro and in vivo. The structure-activity relationship (SAR) studies revealed that most target compounds inhibited the growth of colon cancer cells in vitro, of which 3-(6-chloro-9H-purin-9-yl)dodecan-1-ol (9b) exhibited the most potent effect against the HCT-116 and SW480 cells with IC50 values of 0.89 and 1.15 µM, respectively. Furthermore, all of the (R)-configured acyclic nucleoside derivatives displayed more potent anticancer activity compared to their (S)-counterparts. Mechanistic studies revealed that compound 9b triggered apoptosis in the cancer cell lines via depolarization of the mitochondrial membrane and effectively inhibited colony formation. Importantly, compound 9b inhibited the growth of the SW480 xenograft in a mouse model with low systemic toxicity. These results indicated that acyclic nucleoside compounds are viable as potent and effective anticancer agents, and compound 9b may serve as a promising lead compound that merits further attention in future anticancer drug discovery.

A bio-syncretic phototransistor based on optogenetically engineered living cells.

Human eyes rely on photosensitive receptors to convert light intensity into action potentials for visual perception, and thus bio-inspired photodetectors with bioengineered photoresponsive elements for visual prostheses have received considerable attention by virtue of superior biological functionality and better biocompatibility. However, the current bioengieered photodetectors based on biological elements face a lot of challenges such as slow response time and lack of effective detection of weak bioelectrical signals, resulting in difficulty to perform imaging. Here, we report a human eye-inspired phototransistor by integrating optogenetically engineered living cells and a graphene-based transistor. The living cells, engineered with photosensitive ion channels, channelrhodopsin-2 (ChR2), and thus endowed with the capability of transducing light intensity into bioelectrical signals, are coupled with the graphene layer of the transistor and can regulate the transistor's output. The results show that the photosensitive ion channels enable the phototransistor to output stronger photoelectrical currents with relatively fast response (~25 ms) and wider dynamic range, and demonstrate the transistor owns optical and biological gating with a significant large on/off ratio of 197.5 and high responsivity of 1.37 mA W-1. An artificial imaging system, which mimics the pathway of human visual information transmission from the retina through the lateral geniculate nucleus to the visual cortex, is constructed with the transistor and demonstrate the feasibility of imaging using the bioengineered cells. This work shows a potential that optogenetically engineered cells can be used to develop novel visual prostheses and paves a new avenue for engineering bio-syncretic sensing devices.

IL-6-targeted ultrasmall superparamagnetic iron oxide nanoparticles for optimized MRI detection of atherosclerotic vulnerable plaques in rabbits.

Vulnerable plaques of atherosclerosis (AS) are the main culprit lesion for the serious risk of acute cardiovascular disease (CVD). Therefore, developing new non-invasive methods to detect vulnerable plaques and to evaluate their stability effectively is of great value in the early diagnosis of CVD. IL-6 plays a vital role in the development and rupture of AS. In this study, IL-6-targeted superparamagnetic iron oxide nanoparticles (Anti-IL-6-USPIO) are synthesized by a chemical condensation reaction. An AS model was established by damaging rabbit abdominal aortic intima with Foley's tube in combination with a high cholesterol diet. The results confirm that Anti-IL-6-USPIO have excellent IL-6-targeting ability and usefulness in detecting vulnerable plaques in vitro and in vivo, which may provide a novel, non-invasive strategy for evaluating acute cardiovascular risk or exploiting anti-atherosclerotic drugs.

Development of an image biosensor based on an optogenetically engineered cell for visual prostheses.

Visual prostheses provide blind patients with artificial vision via electrical stimulation of surviving visual cells resulting in partial restoration of vision in many patients. However, high-resolution visual perception, long-term biocompatibility and safety remain the significant challenges of existing visual prostheses. Here, we present a novel method to develop a new visual prosthesis using living cells as integrated electronics and implantable microelectrodes. The living cells modified with channelrhodopsin-2 showed excellent light-sensitive properties and encoded image information with cellular deformations triggered by light stimulation. The photoresponsive properties of the cells were determined using a single pixel imaging system, which indicated that the cells can act as a good light-sensitive biosensor. Additionally, the imaging feasibility of the cells was further validated through successful and clear imaging of several object scenes using the same system. This work represents a step toward the design and use of living cells as an image biosensor for the development of a new generation of high-resolution visual prostheses.

Imaging with Optogenetically Engineered Living Cells as a Photodetector.

Biosyncretic systems integrating biological components with electromechanical devices have recently become a promising technology, in which biological components are used as actuators or sensing elements with higher-level performance than artificial systems. Here, a biosyncretic imaging system using an optogenetically engineered living cell as a photodetector is shown. The photoresponsive properties of the cell, such as spectrum and response range, dynamic characteristics, are measured and indicate that the cell functions as an excellent photodetector. In the system, the cell is directly utilized to generate light-triggered ionic currents, which encode the spatial image information and therefore are used to reconstruct the scenes under the view based on compressive sensing. Imaging with the cell-based photodetector is successfully performed by acquiring high-definition images using the system. The system also displays function superiority to a commercial photodiode, such as wider dynamic responsivity range. This work represents a step toward directly imaging with living materials and paves a new road for the development of future on-body bionic devices.

Development of a novel optogenetic indicator based on cellular deformations for mapping optogenetic activities.

Optogenetic techniques have changed the landscape of neuroscience by offering high temporal and spatial mapping of the activities of genetically defined population of cells with optical controlling tools. The mapping of optogenetic activities demands optogenetic indicators whose optical properties change in response to cellular activities, but the existing optogenetic indicators only specifically characterize limited optogenetic activities. Here, we propose a novel optogenetic indicator based on cellular deformation to characterize the activities of optogenetically engineered cells. The cellular activities triggered by light stimulation lead to changes in the cell membrane structure and result in cellular deformation, which is measured by atomic force microscopy. The deformation recordings of the cells expressing channelrhodopsin-2 (ChR2) and the corresponding control experiments together confirm that the deformation is generated generally when the cells are exposed to light, which is also validated indirectly via the change in the Young's modulus of the cells before and after absorption of photons. The activities of cells expressing different subtypes of opsins were also recorded using the optogenetic indicator of cellular deformation. This study provides a novel and general optogenetic indicator based on cellular deformation for monitoring the activities of optogenetically engineered cells. Moreover, this new optogenetic indicator offers ever-better tools for the applications of optogenetic activity mapping and neural and brain imaging.

A fast imaging method of scanning ion conductance microscopy.

Scanning ion conductance microscopy (SICM) has attracted considerable attention in the biological field as a noninvasive, high-resolution and non-force contact imaging technology. However, the development of improvement to the SICM imaging rate remains a great challenge for applications of rapid or dynamic imaging. In this paper, a fast SICM imaging method is proposed to improve the imaging efficiency via the design of a compressive sampling strategy and a reduction in the reconstruction time of sparse signals using the 2D normalized iterative hard thresholding (2D-NIHT) algorithm. The imaging performance of the method is validated by the simulation of recovery of a random synthetic image, and the superiority of the 2D-NIHT algorithm is also demonstrated by comparison of its reconstruction performance with that of other typical algorithms. The actual imaging performance of the method in SICM is also validated by the imaging of two biological samples, a virus and a living cell, and the results show that the method can duplicate the sample surface topography with high-definition and shorter imaging time. Our study offers a general imaging method for the applications of scanning probe microscopies to realize faster and higher-resolution imaging of biological samples.

Association Between Serum LDL-C and ApoB and SYNTAX Score in Patients With Stable Coronary Artery Disease.

The aim of this study was to examine the relationship between low-density lipoprotein cholesterol (LDL-C) and apolipoprotein (Apo) B levels and the SYNergy between percutaneous coronary intervention with TAXus and cardiac surgery (SYNTAX) score (SS) in patients with stable angina pectoris. We enrolled 594 patients who were suspected to have coronary heart disease (CHD) and who underwent coronary angiography. Patients were divided into 4 groups based on the SS: normal (SS = 0, n = 154), low SS (SS ≤ 22, n = 210), intermediate SS (22 < SS < 32, n = 122), and high SS (SS ≥ 33, n = 63). Positive correlations between lipoprotein (a), LDL-C, ApoB, total cholesterol, and SS were significant ( r = 0.132, 0.632, 0.599, and 0.313, respectively; P < .01), whereas high-density lipoprotein cholesterol (HDL-C), ApoA1, and ApoA1/ApoB levels showed a significant negative correlation ( r = -0.29, -0.344, and -0.561, respectively; P < .01). Multivariate linear regression analysis revealed that LDL-C, ApoB, ApoA1/ApoB, fibrinogen (Fg), and HDL-C levels had an effect on SS (standardized regression coefficients were 0.41, 0.29, -0.12, 0.08, and -0.09, respectively; P < .05). In conclusion, LDL-C, ApoB, ApoA1/ApoB, Fg, and HDL-C levels affected the SS and were predictors of CHD complexity.

Label-free multidimensional information acquisition from optogenetically engineered cells using a graphene transistor.

The optogenetic technique, which allows the manipulation of cellular activity patterns in space and time by light, has transformed the field of neuroscience. However, acquiring multidimensional optogenetic information remains challenging despite the fact that several cellular information detection methods have been proposed. Herein, we present a new method to acquire label-free multidimensional information from optogenetically engineered cells using a graphene transistor. Using a graphene film to form a strong densely packed layer with cells, the cellular action potentials were characterized as light-activated transistor conductance signals, which quantified the multidimensional optogenetic information. Based on this approach, some important cellular optogenetic information, including electrophysiological state, cell concentration, expression levels of opsin and response to variable light intensity, were also precisely detected. Furthermore, the graphene transistor was also used to distinguish cells expressing different channelrhodopsin-2 variants. Our study offers a general detection method of multidimensional optogenetic information for extending the applications of the optogenetic technique and provides a novel sensor for the development of future biological prosthetic devices.

Thermometry of photosensitive and optically induced electrokinetics chips.

Optically induced electrokinetics (OEK)-based technologies, which integrate the high-resolution dynamic addressability of optical tweezers and the high-throughput capability of electrokinetic forces, have been widely used to manipulate, assemble, and separate biological and non-biological entities in parallel on scales ranging from micrometers to nanometers. However, simultaneously introducing optical and electrical energy into an OEK chip may induce a problematic temperature increase, which poses the potential risk of exceeding physiological conditions and thus inducing variations in cell behavior or activity or even irreversible cell damage during bio-manipulation. Here, we systematically measure the temperature distribution and changes in an OEK chip arising from the projected images and applied alternating current (AC) voltage using an infrared camera. We have found that the average temperature of a projected area is influenced by the light color, total illumination area, ratio of lighted regions to the total controlled areas, and amplitude of the AC voltage. As an example, optically induced thermocapillary flow is triggered by the light image-induced temperature gradient on a photosensitive substrate to realize fluidic hydrogel patterning. Our studies show that the projected light pattern needs to be properly designed to satisfy specific application requirements, especially for applications related to cell manipulation and assembly.

Mask-free fabrication of a versatile microwell chip for multidimensional cellular analysis and drug screening.

Cells are frequently studied because they are basic structural, functional, and biological units of living organisms. Extracting features from cellular behaviors can facilitate decision making in medical diagnoses and represents an important aspect in the development of biomedical engineering. Previous studies have just focused on either the individual cell or cell clusters separately, which leads to a great lack of information. Microwell technologies could address the challenges of in vitro cellular studies, from individual cell studies to 3D functional assays, by providing more information from smaller sample volumes and enabling the incorporation of low-cost high-throughput assays in the drug discovery process. To this end, the present study describes an easy-to-use method for fabricating a versatile microwell chip that utilizes a digital micro-mirror device printing system, and the chip can be employed in multidimensional cellular analysis, ranging from the single cell to the 3D spheroid level. The microwell manufacturing process, using a digital mask in place of a conventional physical mask, is based on shadowed light and is full of flexibility. Three different dimensions (single cell (1D), cell monolayer (2D) and cell spheroid (3D)) of one cell type can be formed using a microwell array and the analyses of biological characteristics are achieved separately. Single cells and cell clusters can be controlled via customized geometries of microfabricated selectively adhesive poly(ethylene glycol) diacrylate (PEGDA) wells. The effects of shape on cellular growth and hybrid tissue layers were investigated by peeling off the microwells. Furthermore, 3D multicellular spheroids were successfully established in a controllable and high-throughput format. Preclinical drug screening was investigated and distinct differences were observed in the tolerance response to drug testing between the 2D and 3D conditions. The study results further demonstrate that the high-density microwell chip is an easy-to-use multidimensional cellular analysis and rapid drug screening technique, and it could be easily adapted for a wide range of biological research applications.

High-Throughput Fabrication and Modular Assembly of 3D Heterogeneous Microscale Tissues.

3D hydrogel microstructures that encapsulate cells have been used in broad applications in microscale tissue engineering, personalized drug screening, and regenerative medicine. Recent technological advances in microstructure assembly, such as bioprinting, magnetic assembly, microfluidics, and acoustics, have enabled the construction of designed 3D tissue structures with spatially organized cells in vitro. However, a bottleneck exists that still hampers the application of microtissue structures, due to a lack of techniques that combined high-throughput fabrication and flexible assembly. Here, a versatile method for fabricating customized microstructures and reorganizing building blocks composed of functional components into a combined single geometric shape is demonstrated. The arbitrary microstructures are dynamically synthesized in a microfluidic device and then transferred to an optically induced electrokinetics chip for manipulation and assembly. Moreover, building blocks containing different cells can be arranged into a desired geometry with specific shape and size, which can be used for microscale tissue engineering.