Muscle cell-powered biohybrid robots represent a transformative fusion of biological tissue engineering and robotics, offering unprecedented potential for biomedical applications targeted at drug delivery, regenerative medicine, bioengineered heart patches, lab-on-a-chip devices, biosensors, and soft surgical tools. This review categorizes the currently available examples and further explores advanced biofabrication techniques that drive the development of biohybrid systems, with a focus on 3D bioprinting, electrospinning, micro/nano patterning, self-assembly, and microfluidic devices. These fabrication strategies facilitate precise cell alignment, enhance electrical and mechanical properties, and enable the seamless integration of biological components with engineered structures. By incorporating both cardiomyocytes and skeletal muscle cells, biohybrid robots achieve controlled actuation, autonomous movement, and adaptability to environmental stimuli. Furthermore, we discuss the latest optimization strategies in biofabrication, addressing key challenges such as scalability, biocompatibility, and functional integration. Biohybrid robots, including swimmers, actuators, and pumps, enable targeted drug delivery, assistive devices, and fluid transport in engineered tissues. Their integration with biological systems advances regenerative medicine, disease modeling, drug screening, and soft robotics. This review provides a comprehensive perspective on the state-of-the-art advancements and potential optimization in the fabrication techniques, paving the way for the next generation of biohybrid robotic systems.

Brain organoids have emerged as promising three-dimensional (3D) models that recapitulate key aspects of human brain development, neural circuit formation, and neurological disorders. However, conventional culture systems face critical limitations, including inadequate vascularization, restricted diffusion of nutrients and oxygen, insufficient neuronal maturation, and poor reproducibility, all of which hinder long-term stability and clinical translation. To address these challenges, organoid-on-a-chip technologies have been developed to provide controlled microenvironments, fluidic dynamics, and enhanced tissue integration; nevertheless, significant barriers remain. In recent years, nanomaterials have been increasingly incorporated into brain organoids and chip-based systems to overcome these limitations. Due to their unique structural, electrical, and biochemical properties, nanomaterials can mimic components of the extracellular matrix, promote cellular organization, enhance electrophysiological maturation, and enable advanced sensing modalities. Their integration with organoid-on-a-chip platforms further facilitates vascularization, supports long-term culture, and contributes to the generation of physiologically relevant neural models. This review provides a comprehensive overview of brain organoid technology, the functional roles of nanomaterials in these systems, and recent advances in nanomaterial-based brain organoid-on-a-chip platforms. Additionally, we summarize how these interdisciplinary approaches enhance the modeling of neurological diseases, improve drug evaluation including organoid-based biohybrid robot on-a-chip, and support the development of personalized medicine. Finally, we discuss persisting limitations and outline future directions toward the realization of intelligent, reproducible, and clinically translatable neural platforms. We hope this review will inspire innovative strategies and accelerate progress at the intersection of nanomaterials, organoid biology, and chip-based technologies, thereby advancing personalized and effective treatments in neuroscience and biomedicine.

Exosomes are nanoscale extracellular vesicles secreted by both cancerous and healthy cells that carry a diverse array of biomolecules, including nucleic acids and proteins, reflecting the physiological and pathological states of their cells of origin. This makes them highly promising biomarkers for liquid biopsy-based cancer diagnostics. However, conventional methods for exosome analysis from isolation to detection are often time-consuming, labor-intensive, and lack the sensitivity and specificity required for clinical applications. To address these limitations, nanomaterials are increasingly recognized as powerful tools for both exosome isolation and biomarker detection. Their unique physicochemical properties enable enhanced capture efficiency, precise molecular recognition, and signal amplification. In recent years, there has been increasing interest in developing nanomaterial-based total analysis systems (TAS) that seamlessly integrate exosome isolation and detection into unified, high-throughput diagnostic platforms. Consolidating the fragmented workflow into compact, high-throughput devices, these TAS hold strong promise for clinical and point-of-care (POC) applications. While previous reviews have primarily focused on either isolation or detection strategies, this work provides comprehensive overview dedicated to integrated nanomaterial-based TAS that combines both isolation and detection in a single platform. In this review, we discuss the biological relevance of exosome proteins and miRNAs as cancer biomarkers. We then examine nanomaterial-assisted strategies for exosome isolation and for signal transduction and amplification in exosome biomarker detection. Finally, we highlight newly developed TAS for exosome analysis, most reported in studies published between 2023 and 2025, emphasizing their potential for clinical application and POC applications. This review aims to establish a roadmap for future innovations in exosome-based diagnostics enabled by nanotechnology. To this end, we critically assess the current challenges and outline future directions, providing perspectives for their clinical translation.

Circulating tumor RNA (ctRNA) is a sensitive biomarker for early cancer diagnosis, offering real-time gene expression profiles and tumor-specific signatures missed by circulating tumor DNA. Despite its potential, a simple, rapid, and sensitive ctRNA detection sensor has not yet been developed. For the first time, we present an amplification-free electrochemical biosensor for ctRNA detection by integrating the CRISPR/Cas13a system with a silver ion (Ag⁺)-mediated RNA probe and an Au porous-lattice nanoelectrode (APLNE). A three-cytosine-cytosine (C–C) mismatched RNA duplex was employed as a signal probe, allowing site-specific Ag⁺ intercalation to form stable C–Ag⁺–C coordination complexes (RNA-3Ag⁺) that generate strong redox peaks. To further enhance performance, the APLNE, featuring a highly aligned porous gold nanostructure, was used to increase the effective surface area, improving probe immobilization and electrochemical signals. Upon target recognition, CRISPR/Cas13a cleaved the RNA-3Ag⁺ probe, resulting in a significant signal reduction. This biosensor detects KRAS G12D ctRNA, a key pancreatic cancer mutation, with ultrahigh sensitivity (LOD = 0.5 fM) in just 20 min and demonstrates excellent specificity in complex biological samples. Operating without nucleic acid amplification or toxic redox reagents, this simple, cost-effective, and eco-friendly platform shows strong potential for liquid biopsy-based diagnostics and point-of-care cancer screening. Amplification-free CRISPR/Cas13a-based electrochemical biosensor for ctRNA detection APLNE with high surface area enabling efficient signal transduction A redox signal enhanced by Ag⁺-coordinated C–C mismatched RNA probes Achieve sensitive detection of KRAS G12D ctRNA with LOD = 0.5 fM within 20 mins Demonstrate practical applicability in human serum and cell media

ABSTRACT Rapid and sensitive detection of disease‐related biomarkers is critical for early diagnosis and appropriate treatment. However, conventional analytical methods are often time‐consuming and require complex procedures and skilled personnel. Electrochemical biosensors have therefore attracted considerable attention because they offer high sensitivity, rapid response, low cost and strong potential for point‐of‐care diagnostics. Recent efforts have focused on enhancing analytical performance through electrode interface engineering and signal amplification strategies. In particular, nanomaterials are widely employed to improve interfacial signal transduction and increase the loading density of biorecognition elements. In addition, CRISPR/Cas systems provide highly specific target recognition and inherent signal amplification, enabling the sensitive detection of diverse analytes. More recently, microfluidic technologies have emerged as key components in electrochemical biosensing platforms, enabling precise manipulation of small sample volumes, reduced reagent consumption, accelerated reaction kinetics and automated sample‐to‐answer analysis. They also facilitate multiplexed detection and support the development of compact, portable diagnostic devices. Furthermore, microfluidic platforms enable the integration of multiple analytical steps, including sample preparation, target recognition and signal readout, within a single miniaturised system, thereby improving analytical efficiency while minimising user intervention and operational errors. These advantages make microfluidic‐integrated electrochemical biosensors promising candidates for next‐generation portable diagnostic technologies. In this review, we summarise the fundamental principles of electrochemical biosensors and highlight recent advances in nanomaterial‐based interface design, CRISPR‐assisted electrochemical sensing and microfluidic‐integrated platforms for practical diagnostic applications.

Organoid-on-a-chip (OoC) technology integrates three-dimensional (3D) organoid models with chip platforms and has revolutionized biomedical research by closely replicating key physiological and pathological features of human tissues. However, challenges, such as limited structural stability, poor differentiation, inefficient cellular communication, and suboptimal biochemical signaling, continue to hinder the full potential of the OoC technology. Meanwhile, nanoparticles offer a powerful solution to these limitations by precisely modulating the microenvironment, enhancing biomolecular interactions, and improving the functional integration of organoids within biochips. Moreover, incorporating metallic, polymeric, lipid-based, and inorganic nanoparticles into OoC platforms has facilitated controlled extracellular matrix modulation and stem cell differentiation, significantly improving organoid viability and functional stability. Furthermore, tailored nanoparticle functionalization strategies have enhanced biocompatibility and targeting efficiency, broadening the use of OoCs in biomedical applications. This review comprehensively analyzes nanoparticle-functionalized OoC systems, emphasizing the contributions to drug evaluation, disease modeling, and toxicity assessment. Finally, we discuss key challenges, including biocompatibility concerns, standardization issues, and translational barriers, and explore future directions for the next generation of nanoparticle-enhanced OoC platforms.

A 3D motor neuron (MN) spheroid has been developed to investigate neurodegenerative and neuromuscular junction (NMJ) disease. However, core necrosis and reduced neurogenesis, impairing neural network formation, were observed as the MN spheroid matured. In this study, to enhance neural network formation, a biohybrid MN spheroid composed of neural cells/reduced graphene oxide (rGO)/human umbilical vein endothelial cells (HUVECs) was generated for the first time and applied to 3D biosensing system for MNJ disease. By incorporating rGO and HUVECs at the onset of human neural stem cell (hNSC) culture, rGO and HUVECs were evenly distributed within MN spheroid generated by differentiation of hNSC, which improved oxygen- and nutrient- supply by reduction of core necrosis, and enhanced neurogenesis. The fabricated biohybrid MN spheroid improved neural network formation and electrophysiological signal. This method was also applied to generate biohybrid cerebral organoids from human induced pluripotent stem cells (hiPSCs), emphasizing its versatility for diverse 3D neural models. Then, a 3D NMJ biosensing system was fabricated by positioning the biohybrid MN spheroid with muscle bundles to evaluate its utility in neuromuscular disease modeling. Biohybrid MN spheroids generated from induced pluripotent stem cells of sporadic amyotrophic lateral sclerosis (ALS) patients were used to make NMJ. Reduced contraction of the connected muscle bundle due to ALS could be restored by upon treatment with the bosutinib, ALS drug, demonstrating the potential use for drug screening. The method to generate biohybrid spheroid can be applied to generation of various biohybrid brain organoids, and the proposed 3D NMJ biosensing system can be used to drug screening of diverse neuromuscular diseases.

The development of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas) technology (CRISPR/Cas) as a gene-editing tool has the potential to revolutionize nucleic acid analysis. Recently, CRISPR/Cas systems have demonstrated considerable promise in the development of biosensors for the detection of essential disease biomarkers because they exhibit nonspecific collateral cleavage properties upon target sequence recognition. However, the CRISPR/Cas-based biosensors developed thus far have limitations, such as complicated steps, low sensitivity, low selectivity, and low signal-to-noise ratios. These limitations can be overcome by incorporating the unique characteristics of plasmonic nanomaterials into CRISPR/Cas systems to enhance the signal and improve the sensitivity of these biosensors. From this perspective, current interdisciplinary studies on CRISPR/Cas-based nanobiosensors comprising plasmonic nanomaterials can contribute to the development of highly sensitive CRISPR/Cas-based nanobiosensors. These nanobiosensors can detect attractive disease biomarkers, such as viral nucleic acids, small molecules, and proteins. This review article provides a thorough overview of nanobiosensors that incorporate CRISPR/Cas systems combined with plasmonic nanomaterials to enhance biosensing performance. We believe this review will inspire novel approaches and further innovation in the fields of molecular diagnostics and biomedicine aimed at using CRISPR/Cas systems and plasmonic nanomaterials for more personalized and effective medical treatments.

Recently, flexible electronics have significantly transformed information and communications technology (ICT). In particular, wearable devices, via integration with attachable biosensors, have driven the development of new types of biosensors and diagnostic devices for point-of-care testing (POCT). Moreover, wearable electrochemical biosensors can be applied to diagnose diseases in real time based on the synergistic effect generated from the incorporation of the electrochemical technique. Besides, to improve the sensitivity of electrochemical biosensors while retaining their wearability, novel nanomaterials and nanotechnologies have been introduced. In this review, recent studies on nanotechnology-based wearable electrochemical biosensors for accurate disease diagnosis are discussed. First, widely used techniques for developing flexible electrodes, including nanolithography- and nano/microneedle-based patches, are presented. Next, the latest studies on developing wearable electrochemical biosensors for the diagnosis of diseases such as diabetes and dermatitis are discussed by categorizing the biosensors into nanolithography- and nano/microneedle-based categories. Finally, this review explores the latest research trends on the application of nanotechnology-enabled nanopatterning and nano/microneedle technologies to electrochemical wearable biosensors. This review suggests novel approaches and methods for developing wearable electrochemical biosensors for real-time disease diagnosis under POCT applications.

Photostimulated actuators can be remotely actuated and operated at high speed, making them potential candidates for soft robotics. In this study, a paper-based actuator activated solely by light was developed for weight lifting. A polymer blend, primarily made of agarose-poly(3,4-ethylenedioxythiophene):poly(styrenesulfonic acid) (PEDOT:PSS), as a photosensitizer, MXenes and methylene blue (MB) was employed on a paper substrate to produce a bilayer structure using a paintbrush technique. The resultant paper/polymer bilayer actuator was activated by light and showed effective bending and weight-lifting capabilities, which can contribute to wireless paper soft robotics and biohybrid robots.

Biohybrid robots and biohybrid robot-on-a-chip have been developed for drug screening and toxicity screening the complementing animal experiments. A sensing system is needed to evaluate the response to external stimulation, but until now, only the human motor system-based biohybrid robot-on-a-chip with neuromuscular system has been developed without a sensing system. A human nervous system-based biohybrid robot-on-a-chip with eye function as a sensing system in addition to brain/motor neuron/muscle functions is proposed for the first time. Eye assembloid is fabricated by a combination of thalamic organoid covered with Au nanomesh and four retinal organoids. By a combination of eye assembloid, cerebral organoid, motor neuron spheroid, and muscle bundle on polymer substrate, a human nervous system-based biohybrid robot-on-a-chip is made. When blue light is used to cause retinal damage or a hydroxychloroquine (HCQ), a retinal toxic chemical, is applied to the eye assembloid, they caused a decrease in muscle bundle contraction. These results indicated that electrophysiological signals generated by the eye assembloid are transmitted to the muscle bundle through cerebral organoid and motor neuron spheroids, and thus the proposed system can perform the toxicity screening. Human nervous system-based biohybrid robot-on-a-chip can be applied to drug screening of neurodegenerative diseases and toxicity screening for the complement of animal experiments in the future.

Osteoporosis is a progressive skeletal disorder involving decreased bone density and compromised structural integrity, ultimately heightening fracture risk. While maghemite nanoparticles enhance osteoblast activity, their limited influence on osteoclasts reduces their therapeutic efficacy. To overcome this, we developed ascorbic acid-coated maghemite nanoparticles (AMN) that were further surface-modified with hyaluronic acid and polyethylene glycol for improved biocompatibility and stability. AMN scavenges reactive oxygen species and promotes collagen synthesis, offering a dual-functional approach to bone regeneration. Physicochemical analyses, including zeta potential, UV–Vis spectroscopy, and SEM, confirmed AMN formation and stability. In vitro, AMN significantly enhanced MG-63 cell growth, collagen synthesis, ALP activity, and mineral formation. It also TRAP activity measured during osteoclast induction from RAW 264.7 cell, demonstrating its osteoclast-inhibitory potential. Furthermore, AMN supports the formation of a collagen-rich extracellular matrix, facilitating biomineralization and osteogenesis. In vivo, ovariectomized rats treated with AMN exhibited increased bone volume fraction, thicker trabeculae, and reduced trabecular separation, indicating enhanced osteogenesis (BMP2, RUNX2, COL1, and osteocalcin) and suppressed bone resorption (TRAP, RANKL, and RANK). AMN is a promising therapeutic candidate for osteoporosis as it simultaneously promotes bone formation and inhibits bone loss, with strong potential for clinical applications.

Genomic disorders affecting the central nervous system(CNS)are among the most complex and devastating conditions in human health.Moreover,these disorders,such as Rett syndrome,spinal muscular atrophy,and Fragile X syndrome,are typically caused by mutations in genes essential for neural development,synaptic function,or cellular homeostasis.Despite the genetic diversity involved,these diseases share key pathological features,including progressive neurodegeneration,disruption of neural circuits,and loss of cognitive or motor function.

Human papillomavirus (HPV) is the most common virus for genital tract infections. Cervical cancer ranks as the fourth most prevalent cancer globally, with over 99% of cases in women attributed to HPV infection. This infection continues to pose an ongoing threat to public health. Therefore, the development of rapid, high-throughput, and sensitive HPV detection platforms is important, especially in regions with limited access to advanced medical resources. CRISPR-based biosensors, a promising new method for nucleic acid detection, are now rapidly and widely used in basic and applied research and have received much attention in recent years for HPV diagnosis and treatment. In this review, we discuss the mechanisms and functions of the CRISPR-Cas system, focusing on its applications in HPV diagnostics. The review covers CRISPR technologies such as CRISPR-Cas9, CRISPR-Cas12, and CRISPR-Cas13, along with nucleic acid amplification methods, CRISPR-based signal output systems, and point-of-care testing (POCT) strategies. This comprehensive overview highlights the versatility and potential of CRISPR technologies in HPV detection. We also discuss the numerous CRISPR biosensors developed since the introduction of CRISPR to detect HPV. Finally, we discuss some of the challenges faced in HPV detection by the CRISPR-Cas system.

The clustered regularly interspaced short palindromic repeat-Cas (CRISPR-Cas) system, combined with isothermal nucleic acid amplification, offers a powerful sensing mechanism for transducing sequence-specific information. However, analyzing exosomal miRNAs (Exo-miRs) using the CRISPR-Cas system remains difficult and often involves complex steps, such as external exosome isolation/ enrichment or complex signal amplification and detection. Here, we introduce hourglass-inspired total analysis system (H-TAS) for easy and sensitive colorimetric detection of Exo-miR in one-pot using the CRISPR-Cas12a system with isothermal nucleic amplification. In this device, the entire system operates through a successive configuration of two innovative systems: (i) exosome isolation/enrichment and one-pot isothermal Exo-miR amplification via promoter/antibodyenriched gold-satellite-magnetic nanoparticles and (ii) enhanced colorimetric signal analysis using multi-enzyme-encapsulated nanoparticles and the CRISPR-Cas12a system. Moreover, to facilitate the successive combination and ease of use of the two systems, we developed an hourglass-inspired device capable of performing all the analysis steps to detect Exo-miRs without changing the reaction vial. Consequently, the developed H-TAS can readily proceed with all processes required for Exo-miR detection using only a simple flipping action. As a proof-of-concept demonstration, we applied H-TAS to miR-21 and Exo-miR-21 analysis and confirmed adequate specificity and sensitivity (limit of detection = 0.711 fM, 1.308 fM respectively) within 190 min (sample-to-answer time) without needing specialized instruments. Furthermore, the adaptability of the proposed device was verified by analysis of Exo-miR-21 from various breast cell-derived exosomes and clinical samples. Therefore, H-TAS may be a promising platform for detecting Exo-miRs in a highly sensitive, selective, simple, and time-efficient manner.

Pyocyanin is considered a maker of Pseudomonas aeruginosa (P. aeruginosa) infection. Pyocyanin is among the toxins released by the P. aeruginosa bacteria. Therefore, the development of a direct detection of PYO is crucial due to its importance. Among the different optical techniques, the Raman technique showed unique advantages because of its fingerprint data, no sample preparation, and high sensitivity besides its ease of use. Noble metal nanostructures were used to improve the Raman response based on the surface-enhanced Raman scattering (SERS) technique. Anodic metal oxide attracts much interest due to its unique morphology and applications. The porous metal structure provides a large surface area that could be used as a hard template for periodic nanostructure array fabrication. Porous shapes and sizes could be controlled by controlling the anodization parameters, including the anodization voltage, current, temperature, and time, besides the metal purity and the electrolyte type/concentration. The anodization of aluminum foil results in anodic aluminum oxide (AAO) formation with different roughness. Here, we will use the roughness as hotspot centers to enhance the Raman signals. Firstly, a thin film of gold was deposited to develop gold/alumina (Au/AAO) platforms and then applied as SERS-active surfaces. The morphology and roughness of the developed substrates were investigated using scanning electron microscopy (SEM) and atomic force microscopy (AFM) techniques. The Au/AAO substrates were used for monitoring pyocyanin secreted from Pseudomonas aeruginosa microorganisms based on the SERS technique. The results showed that the roughness degree affects the enhancement efficiency of this sensor. The high enhancement was obtained in the case of depositing a 30 nm layer of gold onto the second anodized substrates. The developed sensor showed high sensitivity toward pyocyanin with a limit of detection of 96 nM with a linear response over a dynamic range from 1 µM to 9 µM.

In this review, we discussed the unique features of carbon nanomaterial-integrated nanocomposites for application in the biomedical field, including for biosensors, drug delivery, and tissue engineering.

Brain organoids are being recognized as valuable tools for drug evaluation in neurodegenerative diseases due to their similarity to the human brain's structure and function. However, a critical challenge is the lack of selective and sensitive electrochemical sensing platforms to detect the response of brain organoids, particularly changes in the neurotransmitter concentration upon drug treatment. This study introduces a 3D concave electrode patterned with a mesoporous Au nanodot for the detection of electrochemical signals of dopamine in response to drugs in brain organoids for the first time. The mesoporous Au nanodot-patterned film was fabricated using laser interference lithography and electrochemical deposition. Then, the film was attached to a polymer-based 3D concave mold to obtain a 3D concave electrode. Midbrain organoids generated from Parkinson's disease (PD) patient-derived iPSCs with gene mutations (named as PD midbrain organoid) or normal midbrain organoids were positioned on the developed 3D concave electrode. The 3D concave electrode showed a 1.4 times higher electrochemical signal of dopamine compared to the bare gold electrode. And the dopamine secreted from normal midbrain organoids or PD midbrain organoids on the 3D concave electrode could be detected electrochemically. After the treatment of PD midbrain organoids with levodopa, the drug for PD, the increase in dopamine level was detected due to the activation of dopaminergic neurons by the drug. The results suggest the potential of the proposed 3D concave electrode combined with brain organoids as a useful tool for assessing drug efficacy. This sensing system can be applied to a variety of organoids for a comprehensive drug evaluation.