Self-propelling magnetic nanorobots capable of intrinsic-navigation in biological fluids with enhanced pharmacokinetics and deeper tissue penetration implicates promising strategy in targeted cancer therapy. Here, multi-component magnetic nanobot designed by chemically conjugating magnetic fe 3 o 4 nanoparticles (NPs), anti-epithelial cell adhesion molecule antibody (anti-EpCAM mAb) to multiwalled carbon nanotubes (CNT) loaded with an anticancer drug, doxorubicin hydrochloride (DOX) is reported. Autonomous propulsion of the nanobots and their external magnetic guidance is enabled by enriching fe 3 o 4 nps with dual catalytic-magnetic functionality. the nanobots propel at high velocities even in complex biological fluids. In addition, the nanobots preferably release DOX in the intracellular lysosomal compartment of human colorectal carcinoma (HCT116) cells by the opening of Fe 3 o 4 np gate. Further, nanobot reduce ex vivo HCT116 tumor spheroids more efficiently than free DOX. The multicomponent nanobot's design represents a more pronounced method in targeting tumors with selfassisted anticancer drug delivery for 'far-reaching' sites in treating cancers. Designing miniaturized and versatile robots in the dimensional-range of a few micrometers or less offer potential for unprecedented biomedical applications, such as refinements in targeted drug delivery platforms 1-7. Miniature robotic systems provide considerable benefits over conventional and micro/nanoparticle-based therapies 8,9. Existing anticancer drug delivery systems demonstrate pharmacokinetic (PK) limitations as they are passive systems driven by the blood fluidics and lack intrinsic navigation for long circulation time, targeting, localized delivery, and tissue penetration 10,11. Furthermore, despite surface functionalization with a specific ligand that allows nanocarriers to increase the active targeting ability; the nanocarriers are unable to guide themselves to a target. Hence, for targeted anticancer delivery of therapeutic payloads to disease sites, drug carriers are desired to possess some distinctive traits, including self-propelling force and velocity, navigational functions, precise cell targeting, drug cargo-towing and finally tissue penetration with the release of drug payload 12-16. Micro/nanomotors with efficient cargo towing and effective penetrating abilities make them excellent delivery vehicles that can meet the necessary features for targeted delivery of therapeutics 6. Chemically propelled micro-/ nanorobots have been widely explored for active drug delivery, and tremendous progresses has been made in the past few years 17. However, designing nanobots for biological functionality is still a challenge as they have some inherent limitations, such as complex preparation technology, difficulty of surface modification, difficulty of motion in biological fluids and depending on the material, poor biocompatibility or biodegradability 6,18,19. Furthermore, none of the reported micro/nanobot system has demonstrated practically useful speed hig...

Nanosized robots with self-propelling and navigating capabilities have become an exciting field of research, attributable to their autonomous motion and specific biomolecular interaction ability for bio-analysis and diagnosis. Here, we report magnesium (Mg)-Fe3O4-based Magneto-Fluorescent Nanorobot (“MFN”) that can self-propel in blood without any other additives and can selectively and rapidly isolate cancer cells. The nanobots viz; Mg-Fe3O4-GSH-G4-Cy5-Tf and Mg-Fe3O4-GSH-G4-Cy5-Ab have been designed and synthesized by simple surface modifications and conjugation chemistry to assemble multiple components viz; (i) EpCAM antibody/transferrin, (ii) cyanine 5 NHS (Cy5) dye, (iii) fourth generation (G4) dendrimers for multiple conjugation and (iv) glutathione (GSH) by chemical conjugation onto one side of Mg nanoparticle. The nanobots propelled efficiently not only in simulated biological media, but also in blood samples. With continuous motion upon exposure to water and the presence of Fe3O4 shell on Mg nanoparticle for magnetic guidance, the nanobot offers major improvements in sensitivity, efficiency and speed by greatly enhancing capture of cancer cells. The nanobots showed excellent cancer cell capture efficiency of almost 100% both in serum and whole blood, especially with MCF7 breast cancer cells.

Here, chemically powered and magnetically guided janus magneto‐fluorescent dual‐engine nanobots (MFDEN) that harvest their energy from the reactions with two different fuels are reported. The Mg‐Fe3O4 Janus nanobots prepared by depositing Fe3O4 hemispherical layer on Mg nanoparticles are propelled efficiently by the thrust of oxygen bubble generated from the reactions of Fe3O4 in H2O2 media, and by hydrogen bubble produced at the partial Mg opening in aqueous media. High speeds and long lifetimes are achieved in complex biological media with H2O2 or with NaHCO3, respectively. For the first time, this study demonstrates the utility of nanobots for efficient isolation of trophoblastic cells from blood. Considering the extreme rarity of these cells in blood, efficient technique that enriches sufficient cells rapidly and with specificity is desired. The nanobots exhibit excellent capture efficiency of almost 100%, both in serum and whole blood for trophoblastic Be‐Wo cells. Furthermore, the high capture efficiency is achieved in 5 min. Feasibility of obtaining an isolated cell based diagnosis from whole blood is demonstrated. Although determining the exact sensitivity and specificity will require more data, this study supports the potential of the multiple fuel‐powered nanorobots for clinical translation of circulating fetal cell based non‐invasive prenatal testing.

Purpose: The field of cancer nanomedicine has made significant progress, but its clinical translation is impeded by many challenges, such as the difficulty in analysing intracellular anticancer drug release by the nanocarriers due to the lack of suitable tools. Here, we propose the development of a combinatorial imaging and analysis technique to evaluate anticancer drug such as doxorubicin HCl (DOX) released by a nanocarrier inside the HCT116 colon cancer cells and its subsequent intracellular accumulation. Procedure: Fluorescent cell images were captured and subjected to combined image analysis and machine learning based procedures to assess and quantify the delivery and retention rate of DOX inside the cancer cells by multifunctional CNT-DOX-Fe3O4nanocarrier. Results: We show that DOX in HCT116 cells was higher for multifunctional CNT-DOX-Fe3O4nanocarrierthan free DOX, indicating efficient and steady release of DOX as well as superior retentive property of the nanocarrier. Initially (1 h and 4 h) the luminance intensity of DOX in the cell cytoplasm delivered by CNT-DOX-Fe3O4nanocarrier was ~0.34 times and ~0.42 times lesser than that of free DOX delivered normally. However, at 24 h and 48 h post treatment the luminance intensity of DOX for CNT-DOX-Fe3O4nanocarrier was ~1.98 times and 1.92 times higher than that of free DOX. Furthermore, the luminance intensity of DOX for CNT-DOX-Fe3O4in the whole cell was ~1.35 times and ~1.62 times higher than that of free DOX at 24h and 48 h, respectively. Conclusions: The high-throughput nature of our image analysis workflow allowed us to automate the process of DOX retention analysis, and enabled us to devise machine learning-based modeling to predict the percentage of anticancer drug retention in cells. The development of models to automatically quantify and predict intra-cellular drug release in cancer cells could benefit personalized treatments by optimizing the design of nanocarriers.

The field of cancer nanomedicine has made significant progress, but its clinical translation is impeded by many challenges, such as the difficulty in analyzing intracellular anticancer drug release by the nanocarriers due to the lack of suitable tools. Here, we propose the development of an image-based strategy involving machine learning (ML) to evaluate anticancer drug such as doxorubicin hydrochloride (DOX) released by a nanocarrier inside the HCT116 colon cancer cells and its subsequent intracellular accumulation. This technique combines fluorescent cell imaging with ML-based image analysis to assess and quantify the delivery of DOX by nanoparticles within them. We show that DOX in HCT116 cells was higher for multifunctional CNT-DOX-Fe 3 O 4 nanocarrier than free DOX, indicating efficient and steady release of DOX as well as superior retentive property of the nanocarrier. Initially (1 and 4 hours), the luminance intensity of DOX in the cell cytoplasm delivered by CNT-DOX-Fe 3 O 4 nanocarrier was ~0.34 and ~0.42 times lesser than that of free DOX delivered normally. However, at 24 and 48 hours posttreatment, the luminance intensity of DOX for CNT-DOX-Fe 3 O 4 nanocarrier was ~1.98 and ~1.92 times higher than that of free DOX. Furthermore, the luminance intensity of DOX for CNT-DOX-Fe 3 O 4 in the whole cell was ~1.35 and ~1.62 times higher than that of free DOX at 24 and 48 hours, respectively. The high-throughput nature of our image analysis workflow allowed us to automate the process of DOX retention analysis and enabled us to devise ML-based modeling to predict the percentage of anticancer drug retention in cells. The development of models to automatically quantify and predict intracellular drug release in cancer cells could benefit personalized treatments by optimizing the design of nanocarriers.