Cardiotoxicity Assessment through a Polymer-Based Cantilever Platform: An Integrated Electro-Mechanical Screening Approach.

Preclinical drug screening for cardiac toxicity has traditionally relied on observing changes in cardiomyocytes' electrical activity, primarily through invasive patch clamp techniques or non-invasive microelectrode arrays (MEA). However, relying solely on field potential duration (FPD) measurements for electrophysiological assessment can miss the full spectrum of drug-induced toxicity, as different drugs affect cardiomyocytes through various mechanisms. A more comprehensive approach, combining field potential and contractility measurements, is essential for accurate toxicity profiling, particularly for drugs targeting contractile proteins without affecting electrophysiology. However, previously proposed platform has significant limitations in terms of simultaneous measurement. The novel platform addresses these issues, offering enhanced, non-invasive evaluation of drug-induced cardiotoxicity. It features eight cantilevers with patterned strain sensors and MEA, enabling real-time monitoring of both cardiomyocyte contraction force and field potential. This system can detect minimum cardiac contraction force of ≈2 µN and field potential signals with 50 µm MEA diameter, using the same cardiomyocytes in measurements of two parameters. Testing with six drugs of varied mechanisms of action, the platform successfully identifies these mechanisms and accurately assesses toxicity profiles, including drugs not inhibiting potassium channels. This innovative approach presents a comprehensive, non-invasive method for cardiac function assessment, poised to revolutionize preclinical cardiotoxicity screening.

Correction: Quantitative assessment of cardiomyocyte mechanobiology through high-throughput cantilever-based functional well plate systems.

Correction for 'Quantitative assessment of cardiomyocyte mechanobiology through high-throughput cantilever-based functional well plate systems' by Jongyun Kim et al., Analyst, 2023, 148, 5133-5143, https://doi.org/10.1039/D3AN01286G.

Quantitative assessment of cardiomyocyte mechanobiology through high-throughput cantilever-based functional well plate systems.

Proper regulation of the in vitro cell culture environment is essential for disease modelling and drug toxicity screening. The main limitation of well plates used for cell culture is that they cannot accurately maintain energy sources and compounds needed during cell growth. Herein, to understand the importance of perfusion in cardiomyocyte culture, changes in contractile force and heart rate during cardiomyocyte growth are systematically investigated, and the results are compared with those of a perfusion-free system. The proposed perfusion system consists of a Peltier refrigerator, a peristaltic pump, and a functional well plate. A functional well plate with 12 wells is made through injection moulding, with two tubes integrated in the cover for each well to continuously circulate the culture medium. The contractile force of cardiomyocytes growing on the cantilever surface is analysed through changes in cantilever displacement. The maturation of cardiomyocytes is evaluated through fluorescence staining and western blot; cardiomyocytes cultured in the perfusion system show greater maturity than those cultured in a manually replaced culture medium. The pH of the culture medium manually replaced at intervals of 3 days decreases to 6.8, resulting in an abnormal heartbeat, while cardiomyocytes cultured in the perfusion system maintained at pH 7.4 show improved contractility and a uniform heart rate. Two well-known ion channel blockers, verapamil and quinidine, are used to measure changes in the contractile force of cardiomyocytes from the two systems. Cardiomyocytes in the perfusion system show greater stability during drug toxicity screening, proving that the perfusion system provides a better environment for cell growth.

Stress induced self-rollable smart-stent-based U-health platform for in-stent restenosis monitoring.

To date, several smart stents have been proposed to continuously detect biological cues, which is essential for tracking patients' critical vital signs and therapy. However, the proposed smart stent fabrication techniques rely on conventional laser micro-cutting or 3D printing technologies. The sensors are then integrated into the stent structure using an adhesive, conductive epoxy, or laser micro-welding process. The sensor packaging method using additional fabrication processes can cause electrical noise, and there is a possibility of sensor detachment from the sent structure after implantation, which may pose a significant risk to patients. Herein, we are demonstrating for the first time a single-step fabrication method to develop a smart stent with an integrated sensor for detecting in-stent restenosis and assessing the functional dynamics of the heart. The smart stent is fabricated using a microelectromechanical system (MEMS)-based micromachining technology. The proposed smart stent can detect biological cues without additional power and wirelessly transmit the signal to the network analyzer. The cytocompatibility of the smart stent is confirmed through a cytotoxicity test by monitoring the cell growth, proliferation, and viability of the cultured cardiomyocytes. The capacitance of the smart stent exhibits an excellent linear relationship with the applied pressure. The exceptional sensitivity of the pressure sensor enabled the proposed smart stent to detect biological cues during in vivo analysis. The preliminary findings confirmed the proposed smart stent's higher level of structural integrity, durability and repeatability. Finally, the practical feasibility of the smart stent is demonstrated by monitoring diastole and systole at various beat rates using a phantom. The results of the phantom study showed a similar pattern to the human model, indicating the potential use of the proposed multifunctional smart stent for real-time applications.

Enhancement of cardiac contractility using gold-coated SU-8 cantilevers and their application to drug-induced cardiac toxicity tests.

Herein, we propose an array of gold (Au)-coated SU-8 cantilevers with microgrooves for improved maturation of cardiomyocytes and describe its applications to drug-induced cardiac toxicity tests. Firstly, we evaluated the effect of cell culture substrates such as polydimethylsiloxane (PDMS), polyimide (PI), and SU-8 on the cardiomyocyte's maturation. Among these, the SU-8 with microgroove structures exhibits improved cardiomyocyte maturation. Further, thin layers of graphene and Au are coated on SU-8 substrates and the effects of these materials on cardiomyocyte maturation are evaluated by analyzing the expression of proteins such as alpha-actinin, Connexin 43 (Cx43), and Vinculin. While both conductive materials enhanced protein expression when compared to bare SU-8, the Au-coated SU-8 substrates demonstrated superior cardiomyocyte maturation. The cantilever structure is constructed using microgroove patterned SU-8 with and without an Au coating. The Au-coated SU-8 cantilever showed maximum displacement of 17.6 ± 0.3 µm on day 21 compared to bare SU-8 (14.2 ± 0.4 µm) owing to improved cardiomyocytes maturation. Verapamil and quinidine are used to characterize drug-induced changes in the contraction characteristics of cardiomyocytes on bare and Au-coated SU-8 cantilevers. The relative contraction forces and beat rates changed according to the calcium and sodium channel related drugs. Matured cardiomyocytes are less influenced by the drugs compared to immature cardiomyocytes and showed reliable IC50 values. These results indicate that the proposed Au-coated SU-8 cantilever array could help improve the accuracy of toxicity screening results by allowing for the use of cardiomyocytes that have been matured on the drug screening platform.

Multi-layered polymer cantilever integrated with full-bridge strain sensor to enhance force sensitivity in cardiac contractility measurement.

In this study, we developed a multi-layered functional cantilever for real-time force measurement of cardiomyocytes in cell culture media. The functional cantilever with a full-bridge circuit configuration was composed of one polydimethylsiloxane (PDMS) and two polyimide (PI) layers, forming two resistive sensors on each upper side of the two PI layers. The PI layers were chemically bonded using an oxygen plasma treatment, with a thin composite layer consisting of Cr/SiO2/PDMS. These greatly improved the force sensitivity and the long-term reliability of the integrated strain sensor operating in liquids. The nanogrooved PDMS top layer bonded on the upper PI layer was employed to further improve the growth of cardiomyocytes on the functional cantilever. The difference in resistance changes and response characteristics was confirmed by evaluating the characteristics of the multi-layered polymer cantilevers with half-bridge and full-bridge circuit configurations. We also employed the cantilever devices to measure the contraction force of cardiomyocytes for 16 days and side effects in real time in human-induced pluripotent stem cells treated with the cardiovascular drug verapamil. The sensor-integrated cantilever devices are expected to be utilized as a novel biomedical sensor for evaluating the mechanobiology of cardiomyocytes, as well as in drug screening tests.

A Wireless Pressure Sensor Integrated with a Biodegradable Polymer Stent for Biomedical Applications.

This paper describes the fabrication and characterization of a wireless pressure sensor for smart stent applications. The micromachined pressure sensor has an area of 3.13 × 3.16 mm² and is fabricated with a photosensitive SU-8 polymer. The wireless pressure sensor comprises a resonant circuit and can be used without the use of an internal power source. The capacitance variations caused by changes in the intravascular pressure shift the resonance frequency of the sensor. This change can be detected using an external antenna, thus enabling the measurement of the pressure changes inside a tube with a simple external circuit. The wireless pressure sensor is capable of measuring pressure from 0 mmHg to 230 mmHg, with a sensitivity of 0.043 MHz/mmHg. The biocompatibility of the pressure sensor was evaluated using cardiac cells isolated from neonatal rat ventricular myocytes. After inserting a metal stent integrated with the pressure sensor into a cardiovascular vessel of an animal, medical systems such as X-ray were employed to consistently monitor the condition of the blood vessel. No abnormality was found in the animal blood vessel for approximately one month. Furthermore, a biodegradable polymer (polycaprolactone) stent was fabricated with a 3D printer. The polymer stent exhibits better sensitivity degradation of the pressure sensor compared to the metal stent.

Biomechanical Characterization of Cardiomyocyte Using PDMS Pillar with Microgrooves.

This paper describes the surface-patterned polydimethylsiloxane (PDMS) pillar arrays for enhancing cell alignment and contraction force in cardiomyocytes. The PDMS micropillar (µpillar) arrays with microgrooves (µgrooves) were fabricated using a unique micro-mold made using SU-8 double layer processes. The spring constant of the µpillar arrays was experimentally confirmed using atomic force microscopy (AFM). After culturing cardiac cells on the two different types of µpillar arrays, with and without grooves on the top of µpillar, the characteristics of the cardiomyocytes were analyzed using a custom-made image analysis system. The alignment of the cardiomyocytes on the µgrooves of the µpillars was clearly observed using a DAPI staining process. The mechanical force generated by the contraction force of the cardiomyocytes was derived from the displacement of the µpillar arrays. The contraction force of the cardiomyocytes aligned on the µgrooves was 20% higher than that of the µpillar arrays without µgrooves. The experimental results prove that applied geometrical stimulus is an effective method for aligning and improving the contraction force of cardiomyocytes.

Coaxial bioprinting of a stentable and endothelialized human coronary artery-sized in vitro model.

Atherosclerosis accounts for two-thirds of deaths attributed to cardiovascular diseases, which continue to be the leading cause of mortality. Current clinical management strategies for atherosclerosis, such as angioplasty with stenting, face numerous challenges, including restenosis and late thrombosis. Smart stents, integrated with sensors that can monitor cardiovascular health in real-time, are being developed to overcome these limitations. This development necessitates rigorous preclinical trials on either animal models or in vitro models. Despite efforts being made, a suitable human-scale in vitro model compatible with a cardiovascular stent has remained elusive. To address this need, this study utilizes an in-bath bioprinting method to create a human-scale, freestanding in vitro model compatible with cardiovascular stents. Using a coaxial nozzle, a tubular structure of human coronary artery (HCA) size is bioprinted with a collagen-based bioink, ensuring good biocompatibility and suitable rheological properties for printing. We precisely replicated the dimensions of the HCA, including its internal diameter and wall thickness, and simulated the vascular barrier functionality. To simplify post-processing, a pumpless perfusion bioreactor is fabricated to culture a HCA-sized model, eliminating the need for a peristaltic pump and enabling scalability for high-throughput production. This model is expected to accelerate stent development in the future.

PANI and graphene/PANI nanocomposite films--comparative toluene gas sensing behavior.

The present work discusses and compares the toluene sensing behavior of polyaniline (PANI) and graphene/polyaniline nanocomposite (C-PANI) films. The graphene-PANI ratio in the nanocomposite polymer film is optimized at 1:2. For this, N-methyl-2-pyrrolidone (NMP) solvent is used to prepare PANI-NMP solution as well as graphene-PANI-NMP solution. The films are later annealed at 230 °C, characterized using scanning electron microscopy (SEM) as well Fourier transform infrared spectroscopy (FTIR) and tested for their sensing behavior towards toluene. The sensing behaviors of the films are analyzed at different temperatures (30, 50 and 100 °C) for 100 ppm toluene in air. The nanocomposite C-PANI films have exhibited better overall toluene sensing behavior in terms of sensor response, response and recovery time as well as repeatability. Although the sensor response of PANI (12.6 at 30 °C, 38.4 at 100 °C) is comparatively higher than that of C-PANI (8.4 at 30 °C, 35.5 at 100 °C), response and recovery time of PANI and C-PANI varies with operating temperature. C-PANI at 50 °C seems to have better toluene sensing behavior in terms of response time and recovery time.

Correction: Enhanced cardiomyocyte structural and functional anisotropy through synergetic combination of topographical, conductive, and mechanical stimulation.

Correction for 'Enhanced cardiomyocyte structural and functional anisotropy through synergetic combination of topographical, conductive, and mechanical stimulation' by Jongyun Kim et al., Lab Chip, 2023, 23, 4540-4551, https://doi.org/10.1039/D3LC00451A.

Enhanced cardiomyocyte structural and functional anisotropy through synergetic combination of topographical, conductive, and mechanical stimulation.

Drug-induced cardiotoxicity, a significant concern in the pharmaceutical industry, often results in the withdrawal of drugs from the market. The main cause of drug-induced cardiotoxicity is the use of immature cardiomyocytes during in vitro drug screening procedures. Over time, several methods such as topographical, conductive, and mechanical stimulation have been proposed to enhance both maturation and contractile properties of these cardiomyocytes. However, the synergistic effects of integrating topographical, conductive, and mechanical stimulation for cardiomyocyte maturation remain underexplored and poorly understood. To address this limitation, herein, we propose a grooved polydimethylsiloxane (PDMS) membrane embedded with silver nanowires (AgNWs-E-PDMS). The proposed AgNWs-E-PDMS membrane enhances the maturation of cardiomyocytes and provides a more accurate evaluation of drug-induced cardiotoxicity. When subjected to 10% tensile stress on the AgNWs-E-PDMS membrane, cardiomyocytes displayed substantial enhancements. Specifically, the contraction force, sarcomere length, and connexin-43 (Cx43) expression are increased by 2.0-, 1.5-, and 2.4-times, respectively, compared to the control state. The practical feasibility of the proposed device as a drug screening platform is demonstrated by assessing the adverse effects of lidocaine on cardiomyocytes. The contraction force and beat rate of lidocaine treated cardiomyocytes cultured on the AgNWs-E-PDMS membrane under mechanical stimulation decreased to 0.9 and 0.64 times their initial values respectively, compared to 0.6 and 0.51 times in the control state. These less pronounced changes in the contraction force and beat rate signify the superior drug response in the cardiomyocytes, a result of their enhanced maturation and growth on the AgNWs-E-PDMS membrane combined with mechanical stimulation.

Implantable Self-Reporting Stents for Detecting In-Stent Restenosis and Cardiac Functional Dynamics.

Despite the increasing number of stents implanted each year worldwide, patients remain at high risk for developing in-stent restenosis. Various self-reporting stents have been developed to address this challenge, but their practical utility has been limited by low sensitivity and limited data collection. Herein, we propose a next-generation self-reporting stent that can monitor blood pressure and blood flow inside the blood arteries. This proposed self-reporting stent utilizes a larger inductor coil encapsulated on the entire surface of the stent strut, resulting in a 2-fold increase in the sensing resolution and coupling distance between the sensor and external antenna. The dual-pressure sensors enable the detection of blood flow in situ. The feasibility of the proposed self-reporting stent is successfully demonstrated through in vivo analysis in rats, verifying its biocompatibility and multifunctional utilities. This multifunctional self-reporting stent has the potential to greatly improve cardiovascular care by providing real-time monitoring and unprecedented insight into the functional dynamics of the heart.

Rapid remodeling observed at mid-term in-vivo study of a smart reinforced acellular vascular graft implanted on a rat model.

BACKGROUND: The poor performance of conventional techniques used in cardiovascular disease patients requiring hemodialysis or arterial bypass grafting has prompted tissue engineers to search for clinically appropriate off-the-shelf vascular grafts. Most patients with cardiovascular disease lack suitable autologous tissue because of age or previous surgery. Commercially available vascular grafts with diameters of < 5 mm often fail because of thrombosis and intimal hyperplasia. RESULT: Here, we tested tubular biodegradable poly-e-caprolactone/polydioxanone (PCL/PDO) electrospun vascular grafts in a rat model of aortic interposition for up to 12 weeks. The grafts demonstrated excellent patency (100%) confirmed by Doppler Ultrasound, resisted aneurysmal dilation and intimal hyperplasia, and yielded neoarteries largely free of foreign materials. At 12 weeks, the grafts resembled native arteries with confluent endothelium, synchronous pulsation, a contractile smooth muscle layer, and co-expression of various extracellular matrix components (elastin, collagen, and glycosaminoglycan). CONCLUSIONS: The structural and functional properties comparable to native vessels observed in the neoartery indicate their potential application as an alternative for the replacement of damaged small-diameter grafts. This synthetic off-the-shelf device may be suitable for patients without autologous vessels. However, for clinical application of these grafts, long-term studies (> 1.5 years) in large animals with a vasculature similar to humans are needed.

Chemo-mechanical joint detection with both dynamic and static microcantilevers for interhomologue molecular identification.

The study presents a novel chemo-mechanical joint-sensing method to distinguish a certain molecule from its homologous chemicals, using both a resonant cantilever for gravimetric sensing and a static cantilever for surface-stress sensing. Homologous amines of trimethylamine (TMA, Me(3)N), dimethylamine (DMA, Me(2)NH), and monomethylamine (MMA, MeNH(2)) are herein used as model objects for investigation. The molecular identification is based on experimental characterizations on both molecule adsorbing capability (by the resonant cantilever) and intermolecular lateral interaction (by the static cantilever). The intensities of the two sets of sensing signals are expected to be in opposite sequence with each other, due to the complementary relationship among the interhomologue molecule structures, i.e., a molecule containing a greater number of methyl substituents must possess a fewer number of nonsubstituted hydrogens. On the basis of the proposed idea, ppm-level vapors of the three amines are sequentially detected by a resonant microcantilever to characterize the molecular adsorption speed and another static cantilever to characterize the intermolecular lateral attraction induced surface stress. From the experiment, a pair of opposite sequence in sensing-signal amplitude has indeed been obtained that verifies the proposed joint-sensing method. In addition, the two sensing signals both show a linear relationship with chemical concentration (at low-concentration range). Further comparison between the two sensing results can help to build a model to identify the molecule among a series of its homologous chemicals by eliminating the influence from concentration. Since a complementary relationship among homologous molecule structures widely exists, the dual-sensing method is promising in on-the-spot rapid molecular identification among homologous chemicals.

Stress-assisted gold micro-wrinkles on a polymer cantilever for cardiac tissue engineering.

Surface topography of devices is crucial for cardiac tissue engineering. In this study, we fabricated a unique cantilever-based device, whose surface was structured with stress-assisted micro-wrinkles. The Au micro-wrinkle patterns on the cantilever surface helped the cardiomyocytes to grow similarly to those in the native cardiac tissues by aligning them and providing them a conductive surface, thereby enhancing the contractile properties of the cells. The patterned Au surface also enhanced the electrical conductivity during cell-to-cell interactions. Additionally, the expression levels of proteins related to intracellular adhesion and contraction significantly increased in the polymer cantilevers with metallic wrinkle patterns. The roles of the polymer cantilever in improving the electrical conductivity and force-sensing properties were confirmed. Thereafter, the cantilever's response to cardiotoxicity was evaluated by introducing drugs known to induce toxicity to cardiomyocytes. The proposed cantilever is a versatile device that may be used to screen drug-induced cardiotoxicity during drug development.

Crack-Based Sensor by Using the UV Curable Polyurethane-Acrylate Coated Film with V-Groove Arrays.

Over the years, several bare metal and crack-based strain sensors have been proposed for various fields of science and technology. However, due to their low gauge factor, metal-based strain sensors have limited practical applications. The crack-based strain sensor, on the other hand, demonstrated excellent sensitivity and a high gauge factor. However, the crack-based strain sensor exhibited non-linear behavior at low strains, severely limiting its real-time applications. Generally, the crack-based strain sensors are fabricated by generating cracks by bending a polymer film on which a metal layer has been deposited with a constant curvature. However, the random formation of cracks produces nonlinear behavior in the crack sensors. To overcome the limitations of the current state of the art, we propose a V-groove-based metal strain sensor for human motion monitoring and Morse code generation. The V-groove crack-based strain sensor is fabricated on polyurethane acrylate (PUA) using the modified photolithography technique. During the procedure, a V-groove pattern formed on the surface of the sensor, and a uniform crack formed over the entire surface by concentrating stress along the groove. To improve the sensitivity and selectivity of the sensor, we generated the cracks in a controlled direction. The proposed strain sensor exhibited high sensitivity and excellent fidelity compared to the other reported metal strain sensors. The gauge factor of the proposed V-groove-induced crack sensor is 10-fold higher than the gauge factor of the reported metal strain sensors. In addition, the fabricated V-groove-based strain sensor exhibited rapid response and recovery times. The practical feasibility of the proposed V-groove-induced crack-based strain sensor is demonstrated through human motion monitoring and the generation of Morse code. The proposed V-groove crack sensor can detect multiple motions in a variety of human activities and is anticipated to be utilized in several applications due to its high durability and reproducibility.

The effect of topographical and mechanical stimulation on the structural and functional anisotropy of cardiomyocytes grown on a circular PDMS diaphragm.

Due to their immature morphology and functional immaturity, cardiomyocytes have limited use as an in vitro disease model of the native heart. Mechanical stimulation induces structural growth in cardiomyocytes in vitro by addressing the electrical-mechanical interactions between the tissues. However, current in vitro models are restricted in their capacity to replicate the milieu observed in natural myocardium. Herein, we proposed a Galinstan strain sensor integrated nanogrooved circular PDMS diaphragm to mimic the native cardiac tissues. The impact of combined topographical and mechanical stimulation on cultured cardiomyocytes at various strain areas on a circular PDMS diaphragm is studied in detail. An inverted microscope is used to image live cells and video acquisition to study the contractility of cultured cardiomyocytes. The structural changes of the cultured cardiomyocytes are investigated by its sarcomere length and connexin-43 (Cx43) expression using immunocytochemistry analysis. Cyclic strain is found to promote structural development in cultured cardiomyocytes, and diaphragms with nano-groove patterns displayed increased contractile activity and gene expression (sarcomere length ∼1.97 ± 0.03 µm and normalized Cx43-1.57) as compared to flat diaphragms (sarcomere length ∼1.82 ± 0.02 µm and normalized Cx43-1.32). The nanogrooved circular diaphragm exhibited distinct stretching mechanisms at various places, with the equibi-axial stretching regions providing the optimal structural growth and formation of natural myocardium at the diaphragm's center. Cardiomyocytes that are more mature have the potential to produce a more realistic in vitro cardiac model for disease modeling and medication development.

MEA-integrated cantilever platform for comparison of real-time change in electrophysiology and contractility of cardiomyocytes to drugs.

Drug-induced cardiotoxicity is a potentially severe side effect that can alter the contractility and electrophysiology of the cardiomyocytes. Cardiotoxicity is generally assessed through animal models using conventional drug screening platforms. Despite significant developments in drug screening platforms, the difficulty in measuring electrophysiology and contractile profile together affects the investigation of cardiotoxicity in potential drugs. Some drugs can prove to be more toxic to contractility than electrophysiology, which demands the need for a reliable, dual, and simultaneous drug screening platform. Herein, we propose the microelectrode array integrated SU-8 cantilever for dual and simultaneous measurement of electrophysiology and contractility of cardiomyocytes. The SU-8 cantilever is integrated with microelectrode array (C-MEA) using conventional photolithographic techniques. Drug tests are conducted to verify the feasibility of the C-MEA platform using three cardiovascular drugs. Clinically recognized drugs, quinidine and verapamil, are used to activate both the hERG channel and the contractile characteristics of cardiomyocytes. The effect of ion channel blockers on the field potential duration (FPD) of the cardiomyocytes is compared with several contractility-based parameters. The contraction-relaxation duration (CRD) profile is relatively close to that of FPD in tested drugs (half-maximal (IC50) toxicities are 1.093 µM (FPD) and 1.924 µM (CRD) for quinidine and 166.2 nM (FPD) and 459.4 nM (CRD) for verapamil). Blebbistatin, a known myosin II inhibitor, primarily affects the contractile profile of cardiomyocytes but not their field potential, with no evident correlation between contractility and field potential profiles. The proposed cantilever-based mechano-electrophysiology measurements platform provides a promising and accurate means to assess cardiotoxicity.

A Comparative Study of an Anti-Thrombotic Small-Diameter Vascular Graft with Commercially Available e-PTFE Graft in a Porcine Carotid Model.

BACKGROUND: We have designed a reinforced drug-loaded vascular graft composed of polycaprolactone (PCL) and polydioxanone (PDO) via a combination of electrospinning/3D printing approaches. To evaluate its potential for clinical application, we compared the in vivo blood compatibility and performance of PCL/PDO + 10%DY grafts doped with an antithrombotic drug (dipyridamole) with a commercial expanded polytetrafluoroethylene (e-PTFE) graft in a porcine model. METHODS: A total of 10 pigs (weight: 25-35 kg) were used in this study. We made a new 5-mm graft with PCL/PDO composite nanofiber via the electrospinning technique. We simultaneously implanted a commercially available e-PTFE graft (n = 5) and our PCL/PDO + 10%DY graft (n = 5) into the carotid arteries of the pigs. No anticoagulant/antiplatelet agent was administered during the follow-up period, and ultrasonography was performed weekly to confirm the patency of the two grafts in vivo. Four weeks later, we explanted and compared the performance of the two grafts by histological analysis and scanning electron microscopy (SEM). RESULTS: No complications, such as sweating on the graft or significant bleeding from the needle hole site, were seen in the PCL/PDO + 10%DY graft immediately after implantation. Serial ultrasonographic examination and immunohistochemical analysis demonstrated that PCL/PDO + 10%DY grafts showed normal physiological blood flow and minimal lumen reduction, and pulsed synchronously with the native artery at 4 weeks after implantation. However, all e-PTFE grafts occluded within the study period. The luminal surface of the PCL/PDO + 10%DY graft in the transitional zone was fully covered with endothelial cells as observed by SEM. CONCLUSION: The PCL/PDO + 10%DY graft was well tolerated, and no adverse tissue reaction was observed in porcine carotid models during the short-term follow-up. Colonization of the graft by host endothelial and smooth muscle cells coupled with substantial extracellular matrix production marked the regenerative capability. Thus, this material may be an ideal substitute for vascular reconstruction and bypass surgeries. Long-term observations will be necessary to determine the anti-thrombotic and remodeling potential of this device.