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.

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.

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.

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.

Primary headaches in children and adolescents - experiences at a single headache center in Korea.

BACKGROUND: Headache is a common complaint in children and adolescents. Recently, an increased prevalence of headache in children and adolescents has been reported. METHODS: We retrospectively reviewed the medical records of children and adolescents attending the Headache Clinic of Daejeon St. Mary's Hospital during the period from January 2005 through December 2016. RESULTS: The study population consisted of 2466 children, aged between 3 and 18 years (mean age: 10.9). Our study showed an increase in the number of patients visiting the hospital with headaches during the past decade. Compared with 2005, the number of patients with headache increased three-fold in 2016. Interestingly, the proportion of boys, preschool children, and other primary headaches revealed a steady and statistically significant increase. CONCLUSION: Due to a steady increase in pediatric headaches, the earlier the problem is recognized and properly diagnosed and a treatment plan is established, the greater the likelihood of a better lifelong outcome. Studies are needed to estimate recent trend in prevalence and to identify the demographic and socioeconomic factors predicting the occurrence of headache.

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.

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.

A Huge Malignant Gastric Gastrointestinal Stromal Tumor in a Young Patient.

Malignant gastrointestinal stromal tumors (GISTs) are rare neoplasms originating from the gastrointestinal tract that rarely occur in patients below 40 years of age. To our knowledge, there have been no previous reports of satellite and metastatic nodules in GIST. We present a case of a young patient with a huge malignant gastric GIST accompanied by spontaneous bleeding and satellite and metastatic nodules, successfully treated surgically, without preoperative chemotherapy administration. A 28-year-old man was admitted to Haeundae Paik Hospital with melena. A huge bulging gastric mass with ulceration and bleeding was observed on endoscopy. A subepithelial lesion on the stomach body, abutting the pancreatic body and tail, with regional lymph node enlargement was confirmed by EUS and CT. Radical total gastrectomy was performed, the invasion surrounding the pancreatic tail and spleen were surgically dissected, and enlarged lymph nodes around the celiac trunk and the common hepatic artery were removed. The pathology results showed a malignant GIST with two satellite nodules and a metastatic tumor nodule at the left paracardial lymph node site. After complete resection of the malignant GIST, adjuvant chemotherapy with imatinib was initiated. Follow-up CT and endoscopy performed 6 months after surgery confirmed no recurrence of the disease.

Gastric Malignant Peripheral Nerve Sheath Tumor in Type 1 Neurofibromatosis.

Gastric malignant peripheral nerve sheath tumors (MPNSTs) are extremely rare spindle cell sarcomas that arise within the peripheral nerves of the gastrointestinal tract. MPNST can present as a mass that may or may not be accompanied by obstruction or bleeding. Type 1 neurofibromatosis (NF) is an autosomal dominant genetic disorder with an incidence of 1 in 2,500-3,000. Plexiform neurofibromas in Type 1 NF can undergo a malignant transformation to MPNSTs. Approximately half of the incidence of MPNST is associated with the NF-1 gene. MPNST behaves aggressively, and radical excisional surgery is important for treatment. Recurrence and metastasis are significant, even after a radical excision. Despite multidisciplinary treatment, the five-year survival rate is only 30-50%. This paper reports the case of a 47-year-old man with Type 1 NF who presented with hemorrhage of a gastric subepithelial lesion. He underwent surgery under the suspicion of a gastrointestinal stromal tumor, but it was diagnosed as MPNST after confirming the histopathological appearance and immunohistochemical profiles. In addition, the large mass invaded the spleen and diaphragm. Radical surgery was performed, and additional chemotherapy was administered. This paper reports the experience of a patient with NF 1 with advanced MPNST discovered due to a subepithelial lesion.

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.

64 PI/PDMS hybrid cantilever arrays with an integrated strain sensor for a high-throughput drug toxicity screening application.

Herein, we propose a novel biosensing platform involving an array of 64 hybrid cantilevers and integrated strain sensors to measure the real-time contractility of the drug-treated cardiomyocytes (CMs). The strain sensor is integrated on the polyimide (PI) cantilever. To improve the strain sensor reliability and construct the engineered cardiac tissue, the nanogroove-patterned polydimethylsiloxane (PDMS) encapsulation layer is bonded on the PI cantilever. The preliminary sensing characteristics demonstrate the superior structural integrity, robustness, enhanced sensitivity, and repeatability of the proposed devices. The long-term durability and biocompatibility of the PI/PDMS hybrid cantilever is verified by evaluating the cell viability and contractility. We also validate the proposed biosensing platform for cardiotoxicity measurement by applying it to two specific cardiovascular drugs: quinidine and verapamil. In response to quinidine and verapamil, the engineered CMs exhibited negative inotropic and chronotropic effects. The fabricated cantilever device successfully detected the quinidine-induced adverse effects in CMs such as early after depolarization (EADs) and Torsade de points (TdP) in real-time. The array of hybrid cantilevers with integrated strain sensors has the potential to satisfy the need for innovative analytic platforms owing to its high throughput and simplified data analysis.

On-stage bioreactor platform integrated with nano-patterned and gold-coated PDMS diaphragm for live cell stimulation and imaging.

Over the years, several in-vitro biosensing platforms have been developed for enhancing the maturation of the cultured cells. However, most of the proposed platforms met with limited success due to its inability for live-cell imaging, complicated fabrication, and not being advantageous from an economic perspective due to a higher price. To overcome the drawbacks of the current state-of-the-art, herein, we developed a next-generation stage-top incubator (STI) incorporated with nano grooves patterned PDMS diaphragm (NGPPD). The proposed device consists of a miniatured STI, the NGPPD functional well plates, and a mechanical stimulator. A thin layer of gold (Au) is deposited on the NGPPD to enhanced myogenic differentiation, cell maturation, and cell-cell interactions. The nano grooves are integrated on the PDMS surface to align the cardiomyocytes in the grooved direction during the culture period. The cardiomyocytes cultivated on the Au-deposited NGPPD are stimulated topographically and mechanically during the cultivation period. The enhanced cardiomyocytes maturation cultured on the Au-deposited NGPPD is experimentally demonstrated using immunofluorescence staining and PCR analysis.

Micro-patterned SU-8 cantilever integrated with metal electrode for enhanced electromechanical stimulation of cardiac cells.

Over the past few years, cardiac tissue engineering has undergone tremendous progress. Various in vitro methods have been developed to improve the accuracy in the result of drug-induced cardiac toxicity screening. Herein, we propose a novel SU-8 cantilever integrated with an electromechanical-stimulator to enhance the maturation of cultured cardiac cells. The simultaneous electromechanical stimulation significantly enhances the contraction force of the cardiomyocytes, thereby increasing cantilever displacement. Fluorescence microscopy analysis was performed to confirm the improved maturation of the cardiomyocytes. After the initial experiments, the contractile behaviors of the cultured cardiomyocytes were investigated by measuring the mechanical deformation of the SU-8 cantilever. Finally, the proposed electromechanical-stimulator-integrated SU-8 cantilever was used to evaluate the adverse effects of different cardiac vascular drugs, i.e., verapamil, lidocaine, and isoproterenol, on the cultured cardiomyocytes. The physiology of the cardiac-drug-treated cardiomyocytes was examined with and without electrical stimulation of the cardiomyocytes. The experimental results indicate that the proposed cantilever platform can be used as a predictive assay system for preliminary cardiac drug toxicity screening applications.

Highly durable crack sensor integrated with silicone rubber cantilever for measuring cardiac contractility.

To date, numerous biosensing platforms have been developed for assessing drug-induced cardiac toxicity by measuring the change in contractile force of cardiomyocytes. However, these low sensitivity, low-throughput, and time-consuming processes are severely limited in their real-time applications. Here, we propose a cantilever device integrated with a polydimethylsiloxane (PDMS)-encapsulated crack sensor to measure cardiac contractility. The crack sensor is chemically bonded to a PDMS thin layer that allows it to be operated very stably in culture media. The reliability of the proposed crack sensor has been improved dramatically compared to no encapsulation layer. The highly sensitive crack sensor continuously measures the cardiac contractility without changing its gauge factor for up to 26 days (>5 million heartbeats), while changes in contractile force induced by drugs are monitored using the crack sensor-integrated cantilever. Finally, experimental results are compared with those obtained via conventional optical methods to verify the feasibility of building a contraction-based drug-toxicity testing system.

Characteristics of and risk factors for colorectal neoplasms in young adults in a screening population.

AIM: To investigate prevalence and risk factors for colorectal neoplasms in adults aged < 50 years, for whom screening is not recommended. METHODS: This cross-sectional study compared prevalence and characteristics of colorectal and advanced adenomas in patients aged < 50 years who underwent colonoscopy screening with subjects aged ≥ 50 years. To evaluate risk factors for colorectal and advanced adenoma in young adults, we used multivariable logistic regression models. Colorectal neoplasm characteristics were evaluated and compared with those in older patients. RESULTS: Among 2819 patients included, prevalences of colorectal adenoma and advanced adenoma were 19.7% and 1.5%, respectively. As patient age increased, so did the prevalence of colorectal neoplasm. However, prevalence of advanced adenoma did not differ between age-groups 45-49 years and ≥ 50 years (OR = 0.43, 95%CI: 0.17-1.07, P = 0.070). In younger age-group (< 50 years), colorectal adenoma was significantly associated with older age, waist circumference (OR = 1.72, 95%CI: 1.15-2.55, P = 0.008), and current smoking (OR = 1.60, 95%CI: 1.07-2.41, P = 0.023). Alcohol consumption was an independent risk factor for colorectal advanced adenoma (OR = 3.69, 95%CI: 1.08-12.54, P = 0.037). Multiple neoplasms and large neoplasms (≥ 1 cm) were more prevalent in subjects ≥ 50 years. CONCLUSION: Current screening strategies for colorectal cancer may need to be amended to account for patient age, especially in young subjects with abdominal obesity, current smoking and alcohol consumption.