Smartphone-based iontophoresis transdermal drug delivery system for cancer treatment.

Cancer is a leading cause of the death worldwide. However, the conventional cancer therapy still suffers from several limitations, such as systemic side effects, poor efficacy, and patient compliance due to limited accessibility to the tumor site. To address these issues, the localized drug delivery system has emerged as a promising approach. In this study, we developed an iontophoresis-based transdermal drug delivery system (TDDS) controlled by a smartphone application for cancer treatment. Iontophoresis, a low-intensity electric current-based TDDS, enhances drug permeation across the skin to provide potential for localized drug delivery and minimize systemic side effects. The fundamental mechanism of our system was modeled using finite element analysis and its performance was corroborated through the flow-through skin permeation tests using a plastic-based microfluidic chip. The results of in vitro cell experiments and skin deposition tests successfully demonstrated that our smartphone-controlled iontophoresis system significantly enhanced the drug permeation for cancer treatment. Therefore, this hand-held smartphone-based iontophoresis TDDS could be a powerful tool for self-administrated anticancer drug delivery applications.

Droplet digital recombinase polymerase amplification for multiplexed detection of human coronavirus.

Since the outbreak of coronavirus 2019 (COVID-19), detection technologies have been attracting a great deal of attention in molecular diagnosis applications. In particular, the droplet digital PCR (ddPCR) has become a promising tool as it offers absolute quantification of target nucleic acids with high specificity and sensitivity. In recent years, the combination of the isothermal amplification strategies has made ddPCR a popular method for on-site testing by enabling amplification at a constant temperature. However, the current isothermal ddPCR assays are still challenging due to inherent non-specific amplification. In this paper, we present a multiplexed droplet digital recombinase polymerase amplification (MddRPA) with precise initiation of the reaction. First, the reaction temperature and dynamic range of reverse transcription (RT) and RPA were characterized by real-time monitoring of fluorescence intensities. Using a droplet-based microfluidic chip, the master mix and the initiator were fractionated and rapidly mixed within well-confined droplets. Due to the high heat transfer and mass transfer of the droplets, the precise initiation of the amplification was enabled and the entire assay could be conducted within 30 min. The concentrations of target RNA in the range from 5 copies per µL to 2500 copies per µL could be detected with high linearity (R2 > 0.999). Furthermore, the multiplexed detection of three types of human coronaviruses was successfully demonstrated with high specificity (>96%). Finally, we compared the performance of the assay with a commercial RT-qPCR system using COVID-19 clinical samples. The MddRPA assay showed a 100% concordance with the RT-qPCR results, indicating its reliability and accuracy in detecting SARS-CoV-2 nucleic acids in clinical samples. Therefore, our MddRPA assay with rapid detection, precise quantification, and multiplexing capability would be an interesting method for molecular diagnosis of viral infections.

Deep Learning-Assisted Droplet Digital PCR for Quantitative Detection of Human Coronavirus.

Since coronavirus disease 2019 (COVID-19) pandemic rapidly spread worldwide, there is an urgent demand for accurate and suitable nucleic acid detection technology. Although the conventional threshold-based algorithms have been used for processing images of droplet digital polymerase chain reaction (ddPCR), there are still challenges from noise and irregular size of droplets. Here, we present a combined method of the mask region convolutional neural network (Mask R-CNN)-based image detection algorithm and Gaussian mixture model (GMM)-based thresholding algorithm. This novel approach significantly reduces false detection rate and achieves highly accurate prediction model in a ddPCR image processing. We demonstrated that how deep learning improved the overall performance in a ddPCR image processing. Therefore, our study could be a promising method in nucleic acid detection technology.

Development of an IoT-integrated multiplexed digital PCR system for quantitative detection of infectious diseases.

For rapid detection of the COVID-19 infection, the digital polymerase chain reaction (dPCR) with higher sensitivity and specificity has been presented as a promising method of point-of-care testing (POCT). Unlike the conventional real-time PCR (qPCR), the dPCR system allows absolute quantification of the target DNA without a calibration curve. Although a number of dPCR systems have previously been reported, most of these previous assays lack multiplexing capabilities. As different variants of COVID-19 have rapidly emerged, there is an urgent need for highly specific multiplexed detection systems. Additionally, the advances in the Internet of Things (IoT) technology have enabled the onsite detection of infectious diseases. Here, we present an IoT-integrated multiplexed dPCR (IM-dPCR) system involving sample compartmentalization, DNA amplification, fluorescence imaging, and quantitative analysis. This IM-dPCR system comprises three modules: a plasmonic heating-based thermal cycler, a multi-color fluorescence imaging set-up, and a firmware control module. Combined with a custom-developed smartphone application built on an IoT platform, the IM-dPCR system enabled automatic processing, data collection, and cloud storage. Using a self-priming microfluidic chip, 9 RNA groups (e.g., H1N1, H3N2, IFZ B, DENV2, DENV3, DENV4, OC43, 229E, and NL63) associated with three infectious diseases (e.g., influenza, dengue, and human coronaviruses) were analyzed with higher linearity (>98%) and sensitivity (1 copy per µL). The IM-dPCR system exhibited comparable analytical accuracy to commercial qPCR platforms. Therefore, this IM-dPCR system plays a crucial role in the onsite detection of infectious diseases.

3D Multicellular Tumor Spheroids in a Microfluidic Droplet System for Investigation of Drug Resistance.

A three-dimensional (3D) tumor spheroid model plays a critical role in mimicking tumor microenvironments in vivo. However, the conventional culture methods lack the ability to manipulate the 3D tumor spheroids in a homogeneous manner. To address this limitation, we developed a microfluidic-based droplet system for drug screening applications. We used a tree-shaped gradient generator to control the cell density and encapsulate the cells within uniform-sized droplets to generate a 3D gradient-sized tumor spheroid. Using this microfluidic-based droplet system, we demonstrated the high-throughput generation of uniform 3D tumor spheroids containing various cellular ratios for the analysis of the anti-cancer drug cytotoxicity. Consequently, this microfluidic-based gradient droplet generator could be a potentially powerful tool for anti-cancer drug screening applications.

Conductive Silver/Carbon Fiber Films for Rapid Detection of Human Coronavirus.

Polymerase chain reaction has gained attention since the outbreak of novel coronavirus in 2019. Due to its high specificity and capability for early detection, it is considered a standard method for the diagnosis of infectious diseases. However, the conventional thermocyclers used for nucleic acid amplification are not suitable for point-of-care testing applications, as they require expensive instruments, high-power consumption, and a long turnaround time. To suppress the widespread of the pandemic, there is an urgent need for the development of a rapid, inexpensive, and portable thermal cycler. Therefore, in this paper, we present a conductive silver/carbon fiber film-based thermal cycler with low power consumption (<5 W), efficient heating (~4.5 °C/s), low cost (

Early medial reconstruction combined with severely injured medial collateral ligaments can decrease residual medial laxity in anterior cruciate ligament reconstruction.

INTRODUCTION: This study aimed to describe an anatomic medial knee reconstruction technique for combined anterior cruciate ligament (ACL) and grade III medial collateral ligament (MCL) injuries and to assess knee function and stability restoration in patients who underwent primary MCL reconstruction compared with primary repair. METHODS: A total of 105 patients who had undergone anatomic ACL reconstruction between 2008 and 2017 were enrolled in this retrospective study and divided into two groups according to concomitant MCL ruptures. Group A included patients with isolated ACL ruptures without MCL injuries. Group B included patients with both ACL and MCL injuries, and it was subdivided into three groups according to the severity of the MCL injury and treatment modality: B-1, grade I or II MCL injury treated conservatively; B-2: grade III MCL injury treated by primary MCL repair; and B-3: grade III MCL injury treated by primary reconstruction. Knee stability was measured via Telos valgus radiography at 6-month and 2-year postoperative. The Lysholm score, Tegner activity level, Likert scales (satisfaction), and return to previous sports were evaluated at 2-year postoperative. RESULTS: At 6-month postoperative, there was no significant difference in medial laxity between the B-2 and B-3 groups. However, at 2-year postoperative, medial laxity were significantly higher both at 30° of flexion (5.2° versus 2.2°, p = 0.020) and at full extension (3.4° versus 1.1°, p < 0.001) in patients in B-2 group compared to those in B-3 group. There were no statistically significant differences between the two groups with respect to Lysholm scores, Tegner activity levels, Likert scales (satisfaction), and returning to previous sports at the 2-year follow-up. CONCLUSION: Primary medial reconstruction combined with severely injured MCL in ACL reconstruction may decrease residual medial laxity more than primary repair. LEVEL OF EVIDENCE: Retrospective observational study, IV.

Effect of biochemical and biomechanical factors on vascularization of kidney organoid-on-a-chip.

Kidney organoids derived from the human pluripotent stem cells (hPSCs) recapitulating human kidney are the attractive tool for kidney regeneration, disease modeling, and drug screening. However, the kidney organoids cultured by static conditions have the limited vascular networks and immature nephron-like structures unlike human kidney. Here, we developed a kidney organoid-on-a-chip system providing fluidic flow mimicking shear stress with optimized extracellular matrix (ECM) conditions. We demonstrated that the kidney organoids cultured in our microfluidic system showed more matured podocytes and vascular structures as compared to the static culture condition. Additionally, the kidney organoids cultured in microfluidic systems showed higher sensitivity to nephrotoxic drugs as compared with those cultured in static conditions. We also demonstrated that the physiological flow played an important role in maintaining a number of physiological functions of kidney organoids. Therefore, our kidney organoid-on-a-chip system could provide an organoid culture platform for in vitro vascularization in formation of functional three-dimensional (3D) tissues.

CuS/rGO-PEG Nanocomposites for Photothermal Bonding of PMMA-Based Plastic Lab-on-a-Chip.

We developed copper sulfide (CuS)/reduced graphene oxide (rGO)-poly (ethylene glycol) (PEG) nanocomposites for photothermal bonding of a polymethyl methacrylate (PMMA)-based plastic lab-on-a-chip. The noncontact photothermal bonding of PMMA-based plastic labs-on-chip plays an important role in improving the stability and adhesion at a high-temperature as well as minimizing the solution leakage from microchannels when connecting two microfluidic devices. The CuS/rGO-PEG nanocomposites were used to bond a PMMA-based plastic lab-on-a-chip in a short time with a high photothermal effect by a near-infrared (NIR) laser irradiation. After the thermal bonding process, a gap was not generated in the PMMA-based plastic lab-on-a-chip due to the low viscosity and density of the CuS/rGO-PEG nanocomposites. We also evaluated the physical and mechanical properties after the thermal bonding process, showing that there was no solution leakage in PMMA-based plastic lab-on-a-chip during polymerase chain reaction (PCR) thermal cycles. Therefore, the CuS/rGO-PEG nanocomposite could be a potentially useful nanomaterial for non-contact photothermal bonding between the interfaces of plastic module lab-on-a-chip.

Plasmonic heating-based portable digital PCR system.

A miniaturized polymerase chain reaction (PCR) system is not only important for medical applications in remote areas of developing countries, but also important for testing at ports of entry during global epidemics, such as the current outbreak of the coronavirus. Although there is a large number of PCR sensor systems available for this purpose, there is still a lack of portable digital PCR (dPCR) heating systems. Here, we first demonstrated a portable plasmonic heating-based dPCR system. The device has total dimensions of 9.7 × 5.6 × 4.1 cm and a total power consumption of 4.5 W, allowing for up to 25 dPCR experiments to be conducted on a single charge of a 20 000 mAh external battery. The dPCR system has a maximum heating rate of 10.7 °C s-1 and maximum cooling rate of 8 °C s-1. Target DNA concentrations in the range from 101 ± 1.4 copies per µL to 260 000 ± 20 000 copies per µL could be detected using a poly(dimethylsiloxane) (PDMS) microwell membrane with 22 080 well arrays (20 µm diameter). Furthermore, the heating system was demonstrated using a mass producible poly(methyl methacrylate) PMMA microwell array with 8100 microwell arrays (80 µm diameter). The PMMA microwell array could detect a concentration from 12 ± 0.7 copies per µL to 25 889 ± 737 copies per µL.

Generation of tumor spheroids using a droplet-based microfluidic device for photothermal therapy.

Despite their simplicity, monolayer cell cultures are not able to accurately predict drug behavior in vivo due to their inability to accurately mimic cell-cell and cell-matrix interactions. In contrast, cell spheroids are able to reproduce these interactions and thus would be a viable tool for testing drug behavior. However, the generation of homogenous and reproducible cell spheroids on a large scale is a labor intensive and slow process compared to monolayer cell cultures. Here, we present a droplet-based microfluidic device for the automated, large-scale generation of homogenous cell spheroids in a uniform manner. Using the microfluidic system, the size of the spheroids can be tuned to between 100 and 130 µm with generation frequencies of 70 Hz. We demonstrated the photothermal therapy (PTT) application of brain tumor spheroids generated by the microfluidic device using a reduced graphene oxide-branched polyethyleneimine-polyethylene glycol (rGO-BPEI-PEG) nanocomposite as the PTT agent. Furthermore, we generated uniformly sized neural stem cell (NSC)-derived neurospheres in the droplet-based microfluidic device. We also confirmed that the neurites were regulated by neurotoxins. Therefore, this droplet-based microfluidic device could be a powerful tool for photothermal therapy and drug screening applications.

Three-dimensional measurement and registration accuracy of a third-generation optical tracking system for navigational maxillary orthognathic surgery.

OBJECTIVE: The purpose of this study was to evaluate the accuracy of an optical tracking system during reference point localization, measurement, and registration of skull models for navigational maxillary orthognathic surgery. STUDY DESIGN: Accuracy was first evaluated on the basis of the position recording discrepancy at a static point and at 2 points of fixed lengths. Ten reference points were measured on a skull model at 7 different locations, and their measurements were compared with predicted positions by using 4 registration methods. Finally, positional tracking of reference points for simulated maxillary surgery was performed and compared with laser scan data. RESULTS: The average linear measurement discrepancy was 0.28 mm, and the mean measurement discrepancy with the 5 registered cranial points was 1.53 mm. The average measurement discrepancy after maxillary surgery was 1.91 mm (for impaction) and 1.56 mm (for advancement). The registration discrepancy in jitter and point registration on the y-axis was significantly greater than on the other axes. CONCLUSIONS: The optical tracking system seems clinically acceptable for precise tracking of the maxillary position during navigational orthognathic surgery, notwithstanding the chance of greater measurement error on the y-axis.

Compensatory dentoalveolar supraeruption and occlusal plane cant after botulinum-induced hypotrophy of masticatory closing muscles in juvenile rats.

OBJECTIVE: The purpose of this study was to investigate changes in the dentoalveolus and occlusal plane associated with the hypotrophy of unilateral masticatory muscles following botulinum toxin (BTX) treatment in the juvenile period of rats. DESIGN: We hypothesized that the loss of functional loading of masticatory muscles and occlusal force invites compensatory dentoalveolar supraeruption, accelerating occlusal cant and skeletal asymmetry. In order to confirm this hypothesis, six-week-old male rats (N = 5) were treated with BTX simultaneously at the unilateral masseter, temporalis, and medial pterygoid muscles, with a booster injection after six weeks for the experimental group. The control group (N = 6) had saline injections on both sides at the same sites and on the same schedule. RESULTS: After 12 weeks, masseter and medial pterygoid muscles on the BTX side showed hypotrophic change. The mandibular structure was asymmetrical, with decreased size and lateral tilting. The maxillary and mandibular molars were supraerupted from the Frankfort plane or mandibular inferior border with lateral tilt. They accompanied downward occlusal plane cant resulting from the supraerupted maxillary and mandibular molars on the BTX side. The dentoalveolar structural changes included diminished alveolar bone density, narrow periodontal ligament space, and disorganized distribution of periodontal collagen fiber. CONCLUSIONS: Unilateral hypotrophy of masticatory muscles affected the growth, symmetry, and structure of the skeletal jaws and dentoalveolus. Our hypothesis about the dentoalveolar compensation, that muscular hypotrophy was closely integrated with dentoalveolar supraeruption and an inclined occlusal plane, was confirmed.

Papillary Cannulation Facilitated by Submucosal Saline Injection into an Intradiverticular Papilla.

Endoscopic retrograde cholangiopancreatography (ERCP) of the intradiverticular papilla with its invisible orifice remains challenging. Several techniques have been introduced to evert the papillary opening to facilitate cannulation. A 79-year-old woman with bile duct stones underwent ERCP, which revealed that the papilla was located inside a large diverticulum and tended to rotate inward with a trial of papillary cannulation. Submucosal papillary injection of 3 cc of normal saline was performed at 3 and 9 o'clock. Eversion and fixation of a papilla in the diverticulum with this technique allowed selective cannulation of the biliary tree. Stones were retrieved after endoscopic papillary balloon dilation without complications. She had an uneventful post-procedural course. Our findings suggest that submucosal saline injection technique is safe and effective for selective cannulation and can be recommended when cannulation is very difficult because of an intradiverticular papilla.

Droplet-based synthesis of homogeneous magnetic iron oxide nanoparticles.

Nanoparticles have gained large interest in a number of different fields due to their unique properties. In medical applications, for example, magnetic nanoparticles can be used for targeting, imaging, magnetically induced thermotherapy, or for any combination of the three. However, it is still a challenge to obtain narrowly dispersed, reproducible particles through a typical lab-scale synthesis when researching these materials. Here, we present a droplet capillary reactor that can be used for the synthesis of magnetic iron oxide nanoparticles. Compared to conventional batch synthesis, the particles synthesized in our droplet reactor have a narrower size distribution and a higher reproducibility. Furthermore, we demonstrate how the particle size can be changed from 5.2 ± 0.9 nm to 11.8 ± 1.7 nm by changing the reaction temperature and droplet residence time in the droplet capillary reactor.

Interstitial Nephritis Caused by Anorexia Nervosa in Young Male; A Case Report and Literature Review.

Severe eating disorders characterized by repetitive episodes of purging and vomiting can occasionally trigger acute kidney injury. However, interstitial nephritis induced by episodes of repeated vomiting has rarely been reported, and the pathophysiology of this entity remains unknown. A 26-year-old man was admitted to our hospital because of known hypokalemia. His serum electrolyte profile showed: sodium 133 mEq/L, potassium 2.6 mEq/L, chloride 72 mEq/L, total carbon dioxide 50 mEq/L, blood urea nitrogen/creatinine ratio (BUN/Cr) 21.9/1.98 mg/dL, and magnesium 2.0 mg/dL. Arterial blood gas analysis showed: pH 7.557, partial pressure of carbon dioxide 65.8 mmHg, and bicarbonate 58.5 mEq/L. His urinary potassium concentration was 73.2 mEq/L, and Cr was 111 mg/dL. Renal biopsy revealed acute tubular necrosis and tubulointerstitial nephritis with a few shrunken glomeruli. Repeated psychogenic vomiting may precipitate acute kidney injury and interstitial nephritis secondary to volume depletion and hypokalemia. Serum electrolyte levels and renal function should be carefully monitored in patients diagnosed with eating disorders to prevent tubular ischemia and interstitial nephritis.

Skeletal unit construction of rat mandible based on the masticatory muscle anatomy and double microcomputed tomography.

This study aimed to divide the mandible into skeletal units based on three-dimensional (3D) muscular anatomy with microcomputed tomography (micro-CT) of Sprague-Dawley rat. Five normal rats were micro-CT scanned at 12 weeks of age before and after contrast enhancements for the masticatory muscles. Three-dimensional reconstruction of the mandible was performed from the initial micro-CT images, followed by segmentation of the masticatory muscles using the second enhanced micro-CT data. Bone and muscle models were superimposed based on the teeth and bony structures to evaluate muscular orientation and attachment. The mandible was divided into skeletal units using the bony structures and muscle attachments. The mandibular foramen and mental foramen were adopted as the reference points based on their anatomical and developmental significance. The skeletal units consisted of the condylar, coronoid, angular, body and symphyseal units. Further evaluation of these units in relation to development, growth, and other biology and medicine will be helpful in elucidating their biological identities.

Dual-nozzle microfluidic droplet generator.

The droplet-generating microfluidics has become an important technique for a variety of applications ranging from single cell analysis to nanoparticle synthesis. Although there are a large number of methods for generating and experimenting with droplets on microfluidic devices, the dispensing of droplets from these microfluidic devices is a challenge due to aggregation and merging of droplets at the interface of microfluidic devices. Here, we present a microfluidic dual-nozzle device for the generation and dispensing of uniform-sized droplets. The first nozzle of the microfluidic device is used for the generation of the droplets, while the second nozzle can accelerate the droplets and increase the spacing between them, allowing for facile dispensing of droplets. Computational fluid dynamic simulations were conducted to optimize the design parameters of the microfluidic device.

Three-dimensional growth pattern of the rat mandible revealed by periodic live micro-computed tomography.

OBJECTIVE: We wanted to evaluate the three-dimensional (3D) mandibular growth of Sprague-Dawley rats from 4th to 16th postnatal weeks with periodic and live micro-computed tomographic scanning. DESIGN: Twenty Sprague-Dawley rats were used for micro-CT scanning from 4th to 16th postnatal weeks. After 3D reconstruction of rat mandible, we performed the linear and angular measurements and the superimposition of the 3D models to evaluate the mandibular growth of rat. RESULTS: The results showed that the growth direction of the condylar and coronoid regions was superior primarily and posterior secondarily, while the condyle had minimal lateral growth. Moreover, the angular region was growing mainly toward the posterior and lateral direction, while the body and symphysis maintained small, incremental anterior-posterior growth. CONCLUSIONS: We could evaluate the amount, rate, and direction of growth using the mandibular skeletal unit. Some reference points and measurements were more relevant in properly characterizing 3D growth of the mandible. Their growth rates were the greatest between 4th and 8th postnatal weeks, a period which seems most appropriate for studies of rat mandible growth.

Positional symmetry of porion and external auditory meatus in facial asymmetry.

BACKGROUND: The porion (Po) is used to construct the Frankfort horizontal (FH) plane for cephalometrics, and the external auditory meatus (EAM) is to transfer and mount the dental model with facebow. The classical assumption is that EAM represents Po by the parallel positioning. However, we are sometimes questioning about the possible positional disparity between Po and EAM, when the occlusal cant or facial midline is different from our clinical understandings. The purpose of this study was to evaluate the positional parallelism of Po and EAM in facial asymmetries, and also to investigate their relationship with the maxillary occlusal cant. METHODS: The 67 subjects were classified into three groups. Group I had normal subjects with facial symmetry (1.05 ± 0.52 mm of average chin deviation) with minimal occlusal cant (<1.5 mm). Asymmetry group II-A had no maxillary occlusal cant (average 0.60 ± 0.36), while asymmetry group II-B had occlusal cant (average 3.72 ± 1.47). The distances of bilateral Po, EAM, and mesiobuccal cusp tips of the maxillary first molars (Mx) from the horizontal orbital plane (Orb) and the coronal plane were measured on the three-dimensional computed tomographic images. Their right and left side distance discrepancies were calculated and statistically compared. RESULTS: EAM was located 10.3 mm below and 2.3 mm anterior to Po in group I.  The vertical distances from Po to EAM of both sides were significantly different in group II-B (p = 0.001), while other groups were not. Interside discrepancy of the vertical distances from EAM to Mx in group II-B also showed the significant differences, as compared with those from Po to Mx and from Orb to Mx. CONCLUSIONS: The subjects with facial asymmetry and prominent maxillary occlusal cant tend to have the symmetric position of Po but asymmetric EAM. Some caution or other measures will be helpful for them to be used during the clinical procedures.