Comparison of the performance of the PANArray™ HPV test and DNA chip test for genotyping of human papillomavirus in cervical swabs

송혜정,이정원,김병기,송상용,배덕수,김대식 한국바이오칩학회 2010 BioChip Journal Vol.4 No.3

The aim of this study was to evaluate the recently developed PANArray™ Human papilloma virus (HPV) kit for detection and genotyping of 19 high-risk and 13 low-risk HPV types, and compare it with the commercially available DNA chip kit geno-typing of 24 HPVs. We llected cervical swabs from 741 patients with various stages of invasive cervical carcinoma being treated at the Samsung Medical Center. The overall HPV positivity rate was 73% using PANArray™ HPV and 72.1% with the DNA chip, and no statistically significant differences were found with respect to the cytology grade. Comparing the results of the two chips, concordant results were found in 637/741 samples (85.9%), compatible in 69/741 (9.3%) and discordant in 35/741(4.7%). Type-specific sequencing analysis of all samples revealed a 99.7% confirmation of PANArray™ HPV genotyping results, compared to 91.0% of DNA chip. The PANArray™ HPV test thus proved to be highly sensitive and accurate even when multiple HPV infections were present.

Identification of Potential Allergens of Atractylodes japonica and Addition of Panels for Allergic Diseases

지원아,고현민,권승원,정우상,장형진 한국바이오칩학회 2023 BioChip Journal Vol.17 No.1

The use of herbal medicines is increasing significantly worldwide. However, studies on side effects such as allergic reactions of herbal medicines are lacking. To provide an effective prescription, a diagnostic test to determine whether an allergic reaction is induced in the patient must be preceded. In this study, the purpose of this study is to investigate whether Atractylodes japonica extract, used as herbal medicine, binds to human serum IgE and induces an allergic reaction as an immune response complex, and to add a panel of allergens to the Allergy Q kit. First, using the Allergy-Q test kit, 253 sera samples were screened and 4 samples each from positive and negative serogroups were randomly selected. AJ extracts were separated by size through sodium dodecyl sulfate–polyacrylamide gel electrophoresis and stained with Coomassie blue dye. We then used Western blotting and in-gel digest to identify proteins that bind to human serum IgE, and predicted six proteins, including putative chitinases, proteasomes, and hypothetical proteins, using MASCOT program matching. Consequently, we demonstrated the potential of AJ as an allergen by confirming the sensitivity between AJ-derived proteins and human IgE.

Polydopamine Nanoparticle-Incorporated Fluorescent Hydrogel for Fluorescence Imaging-Guided Photothermal Therapy of Cancers

이경관,이상천,김홍기,이창수 한국바이오칩학회 2023 BioChip Journal Vol.17 No.1

In this study, we report a fluorescent polydopamine nanoparticle-incorporated hydrogel (FPNP-hydrogel) for fluorescence imaging-guided photothermal cancer therapy. The FPNP-hydrogel was prepared by combining fluorescent polydopamine nanoparticles (FPNPs) with polyvinyl alcohol (PVA) using a simple method. The unique optical properties of the FPNPs facilitated the FPNP-hydrogel the excellent biocompatibility, strong fluorescence, and high photostability, showing an availability of fluorescence imaging in BALB/c nude mice. When combined with near-infrared laser irradiation, the FPNP-hydrogel noticeably reduced tumor volumes in 4T1 tumor-bearing mice within 14 days without inducing the inflammatory response in major organs. In conclusion, the results suggest that multifunctional hydrogels with excellent fluorescence properties and high photothermal conversion efficiency could be useful for designing novel therapeutic methods for cancer treatment.

Development of Tumor-Vasculature Interaction on Chip Mimicking Vessel Co-Option of Glioblastoma

배진승,김민혁,한석규,박성수 한국바이오칩학회 2023 BioChip Journal Vol.17 No.1

Vessel co-option (VC) differs from angiogenesis in that tumor cells grow toward blood vessels. Through VC, tumor cells can receive relatively more nutrients and oxygen from blood vessels. Despite its clinical significance, VC is relatively less studied compared to angiogenesis because of difficulties in longitudinal observation of VC in vivo and lack of proper VC models in vitro. A needle template method in which microchannels are formed in hydrogel by needles was used to form blood vessels and mimic angiogenesis. However, it has not yet been used to mimic VC. In this study, we report the development of VC on chip based on the needle template method. On the VC on chip, the effect of distance between spheroids and blood vessels on VC induction was investigated by seeding glioblastoma (GBM) spheroids 50 and 250 μm from the preformed blood vessels. Irrespective of distance, cancer cells from the spheroids grew toward the blood vessels but did not penetrate the vessels, indicating that GBM cells showed VC-like behavior. These results suggest that our chip could recapitulate VC in GBM.

Recent Advances in Electrochemical and Optical Biosensors for Cancer Biomarker Detection

손민형,박석원,사공희연,정윤경 한국바이오칩학회 2023 BioChip Journal Vol.17 No.1

Cancer is a universal disease with a high mortality rate and caused by an uncontrolled growth of abnormal cells. Each cancer has its unique molecular change such as up/down-regulations of biological molecules in cancer cells. These biological molecules have been identified as biomarkers and used as a target analyte for cancer diagnosis. Since the level of cancer biomarkers is very insufficient at an early stage cancer, there is a need for technological advances that can be more accurate, ultra-sensitive, trustworthy, and selective to biomarkers. This review summarizes evaluation methods and recent advances in electrochemical and optical (fluorescent and colorimetric) biosensors to detect various cancer biomarkers as molecular targets. Electrochemical methods demonstrate rapid and ultra-sensitive detection of cancer biomarkers. And, fluorescent biosensors present improved target detection sensitivity by signal amplification strategy as well as provide the possibility to sense multiple biomarker targets with multiple fluorescent probes. Colorimetric biosensors easily and quickly detect cancer biomarkers by the naked eye for use as a point-of-care testing (POCT) platform. These biosensors can be further improved by understanding the complication of cancer cells and the molecular alterations in cancer progression for early diagnosis of cancer and patient prognosis.

Fabrication of Cell Spheroids for 3D Cell Culture and Biomedical Applications

박세연,홍혜진,이현종 한국바이오칩학회 2023 BioChip Journal Vol.17 No.1

The significance of 3D cell culture—i.e., the maintenance of cell characteristics by mimicking the natural environment—has been increasing, and various 3D cell culture methods have been proposed. 3D cell culture is a widely used cell spheroid culture method, which is a representative method of matrix-free 3D cell culture. In contrast with that in conventional 2D cell culture, cells aggregate in a spherical shape with abundant cell–cell and cell–matrix interactions that more closely resemble the in vivo environment in a 3D culture. In a cell spheroid culture, the viability and functionality of spheroids change based on the cell type and morphology, and their controllable characteristics differ depending on the fabrication method used. Moreover, the mass production and size controllability of spheroids are dependent on the fabrication method used;therefore, the spheroid utilization method is also different. In this review, representative cell spheroid fabrication methods are examined, and the possibility of standardization is discussed so that these methods can be widely used. In addition, the biomedical applications of spheroids and their potential for future development are explored.

Organ-On-A-Chip Technology: An In-depth Review of Recent Advancements and Future of Whole Body-on-chip

Rishab Driver,Shweta Mishra 한국바이오칩학회 2023 BioChip Journal Vol.17 No.1

One of the leading challenges facing the pharmaceutical industry today is the ‘high attrition rate problem’, with a large number of drug candidates being abandoned before reaching the clinical stage. The high failure rate is not only costly for the pharmaceutical industry but also hinders the access of patients to better treatment options. This ever-widening gap can be attributed to the current in-vitro and animal testing techniques for predicting human responses to drug candidates. Twodimensional (2D) in-vitro cell culture fails to recapitulate key physical and biochemical cues while phylogenetic differences between animal models and humans in key physiological systems make data generated by these platforms unreliable. There have been several solutions proposed but one that has been gaining a lot of prominence are microfluidic-based microsystems able to mimic organs, called ‘Organ-On-A-Chip’ devices and is built upon the principles of microfluidics, material science, and cell biology. Here we delve into the ongoing research involving this platform demonstrating its key strengths and the application of this technology in several key stages of drug discovery such as target identification and high throughput screening. We also discuss the potential, future, and key obstacles that lie in the universal adoption of this technology.

Analysis of genomic profile in mouse lymphoma L5178Y cells exposed to food colorant gardenia yellow

Md. Mujibur Rahman,서영록 한국바이오칩학회 2010 BioChip Journal Vol.4 No.4

Gardenia yellow is renowned for its application as natural food colorant and traditional Chinese medicine. The consumption of this ingredient has been increasing in the production of several food products and the treatment of hepatic and inflammatory diseases. Safety issue is of prime importance for the gardenia yellow consumer. To evaluate the biological responses imposed by gardenia yellow, we have performed transcriptomic analysis using microarray in mouse lymphoma L5178Y cell line to gain insight into possible mechanisms of food additive gardenia yellow by which interaction in the lymphocyte cells can not be excluded. Here, the L5178Y cells were treated with gardenia yellow at non-lethal dose (5,000 mg/L). In this paper we firstly demonstrated the gene expression profile in the L5178Y cells in response to gardenia yellow. We have identified 36 mRNAs differentially expressed in the L5178Y cells against gardenia yellow. The altered expression level of 11 genes (Cd22, Fos, Gmip, Hras1,Il4, Il7r, Il10, Ndst1, Osgin1, Pik3r2, and Tnfsf4), whose products are associated with cellular processes including immune responses, signal transduction, cell proliferation, and apoptosis, was subsequently confirmed by real-time quantative reverse transcription PCR (qRT-PCR). Using Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database, we have also categorized these differentially expressed genes into particular pathway in accordance with gardenia yellowmediated biological processes including immunological and inflammatory responses, oxidative stress-sensitive defense, and cell proliferation. Our array analysis experiments led to the novel discovery that gardenia yellow is capable of interfering immune system and cell proliferation at transcription level, even at non-lethal dose. Our study unraveled the virtue of gene expression profiling in food toxicogenomics in terms of providing plausible possibility on the interaction of food additive gardenia yellow in correlation to pathology. Furthermore, the identified genes could be implicated as potential biomarkers for safety evaluation and health harmful effects of the gardenia yellow.

Flexible Capillary Microfluidic Devices Based on Surface-Energy Modified Polydimethylsiloxane and Polymethylmethacrylate with Room-Temperature Chemical Bonding

Lai Thi Ngoc Huyen,홍석주,TranQuangTrung,Montri Meeseepong,김아리,이내응 한국바이오칩학회 2023 BioChip Journal Vol.17 No.1

Polydimethylsiloxane (PDMS) has been widely used for the rapid prototyping of microfluidic devices for biosensor cartridges. However, using PDMS to prototype capillary-driven microfluidic devices is often limited by the difficulty of maintaining the surface energy of surface-treated PDMS for an extended period in addition to the degradation of the biosensing elements during the bonding process at elevated temperature. Herein, prototyping of a flexible capillary microfluidic channel (FCMC) device based on the room-temperature bonding of the surface energy-modified PDMS (m-PDMS) microfluidic channel and a thermoplastic lid, polymethylmethacrylate (PMMA), is introduced for prolonged control of passive liquid flow characteristics. The m-PDMS was fabricated by blending polydimethylsiloxane-ethylene oxide (60–70%) block copolymer (PDMS-b-PEO) additive with pre-PDMS, of which the water contact angles could be controlled between 38.5° and 78.5° by adjusting the ratio of the two components. Room-temperature bonding of the m-PDMS and PMMA sheets functionalized by 3-glycidoxypropyltrimethoxysilane and aminopropyltriethoxysilane, respectively, was introduced to fabricate the FCMC devices via the formation of a stable linker epoxy-amine without the requirement of elevated temperatures. The FCMC device possessed longevity to passively drive liquid in the channel for 2 months under ambient conditions due to the prolonged stable hydrophilicity of m-PDMS. The proposed approaches provide great potential for prototyping passive microfluidic devices for biosensor cartridge applications.

Direct Reverse Transcription Real-Time PCR of Viral RNA from Saliva Samples Using Hydrogel Microparticles

Emmanuel George Kifaro,김미정,정승원,노진용,송창선,Gerald Misinzo,김상경 한국바이오칩학회 2022 BioChip Journal Vol.16 No.4

In recent decades “saliva” has emerged as an important non-invasive biofluid for diagnostic purposes in both human and animal health sectors. However, with the rapid evolution of molecular detection technologies, the limitation has been the lack of an efficient method for the facile amplification of target RNA from such a complex matrix. Herein, we demonstrate the novel application of hydrogel microparticles of primer-immobilized networks (PIN) for direct quantitative reverse transcription PCR (dirRT-qPCR) of viral RNA from saliva samples without prior RNA purification. Each of these highly porous PIN particles operates as an independent reactor. They filter in micro-volumes of the analyte solution. Viral RNA is captured and converted to complementary DNA (cDNA) through the RT step using covalently incorporated RT primers. The PIN with cDNA of the viral target will be ready for subsequent highly specific qPCR. Preceded by heat-treatment for viral lysis, we were able to conduct PIN dirRT-qPCR with 95% efficiency of the matrix (M) gene for influenza A virus (IAV) and 5’ untranslated region (5’ UTR) for chicken coronavirus spiked into saliva samples. The addition of reverse transcriptase enzyme (RTase) and 10% dilution of the matrix improved the assay sensitivity considerably. PIN particles’ compatibility with microfluidic PCR chip technology has significantly reduced total sample processing time to 50 min, instead of an average of 120 min that are normally used by other assays. We anticipate this technology will be useful for other viral RNA targets by changing the incorporated RT primer sequences and can be adapted for onsite diagnostics.