ABSTRACT
Monocarboxylate transporter-1 (MCT-1) inhibitors were screened from the Fv-antibody library, which contained complementary determining region 3 with randomized amino acid sequences (11 residues) through site-directed mutagenesis. Fv-antibodies against MCT-1 were screened from the autodisplayed Fv-antibody library. Two clones were screened, and the binding affinity (KD) against MCT-1 was estimated using flow cytometry. The screened Fv-antibodies were expressed as soluble fusion proteins (Fv-1 and Fv-2) and the KD for MCT-1 was estimated using the SPR biosensor. The inhibitory activity of the expressed Fv-antibodies was observed in HEK293T and Jurkat cell lines by measuring intracellular pH and lactate accumulation. The level of cell viability in HEK293T and Jurkat cell lines was decreased by the inhibitory activity of the expressed Fv-antibodies. The binding properties of the Fv-antibodies to MCT-1 were analyzed using molecular docking simulations. Overall, the results showed that the screened Fv-antibodies against MCT-1 from the Fv-antibody library had high binding affinity and inhibitory activity against MCT-1, which could be used as potential therapeutic drug candidates for the MCT-1 inhibitor.
Subject(s)
Antibodies , Carrier Proteins , Humans , Molecular Docking Simulation , HEK293 Cells , Amino Acid Sequence , Gene LibraryABSTRACT
Wearable devices that can mechanically conform to human skin are a necessity for reliable monitoring and decoding of biomechanical activities through skin. Most inorganic piezoelectrics, however, lack deformability and damage tolerance, impeding stable motion monitoring. Here, we present an air-permeable fabric-based ZnO nanogenerator with mechanical adaptivity to diverse deformations for wearable piezoelectric sensors, collecting biomechanical health data. We fabricate ZnO nanorods incorporated throughout the entire nylon fabric, with a strategically positioned neutral mechanical plane, for bending-sensitive electronics (2.59 µA mm). Its hierarchically interlocked geometry also permits sensitive tactile sensing (0.15 nA kPa-1). Various physiological information about activities, including pulse beating, breathing, saliva swallowing, and coughing, is attained using skin-mounted sensors. Further, the pyroelectric sensing capability of a mask-attached device is demonstrated by identifying specific respiratory patterns. Our wearable healthcare sensors hold great promise for real-time monitoring of health-related vital signs, informing individuals' health status without disrupting their daily lives.
Subject(s)
Biosensing Techniques , Wearable Electronic Devices , Zinc Oxide , Humans , Skin , MotionABSTRACT
Rapid spread of infectious diseases is a global threat and has an adverse impact on human health, livelihood, and economic stability, as manifested in the ongoing coronavirus disease 2019 (COVID-19) pandemic. Even though people wear a face mask as protective equipment, direct disinfection of the pathogens is barely feasible, which thereby urges the development of biocidal agents. Meanwhile, repetitive respiration generates temperature variation wherein the heat is regrettably wasted. Herein, a biocidal ZnO nanorod-modified paper (ZNR-paper) composite that is 1) integrated on a face mask, 2) harvests waste breathing-driven thermal energy, 3) facilitates the pyrocatalytic production of reactive oxygen species (ROS), and ultimately 4) exhibits antibacterial and antiviral performance is proposed. Furthermore, in situ generated compressive/tensile strain of the composite by being attached to a curved mask is investigated for high pyroelectricity. The anisotropic ZNR distortion in the bent composite is verified with changes in ZnO bond lengths and OZnO bond angles in a ZnO4 tetrahedron, resulting in an increased polarization state and possibly contributing to the following pyroelectricity. The enhanced pyroelectric behavior is demonstrated by efficient ROS production and notable bioprotection. This study exploring the pre-strain effect on the pyroelectricity of ZNR-paper might provide new insights into the piezo-/pyroelectric material-based applications.
Subject(s)
COVID-19 , Zinc Oxide , Humans , COVID-19/prevention & control , Zinc Oxide/chemistry , Masks , Reactive Oxygen Species , RespirationABSTRACT
Nanostructuresâcoupled with mass spectrometryâhave been intensively investigated to improve the detection sensitivity and reproducibility of small biomolecules in laser desorption/ionization mass spectrometry (LDI-MS). However, the impact of laser-induced shock wave on the ionization of the nanostructures has rarely been reported. Herein, we systematically elucidate the laser shock wave effect on the ionization in terms of the in situ development of atomic defects and piezoelectricity in two-dimensional graphitic carbon nitride nanosheets (g-C3N4 NS) by short laser pulses. The mass analysis results of immunosuppressive drugs verify the enhanced LDI-MS performance, structurally originating from anisotropic lattice distortions in g-C3N4 NS, i.e., in-plane extension (contraction) and out-of-plane contraction (extension) that modulate the charge carrier motion. Along with the experimental investigations, density functional theory calculations on Mulliken charges and dipole moments demonstrate the contribution of defect and piezoelectricity to the ionization. The results of this study provide a mechanistic understanding of the underlying ionization processes, which is crucial for revealing the full potential of laser shock waves in LDI-MS.
Subject(s)
Graphite , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Reproducibility of Results , Graphite/chemistry , LasersABSTRACT
Short and medium chain acylcarnitines have been used for the diagnosis of various fatty acid oxidation and organic acid disorders. This report presents a multiplex and quantitative analysis of acylcarnitines using MALDI-TOF MS based on a parylene matrix chip. The parylene matrix chip was fabricated by the deposition of a nanoporous film of parylene on an organic matrix array, which reduced the number of mass peaks from the organic matrix in the low m/z range. Quantitative analysis was possible using the parylene matrix chip because of the formation of nano-sized sample crystals on the nanoporous parylene film. Seven acylcarnitines were quantitatively analyzed using the chip; the method detection range included the cut-off values for metabolic disorders. The seven acylcarnitines of different concentrations were simultaneously detected using the parylene matrix chip and the interference from the mixed carnitines was estimated. Real L-carnitine (C0) samples were analyzed using serial dilution, and the recoveries were calculated by comparisons with a standard curve.
Subject(s)
Carnitine , Xylenes , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Xylenes/chemistryABSTRACT
Inhibitors for monoamine oxidase-B (MAO-B) were screened from an FV library with a randomized complementarity-determining region 3 (CDR3) region using a monoclonal antibody against dopamine. As the first step, the FV library was expressed on the outer membrane of E. coli by site-directed mutagenesis of the randomized CDR3 region. Among the FV library, variants with a binding affinity to monoclonal antibodies against dopamine were screened and cloned. From the comparison of the binding activity of the screened clones to a control clone with a modified FV antibody (only with CDR1 and CDR2), the CDR3 regions of screened clones were determined to directly interact with the monoclonal antibody against dopamine. These CDR3 sequences were then synthesized as mimotopes (mimicking peptides) of dopamine. The inhibitory activity of two mimotopes against MAO-B was analyzed using HeLa cells overexpressing MAO-B, as well as using activated human astrocytes; their inhibitory activity was compared to that of a commercial inhibitor of MAO-B, selegiline. The inhibition efficiency of the two mimotopes (in comparison with selegiline) was estimated to be 67.2% and 69.4% in the HeLa cells and 64.4% and 58.0% in the human astrocytes. The gene expression pattern in astrocytes after treatment with the two mimotopes was also analyzed and compared with that in the human astrocytes treated with selegiline. Finally, the interaction between two mimotopes and MAO-B was analyzed using docking simulation, and the candidate regions of MAO-B for the interaction with each mimotope were explored through the docking simulation.
Subject(s)
Monoamine Oxidase , Selegiline , Antibodies, Monoclonal , Dopamine/metabolism , Escherichia coli/metabolism , HeLa Cells , Humans , Monoamine Oxidase/genetics , Monoamine Oxidase/metabolism , Monoamine Oxidase Inhibitors/pharmacology , Peptides , Selegiline/pharmacologyABSTRACT
Fundamental properties of nanostructured substrates govern the performance of laser desorption/ionization mass spectrometry (LDI-MS); however, limited studies have elucidated the desorption/ionization mechanism based on the physicochemical properties of substrates. Herein, the enhancement in desorption/ionization is investigated using a hybrid matrix of Au nanoisland-functionalized ZnO nanotubes (AuNI-ZNTs). The underlying origin is explored in terms of the photo-electronic and -thermal properties of the matrix. This is the first study to report the effect of laser-induced surface restructuring/melting phenomenon on the LDI-MS performance. AuNI plays a central role as a photothermal nanofurnace, which facilitates the internal energy transfer from the AuNI to the adsorbed analytes by reconstruction in the structurally dynamic AuNI and therefore favors the desorption process. Moreover, piezoelectricity is driven in situ in the AuNI-ZNT hybrid, which modulates the overall band structure and thereby promotes the ionization process. Ultimately, high LDI-MS performance is demonstrated by analyzing small metabolites of fatty acids and monosaccharides, which are challenged to be detected in conventional LDI-MS. This study emphasizing the understanding of matrix properties can provide insights into the design and development of a novel nanomaterial as an efficient LDI matrix. Furthermore, the developed hybrid matrix can overcome the major hurdles existing in conventional LDI-MS.
Subject(s)
Nanostructures , Lasers , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , SpectrophotometryABSTRACT
Patients with odontogenic cysts and tumors may have to undergo serious surgery unless the lesion is properly detected at the early stage. The purpose of this study is to evaluate the diagnostic performance of the real-time object detecting deep convolutional neural network You Only Look Once (YOLO) v2-a deep learning algorithm that can both detect and classify an object at the same time-on panoramic radiographs. In this study, 1602 lesions on panoramic radiographs taken from 2010 to 2019 at Yonsei University Dental Hospital were selected as a database. Images were classified and labeled into four categories: dentigerous cysts, odontogenic keratocyst, ameloblastoma, and no cyst. Comparative analysis among three groups (YOLO, oral and maxillofacial surgeons, and general practitioners) was done in terms of precision, recall, accuracy, and F1 score. While YOLO ranked highest among the three groups (precision = 0.707, recall = 0.680), the performance differences between the machine and clinicians were statistically insignificant. The results of this study indicate the usefulness of auto-detecting convolutional networks in certain pathology detection and thus morbidity prevention in the field of oral and maxillofacial surgery.
ABSTRACT
It has been challenging to detect small analytes in both positive and negative ion modes using organic matrices in conventional matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Herein, TiO2 nanowires are presented as a solid matrix to form dual ions of analytes regardless of their chemical properties and to demonstrate versatile applicability in LDI-MS.
ABSTRACT
Recently, the parylene-matrix chip was developed for quantitative analysis of small molecules less than 1 kDa. In this study, MALDI-TOF MS based on the parylene-matrix chip was performed to clinically diagnose intrahepatic cholangiocarcinoma (IHCC) and colorectal cancer (CRC). The parylene-matrix chip was applied for the detection of small cancer biomarkers, including N-methyl-2-pyridone-5-carboxamide (2PY), glutamine, lysophosphatidylcholine (LPC) 16:0, and LPC 18:0. The feasibility of MALDI-TOF MS based on the parylene-matrix chip was confirmed via analysis of spot-to-spot and shot-to-shot reproducibility. Serum metabolite markers of IHCC, N-methyl-2-pyridone-5-carboxamide (2PY), and glutamine were quantified using MALDI-TOF MS based on the parylene-matrix chip. For clinical diagnosis of CRC, two water-insoluble (barely soluble) biomarkers, lysophosphatidylcholine (LPC) 16:0 and LPC 18:0, were quantified. Finally, glutamine and LPC 16:0 were simultaneously detected at a range of concentrations in sera from colon cancer patients using the parylene-matrix chip. Thus, this method yielded high-throughput detection of cancer biomarkers for the mixture samples of water-soluble analytes (2PY and glutamine) and water-insoluble analytes (LPC 16:0 and LPC 18:0).
Subject(s)
Bile Duct Neoplasms/blood , Biomarkers, Tumor/blood , Cholangiocarcinoma/blood , Colorectal Neoplasms/blood , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Bile Duct Neoplasms/diagnosis , Cholangiocarcinoma/diagnosis , Colorectal Neoplasms/diagnosis , Equipment Design , Glutamine/blood , Humans , Limit of Detection , Lysophosphatidylcholines/blood , Lysophosphatidylcholines/chemistry , Polymers , Pyridones/blood , Reproducibility of Results , Solubility , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/instrumentation , XylenesABSTRACT
PURPOSE: To compare the surgical outcomes of peritoneal irrigation versus suction alone during laparoscopic appendectomy and to identify the risk factors of surgical site infection in patients with uncomplicated acute appendicitis. METHODS: Data from patients with uncomplicated acute appendicitis between January 2014 and March 2016 were reviewed. We compared the irrigation and suction alone groups with regard to the following parameters: postoperative complication incidence rate, length of hospital stay, operation time, time to flatus, time to diet commencement, and duration of postoperative antibiotic. RESULTS: A total of 578 patients underwent laparoscopic appendectomy for uncomplicated acute appendicitis. Twenty-five patients were excluded from the analysis because of need for drain insertion, loss to follow-up, simultaneous surgery for another indication, presence of an appendix tumor, or pregnancy. A total of 207 patients (37.4%) had undergone irrigation, and 346 patients (62.6%) received suction alone during laparoscopic appendectomy. The preoperative fever rate was significantly higher in the irrigation group than in the suction alone group. Operative time was also significantly longer in the irrigation group than in the suction alone group (53.8 ± 18.5 minutes vs. 57.8 ± 21.4 minutes, P = 0.027). The postoperative complication rate was higher in the irrigation group than in the suction alone group (4.5% vs. 12.6%, P = 0.001). Multiple logistic regression analysis showed that irrigation and preoperative fever were risk factors for surgical site infection after laparoscopic appendectomy for uncomplicated acute appendicitis. CONCLUSION: There is no advantage to irrigating the peritoneal cavity over suction alone during laparoscopic appendectomy for uncomplicated acute appendicitis. Irrigation may actually prolong the operative time and therefore be detrimental.
ABSTRACT
Hypotonia, Ataxia, and Delayed Development Syndrome (HADDS) is an autosomal-dominant, extremely rare neurodevelopmental disorder caused by the heterozygous EBF3 gene mutation. EBF3 is located on chromosome 10q26.3 and acts as a transcription factor that regulates neurogenesis and differentiation. This syndrome is characterized by dysmorphism, cerebellar hypoplasia, urogenital anomaly, hypotonia, ataxia, intellectual deficit, and speech delay. The current report describes a 3-year-old Korean male carrying a de novo EBF3 mutation, c.589A>G (p.Asn197Asp), which was identified by whole exome sequencing. He manifested facial dysmorphism, hypotonia, strabismus, vermis hypoplasia, and urogenital anomalies, including vesicoureteral reflux, cryptorchidism, and areflexic bladder. This is the first report of a case of HADDS cause by an EBF3 mutation in the Korean population.
ABSTRACT
Hypotonia, Ataxia, and Delayed Development Syndrome (HADDS) is an autosomal-dominant, extremely rare neurodevelopmental disorder caused by the heterozygous EBF3 gene mutation. EBF3 is located on chromosome 10q26.3 and acts as a transcription factor that regulates neurogenesis and differentiation. This syndrome is characterized by dysmorphism, cerebellar hypoplasia, urogenital anomaly, hypotonia, ataxia, intellectual deficit, and speech delay. The current report describes a 3-year-old Korean male carrying a de novo EBF3 mutation, c.589A>G (p.Asn197Asp), which was identified by whole exome sequencing. He manifested facial dysmorphism, hypotonia, strabismus, vermis hypoplasia, and urogenital anomalies, including vesicoureteral reflux, cryptorchidism, and areflexic bladder. This is the first report of a case of HADDS cause by an EBF3 mutation in the Korean population.
ABSTRACT
Purpose@#To compare the surgical outcomes of peritoneal irrigation versus suction alone during laparoscopic appendectomy and to identify the risk factors of surgical site infection in patients with uncomplicated acute appendicitis. @*Methods@#Data from patients with uncomplicated acute appendicitis between January 2014 and March 2016 were reviewed. We compared the irrigation and suction alone groups with regard to the following parameters: postoperative complication incidence rate, length of hospital stay, operation time, time to flatus, time to diet commencement, and duration of postoperative antibiotic. @*Results@#A total of 578 patients underwent laparoscopic appendectomy for uncomplicated acute appendicitis. Twenty-five patients were excluded from the analysis because of need for drain insertion, loss to follow-up, simultaneous surgery for another indication, presence of an appendix tumor, or pregnancy. A total of 207 patients (37.4%) had undergone irrigation, and 346 patients (62.6%) received suction alone during laparoscopic appendectomy. The preoperative fever rate was significantly higher in the irrigation group than in the suction alone group. Operative time was also significantly longer in the irrigation group than in the suction alone group (53.8 ± 18.5 minutes vs. 57.8 ± 21.4 minutes, P = 0.027). The postoperative complication rate was higher in the irrigation group than in the suction alone group (4.5% vs. 12.6%, P = 0.001). Multiple logistic regression analysis showed that irrigation and preoperative fever were risk factors for surgical site infection after laparoscopic appendectomy for uncomplicated acute appendicitis. @*Conclusion@#There is no advantage to irrigating the peritoneal cavity over suction alone during laparoscopic appendectomy for uncomplicated acute appendicitis. Irrigation may actually prolong the operative time and therefore be detrimental.
ABSTRACT
In this work, medical diagnosis of sepsis was conducted via quantitative analysis of lysophosphatidylcholine 16:0 (LPC 16:0) by using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry based on a parylene-matrix chip. In the first step, specific mass peaks for the diagnosis of sepsis were searched by comparing MALDI-TOF mass spectra of sepsis patient sera with healthy controls and pneumonia patient sera. Two mass peaks at m/z = 496.3 and 518.3 were chosen as those that are specifically different for sepsis sera to compare with healthy controls and pneumonia patient sera. These mass peaks were identified to be protonated and sodium adducts of LPC 16:0 by using tandem mass spectra (MS2 and MS3) of purely synthesized LPC 16:0 and extracted LPC 16:0 from a healthy control and a sepsis patient. In the next step, a standard curve for LPC 16:0 for the quantitative analysis of LPC 16:0 with MALDI-TOF MS based on the parylene-matrix chip was prepared, and the statistical correlation to the LC-MS analysis results was demonstrated by using the Bland-Altman test and Passing-Bablok regression. Finally, MALDI-TOF MS based on the parylene-matrix chip was used for the quantification of LPC 16:0 with sera from patients with severe sepsis and septic shock (n = 143), pneumonia patients (n = 12), and healthy sera (n = 31). The sensitivity and the selectivity of medical diagnosis of sepsis was estimated to be 97.9% and 95.5% by using MALDI-TOF MS based on the parylene-matrix chip, respectively.
Subject(s)
Lysophosphatidylcholines/blood , Polymers/chemistry , Sepsis/diagnosis , Xylenes/chemistry , Adult , Aged , Aged, 80 and over , Biomarkers/blood , Female , Humans , Limit of Detection , Male , Middle Aged , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/instrumentation , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methodsABSTRACT
Crystals of monosodium urate monohydrate (MSU) and calcium pyrophosphate dihydrate (CPPD) are known to induce arthropathic diseases called gout and pseudogout, respectively. These crystals are deposited in various joints or tissues, causing severe pain. Correct identification of crystals is crucial for the appropriate treatment of gout and pseudogout, which exhibit very similar symptoms. Herein, a novel approach of laser desorption/ionization time-of-flight (LDI-ToF) mass spectrometry (MS) was introduced to analyze MSU and CPPD crystals with three different types of nanostructured TiO2 materials including TiO2 nanoparticles (P25), TiO2 nanowires synthesized from wet-corrosion method, and the mixture of P25 and TiO2 nanowires (P25/TiO2 nanowires) as inorganic solid matrices. Furthermore, the feasibility of LDI-ToF MS based on these TiO2 nanostructures for the analysis of the two arthropathy-related crystals was tested using spiked samples in synovial fluid at known crystal concentrations. The mass analysis results of MSU and CPPD crystals demonstrated that (1) the electrostatic interaction between analytes and solid matrices was key for the analyte ionization and (2) LDI-ToF MS with nanostructured TiO2 materials has the potential to be a practical approach for the diagnosis of gout and pseudogout.
Subject(s)
Calcium Pyrophosphate/analysis , Gout , Nanostructures/chemistry , Titanium/chemistry , Uric Acid/analysis , Crystallization , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Time FactorsABSTRACT
A combination nanostructured matrix with metal Au nanoislands and semiconductor TiO2 nanowires is presented to enhance both desorption and ionization efficiency in laser desorption/ionization (LDI) mass spectrometry. The heterostructure of Au nanoislands on TiO2 nanowires was fabricated via (1) TiO2 nanowire synthesis through a modified wet-corrosion method and (2) Au nanoisland formation through thermal annealing of a sputtered Au layer on the TiO2 nanowires. Herein, the synergistic effect of this heterostructure for highly efficient ion production was experimentally elucidated in terms of the formation of high temperature on the surface of Au and the creation of a Schottky barrier at the Au-TiO2 interface. Finally, four types of immunosuppressors were analyzed to demonstrate the improved ionization performance of the heterostructure for LDI mass spectrometry.
ABSTRACT
RATIONALE: Magnetic particles coated with gold nanoparticles (Au NPs) (Au-MAGs) were developed and used (1) for sample concentration and (2) as a solid matrix for laser adsorption/desorption mass spectrometry (LDI-MS). METHODS: The Au-MAGs were prepared by (1) coating polystyrene on iron oxide NPs (PS-MNPs), (2) coating poly-l-lysine on the PS-MNPs (PLL-coated PS-MNPs), and (3) coating negatively charged Au NPs on the PLL-coated PS-MNPs (Au-MAGs). RESULTS: The Au-MAGs were used to concentrate the target analyte by means of electrostatic interactions between the positively charged GHP9 and the negatively charged Au-MAGs as well as selective interactions (such as gold-sulfur interactions) between glutathione (GSH) and Au-MAGs. Then, the concentrated analyte could be ionized for LDI-MS. CONCLUSIONS: The Au-MAGs were demonstrated (1) to concentrate the target analyte in a sample solution, tested by electrostatic interactions and selective interactions between gold and sulfur and (2) to ionize the concentrated analyte for LDI-MS.
ABSTRACT
In this work, TiO2 nanowires synthesized from a wet-corrosion process were presented for peptide sequencing by photocatalytic reaction with UV radiation. For the photocatalytic decomposition of peptides, the peptide sample was dropped on a target plate containing synthesized TiO2 nanowire zones and UV-irradiated. Subsequently, the target plate was analyzed by laser desorption/ionization time-of-flight (LDI-TOF) mass spectrometry using the synthesized TiO2 nanowires as a solid matrix. The feasibility of peptide sequencing based on the photocatalytic reaction with the synthesized TiO2 nanowires was demonstrated using six types of peptides GHP9 (G1-H-P-Q-G2-K1-K2-K3-K4, 1006.59 Da), BPA-1(K1-S1-L-E-N-S2-Y-G1-G2-G3-K2-K3-K4, 1394.74 Da), PreS1(F1-G-A-N1-S-N2-N3-P1-D1-W-D2-F2-N4-P2-N5, 1707.68 Da), HPQ peptide-1 (G-Y-H-P-Q-R-K, 884.45 Da), HPQ peptide-2 (K-R-H-P-Q-Y-G, 884.45 Da), and HPQ peptide-3 (R-Y-H-P-Q-G-K, 884.45 Da). The identification of three different peptides with the same molecular weight was also demonstrated by using the synthesized TiO2 nanowires for their photocatalytic decomposition as well as for LDI-TOF mass spectrometry as a solid-matrix.
Subject(s)
Mass Spectrometry/methods , Nanowires/chemistry , Peptides , Sequence Analysis, Protein , Titanium/chemistry , Corrosion , Peptides/chemistry , Peptides/genetics , Sequence Analysis, Protein/instrumentation , Sequence Analysis, Protein/methodsABSTRACT
PURPOSE: In this study, the retrospective radiographic study is executed to evaluate amount of bone loss of various conditions in patients using customized abutment for 4 years of follow-up. MATERIALS AND METHODS: The subjects of this study were implant fixed dental prosthesis using CAD/CAM customized abutments. CAD/CAM customized abutment and fixed dental prosthesis were manufactured by the Prosthodontics Department of Chosun University Dental Hospital from August 1, 2011 to July 31, 2012. Radiological assessments were performed on the patients who were treated by the fixed prosthodontics. After each treatment, a retrospective study was performed for a total of 4 years at 3 months, 6 months, 1 year, 2 years, 3 years, and 4 years. RESULTS: As a result of the study, the customized abutment using CAD/CAM showed less bone loss than the results of existing research. There was no statistically significant differences at alveolar bone loss between splinting group and non-splinting group (respectively 0.27 mm, 0.5 mm). Also, there were statistically significant differences at alveolar bone loss in mx. anterior, mx. posterior, mn. anterior and mn. posterior part (respectively 1.37 mm, 0.39 mm, 0.00 mm, 0.30 mm). CONCLUSION: The customized abutment using CAD/CAM showed less bone loss than the results of existing research, there were statistically significant differences at alveolar bone loss in implant positions.