• Journal of the Optical Society of Korea
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• v.15 no.1
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• pp.61-67
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• 2011

The confocal laser scanning microscopy and two-photon microscopy was implemented based on a single laser source and an objective lens. We imaged and compared the morphology of identical sites of ex vivo human skin using both microscopes. The back-scattering emission from the sample provided the contrast for the confocal microscopy. The intrinsic autofluorescence and the second harmonic generation were used as the luminescence source for the two-photon microscopy. The wavelength of the Ti:Sapphire laser was tuned at 710 nm, which corresponds to the excitation peak of NADH and FAD in skin tissue. The various cell layers in the epidermis and the papillary dermis were clearly distinguished by both imaging modalities. The two-photon microscopy more clearly visualized the intercellular region and the nucleus of the cell compared to the confocal microscopy. The fibrous structures in the dermis were more clearly resolved by the confocal microscopy. Numerous cells in papillary dermal layer, as deep as $100\;{\mu}m$, were observed in both CLSM and two-photon microscopy. While most previous studies focused on fibrous structure imaging (collagen and elastin fiber) in the dermis, we demonstrated that the combined imaging with the CLSM and two-photon microscopy can be applied for the non-invasive study of the population, distribution and metabolism of papillary dermal cells in skin.

• Restorative Dentistry and Endodontics
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• v.44 no.3
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• pp.25.1-25.10
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• 2019

Objective: The aim of this study was to evaluate discoloration of teeth undergoing regenerative endodontic procedures (REPs) using blood clot or platelet-rich fibrin (PRF) as the scaffolds and different calcium silicate-based materials as the intracanal coronal barriers in an ex vivo model. Materials and Methods: Forty-eight bovine incisors were prepared and disinfected using 1 mg/mL double antibiotic paste (DAP). The specimens were then randomly divided into 2 groups (n = 24) according to the scaffolds (blood or PRF). After placement of scaffolds each group was divided into 2 subgroups (n = 12) according to the intracanal coronal barriers (ProRoot MTA or Biodentine). The pulp chamber walls were sealed with dentin bonding agent before placement of DAP and before placement of scaffolds. The color changes (${\Delta}E$) were measured at different steps. The data were analyzed using 2-way analysis of variance. Results: Coronal discoloration induced by DAP was not clinically perceptible (${\Delta}E{\leq}3.3$). Regarding the type of the scaffold, coronal discoloration was significantly higher in blood groups compared with PRF groups at the end of REP and after 1 month (p < 0.05). However, no significant difference was found between PRF and blood clot after 6 months (p > 0.05). Considering the type of intracanal coronal barrier, no significant difference existed between ProRoot MTA and Biodentine (p > 0.05). Conclusions: With sealing the dentinal tubules of pulp chamber with a dentin bonding agent and application of DAP as an intracanal medicament, coronal color change of the teeth following the use of PRF and blood sealed with either ProRoot MTA or Biodentine was not different at 6-month follow-up.

• Journal of Chest Surgery
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• v.54 no.3
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• pp.186-190
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• 2021

Background: Pre-lifting of the sternum marked a major turning point in pectus excavatum repair. The author developed the crane technique in 2002 and successfully applied it to more than 2,000 cases using sternal wire stitching. However, blind sternal suturing limited the use of the wire-stitch crane. We propose a novel screw for sternal lifting as a new tool for the crane technique. Methods: We developed a screw system strong enough to withstand the pressure needed for sternum lifting. The screw was designed to have a broader thread to hold the bony tissue securely. The screw's sustaining power was tested using the torsion, driving torque, and axial pull-out tests in a polyurethane block and ex-vivo porcine sternum. Results: The screws were easily driven into the sternum, and the head of the screw was connectable to the table-mounted retractor. In the torsion test, the 2° offset torsional yield was 4.53 N·m (reference value, 1 N·m). In the polyurethane block driving torque test, the maximum torque was 0.98 N·m (reference value, 0.70 N·m). The axial pull-out test was 446 N (reference value, 100 N). The maximum pull-out resistance in the ex-vivo porcine sternum model was 1,516 N. Conclusion: The screw crane was strong enough to sustain the chest wall weight to be lifted. Thus, the screws could effectively replace the sternal wire stitching in crane pre-lifting of the sternum. We expect that application of the screw-crane will be easy and that it will improve the safety and success rate of pectus repair surgery.

• Molecules and Cells
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• v.45 no.7
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• pp.479-494
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• 2022

Human mesenchymal stem cells (MSCs) are multipotent stem cells that have been intensively studied as therapeutic tools for a variety of disorders. To enhance the efficacy of MSCs, therapeutic genes are introduced using retroviral and lentiviral vectors. However, serious adverse events (SAEs) such as tumorigenesis can be induced by insertional mutagenesis. We generated lentiviral vectors encoding the wild-type herpes simplex virus thymidine kinase (HSV-TK) gene and a gene containing a point mutation that results in an alanine to histidine substitution at residue 168 (TK(A168H)) and transduced expression in MSCs (MSC-TK and MSC-TK(A168H)). Transduction of lentiviral vectors encoding the TK(A168H) mutant did not alter the proliferation capacity, mesodermal differentiation potential, or surface antigenicity of MSCs. The MSC-TK(A168H) cells were genetically stable, as shown by karyotyping. MSC-TK(A168H) responded to ganciclovir (GCV) with an half maximal inhibitory concentration (IC₅₀) value 10-fold less than that of MSC-TK. Because MSC-TK(A168H) cells were found to be non-tumorigenic, a U87-TK(A168H) subcutaneous tumor was used as a SAE-like condition and we evaluated the effect of valganciclovir (vGCV), an oral prodrug for GCV. U87-TK(A168H) tumors were more efficiently ablated by 200 mg/kg vGCV than U87-TK tumors. These results indicate that MSC-TK(A168H) cells appear to be pre-clinically safe for therapeutic use. We propose that genetic modification with HSV-TK(A168H) makes allogeneic MSC-based ex vivo therapy safer by eliminating transplanted cells during SAEs such as uncontrolled cell proliferation.

• Medical Lasers
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• v.10 no.3
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• pp.161-169
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• 2021

Background and Objectives Fractional microneedle radiofrequency systems are popular options to increase elasticity in aging skin. Laser-assisted drug delivery is a promising method for the epidermal injection of topically applied drugs and cosmetic ingredients. This study assesses the safety and efficacy of topical delivery of L-ascorbic acid, vitamin E, and ferulic acid after fractional microneedle radiofrequency treatment for reducing photodamage. Materials and Methods In this prospective, single-center, split-face, controlled pilot study, six women (mean age, 48.0 years; range, 35-57 years; Fitzpatrick skin types III and IV) exhibiting mild to moderate photodamage, underwent a single session of fractional microneedle radiofrequency treatment. The patients were instructed to apply the antioxidant formulation to only one side of the face. Patients were evaluated 3 days, 7 days, and 4 weeks thereafter, using three-dimensional imaging and ultrasound. Ex vivo, the full-thickness human skin was used for molecular and histological evaluation. Statistical analysis was achieved by applying t-tests, Mann-Whitney U tests, and one-way analyses of variance. Results Compared to the untreated side, the antioxidant-treated side exhibited a significant increase in dermal thickness (10.32% vs. 17.54%, p < 0.05), but not in skin elasticity (4.76% vs. 4.69%, p > 0.05). The difference in erythema between the sides was statistically not significant (p > 0.05). In the ex vivo model, expression of FGF2 in the skin was significantly increased after application of the antioxidant formulation, as compared to results obtained subsequent to fractional microneedle radiofrequency treatment only (p < 0.01). Conclusion This study demonstrates that for the treatment of photodamaged skin, laser-assisted delivery of the antioxidant formulation is a safe and effective adjuvant modality following fractional microneedle radiofrequency.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.4
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• pp.255-262
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• 2022

Oxytocin is a neuropeptide produced primarily in the hypothalamus and plays an important role in the regulation of mammalian birth and lactation. It has been shown that oxytocin has important cardiovascular protective effects. Here we investigated the effects of oxytocin on vascular reactivity and underlying the mechanisms in human umbilical vein endothelial cells (HUVECs) in vitro and in rat aorta ex vivo. Oxytocin increased phospho-eNOS (Ser 1177) and phospho-Akt (Ser 473) expression in HUVECs in vitro and the aorta of rat ex vivo. Wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase (PI3K), inhibited oxytocin-induced Akt and eNOS phosphorylation. In the rat aortic rings, oxytocin induced a biphasic vascular reactivity: oxytocin at low dose (10^-9-10^-8 M) initiated a vasorelaxation followed by a vasoconstriction at high dose (10^-7 M). L-NAME (a nitric oxide synthase inhibitor), endothelium removal or wortmannin abolished oxytocin-induced vasorelaxation, and slightly enhanced oxytocin-induced vasoconstriction. Atosiban, an oxytocin/vasopressin 1a receptor inhibitor, totally blocked oxytocin-induced relaxation and vasoconstriction. PD98059 (ERK1/2 inhibitor) partially inhibited oxytocin-induced vasoconstriction. Oxytocin also increased aortic phospho-ERK1/2 expression, which was reduced by either atosiban or PD98059, suggesting that oxytocin-induced vasoconstriction was partially mediated by oxytocin/V1aR activation of ERK1/2. The present study demonstrates that oxytocin can activate different signaling pathways to cause vasorelaxation or vasoconstriction. Oxytocin stimulation of PI3K/eNOS-derived nitric oxide may participate in maintenance of cardiovascular homeostasis, and different vascular reactivities to low or high dose of oxytocin suggest that oxytocin may have different regulatory effects on vascular tone under physiological or pathophysiological conditions.

• Journal of the Korean Society of Industry Convergence
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• v.25 no.3
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• pp.365-378
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• 2022

Complete blood cell count(CBC) is a technique that counts leukocytes for each type of blood cell being analyzed. The principle is that laser is incident to ex vivo flowing leukocytes in a microcapillary tube and scattered light occurs by laser and leukocytes. By collecting the scattered light, we can count the types of cells because different cells generate different light-scattering patterns. However, the technique has an intrinsic limitation, scattering pattern is shown in a wide range region in the resulting, which makes it difficult to accurate analyze and use fluorescent dyes. To overcome this limitation, a new design of CBC with a dual laser, which irradiates with orthogonal angles for collecting quad-scattering information was proposed. Before development, the scattering difference depending on wavelength must be investigated to only catch up to the scattered signal by angles. Some studies, which focused on simple particles, have been conducted to theoretically and experimentally investigate different scatterings by wavelength. In this study, we propose an optical system for measuring scattered light and investigate a complex particle. As a result, the green laser made strong scattering signals in both the forward and side direction: 10% and 30%, respectively.

• Korean Journal of Transplantation
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• v.28 no.3
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• pp.154-159
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• 2014

Background: Lung transplantation (LTx) is a life-saving treatment for patients with end-stage lung disease; however, the shortage of donor lungs has been a major limiting factor to increasing the number of LTx. Growing experience following LTx using donor lungs after cardiac death (DCD) has been promising, although concerns remain. The purpose of this study was to develop a DCD lung harvest model using an ex vivo lung perfusion (EVLP) system and to assess the function of presumably damaged lungs harvested from the DCD donor in pigs. Methods: The 40 kg pigs were randomly divided into the control group with no ischemic lung injury (n=5) and the study group (n=5), which had 1 hour of warm ischemic lung injury after cardiac arrest. Harvested lungs were placed in the EVLP circuit and oxygen capacities (OC), pulmonary vascular resistance (PVR), and peak airway pressure (PAP) were evaluated every hour for 4 hours. At the end of EVLP, specimens were excised for pathologic review and wet/dry ratio. Results: No statistically significant difference in OC (P=0.353), PVR (P=0.951), and PAP (P=0.651) was observed in both groups. Lung injury severity score (control group vs. study group: 0.700±0.303 vs. 0.870±0.130; P=0.230) and wet/dry ratio (control group vs. study group: 5.89±0.97 vs. 6.20±0.57; P=0.560) also showed no statistically significant difference between the groups. Conclusions: The function of DCD lungs assessed using EVLP showed no difference from that of control lungs without ischemic injury; therefore, utilization of DCD lungs can be a new option to decrease the number of deaths on the waiting list.

• Journal of Veterinary Clinics
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• v.39 no.6
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• pp.353-359
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• 2022

Several suture patterns can be used for cystotomy closure, and a continuous suture pattern is the most commonly used. In this study, the fluid-tight ability and other suitabilities of continuous appositional sutures, such as the simple continuous suture pattern (SC), running suture pattern (RN), and Ford interlocking suture pattern (FI), were compared for cystotomy closure. Cystotomy closure was performed using each suture method in 10 cases of ex vivo swine bladders in each group. Suture time, leakage site, suture length, bursting pressure (BP), bursting volume (BV), and circular bursting wall tension (CBWT) were measured. Suture time and suture length were the shortest in RN and the longest in FI. Leakage occurred in two places: the incision line directly and the hole made by the suture. Leakage occurred through the incision line in 4 bladders of the RN group and 2 bladders of the FI group, but not in the SC group, and in the rest of the bladders, leakage occurred through the suture hole. The values of BP, BV, and CBWT increased in the order of FI, SC, and RN. Suture time and suture length can be considered as factors related to healing and side effects. In this study, leakage through the incision was found in a less appositional area; therefore, leakage through the hole could be considered an indicator of better apposition. Good apposition is one of the conditions required for ideal cystotomy closure. The bursting strength representing the fluid-tight ability can be expressed as the CBWT. RN is expected to be efficient and cause a small degree of foreign body reaction; however, it is expected to be less stable. FI has the greatest fluid-tightness ability, but it has been proposed that side effects due to foreign body reactions most frequently occur in FI. In conclusion, SC, which is expected to have a sufficient degree of fluid-tightness and appropriate recovery, is preferable to other continuous appositional suturing methods for cystotomy closure.

• Korean Journal of Radiology
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• v.23 no.2
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• pp.189-201
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• 2022

Objective: To compare the outcomes of radiofrequency ablation (RFA) using dual switching monopolar (DSM), switching bipolar (SB), and combined DSM + SB modes at two different interelectrode distances (25 and 20 mm) in an ex vivo study, which simulated ablation of a 2.5-cm virtual hepatic tumor. Materials and Methods: A total of 132 ablation zones were created (22 ablation zones for each protocol) using three separable clustered electrodes. The performances of the DSM, SB, and combined DSM + SB ablation modes were compared by evaluating the following parameters of the RFA zones at two interelectrode distances: shape (circularity), size (diameter and volume), peritumoral ablative margins, and percentages of the white zone at the midpoint of the two electrodes (ablative margin at midpoint, AMm) and in the electrode path (ablative margin at electrode path, AMe). Results: At both distances, circularity was the highest in the SB mode, followed by the DSM + SB mode, and was the lowest in the DSM mode. The circularity of the ablation zone showed a significant difference among the three energy groups (p < 0.001 and p = 0.002 for 25-mm and 20-mm, respectively). All size measurements, AMm, and AMe were the greatest in the DSM mode, followed by the DSM + SB mode, and the lowest were with the SB mode (all statistically significant). The white zone proportion in AMm and AMe were the greatest in the SB mode, followed by the DSM + SB mode and DSM in general. Conclusion: DSM and SB appear to be complementary in creating an ideal ablation zone. RFA with the SB mode can efficiently eradicate tumors and create a circular ablation zone, while DSM is required to create a sufficient ablative margin and a large ablation zone.