• The Korean Journal of Pain
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• v.34 no.1
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• pp.19-26
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• 2021

Background: Prolotherapy is a proliferation therapy as an alternative medicine. A combination of dextrose solution and lidocaine is usually used in prolotherapy. The concentrations of dextrose and lidocaine used in the clinical field are very high (dextrose 10%-25%, lidocaine 0.075%-1%). Several studies show about 1% dextrose and more than 0.2% lidocaine induced cell death in various cell types. We investigated the effects of low concentrations of dextrose and lidocaine in fibroblasts and suggest the optimal range of concentrations of dextrose and lidocaine in prolotherapy. Methods: Various concentrations of dextrose and lidocaine were treated in NIH-3T3. Viability was examined with trypan blue exclusion assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Migration assay was performed for measuring the motile activity. Extracellular signal-regulated kinase (Erk) activation and protein expression of collagen I and α-smooth muscle actin (α-SMA) were determined with western blot analysis. Results: The cell viability was decreased in concentrations of more than 5% dextrose and 0.1% lidocaine. However, in the concentrations 1% dextrose (D1) and 0.01% lidocaine (L0.01), fibroblasts proliferated mildly. The ability of migration in fibroblast was increased in the D1, L0.01, and D1 + L0.01 groups sequentially. D1 and L0.01 increased Erk activation and the expression of collagen I and α-SMA and D1 + L0.01 further increased. The inhibition of Erk activation suppressed fibroblast proliferation and the synthesis of collagen I. Conclusions: D1, L0.01, and the combination of D1 and L0.01 induced fibroblast proliferation and increased collagen I synthesis via Erk activation.

• Clinical and Experimental Reproductive Medicine
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• v.49 no.4
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• pp.259-269
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• 2022

Objective: Animal-free scaffolds have emerged as a potential foundation for consistent, chemically defined, and low-cost materials. Because of its good potential for high biocompatibility with reproductive tissues and well-characterized scaffold design, we investigated whether polyglycolic acid (PGA) could be used as an animal-free scaffold instead of natural fibrin-agarose, which has been used successfully for three-dimensional human endometrial cell culture. Methods: Isolated primary endometrial cells was cultured on fibrin-agarose and PGA polymers and evaluated various design parameters, such as scaffold porosity and mean fiber diameter. Cytotoxicity, scanning electron microscopy (SEM), and immunostaining experiments were conducted to examine cell activity on fabricated scaffolds. Results: The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay and SEM results showed that endometrial cells grew and proliferated on both scaffolds. Immunostaining showed cytokeratin and vimentin expression in seeded cells after 7 days of culture. On both scaffolds, an epithelial arrangement of cultured cells was found on the top layer and stromal arrangement matrix on the bottom layer of the scaffolds. Therefore, fibrin-agarose and PGA scaffolds successfully mimicked the human endometrium in a way suitable for in vitro analysis. Conclusion: Both fibrin-agarose and PGA scaffolds could be used to simulate endometrial structures. However, because of environmental and ethical concerns and the low cost of synthetic polymers, we recommend using PGA as a synthetic polymer for scaffolding in research instead of natural biomaterials.

• Annals of Hepato-Biliary-Pancreatic Surgery
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• v.27 no.4
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• pp.342-349
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• 2023

Backgrounds/Aims: Liver organoids have emerged as a powerful tool for studying liver biology and disease and for developing new therapies and regenerative medicine approaches. For organoid culture, Matrigel, a type of extracellular matrix, is the most commonly used material. However, Matrigel cannot be used for clinical applications due to the presence of unknown proteins that can cause immune rejection, batch-to-batch variability, and angiogenesis. Methods: To obtain human primary hepatocytes (hPHs), we performed 2 steps collagenase liver perfusion protocol. We treated three small molecules cocktails (A83-01, CHIR99021, and HGF) for reprogramming the hPHs into human chemically derived hepatic progenitors (hCdHs) and used hCdHs to generate liver organoids. Results: In this study, we report the generation of liver organoids in a collagen scaffold using hCdHs. In comparison with adult liver (or primary hepatocyte)-derived organoids with collagen scaffold (hALO_C), hCdH-derived organoids in a collagen scaffold (hCdHO_C) showed a 10-fold increase in organoid generation efficiency with higher expression of liver- or liver progenitor-specific markers. Moreover, we demonstrated that hCdHO_C could differentiate into hepatic organoids (hCdHO_C_DM), indicating the potential of these organoids as a platform for drug screening. Conclusions: Overall, our study highlights the potential of hCdHO_C as a tool for liver research and presents a new approach for generating liver organoids using hCdHs with a collagen scaffold.

• International Journal of Oral Biology
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• v.38 no.4
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• pp.161-167
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• 2013

Human dental pulp stem cells (DPSCs) are multi-potent mesenchymal stem cells that have several differentiation potentials. An understanding of thetissues that differentiate from these cells can provide insights for future regenerative therapeutics and tissue engineering strategies. The mesiodens is the most frequent form of supernumerary tooth from which DPSCs can differentiate into several lineages similar to cells from normal deciduous teeth. Recently, it has been shown that nanoscale structures can affect stem cell differentiation. In our presentstudy, we investigated the effects of a 250-nm nanoscale ridge/groove pattern array on the osteogenic and adipogenic differentiation of dental pulp cells from mesiodenscontaining human DPSCs. To this end, the expression of lineage specific markers after differentiation induction was analyzed by lineage specific staining and RT-PCR. The nanoscale pattern arrayed surface showed apositive effect on the adipogenic differentiation of DPSCs. There was no difference between nanoscale pattern arrayed surface and conventional surface groups onosteogenic differentiation. In conclusion, the nanoscale ridge/groove pattern arrayed surface can be used to enhance the adipogenic differentiation of DPSCs derived from mesiodens. This finding provides an improved understanding of the effects of topography on cell differentiation as well as the potential use of supernumerary tooth in regenerative dental medicine.

• Journal of Dental Anesthesia and Pain Medicine
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• v.18 no.2
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• pp.105-110
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• 2018

Background: A stellate ganglion block (SGB) causes increased blood flow in the maxillofacial region, exhibiting the potential for regenerative effects in damaged tissue. The focus of this study was to understand the efficacy of SGB for regenerative effects against nerve damage. A rat model of the superior cervical ganglion block (SCGB) was created instead of SGB, and facial blood flow, as well as sympathetic nervous system function, were measured. Methods: A vertical incision was made on the left side of the neck of a Wistar rat, and a 5-mm resection of the superior cervical ganglion was performed at the back of the bifurcation of the internal and external branches of the left common carotid artery. Blood flow in the skin at the mandibular angle and mean facial temperature were measured using a laser-Doppler blood flow meter and a thermographic camera, respectively, over a 5-week period after the block. In addition, the degree of ptosis and miosis were assessed over a period of 6 months. Results: The SCGB rat showed significantly higher blood flow at the mandibular angle on the block side (P < 0.05) for 3 weeks, and significantly higher skin temperature (P < 0.05) for 1 week after the block. In the SCGB rat, ptosis and miosis occurred immediately after the block, and persisted even 6 months later. Conclusions: SCGB in rats can cause an increase in the blood flow that persists over 3 weeks.

• Archives of Plastic Surgery
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• v.48 no.5
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• pp.559-567
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• 2021

The potential to differentiate into different cell lines, added to the easy and cost-effective method of extraction, makes adipose-derived stem cells (ADSCs) an object of interest in lymphedema treatment. Our study's goal was to conduct a comprehensive systematic review of the use of ADSCs in lymphatic tissue engineering and regeneration. On July 23, 2019, using PubMed/MEDLINE, Cochrane Clinical Answers, Cochrane Central Register of Controlled Trials, and Embase databases, we conducted a systematic review of published literature on the use of ADSCs in lymphatic tissue engineering and regeneration. There were no language or time frame limitations, and the following search strategy was applied: ((Adipose stem cell) OR Adipose-derived stem cell)) AND ((Lymphedema) OR Breast Cancer Lymphedema). Only original research manuscripts were included. Fourteen studies fulfilled the inclusion criteria. Eleven studies were experimental (in vitro or in vivo in animals), and only three were clinical. Publications on the topic demonstrated that ADSCs promote lymphangiogenesis, and its effect could be enhanced by modulation of vascular endothelial growth factor-C, interleukin-7, prospero homeobox protein 1, and transforming growth factor-β1. Pilot clinical studies included 11 patients with breast cancer-related lymphedema, and no significant side effects were present at 12-month follow-up. Literature on the use of ADSCs in lymphatic tissue engineering and regeneration demonstrated promising data. Clinical evidence is still in its infancy, but the scientific community agrees that ADSCs can be useful in regenerative lymphangiogenesis. Data collected in this review indicate that unprecedented advances in lymphedema treatment can be anticipated in the upcoming years.

• Journal of Yeungnam Medical Science
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• v.39 no.4
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• pp.322-331
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• 2022

Background: The combined use of biomaterials for regeneration may have great biological relevance. This study aimed to compare the regenerative potential of biphasic calcium phosphate (BCP) alone and with growth factor enamel matrix derivatives (EMDs) for the regeneration of intrabony defects at 1 year. Methods: This randomized controlled trial included 40 sites in 29 patients with stage II/III periodontitis and 2/3 wall intrabony defects that were treated with BCP alone (control group) or a combination of BCP and EMD (test group). BCP alloplastic bone grafts provide better bio-absorbability and accelerate bone formation. EMDs are commercially available amelogenins. Mean values and standard deviations were calculated for the following parameters: plaque index (PI), papillary bleeding index (PBI), vertical probing pocket depth (V-PPD), vertical clinical attachment level (V-CAL), and radiographic defect depth (RDD). Student paired and unpaired t-tests were used to compare the data from baseline to 12 months for each group and between the groups, respectively. The results were considered statistically significant at p<0.05. Results: At 12 months, the PI and PBI scores of the control and test groups were not significantly different (p>0.05). The mean V-PPD difference, V-CAL gain, and RDD difference were statistically significant in both groups at 12 months (p<0.001 for all parameters). Intergroup comparisons showed that the mean V-PPD reduction (2.13±1.35 mm), V-CAL gain (2.53±1.2 mm), and RDD fill (1.33±1.0 mm) were statistically significant between the groups at 12 months (p<0.001 for all parameters). Conclusion: BCP and EMDs combination is a promising modality for the regeneration of intrabony defects.

• Journal of Society of Preventive Korean Medicine
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• v.5 no.2
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• pp.122-128
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• 2001

This study was carried out to evaluate antitoxic effects of Houttuynia cordata extract on cadmium by colorimetric methods. The antitoxic activity ofc in NIH 3T3 fibroblasts was evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) and SRB (sulforhodamine B protein) assays. The light microscopic study was carried out to observe morphological changes of the treated cells. The concentration of 10-2 mg/ml of Houttuynia cordata extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were antitoxic and tend to regenerate. These results suggest that the ethyl acetate extract of Houttuynia cordata retains a potential antitoxic activity.

• Journal of Veterinary Clinics
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• v.23 no.2
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• pp.197-201
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• 2006

A 2-year-old female mongrel dog was referred to the Veterinary Medical Center of Chungbuk National University due to anorexia, depression and weight loss after the administration of exogenous estrogen for contraception in the local hospital. Although the dog was medicated with antibiotics and fluid for two weeks in the local hospital, there was a deterioration in the patient's general condition. Therefore the dog was referred to our Veterinary Medical Center. Hematological and biochemical analysis showed non-regenerative anemia, thrombocytopenia, severely immature leukocytosis, hyperbilirubinemia, hyperglycemia and elevated serum alkaline phosphatase, gamma-glutamyltransferase, and lactate dehydrogenase. Abdominal ultrasonography showed double line of gallbladder and irregular hypertrophic wall of uterus. On the basis of these examination's results and history taking, final diagnosis was exogenous estrogen-induced myelotoxicosis with cholecystitis and endometritis. The symptomatic therapy for estrogen toxicosis was performed for 26 days after the final diagnosis. However, the symptom had no response to therapy except endometritis and anemia. On day 135 after administration of estrogen, platelet and neutrophil count were normalized, but monocytic leukocytosis was occurred.

• Molecules and Cells
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• v.46 no.4
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• pp.209-218
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• 2023

In induced pluripotent stem cells (iPSCs), pluripotency is induced artificially by introducing the transcription factors Oct4, Sox2, Klf4, and c-Myc. When a transgene is introduced using a viral vector, the transgene may be integrated into the host genome and cause a mutation and cancer. No integration occurs when an episomal vector is used, but this method has a limitation in that remnants of the virus or vector remain in the cell, which limits the use of such iPSCs in therapeutic applications. Chemical reprogramming, which relies on treatment with small-molecule compounds to induce pluripotency, can overcome this problem. In this method, reprogramming is induced according to the gene expression pattern of extra-embryonic endoderm (XEN) cells, which are used as an intermediate stage in pluripotency induction. Therefore, iPSCs can be induced only from established XEN cells. We induced XEN cells using small molecules that modulate a signaling pathway and affect epigenetic modifications, and devised a culture method which can produce homogeneous XEN cells. At least 4 passages were required to establish morphologically homogeneous chemically induced XEN (CiXEN) cells, whose properties were similar to those of XEN cells, as revealed through cellular and molecular characterization. Chemically iPSCs derived from CiXEN cells showed characteristics similar to those of mouse embryonic stem cells. Our results show that the homogeneity of CiXEN cells is critical for the efficient induction of pluripotency by chemicals.