• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.30 no.4
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• pp.308-315
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• 2004

Aim: The aim of this study was to investigate the mechanism of promoted skin wound healing in skin defects with primary cultured oral mucosal keratinocytes. Materials and methods: Thirty adult female nude mice weighing $20{\pm}2g$ were used for the experiment. Primary cultured and suspended oral mucosal keratinocytes, labeled with BrdU, were scattered onto $1.5cm{\times}1.5cm$ sized full thickness skin defects in the experimental group(N=15), and no grafts were placed the control group(N=15). They were sacrificed at 3 days, 1 week and 2 weeks after the treatment respectively. Histological examination of each wounds were performed to review the healing progress on measuring the length from the wound margin to regenerating epithelial front. The role of keratinocytes were assessed by double immunohistochemical staining with Anti-BrdU and Anti-cytokeratin AE1/3. Results: In the experimental group the wound was completely covered with regenerating epithelia in 2 weeks, but partially regenerated in the control group. The immunohistochemical studies unexpectedly reveal that most of regenerating epithelial cells were induced from marginal epithelium of the margin, not from the scattered keratinocytes. Conclusion: We could successfully confirm that graft of primary cultured oral mucosal keratinocytes promotes the regeneration of skin defects.

• Journal of Korean Neurosurgical Society
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• v.37 no.2
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• pp.129-136
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• 2005

Objective: Currently available therapies for human malignant gliomas have limited efficacy. Toxoplasma gondii, an obligate intracellular protozoan parasite, and Quil-A are nonspecific, potent immune stimulants. T. gondii is shown to have antitumor activity in some types of cancers. Therefore, this study is undertaken to evaluate the antitumor effect of Toxoplasma lysate antigen (TLA), alone or in combination with Quil-A, on human glioma U373MG and U87MG cells. Methods: The in vitro effects of TLA alone or in combination with Quil-A on the proliferation, invasion, and apoptosis of glioma cells were tested using MTT, Matrigel invasion, and DNA fragmentation assays, and the in vivo effects on the growth of gliomas were evaluated in athymic nude mice transplanted with glioma cells. Results: Treatment with TLA resulted in the suppressed proliferation and invasion of both U373MG and U87MG cells, in a dose-dependent manner. In addition, at high concentration, TLA induced glioma cell apoptosis. When TLA was administered in the mouse glioma model, malignant glioma growth was decreased. The combined treatment of TLA with Quil-A significantly inhibited the proliferation and invasion of cultured cells as well as tumor mass of implanted mice. Conclusion: TLA inhibits the proliferation and invasion of glioma cells in vitro and in vivo, and these antitumor effects of TLA are significantly enhanced by the addition of Quil-A.

• Biomedical Science Letters
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• v.19 no.4
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• pp.295-302
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• 2013

Some of the pituitary adenomas are invasive and spread into neighboring tissues. In previous studies, the invasion of pituitary adenomas is thought to be associated with epithelial-mesenchymal transition (EMT). In addition to that, we thought that mesenchymal stem cells (MSCs) exist in relevant microenvironment in pituitary adenoma. However, it has been little known about the existence of MSCs from pituitary adenoma. So we investigated whether mesenchymal stem-like cells (MSLCs) can be isolated from the pituitary adenoma specimen. We isolated and cultured candidate MSLCs from the fresh pituitary adenoma specimen with the same protocols used in culturing bone marrow derived MSCs (BM-MSCs). The cultured candidate MSLCs were analyzed by fluorescence-activated cell sorting (FACS) for surface markers associated with MSCs. Candidate MSLCs were exposed to mesenchymal differentiation conditions to determine the mesenchymal differentiation potential of these cells. To evaluate the tumorigenesis of candidate MSLCs from pituitary adenoma, we implanted these cells into the brain of athymic nude mice. We isolated cells resembling BM-MSCs named pituitary adenoma stroma mesenchymal stem-like cells (PAS-MSLCs). PAS-MSLCs were spindle shaped and had adherent characteristics. FACS analysis identified that the PAS-MSLCs had a bit similar surface markers to BM-MSCs. Isolated cells expressed surface antigen, positive for CD105, CD75, and negative for CD45, NG2, and CD90. We found that these cells were capable of differentiation into adipocytes, osteocytes and chondrocytes. Tumor was not developed in the nude mice brains that were implanted with the PAS-MSLCs. In this study, we showed that MSLCs can be isolated from a pituitary adenoma specimen which is not tumorigenic.

• Tuberculosis and Respiratory Diseases
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• v.58 no.2
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• pp.120-128
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• 2005

Background : Insulin-like growth factor binding protein (IGFBP)-3 regulates non-small cell lung cancer(NSCLC) cell proliferation in vitro and in vivo by inhibiting IGF-mediated signaling pathways. To have better strategies for the treatment of lung cancer, we analyzed the combining effects of adenovirus expressing IGFBP-3 (Ad5CMV-BP3) and SCH66336, a farnesyl transferase inhibitor (FTI) designed to block Ras-mediated proliferative signaling pathways. Methods : To measure the combining effects of Ad5CMV-BP3 and SCH66336 on the proliferation of NSCLC cells, human NSCLC cell lines (H1299, H596, A549, H460, and H358), SCH66336, recombinant adenovirus expressing IGFBP-3 (Ad5CMV-BP3) and athymic nude mice were used in these experiments. Results : The combination of Ad5CMV-BP3 and SCH66336 produced a synergistic enhancement in antiproliferative effects over a range of clinically achievable concentrations in a variety of NSCLC cell lines. Furthermore, we observed a significant reduction in growth of NSCLC xenograft induced in athymic nude mice. Conclusion : In conclusion, this study demonstrated for the first time that the FTI SCH66336 synergizes with IGFBP-3 and enhances its apoptotic activity in NSCLC cells in vitro and in vivo. The combined treatment of Ad5CMV-BP3 and SCH66336 raises the possibility of using this regimen in clinic for the treatment of NSCLC.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.35 no.6
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• pp.353-359
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• 2013

Purpose: Extensive research is actively ongoing for development of an ideal bone substitute that meets the gold standard. Tooth was selected as a donor site for evaluation of potentials in bone substitutes based on its similar chemical compositions to alveolar bone. Previous studies have evaluated inorganic components of autogenous tooth bone graft material (AutoBT) and osteoconductivity. In continuation from the previous studies, the current study was conducted for analysis of organic components and evaluation of osteoinductivity of AutoBT. Methods: Forty-six extracted teeth were collected from actual patients (Korea Tooth Bank, R&D Institute). Extracted teeth were processed into AutoBT and implanted in dorsal subcutaneous muscular tissues of 15 athymic mice. Biopsy samples were harvested at two, five, and eight weeks. The Bradford assay, sodium dodecyl sulphate polyacrylamide gradient gel, and western blotting were performed for investigation of organic contents of AutoBT. Results: Histology analyses showed signs of new bone formation as early as two weeks. Results of the Bradford assay indicated the existence of noncollagenous proteins (NCP). 0.29% (2.89 mg/g) of proteins were extracted by weight in the root portion of AutoBT; 0.02% (0.029 mg/g) and 1.79% (17.93 mg/g) of proteins were measured by weight in crown and block-form of AutoBT, respectively. However, recombinant human bone morphogenetic protein-2 was not observed in AutoBT. Conclusion: Within the limitation of the current study, AutoBT induced new bone formation by NCP embedded in dentin.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.356-358
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• 2003

The use of cultured epidermal sheet grafts for large full-thickness wound has been tempered by weak points such as long culture periods, difficulty in handling the fragile sheets, high costs and the detachment of the skin cells from the culture dishes by enzymatic digestion. To overcome the drawback of epidermal sheet grafts, we have developed a transplantation method to spray the cultured human keratinocytes with the mixtures of rhEGF and fibrin gel matrix to the full-thickness wounds on the dorsum of the athymic mice to regenerate epithelial layers. Wound biopsies were retrieved at 7, 14 and 21 days after transplantation and retrieved biopsies were analyzed by histology and immunohistochemistry. Transplanted keratinocytes and EGF-fibriin gel accelerated wound regeneration compared with control groups. The technique developed in this study overcomes the drawbacks of the current cultured epidermal sheet grafts and accelerates epidermal wound healing.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.259-264
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• 2005

Transplantation of cultured chondrocytes can regenerate cartilage tissues in cartilage defects in humans. However, this method requires a long culture period to expand chondrocytes to a large number of cells for transplantation. In addition, chondrocytes may dedifferentiate during long-term culture. These problems can potentially be overcome by the use of undifferentiated or partially developed cartilage precursor cells derived from neonatal cartilage, which, unlike chondrocytes from adult cartilage, have the capacity for rapid in vitro cell expansion and may retain their differentiated phenotype during long-term culture. The purpose of this study was to compare the cell growth rate and phenotypic modulation during in vitro culture between adult chondrocytes and neonatal chondrocytes, and to demonstrate the feasibility of regenerating cartilage tissues in vivo by transplantation of neonatal chondrocytes expanded in vitro and seeded onto polymer scaffolds. When cultured in vitro, chondrocytes isolated from neonatal (immediately postpartum, 2 h of age) rats exhibited much higher growth rate than chondrocytes isolated from adult rats. After 5 days of culture, more neonatal chondrocytes were in the differentiated state than adult chondrocytes. Cultured neonatal chondrocytes were seeded onto biodegradable polymer scaffolds and transplanted into athymic mice's subcutaneous sites. Four weeks after implantation, neonatal chondrocyte-seeded scaffolds formed white cartilaginous tissues. Histological analysis of the implants with hematoxylin and eosin showed mature and well-formed cartilage. Alcian blue/ safranin-O staining and Masson's trichrome staining indicated the presence of highly sulfated glycosarninoglycans and collagen, respectively, both of which are the major extracellular matrices of cartilage. Immunohistochemical analysis showed that the collagen was mainly type II, the major collagen type in cartilage. These results showed that neonatal chondrocytes have potential to be a cell source for cartilage tissue engineering.

• Korean Journal of Head & Neck Oncology
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• v.12 no.2
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• pp.181-187
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• 1996

We have characterized 4 human squamous carcinoma cell lines established from the larynx and hypopharynx area. All the cell lines were cultured in RPMI-1640 medium. During the growth they showed monolayer adherence pattern in culture flask. They showed tonofilament on transmission electromicroscopy which is characteristic of squamous cell epithelium. DNA finger-printing using Hinf-l proved them to be originated from different beings. Flow cytometric analysis revealed them to show aneuploidy. Immunohistochemical staining for cytokeratin was done using CK1, CK8.13, CK19 and CAM5.2 antibody, and produced various patterns of positivity. All the cell lines showed varying degrees of tumorigenecity in athymic nude mice when injected subcutaneously, but only heterotransplanted SNU-1041 cell line showed continuous tumor growth. Histopathologic findings of the heterotransplanted tumors were identical to those of the original tumors of patients. This study suggests that establishment of many different squamous cell lines might bestow great capability in researches of the head and neck cancer.

• Archives of Plastic Surgery
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• v.44 no.5
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• pp.361-369
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• 2017

As the popularity of fat grafting research increases, animal models are being used as the source of pre-clinical experimental information for discovery and to enhance techniques. To date, animal models used in this research have not been compared to provide a standardized model. We analyzed publications from 1968-2015 to compare published accounts of animal models in fat grafting research. Data collected included: species used, graft characteristics (donor tissue, recipient area, amount injected, injection technique), time of sacrifice and quantification methods. Mice were most commonly used (56% of studies), with the "athymic nude" strain utilized most frequently (44%). Autologous fat was the most common source of grafted tissue (52%). Subcutaneous dorsum was the most common recipient site (51%). On average, $0.80{\pm}0.60mL$ of fat was grafted. A single bolus technique was used in 57% of studies. Fat volume assessment was typically completed at the end of the study, occurring at less than 1 week to one year. Graft volume was quantified by weight (63%), usually in conjunction with another analysis. The results demonstrate the current heterogeneity of animal models in this research. We propose that the research community reach a consensus to allow better comparison of techniques and results. One example is the model used in our laboratory and others; this model is described in detail. Eventually, larger animal models may better translate to the human condition but, given increased financial costs and animal facility capability, should be explored when data obtained from small animal studies is exhausted or inconclusive.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.5
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• pp.2009-2014
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• 2012

Objective: Taspine, isolated from Radix et Rhizoma Leonticis has demosntrated potential proctiective effects against cancer. Tas13D, a novel taspine derivative synthetized by structure-based drug design, have been shown to possess interesting biological and pharmacological activities. The current study was designed to evaluate its antiproliferative activity and underlying mechanisms. Methods: Antiproliferative activity of tas13D was evaluated by xenograft in athymic mice in vivo, and by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and cell migration assays with human liver cancer (SMMC-7721) cell lines in vitro. Docking between tas13D and VEGFR and EGFR was studied by with a Sybyl/Surflex module. VEGF and EGF and their receptor expression was determined by ELISA and real-time PCR methods, respectively. Results: Our present study showed that tas13D inhibited SMMC-7721 xenograft tumor growth, bound tightly with the active site of kinase domains of EGFR and VEGFR, and reduced SMMC-7721 cell proliferation (IC=34.7 ${\mu}mol/L$) and migration compared to negative controls. VEGF and EGF mRNAs were significantly reduced by tas13D treatment in a dose-dependent manner, along with VEGF and EGF production. Conclusion: The obtained results suggest that tas13D inhibits tumor growth and cell proliferation by inhibiting cell migration, downregulating mRNA expression of VEGF and EGF, and decreasing angiogenic factor production. Tas13D deserves further consideration as a chemotherapeutic agent.