• Title/Summary/Keyword: Cell- and tissue-based therapy

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Proteomic Analysis of the Hydrophobic Fraction of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood

  • Jeong, Ju Ah;Lee, Yoon;Lee, Woobok;Jung, Sangwon;Lee, Dong-Seong;Jeong, Namcheol;Lee, Hyun Soo;Bae, Yongsoo;Jeon, Choon-Ju;Kim, Hoeon
    • Molecules and Cells
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    • v.22 no.1
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    • pp.36-43
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    • 2006
  • Mesenchymal stem cells (MSCs) are promising candidates for cell therapy and tissue engineering, but their application has been impeded by lack of knowledge of their core biological properties. In order to identify MSC-specific proteins, the hydrophobic protein fraction was individually prepared from two different umbilical cord blood (UCB)-derived MSC populations; these were then subjected to two-dimensional (2D) gel electrophoresis and peptide mass fingerprinting matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF)-mass spectrometry (MS). Although the 2D gel patterns differed somewhat between the two samples, computer-assisted image analysis identified shared protein spots. 35 spots were reliably identified corresponding to 32 different proteins, many of which were chaperones. Based on their primary sub-cellular locations the proteins could be grouped into 6 categories: extracellular, cell surface, endoplasmic reticular, mitochondrial, cytoplasmic and cytoskeletal proteins. This map of the water-insoluble proteome may provide valuable insights into the biology of the cell surface and other compartments of human MSCs.

Cell-Specific Targeting of Texas Red with Anti-Ep-CAM Antibody

  • Lee, Soo-Chul;Tae, Gun-Sik
    • Journal of Photoscience
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    • v.12 no.3
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    • pp.123-127
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    • 2005
  • The polyclonal antibody was generated against the peptide fragment of 62 amino acid residues (D 181-T242) near the COOH-terminal region of the extracellular domain of epithelial-cell adhesion molecule (Ep-CAM) and shown to be able to recognize Ep-CAM in competitive ELISA. Then, sulforhodamine 101 acid chloride (so called Texas red), a fluorescence dye, was conjugated to the affinity-purified anti-Ep-CAM antibody utilizing the reaction between the aliphatic amines of antibody and the sulfonyl chloride of Texas red. The molar ratio of Texas red to antibody was estimated to be approximately 1.86 by measuring optical densities at 280 nm and 596 nm, implying that the two molecules of Texas red at most were conjugated to antibody. The anti-Ep-CAM antibody-Texas red conjugate was then used for immunohistochemistry of CT-26 murine colon carcinoma cells. Based upon the fluorescence microscope images, anti-Ep-CAM antibody is able to deliver Texas red specifically to the surface of CT-26 cells on which Ep-CAM was actively expressed. This result indicates that anti-Ep-CAM antibody could be useful for the tissue-specific delivery of photosensitizers via antigen-antibody interaction.

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RECONSTRUCTION OF MIDFACIAL DEFECTS UTILIZING RECTUS ABDOMINIS FREE FLAP : REPORT OF 2 CASES (유리 복직근 피판을 이용한 중안모 결손부 재건의 2 치험례)

  • Lee, Seong-Geun;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.22 no.1
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    • pp.98-104
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    • 2000
  • Expansion in the scope and technique of head and neck tumor resection during the past two decades has paralleled precise tumor localization with advanced radiographic imaging and the availability of microvascular free tissue transfer. Especially, the defect reconstruction utilizing free flap results in improvement of patient survival due to decrease of local recurrence by wide resection of cancer. The rectus abdominis free flap has been used widely in reconstruction of the breast and extremities. However, the report of cases on its applications in the head and neck, based on the deep inferior epigastric artery and vein, is rare. This flap is one of the most versatile soft-tissue flaps. The deep inferior epigastric artery and vein are long and large-diameter vessels that are ideal for microvascular anastomosis. The skin area that can be transferred is probably the largest of all flaps presently in use. The versatility of the donor site is due to the ability to transfer large areas of skin with various thickness and amounts of underlying muscle. This article is to report reconstruction of midface defects utilizing the rectus abdominis free flap in 2 patients with maxillary squamous cell carcinoma and discuss briefly considerations in flap design and orbital exenteration, and healing of irradiated recipient site by hyperbaric oxygen therapy with literature review.

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Effects of Adenoviral Gene Transduction on the Stemness of Human Bone Marrow Mesenchymal Stem Cells

  • Marasini, Subash;Chang, Da-Young;Jung, Jin-Hwa;Lee, Su-Jung;Cha, Hye Lim;Suh-Kim, Haeyoung;Kim, Sung-Soo
    • Molecules and Cells
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    • v.40 no.8
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    • pp.598-605
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    • 2017
  • Human mesenchymal stem cells (MSCs) are currently being evaluated as a cell-based therapy for tissue injury and degenerative diseases. Recently, several methods have been suggested to further enhance the therapeutic functions of MSCs, including genetic modifications with tissue- and/or diseasespecific genes. The objective of this study was to examine the efficiency and stability of transduction using an adenoviral vector in human MSCs. Additionally, we aimed to assess the effects of transduction on the proliferation and multipotency of MSCs. The results indicate that MSCs can be transduced by adenoviruses in vitro, but high viral titers are necessary to achieve high efficiency. In addition, transduction at a higher multiplicity of infection (MOI) was associated with attenuated proliferation and senescence-like morphology. Furthermore, transduced MSCs showed a diminished capacity for adipogenic differentiation while retaining their potential to differentiate into osteocytes and chondrocytes. This work could contribute significantly to clinical trials of MSCs modified with therapeutic genes.

A CLINICAL STUDY ON SUPERIORLY BASED PLATYSMA MYOCUTANEOUS CERVICAL FLAP FOR RECONSTRUCTION FOLLOWING INTRAORAL SOFT TISSUE CANCER SURGERY (구강내 연조직 암 절제후 상부기조 광경근 근피부 경부 피판을 이용한 구강내 재건에 관한 임상적 연구)

  • Park, Bong-Wook;Byun, June-Ho;Shin, Hee-Suk;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.30 no.1
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    • pp.83-91
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    • 2008
  • The goal of reconstruction following ablative therapy for intraoral cancer is the restoration of form and function to permit a return to activities of daily life. Traditional reconstruction includes split thickness skin grafts, myocutaneous flaps and, more recently, various free flaps. Free flaps provide higher level of functional recovery relative to that seen with other techniques but require the complexity of the technique and microvascular anastomosis and thus, extended surgical time and occasionally a second team for harvesting. The platysma myocutaneous cervical flap is a possible alternative for intraoral reconstruction. It is thin and pliable like the tissue provided by the radial forearm free flap. It can be harvested with enough tissue to close most head and neck ablative defects. There is virtually no donor site morbidity involved. This study evaluated 7 patients affected by intraoral squamous cell carcinoma (SCC). All patients underwent the resection of intraoral SCC with neck dissection and subsequent intraoral reconstruction with the superiorly based platysma myocutaneous cervical flap. Flap-related complications occurred in 3 patients. Adjuvant radiation therapy was performed in 3 patients. Average follow-up was 24.1 months after surgery, with a range of 8 to 42 months. All patients presented self assessment of discomfort associated with intraoral recipient sites and cervical donor sites. However, the neck function measured by two-inclinometer technique was within the normal range during relatively long term follow-up period. Our study concluded that superiorly based platysma myocutaneous cervical flap is good alternative to free flaps, especially for relatively smaller defects and for the defects appropriate for the rotation arc of the flap.

Preparation of Microspheres Encapsulating a Recombinant TIMP-1 Adenovirus and their Inhibition of Proliferation of Hepatocellular Carcinoma Cells

  • Xia, Dong;Yao, Hui;Liu, Qing;Xu, Liang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6363-6368
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    • 2012
  • Objective: The study aim was to prepare poly-DL-lactide-poly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 (TIMP-1) in an adenovirus to investigate its inhibition on the proliferation of hepatocellular carcinoma cells HepG2. Methods: Microspheres were prepared by encapsulating the recombinant TIMP-1 adenovirus into biodegradable PELA. The particle size, viral load, encapsulation efficiency and in-vitro release were measured. Microspheres were used to infect HepG2 cells, then infection efficiency was examined under a fluorescent microscope and ultrastructural changes assessed by TEM. Expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR and proliferation by MTT and cell growth curve assays. Results: We successfully prepared microspheres encapsulating recombinant TIMP-1 adenovirus with a diameter of $1.965{\mu}m$, an encapsulation efficiency of 60.0%, a viral load of $10.5{\times}10^8/mg$ and approximate 60% of virus release within 120 h, the total releasing time of which was longer than 240 h. The microspheres were confirmed to be non-toxic with blank microspheres. Infected HepG2 cells could stably maintain in-vitro expression of TIMP-1, with significantly effects on biological behaviour Conclusion: PELA microspheres encapsulating a recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for polymer/chemistry-based gene therapy of hepatocellular carcinomas.

A study of the effects of PDGF-BB on the characteristics of bone stromal and periodontal ligament cells (혈소판유래성장인자-BB가 골간질세포와 치주인대세포의 성상에 미치는 영향)

  • Kwon, Young-Hyuk;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.949-965
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    • 1996
  • The main goal of periodontal therapy is to restore the lost periodontal tissue and establish the attachment appratus. Current acceptable therapeutic techniques are included : removal of diseased soft tissue, demineralization of exposed root surface, using the barrier membrane for preventing the downgrowth of gingival epithelial cell, insertion of graft materials as a scaffolding action, and biological mediators for promoting the cell activity. The latest concept one among them has been studied which based on the knowledge of cellular biology of destructed tissue. Platelet-derived growth factor(PDGF) is one of the polypeptide growth factor which have been reported as a biological mediator to regulate activities of wound healing progress including cell proliferation, migration, and metabolism. The purposes of this study is to evaluate the influences of the PDGF as biological mediator to periodontal ligament and bone marrow cell. Both right and left maxillary first molar were extracted from rat which had treated with 0.4% ${\beta}-Aminopropionitril$ for 5 days, and feeded until designed date to sacrifice under anesthesisa. Periodontal ligament were removed from the extracted socket of the rat, and cultured with Dulbecco's Modified Essential Medium(DMEM) contained with 10% Fetal Bovine Serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. Bone marrow cell were culture from bone marrow suspension with which washed out from femur with same medium. The study was performed to evaluate the effect of PDGF to periodontal ligament and bone cell, cell proliferation rate, total protein synthesis, and alkaline phosphatase activity of rat periodontal ligament(PDL) cell and bone stromal(RBS) cell in vitro. The effects of growth factors on both cells were measured at 3, 5th day after cell culture with (control group) or without growth factors(experimental group). The results were as follows: 1. The tendency of cell proliferation under the influence of PDGF showed more rapid proliferation pattern than control at 3 and 5 days after inoculation. 2. The activity of Alkaline phosphatase revealed 14, 80% increased respectively at 3, 5 days culture than control group. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(P<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P<0.05). 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group l(PDL 100%), 2(PDL 70%:GF 30%), and 3(PDL 50%:GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30%:GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05). From the above results, it is assumed that the PDGF on PDL cells and RMB cell culture. GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density. In conclusion, platelet-derived growth factor can promote rapid osteogenesis during early stage of periodontal tissue regeneration.

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Correlation between EGFR Gene Mutations and Lung Cancer: a Hospital-Based Study

  • Kavitha, Matam;Iravathy, Goud;Adi Maha, Lakshmi M;Ravi, V;Sridhar, K;Vijayanand, Reddy P;Chakravarthy, Srinivas;Prasad, SVSS;Tabassum, Shaik Nazia;Shaik, Noor Ahmad;Syed, Rabbani;Alharbi, Khalid Khalaf;Khan, Imran Ali
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7071-7076
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    • 2015
  • Epidermal growth factor receptor (EGFR) is one of the targeted molecular markers in many cancers including lung malignancies. Gefitinib and erlotinib are two available therapeutics that act as specific inhibitors of tyrosine kinase (TK) domains. We performed a case-control study with formalin-fixed paraffin-embedded tissue blocks (FFPE) from tissue biopsies of 167 non-small cell lung carcinoma (NSCLC) patients and 167 healthy controls. The tissue biopsies were studied for mutations in exons 18-21 of the EGFR gene. This study was performed using PCR followed by DNA sequencing. We identified 63 mutations in 33 men and 30 women. Mutations were detected in exon 19 (delE746-A750, delE746-T751, delL747-E749, delL747-P753, delL747-T751) in 32 patients, exon 20 (S786I, T790M) in 16, and exon 21 (L858R) in 15. No mutations were observed in exon 18. The 63 patients with EFGR mutations were considered for upfront therapy with oral tyrosine kinase inhibitor (TKI) drugs and have responded well to therapy over the last 15 months. The control patients had no mutations in any of the exons studied. The advent of EGFR TKI therapy has provided a powerful new treatment modality for patients diagnosed with NSCLC. The study emphasizes the frequency of EGFR mutations in NSCLC patients and its role as an important predictive marker for response to oral TKI in the south Indian population.

The Effects of NEES on PARP Expression and Cell Death in Rat Cerebral Cortex After Ischemic Injury

  • Kim, Sung-Won;Lee, Jung-Sook;Um, Ki-Mai;Kim, Ji-Sung;Lee, Suk-Hee;Choi, Yoo-Rim;Kim, Nyeon-Jun;Kim, Bo-Kyoung;Cho, Mi-Suk;Park, Joo-Hyun;Kim, Soon-Hee
    • Journal of International Academy of Physical Therapy Research
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    • v.1 no.2
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    • pp.107-112
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    • 2010
  • The majority of strokes are caused by ischemia and result in brain tissue damage, leading to problems of the central nervous system including hemiparesis, dysfunction of language and consciousness, and dysfunction of perception. The purpose of this study was to investigate the effects of Poly(ADP-ribose) polymerase(PARP) on necrosis in neuronal cells that have undergone needle electrode electrical stimulation(NEES) prior to induction of ischemia. Ischemia was induced in male SD rats(body weight 300g) by occlusion of the common carotid artery for 5 min, after which the blood was reperfused. After induction of brain ischemia, NEES was applied to Zusanli(ST 36), at 12, 24 and 48 hours. Protein expression was investigated using immuno-reactive cells, which react to PARP antibodies in cerebral nerve cells, and Western blotting. The results were as follows: In the cerebral cortex, the number of PARP reactive cells after 24 hours significantly decreased(p<.05) in the NEES group compared to the GI group. PARP expression after 24 hours significantly decreased(p<.05) in the NEES group compared to the GI group. As a result, NEES showed the greatest effect on necrosis-related PARP immuno-reactive cells 24 hours after ischemia, indicating necrosis inhibition, blocking of neural cell death, and protection of neural cells. Based on the results of this study, NEES can be an effective method of treating dysfunction and improving function of neuronal cells in brain damage caused by ischemia.

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Biodistribution and PET imaging of [18F]FMISO in mousecolon cancer xenografted mice

  • Seelam, Sudhakara Reddy;Lee, Ji Youn;Kim, Young Joo;Lee, Yun-Sang;Jeong, Jae Min
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.1 no.2
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    • pp.137-144
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    • 2015
  • Hypoxia is an important adverse prognostic factor for tumor progression and is a major cause of failure of radiation therapy. In case of short-term hypoxia, the metabolism can recover to normal, but if hypoxia persists, it causes irreversible cell damage and finally leads to death. So a hypoxia marker would be very useful in oncology. In particular, 2-nitroimidazole can be reduced to form a reactive chemical species, which can bind irreversibly to cell components in the absence of sufficient oxygen, thus, the development of radiolabeled nitroimidazole derivatives for the imaging of hypoxia remains an active field of research to improve cancer therapy result. 2-nitroimidazole based hypoxia marker, [$^{18}F$]FMISO holds promise for the evaluation of tumor hypoxia by Positron emission tomography (PET), at both global and local levels. In the present study, [$^{18}F$]FMISO was synthesized using an automatic synthesis module with high radiochemical purity (>99%) in 60 min. Immunohistochemical analysis using pimonidazole confirmed the presence of hypoxia in xenografted CT-26 tumor tissue. A biodistribution study in CT-26 xenografted mice showed that the increased tumor-to-muscle ratio and tumor-to-blood ratios from 10 to 120 min post-injection. In the PET study, [$^{18}F$]FMISO also showed increased tumor-to-muscle ratios from 10 to 120 min post-injection. In conclusion, this study demonstrates the feasibility and utility of [$^{18}F$]FMISO for imaging hypoxiain mouse colon cancer model using small animal PET.