• Title/Summary/Keyword: mesenchymal cell

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The Candidate Tumor Suppressor Gene SLC8A2 Inhibits Invasion, Angiogenesis and Growth of Glioblastoma

  • Qu, Mingqi;Yu, Ju;Liu, Hongyuan;Ren, Ying;Ma, Chunxiao;Bu, Xingyao;Lan, Qing
    • Molecules and Cells
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    • v.40 no.10
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    • pp.761-772
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    • 2017
  • Glioblastoma is the most frequent and most aggressive brain tumor in adults. Solute carrier family 8 member 2 (SLC8A2) is only expressed in normal brain, but not present in other human normal tissues or in gliomas. Therefore, we hypothesized that SLC8A2 might be a glioma tumor suppressor gene and detected the role of SLC8A2 in glioblastoma and explored the underlying molecular mechanism. The glioblastoma U87MG cells stably transfected with the lentivirus plasmid containg SLC8A2 (U87MG-SLC8A2) and negative control (U87MG-NC) were constructed. In the present study, we found that the tumorigenicity of U87MG in nude mice was totally inhibited by SLC8A2. Overexpression of SLC8A2 had no effect on cell proliferation or cell cycle, but impaired the invasion and migration of U87MG cells, most likely through inactivating the extracellular signal-related kinases (ERK)1/2 signaling pathway, inhibiting the nuclear translocation and DNA binding activity of nuclear factor kappa B ($NF-{\kappa}B$), reducing the level of matrix metalloproteinases (MMPs) and urokinase-type plasminogen activator (uPA)-its receptor (uPAR) system (ERK1/2-$NF-{\kappa}B$-MMPs/uPA-uPAR), and altering the protein levels of epithelial to mesenchymal transitions (EMT)-associated proteins E-cardherin, vimentin and Snail. In addition, SLC8A2 inhibited the angiogenesis of U87MG cells, probably through combined inhibition of endothelium-dependent and endothelium-nondependent angiogenesis (vascular mimicry pattern). Totally, SLC8A2 serves as a tumor suppressor gene and inhibits invasion, angiogenesis and growth of glioblastoma.

Clinical use of Platelet Rich Fibrin(PRF) in Various Hard and Soft Tissue Defect : Case Report (혈소판 유래 섬유소(Platelet Rich Fibrin(PRF))를 이용한 골결손부의 치료 : 증례보고)

  • Chee, Young-Deok;Go, Seo-Wook
    • Journal of Dental Rehabilitation and Applied Science
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    • v.23 no.4
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    • pp.303-312
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    • 2007
  • Recently several studies have been developed not only to apply bone materials to bony defect, but also to use osteogenic and osteoinductive materials to form bone more effectively. In 1998 Mark et al applied gel formation of PRP(platelet-rich plasma) in bony transplantation for mandibular reconstruction as one of the method of stimulating bone formation in maxillofacial area, which is contain of varies growth factors. After he reported that PRP accelerate bone formation, which is used in varies bone transplantation and augmentation with a good result. Especially there are amount of growth factors in PRP, and PRP increase angiogenesis, cell division, and mesenchymal cell growth. Moreover it is capable of osteoconduction, hemostatitis, anti-infection, forming the shape at transplantation, ease of handling, and recipient site stability. So it is known that success rate is high in bone transplantation. However PRP need tissue adhesive to make plasma to solid form. Thrombin and calcium chloride, component of PRP, is extracted from autogenic donor. So it is expensive to extract and there is possibility of hepatitis, AIDS, and hematogenous metastasis. After all, tissue adhesive have the limitation and danger of use. So we are willing to introduce that we had get some idea after using PRF(platelet-rich fibrin) in the various hard and soft tissue bony defect, which is self extracted simply and contain growth factors.

Osteogenic Gene Expression on Anodizing Titanium Surface (양극산화 처리된 타이타늄 표면에서 골형성 유전자 발현)

  • Kim, Won-Seok;Kim, Young-Seok;Jeon, Seong-Bae;Jun, Sang-Ho;Lee, Eui-Suk;Jang, Hyon-Seok;Kwon, Jong-Jin;Rim, Jae-Suk
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.34 no.2
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    • pp.91-99
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    • 2012
  • Purpose: The purpose of this study was to evaluate the expression of osteogenic genes associated with bone regeneration on anodizing titanium surface. Methods: $20{\times}20{\times}1$ (mm) commercially pure titanium plate was made, one group was pure titanium, second group was punched, and last group was punched and anodized by electrochemical method. Through the osteogenic cell culture model, the expression of extracellular matrix proteins, such as bone morphogenetic protein-2, bone sialoprotein, aggrecan, osteocalcin, Alkaline phosphatase, collagen I had been evaluated by Real-time polymerase chain reaction, and the morphology of growing cells was evaluated by scanning electron microscopy. Results: The attachment of mesenchymal stem cell was even and well-oriented on all Ti surfaces. The osteogene expression was increased on punching groups but, decreased on anodizing surfaces in 3 week samples. Conclusion: Punched anodizing Ti has possibility be using as a dental implant material, but further in vivo study would be needed.

CLINICOPATHOLOGIC AND IMMUNOHISTOCHEMICAL STUDY IN PLEOMORPHIC ADENOMA (다형성 선종의 임상병리 및 면역조직화학적 연구)

  • Kim, Kyung-Wook;Han, Se-Jin;Lee, Sang-Gu
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.4
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    • pp.384-390
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    • 2006
  • Pleomorphic adenoma is the most common salivary benign tumor, constituting over 60% of parotid gland tumor, 25% of sublingual gland tumor and 50% of minor salivary gland tumor. It is somewhat more frequent in the fourth to sixth decades. The recurrent rate which enforces only a simple enucleation is very high $(20{\sim}45%)$. Histologically, it contains the epithelial cell, the myo-epithelial cell and mesenchymal ingredient, which is various aspect. We analyzed clinicopathologically and immunohistochemically the patients(34 cases) who are diagnosed with pleomorphic adenoma in Dept. of Oral & Maxillofacial Surgery, College of Dentisty, Dankook university since 1998. The results are as follow: 1. The incidence of the tumor was most frequent in age 30 to 50. The ratio of male to female was 1:1.43. 2. The most chief complain was a painless mass(94.1%) and the duration time was more than decade in 18 cases(52.9%). 3. Palate(soft & hard palate) was the most occurred site(64.7%). In major salivary glands, the parotid gland was the most frequent site(17.6%). 4. The tumor size was 2 to 3cm on the average. Most of tumors were with capsule(91.2%). 5. Surgical excision was a main treatment method(20 cases, 58.8%) and 14 cases were excised with a glandectomy, 1 case was treated with a partial maxillectomy. Only 1 case of all cases was recurred. 6. Histopathologically, 9 cases(26.5%) were cellular type, 11 cases(32.4%) were intermediate(classic) type and 14 cases(41.1%) were myxoid type. 7. Immunohistochemically, the specimen of all tumors reacted positively to cytokeratin and vimentin marker.

ANTI-INFLAMMATORY EFFECTS OF PPARγ ON HUMAN DENTAL PULP CELLS (치수세포에서 PPARγ의 항 염증작용에 관한 연구)

  • Kim, Jeong-Hee
    • Restorative Dentistry and Endodontics
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    • v.31 no.3
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    • pp.203-214
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    • 2006
  • Dental pulp is a loose, mesenchymal tissue almost entirely enclosed in the dentin. It consists of cells, ground substance, and neural and vascular supplies. Damage to the dental pulp by mechanical, chemical, thermal, and microbial irritants can provoke various types of inflammatory response. Pulpal inflammation leads to the tissue degradation, which is mediated in part by Matrix metalloproteinase leads to accelerate extracellular matrix degradation with pathological pathway We have now investigated the induction of MMPs and inflammatory cytokines by Lipopolysaccharide (LPS) control of inflammatory mediators by peroxisome proliferator-activated receptors (PPARs). Human dental pulp cells exposed to various concentrations of LPS ($1-10{\mu}g/ml$) revealed elevated levels of MMP-2 and MMP-9 at 24 hrs of culture. LPS also stimulated the production of ICAM-1, VCAM-1, $IL-1{\beta},\;and\;TNF-{\alpha}$. Adenovirus $PPAR{\gamma}\;(Ad/PPAR{\gamma})\;and\;PPAR{\gamma}$ agonist rosiglitazone reduced the synthesis of MMPs, adhesion molecules and pro-inflammatory cytokines. The inhibitory effect of $Ad/PPAR{\gamma}$ was higher than that of $PPAR{\gamma}$ agonist. These result offer new insights in regard to the anti-inflammatory potential of $PPAR{\gamma}$ in human dental pulp cell.

Development and Evaluation of Hyaluronic Acid-Based Hybrid Bio-Ink for Tissue Regeneration

  • Lee, Jaeyeon;Lee, Se-Hwan;Kim, Byung Soo;Cho, Young-Sam;Park, Yongdoo
    • Tissue Engineering and Regenerative Medicine
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    • v.15 no.6
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    • pp.761-769
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    • 2018
  • BACKGROUND: Bioprinting has recently appeared as a powerful tool for building complex tissue and organ structures. However, the application of bioprinting to regenerative medicine has limitations, due to the restricted choices of bio-ink for cytocompatible cell encapsulation and the integrity of the fabricated structures. METHODS: In this study, we developed hybrid bio-inks based on acrylated hyaluronic acid (HA) for immobilizing bio-active peptides and tyramine-conjugated hyaluronic acids for fast gelation. RESULTS: Conventional acrylated HA-based hydrogels have a gelation time of more than 30 min, whereas hybrid bio-ink has been rapidly gelated within 200 s. Fibroblast cells cultured in this hybrid bio-ink up to 7 days showed >90% viability. As a guidance cue for stem cell differentiation, we immobilized four different bio-active peptides: BMP-7-derived peptides (BMP-7D) and osteopontin for osteogenesis, and substance-P (SP) and Ac-SDKP (SDKP) for angiogenesis. Mesenchymal stem cells cultured in these hybrid bio-inks showed the highest angiogenic and osteogenic activity cultured in bio-ink immobilized with a SP or BMP-7D peptide. This bio-ink was loaded in a three-dimensional (3D) bioprinting device showing reproducible printing features. CONCLUSION: We have developed bio-inks that combine biochemical and mechanical cues. Biochemical cues were able to regulate differentiation of cells, and mechanical cues enabled printing structuring. This multi-functional bio-ink can be used for complex tissue engineering and regenerative medicine.

LINC00174 Facilitates Proliferation and Migration of Colorectal Cancer Cells via MiR-3127-5p/ E2F7 Axis

  • Ma, Yuhong;Li, Yuzhen;Tang, Yuanyuan;Tang, Ning;Wang, Dengke;Li, Xiaofei
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1098-1108
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    • 2021
  • The literature indicates that LINC00174 promotes the growth of colorectal cancer (CRC) cells, but its research needs to be enriched. We tried to explore the function and mechanism of LINC00174 in CRC cell proliferation and migration. Bioinformatics analysis predicted the binding relationship and expressions of lncRNA, miRNA and mRNA. Clinical study analyzes the relationship between LINC00174 and clinical data characteristics of CRC patients. The expressions of LINC00174, miR-3127-5p and E2F7 were verified by RT-qPCR, and the combination of the two was verified by dual luciferase analysis and RNA immunoprecipitation as needed. Western blot was used to detect the expression of EMT-related protein and E2F7 protein. Functional experiments were used to evaluate the function of the target gene on CRC cells. LINC00174 was up-regulated in CRC clinical samples and cells and was related to the clinical characteristics of CRC patients. High-expression of LINC00174, contrary to the effect of siLINC00174, promoted cell viability, proliferation, migration and invasion, up-regulated the expressions of N-Cadherin, Vimentin, E2F7, and inhibited the expression of E-Cadherin. MiR-3127-5p was one of the targeted miRNAs of LINC00174 and was down-regulated in CRC samples. In addition, miR-3127-5p mimic partially reversed the malignant phenotype of CRC cells induced by LINC00174. Besides, E2F7 was a target gene of miR-3127-5p, and LINC00174 repressed miR-3127-5p to regulate E2F7. Our research reveals that LINC00174 affected the biological characteristics of CRC cells through regulated miR-3127-5p/ E2F7 axis.

Repopulation of autophagy-deficient stromal cells with autophagy-intact cells after repeated breeding in uterine mesenchyme-specific Atg7 knockout mice

  • Ji-Eun Oh;Sojung Kwon;Hyunji Byun;Haengseok Song;Hyunjung Jade Lim
    • Clinical and Experimental Reproductive Medicine
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    • v.50 no.3
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    • pp.170-176
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    • 2023
  • Objective: Autophagy is highly active in ovariectomized mice experiencing hormone deprivation, especially in the uterine mesenchyme. Autophagy is responsible for the turnover of vasoactive factors in the uterus, which was demonstrated in anti-Müllerian hormone receptor type 2 receptor (Amhr2)-Cre-driven autophagy-related gene 7 (Atg7) knockout (Amhr-Cre/Atg7f/f mice). In that study, we uncovered a striking difference in the amount of sequestosome 1 (SQSTM1) accumulation between virgin mice and breeder mice with the same genotype. Herein, we aimed to determine whether repeated breeding changed the composition of mesenchymal cell populations in the uterine stroma. Methods: All female mice used in this study were of the same genotype. Atg7 was deleted by Amhr2 promoter-driven Cre recombinase in the uterine stroma and myometrium, except for a triangular stromal region on the mesometrial side. Amhr-Cre/Atg7f/f female mice were divided into two groups: virgin mice with no mating history and aged between 11 and 12 months, and breeder mice with at least 6-month breeding cycles with multiple pregnancies and aged around 12 months. The uteri were used for Western blotting and immunofluorescence staining. Results: SQSTM1 accumulation, representing Atg7 deletion and halted autophagy, was much higher in virgin mice than in breeders. Breeders showed reduced accumulation of several vasoconstrictive factors, which are potential autophagy targets, in the uterus, suggesting that the uterine stroma was repopulated with autophagy-intact cells during repeated pregnancies. Conclusion: Multiple pregnancies seem to have improved the uterine environment by replacing autophagy-deficient cells with autophagy-intact cells, providing evidence of cell mixing.

In vitro effects of monophosphoryl lipid A and Poly I:C combination on equine cells

  • Dong-Ha Lee;Eun-bee Lee;Jong-pil Seo ;Eun-Ju Ko
    • Journal of Veterinary Science
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    • v.24 no.3
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    • pp.37.1-37.14
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    • 2023
  • Background: Toll-like receptor (TLR) agonists have been used as adjuvants to modulate immune responses in both animals and humans. Objectives: The objective of this study was to evaluate the combined effects of the TLR 4 agonist monophosphoryl lipid A (MPL) and the TLR 3 agonist polyinosinic:polycytidylic acid (Poly I:C) on equine peripheral blood mononuclear cells (PBMCs), monocyte-derived dendritic cells (MoDCs), and bone marrow-derived mesenchymal stromal cells (BM-MSCs). Methods: The PBMCs, MoDCs, and BM-MSCs collected from three mixed breed horses were treated with MPL, Poly I:C, and their combination. The mRNA expression of interferon gamma (IFN-γ), interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-12p40, tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), and monocyte chemoattractant protein-1 (MCP-1) was determined using real-time polymerase chain reaction. Results: The combination of MPL and Poly I:C significantly upregulated immunomodulatory responses in equine cells/ without cytotoxicity. The combination induced greater mRNA expression of pro-inflammatory cytokines IFN-γ and IL-6 than MPL or Poly I:C stimulation alone in PBMCs. In addition, the combination induced significantly higher mRNA expression of IL-1β, IL-6, and IL-12p40 in MoDCs, and IL-8, MCP-1, and VEGF in BM-MSCs compared to stimulation with a single TLR agonist. Conclusions: The combination of MPL and Poly I:C can be used as a potential adjuvant candidate for vaccines to aid in preventing infectious diseases in horses.

Evaluation of the regenerative capacity of stem cells combined with bone graft material and collagen matrix using a rabbit calvarial defect model

  • Jun-Beom Park;InSoo Kim;Won Lee;Heesung Kim
    • Journal of Periodontal and Implant Science
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    • v.53 no.6
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    • pp.467-477
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    • 2023
  • Purpose: The purpose of this study was to evaluate the regenerative capacity of stem cells combined with bone graft material and a collagen matrix in rabbit calvarial defect models according to the type and form of the scaffolds, which included type I collagen matrix and synthetic bone. Methods: Mesenchymal stem cells (MSCs) were obtained from the periosteum of participants. Four symmetrical 6-mm-diameter circular defects were made in New Zealand white rabbits using a trephine drill. The defects were grafted with (1) group 1: synthetic bone (β-tricalcium phosphate/hydroxyapatite [β-TCP/HA]) and 1×105 MSCs; (2) group 2: collagen matrix and 1×105 MSCs; (3) group 3: β-TCP/HA, collagen matrix covering β-TCP/HA, and 1×105 MSCs; or (4) group 4: β-TCP/HA, chipped collagen matrix mixed with β-TCP/HA, and 1×105 MSCs. Cellular viability and cell migration rates were analyzed. Results: Uneventful healing was achieved in all areas where the defects were made at 4 weeks, and no signs of infection were identified during the healing period or at the time of retrieval. New bone formation was more evident in groups 3 and 4 than in the other groups. A densitometric analysis of the calvarium at 8 weeks post-surgery showed the highest values in group 3. Conclusions: This study showed that the highest regeneration was found when the stem cells were applied to synthetic bone along with a collagen matrix.