• Title/Summary/Keyword: Stromal cell

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Osteogenic Differentiation of Bone Marrow Stem Cells Using Thermo-Sensitive Hydrogels (온도감응성 수화젤을 이용한 골수간엽줄기세포의 골분화 유도)

  • Kim, Sun-Kyung;Hyun, Hoon;Kim, Soon-Hee;Yoon, Sun-Jung;Kim, Moon-Suk;Rhee, John-M.;Khang, Gil-Son;Lee, Hai-Bang
    • Polymer(Korea)
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    • v.30 no.3
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    • pp.196-201
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    • 2006
  • Poly (ethylene glycol)-based diblock and triblock thermo- sensitive polyester copolymers were investigated for application on tissue engineering and injectable biomaterials in drug delivery system due to their nontoxicity, biocompatibility and biodegradability. We synthesized the diblock copolymers consisting of methoxy poly (ethylene glycol) (MPEG) (Mn=750 g/mole) and poly $(\varepsilon-caprolactone)$ (PCL) by ring opening polymerization of $\varepsilon-CL$ with MPEG as an initiator in the presence of HCl $Et_2O$. The effect of diblock copolymers on in vivo osteogenic differentiation of rat bone marrow stromal cells (BMSCS) with and without the presence of osteogenic supplements (dexamethasone) was investigated. Thin sections were cut from paraffin embedded tissues and histological sections were stained by H&E, von Kossa, and immunohistochemical staining for osteocalcin. In conclusion, dexamethasone containing thermo- sensitive hydrogel might be improved osteogenic differentiation of BMSCs. We expect the osteoinduction effect to be excellent when it uses stem cell or other osteogenic materials.

Clinical and Histopathological Findings in Mice Heavily infected with Fibricola seoulensis (Fibricolu seoulensis 중감염 마우스의 임상적 및 조직병리학적 소견)

  • Sun Huh;Jong-Yil Chai;Sung-Tae HONG;Soon-Hyung Lee
    • Parasites, Hosts and Diseases
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    • v.26 no.1
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    • pp.45-54
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    • 1988
  • To observe the clinical course and intestinal histopathology in heavy infection of Fibricola seoulensis, an experimental study was performed in mice. Clinical, experimental infection with 1, 000 metacercariae. On the 11th day after infection, the mice began to die and all of the infected mice were dead by the 16th day. The infected mice showed gradual weight loss. Occult blood was detected after the 10th day. Diarrhea accurred after the 9th day and was recognized in all of the infected mice on the 12th day. Hemoglobin and mean corpuscular hemoglobin decreased significantily after the 12th day, and mean corpuscular hemoglobin concentration decreased in all infected mice. The histopathological changes were more marked in the duodenum than in the jejunum or ileum. Major changes were villous atrophy and crypt hyperplasia, with decreased villus/crypt ratio, inflammatory cell infiltration and stromal edema. The present results suggest that the cause of death of mice heavily infected with F. seoulensis should be malnutrition and severe fluid loss due to malabsorption, together with intestinal bleeding.

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The Effect of Interleukin $1-{\beta}$, Platelet Derived Growth Factor-BB and Transforming Growth $Factor-{\beta}$ on the expression of PDLs17 mRNA in the Cultured Human Periodontal Ligament Fibroblasts (($IL-1{\beta}$), PDGF-BB 그리고 $TGF-{\beta}$가 사람 배양 치주인대 섬유모세포의 PDLs17 mRNA의 발현에 미치는 영향)

  • Lirn, Ki-Jung;Han, Kyung-Yoon;Kirn, Byung-Ock;Yeorn, Chang-Yeob;Park, Joo-Cheol
    • Journal of Periodontal and Implant Science
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    • v.31 no.4
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    • pp.787-801
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    • 2001
  • The molecular mechanisms control the function of PDL(periodonta1 ligament) cells and/or fibroblasts remain unclear. PDLsl7, PDL-specific gene, had previousely identified the cDNA for a novel protein from cultured PDL fibroblasts using subtraction hybridization between gingival fibroblasts and PDL fibroblasts. The purpose of this study was to determine the regulation by growth factors and cytokines on PDLsl7 gene expression in cultured human periodontal ligament cells and observe the immunohistochemical localization of PDLsl7 protein in various tissues of mouse. Primary PDL fibroblasts isolated by scraping the root of the extracted human mandibular third molars. The cells were incubated with various concentration of human recombinant $IL-1{\beta}$, PDGF-BB and TGF\;${\beta}$ for 48h nd 2 weeks. At each time point total RNA was extracted and the levels of transcription ere assessed by reverse transcription-polymerase chain reaction (RT-PCR assay). polyclonal antiserum raised against PDLsl7 peptides, CLSVSYNRSYQINE and SEAVHETDLHDGC, were made, and stained the tooth, periodontium, developing bone, bone marrow and mid-palatal suture of the mouse. The results were as follows. 1. PDLsl7 mRNA levels were increased in response to PDGF (10ng/ml) and $TGF\;{\beta}$(20ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF{\beta}$for 48 h. 2. PDLsl7 was up-regulated only by $TGF{\beta}$(20 ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF\;{\beta}$ for 2 weeks and unchanged by the other stimulants. 3. PDLsl7 was a novel protein coding the 142 amino acid peptides in the ORF and the nucleotide sequences of the obtained cDNA from RT-PCR was exactly same as the nucleotides of the database. 4. Immunohistochemical analysis showed that PDLsl7 is preferentially expressed in the PDL, differentiating osteoblast-like cells and stromal cells of the bone marrow in the adult mouse. 5. The expression of PDLsl7 protein was barely detectable in gingival fibroblasts, hematopoetic cells of the bone marrow and mature osteocytes of the alveolar bone. These results suggest that PDLsl7 might upregulated by PDGF-BB or $TGF{\beta}$ and acts at the initial stage of differentiation when the undifferentiated mesenchymal cells in the bone marrow and PDL differentiate into multiple cell types. However, more research needs to be performed to gain a better understanding of the exact function of PDLsl7 during the differentiation of bone marrow mesenchymal and PDL cells.

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The effect of human mesenchymal stem cell injection on pain behavior in chronic post-ischemia pain mice

  • Yoo, Sie Hyeon;Lee, Sung Hyun;Lee, Seunghwan;Park, Jae Hong;Lee, Seunghyeon;Jin, Heecheol;Park, Hue Jung
    • The Korean Journal of Pain
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    • v.33 no.1
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    • pp.23-29
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    • 2020
  • Background: Neuropathic pain (NP) is considered a clinically incurable condition despite various treatment options due to its diverse causes and complicated disease mechanisms. Since the early 2000s, multipotent human mesenchymal stem cells (hMSCs) have been used in the treatment of NP in animal models. However, the effects of hMSC injections have not been studied in chronic post-ischemia pain (CPIP) mice models. Here, we investigated whether intrathecal (IT) and intrapaw (IP) injections of hMSCs can reduce mechanical allodynia in CPIP model mice. Methods: Seventeen CPIP C57/BL6 mice were selected and randomized into four groups: IT sham (n = 4), IT stem (n = 5), IP sham (n = 4), and IP stem (n = 4). Mice in the IT sham and IT stem groups received an injection of 5 μL saline and 2 × 104 hMSCs, respectively, while mice in the IP sham and IP stem groups received an injection of 5 μL saline and 2 × 105 hMSCs, respectively. Mechanical allodynia was assessed using von Frey filaments from pre-injection to 30 days post-injection. Glial fibrillary acidic protein (GFAP) expression in the spinal cord and dorsal root ganglia were also evaluated. Results: IT and IP injections of hMSCs improved mechanical allodynia. GFAP expression was decreased on day 25 post-injection compared with the sham group. Injections of hMSCs improved allodynia and GFAP expression was decreased compared with the sham group. Conclusions: These results suggested that hMSCs may be also another treatment modality in NP model by ischemia-reperfusion.

Characterization of the Immune Regulation Function of Fibroblastic Reticular Cells Originating from Lymph Node Stroma (림프절 스트로마 유래 fibroblastic reticular cell의 면역조절 기능에 대한 특성 규명)

  • Lee, Jong-Hwan
    • Journal of Life Science
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    • v.26 no.7
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    • pp.789-795
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    • 2016
  • A lymph node (LN) is one of the secondary lymphoid organs. An LN consists of a complicated 3 dimensional frame structure and several stromal cells. Fibroblastic reticular cells (FRC) are distributed in the T zone for interaction with T cells. FRC secrete homing chemokines such as CCL19 and CCL21. Moreover, FRC play a pivotal role in the production of extracellular matrix (ECM) into LN for ECM reorganization against pathogen infections. However, not much is known about the involvement of the immune reaction of FRC. The present report is for the characterization of FRC on immune response. For this, FRC were positioned in several infected situations such as co-culture with macrophage, lipopolysaccharide (LPS), and TNFα stimulation. When a co-culture between FRC and macrophage was performed, a morphological change in FRC was observed, and empty space between FRCs was created by this change. The soluble ICAM-1 protein level was up-regulated by co-culturing with Raw264.7 and the treatment of the ROCK inhibitor Y27632. The activity of matrix metalloproteinase (MMP) was up-regulated by LPS onto FRC. Furthermore, the inflammatory cytokine TNFα regulated the expression of ECM in FRC by a gene chip assay. Collectively, it suggests that FRC are involved in immune reactions.

Regulatory Dendritic Cells Induced by Mesenchymal Stem Cells Ameliorate Dextran Sodium Sulfate-Induced Chronic Colitis in Mice

  • Jo, Hannah;Eom, Young Woo;Kim, Hyun-Soo;Park, Hong Jun;Kim, Hee Man;Cho, Mee-Yon
    • Gut and Liver
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    • v.12 no.6
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    • pp.664-673
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    • 2018
  • Background/Aims: Regulatory dendritic cells (rDCs), which can be induced by mesenchymal stem cells (MSCs), play an important role in inducing and maintaining homeostasis of regulatory T cells and exhibit anti-inflammatory functions. In this study, we investigated whether MSCs could differentiate DCs into rDCs and compared the therapeutic effects of rDCs and MSCs on dextran sodium sulfate (DSS)-induced chronic colitis mice. Methods: Immature DCs (imDCs) and lipopolysaccharide (LPS)-treated mature DCs (mDCs) were co-cultured with MSCs for 48 hours, and then the profiles of surface markers and cytokines and regulatory roles of these DCs for primary splenocytes were analyzed. In addition, the therapeutic effects of MSCs and DCs co-cultured with MSCs were compared in chronic colitis mice. Results: After co-culture of imDCs (MSC-DCs) or LPS-treated mDCs (LPS+MSC-DCs) with MSCs, the expression of CD11c, CD80, CD86, interleukin 6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and interferon-${\gamma}$ (IFN-${\gamma}$), was decreased, but that of CD11b, IL-10, and transforming growth factor-${\beta}$ (TGF-${\beta}$) was increased. Furthermore, MSC-DCs and LPS+MSC-DCs induced the expression of CD4, CD25, and Foxp3 in primary splenocytes isolated from mice. In DSS-induced colitis mice, MSCs and MSC-DCs increased colon length, body weight, and survival rate and induced histological improvement. Moreover, in the colon tissues, the expression of IL-6, TNF-${\alpha}$, and IFN-${\gamma}$ decreased, but that of IL-10, TGF-${\beta}$, and Foxp3 increased in the MSC- and MSC-DC-injected groups. Conclusions: Our data suggest that MSCs differentiate DCs into rDCs, which ameliorate chronic colitis. Thus, rDCs stimulated by MSCs may be therapeutically useful for the treatment of chronic inflammatory diseases.

Effect of Oral Administration of Houttuynia Cordata Extract on Benign Prostatic Hyperplasia (전립선비대증의 어성초추출물에 의한 경구투여 효과)

  • Song, Won-Yeong;Choi, Jeong-Hwa
    • Journal of Life Science
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    • v.29 no.6
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    • pp.705-711
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    • 2019
  • Benign prostatic hyperplasia (BPH) is the most common urogenital disorder in men, benign tumor and is a typical disease deteriorating the quality of old men's lives, and its prevalence increases with age. Though the molecular pathogenesis of BPH has not yet been clearly revealed, it is known that the variation and aging of the endocrine including sex hormone may cause BPH. Especially the hypertrophy of the prostate cell by the formation of the excessive dihydrotestosterone (DHT) is estimated to cause BPH. If testosterone exists excessively in blood, a lot of DHT is produced in prostate by $5{\alpha}-reductase$. Thus, in this study we tried to analyze haematological change and histopathological change by using the model rat with BPH caused by hypodermic injection of testosterone to prove the effect of Houttuynia cordata extracts on BPH. Rats were divided into four experimental groups: no treatment group (N), the testosterone injection and D.W treatment group (DO), the testosterone injection and Houttuynia cordata treatment group (HO) and testosterone injection and finasteride treatment group (FO). Prostate weight, volume and weight ratio in the HO and FO groups were significantly lower than the DO group. Testosterone and DHT levels in the HO group were significantly lower than the DO group. The HO and FO groups showed trophic symptoms and were lined by flattened epithelial cells, thus, the stromal proliferation is relatively low as compared to the DO group. These results suggest that Houttuynia cordata may control benign prostatic hyperplasia.

Effects of Bojungikgi-Tang on the Development of Experimentally-induced Endometriosis in the Rats (보중익기탕(補中益氣湯)이 실험적으로 유발된 흰쥐의 자궁내막증(子宮內膜症) 발달에 미치는 영향)

  • Lee, Jeong-Gu;Lee, Dong-Nyung;Seo, Il-Bock;Kim, Hyung-Jun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.3
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    • pp.1-24
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    • 2009
  • Purpose: This study was performed to investigate the effects of Bojungikgi-Tang on the development of experimentally-induced endometriosis in rats. Methods: Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and we divided the rats into three groups: (1) sham-operated group (n=8), (2) surgically induced endometriosis and untreated control group(n=8), (3) surgically induced endometriosis and Bojungikgi-Tang treated group. Bojungikgi-Tang was orally administrated for 15 days after operation. Then we measured the body weight, the volume of endometriotic implants, the weight of uterus and ovary, and analysed the concentration of cytokines(MCP-1, TNF-${\alpha}$, IL-1${\beta}$, IL-6) in peritoneal fluids. Histopathology, immunohistochemistry for COX-2 and VEGF, and histochemistry for mast cell in transplanted uterine tissue were also performed. Results: 1. The volume($mm^3$) of endometriotic implants in Bojungikgi-Tang treated group(129${\pm}$39) was significantly decreased(p<0.05) compared with control group(222${\pm}$109). 2. The concentration(pg/$m{\ell}$) of MCP-1 in peritoneal fluids in Bojungikgi-Tang treated group(1012${\pm}$248) was significantly decreased(p<0.05) compared with control group(1413${\pm}$346). 3. The concentration(pg/$m{\ell}$ of TNF-${\alpha}$ in peritoneal fluids in Bojungikgi-Tang treated group(887${\pm}$196) was significantly decreased(p<0.05) compared with control group(1126${\pm}$140). 4. Histopathologically, proliferation of endometriotic epithelia, infiltration of inflammatory cells and angiogenesis in transplanted uterine tissue of Bojungikgi-Tang treated group were weakly observed than those of control group. 5. The percentage of positive epithelial layers for COX-2 in Bojungikgi-Tang treated group(56${\pm}$17) was significantly decreased(p<0.05) compared with control group(75${\pm}$16). 6. The VEGF expression of endometriotic epithelia, neovascular endothelia and stromal cells in transplanted uterine tissue of Bojungikgi-Tang treated group were weakly observed than those of control group. Conclusion: On the basis of these results, we concluded that Bojungikgi-Tang has inhibiting effects on the development of transplanted uterine tissue. And these effects may be related with decreased production of MCP-1, TNF-${\alpha}$, and decreased expression of COX-2 and VEGF infiltration by administration of Bojungikgi-Tang.

Effects of an in vitro vitamin D treatment on the inflammatory responses in visceral adipose tissue from Ldlr-/- mice

  • Deok Hoon Kwon;Jungwon Hwang;Hyeyoung You;Na Young Kim;Ga Young Lee;Sung Nim Han
    • Nutrition Research and Practice
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    • v.18 no.1
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    • pp.19-32
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    • 2024
  • BACKGROUND/OBJECTIVES: Atherosclerosis is associated with increased inflammation in the visceral adipose tissue (VAT). Vitamin D has been reported to modulate the inflammatory responses of stromal vascular cells (SVCs) and adipocytes in adipose tissue, but the role of vitamin D in atherosclerosis biology is unclear. This study examined the effects of in vitro 1,25-dihydroxyvitamin D3 (1,25[OH]2D3) treatment on the inflammatory responses of SVCs and adipocytes from atherosclerotic mice. MATERIALS/METHODS: C57BL/6J (B6) mice were divided randomly into 2 groups and fed a 10% kcal fat control diet (control group, CON) or 41% kcal fat, 0.21% cholesterol (high fat + cholesterol, HFC) diet (obese group, OB), and B6.129S7-Ldlrtm1Her/J (Ldlr-/-) mice were fed a HFC diet (obese with atherosclerosis group, OBA) for 16 weeks. SVCs and adipocytes isolated from VAT were pre-incubated with 1,25(OH)2D3 for 24 h and stimulated with lipopolysaccarides for the next 24 h. Proinflammatory cytokine production by adipocytes and SVCs, the immune cell population in SVCs, and the expression of the genes involved in the inflammatory signaling pathway in SVCs were determined. RESULTS: The numbers of total macrophages and SVCs per mouse were higher in OB and OBA groups than the CON group. The in vitro 1,25(OH)2D3 treatment significantly reduced macrophages/SVCs (%) in the OBA group. Consistent with this change, the production of interleukin-6 and monocyte chemoattractant protein 1 (MCP-1) by SVCs from the OBA group was decreased by 1,25(OH)2D3 treatment. The 1,25(OH)2D3 treatment significantly reduced the toll-like receptor 4 and dual-specificity protein phosphatase 1 (also known as mitogen-activated protein kinase phosphatase 1) mRNA levels in SVCs and MCP-1 production by adipocytes from all 3 groups. CONCLUSIONS: These findings suggest that vitamin D can attribute to the inhibition of the inflammatory response in VAT from atherosclerotic mice by reducing proinflammatory cytokine production.

Potential Reproductive Toxicity Study of p53 Expressing Adenoviral Vector in Mice (아데노바이러스 유전자치료벡터의 생식독성 연구)

  • Rhee, Gyu-Seek;Kwack, Seung-Jun;Kim, Soon-Sun;Lee, Rhee-Da;Seok, Ji-Hyun;Chae, Soo-Young;Chung, Soo-Youn;Kim, Seung-Hee;Lee, Seung-Hoon;Park, Kui-Lea
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.151-158
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    • 2007
  • The possibility of inadvertent introduction of therapeutic gene expressing viral vectors has raised safety concerns about germ-line infection. Particularly, for indications such as prostate cancer and ovarian cancer, the proximity of the point of viral administration to organs of the reproductive system raises concerns regarding inadvertent germ-line transmission of genes carried by the virus vector. To evaluate the safety of in vivo adenovirus mediated gene transfer, we explored the biodistribution, persistance and potential germ-line transmission of p53-expressing adenovirus (Ad-CMV-p53). Both male and female Balb/c mice were injected with $1{\times}10^9$ PFU of Ad-CMV-p53. The PCR analysis showed that there were detectable vector sequences in liver, kidney, spleen, seminal vesicle, epididymis, prostate, ovary, and uterus. The RT-PCR analysis for detecting inserted gene, p53 showed that Ad-CMV-p53 viral RNA were present in spleen, prostate and ovary. Direct injected male and female mice of adenovirus vector into testis and ovary were mated and their of offspring were evaluated for germ-line transmission of the adenoviral vector. The PCR and RT-PCR analysis showed no evidence of germline transmission, although vector sequences were detected in DNA extracted from gonadal tissues. Real-time PCR result confirmed a significant decrease of adenovirus in gonad tissues 1 week after injection. We have also analysed the cell specific localization of viral DNA in gonad tissues by using in-situ PCR. Positive signals were detected in interstitial tissue but not in seminiferous tubule in sperm. In the case of ovary, adenovirus signal were localized to the stromal tissue, but no follicular signals were observed. Together, these data provide strong evidence that the risk of the Inadvertent germ-line transmission of vector sequences following intraperitoneal or direct injection into genito-urinary system of adenovirus is extremely low.