• Title/Summary/Keyword: Hematoxylin & Eosin(H&E) staining

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Study on an Effective Decellularization Technique for Cardiac Valve, Arterial Wall and Pericardium Xenographs: Optimization of Decellularization (이종 심장 판막 및 대혈관 이식편과 심낭에서 효과적인 탈세포화 방법에 관한 연구: 탈세포화의 최적화)

  • Park, Chun-Soo;Kim, Yong-Jin;Sung, Si-Chan;Park, Ji-Eun;Choi, Sun-Young;Kim, Woong-Han;Kim, Kyung-Hwan
    • Journal of Chest Surgery
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    • v.41 no.5
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    • pp.550-562
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    • 2008
  • Background: We attempted to reproduce a previously reported method that is known to be effective for decellularization, and we sought to find the optimal condition for decellularization by introducing some modifications to this method. Material and Method: Porcine semilunar valves, arterial walls and pericardium were processed for decellularization with using a variety of combinations and concentrations of decellularizing agents under different conditions of temperature, osmolarity and incubation time. The degree of decellularization and the preservation of the extracellular matrix were evaluated by staining with hematoxylin and eosin and with alpha-Gal and DAPI in some of the decellularized tissues. Result: Decellularization was achieved in the specimens that were treated with sodium deoxycholate, sodium dodesyl sulfate, Triton X-100 and sodium dodesyl sulfate with Triton X-100 as single-step methods, and this was also achieved in the specimens that were treated with hypotonic solution ${\rightarrow}$ Triton X-100 ${\rightarrow}$ sodium dodesyl sulfate, sodium deoxycholate ${\rightarrow}$ hypotonic solution ${\rightarrow}$ sodium dodesyl sulfate, and hypotonic solution sodium dodesyl sulfate as multi-step methods. Conclusion: Considering the number and the amount of the chemicals that were used, the incubation time and the degree of damage to the extracellular matrix, a single-step method with sodium dodesyl sulfate and Triton X-100 and a multi-step method with hypotonic solution followed by sodium dodesyl sulfate were both relatively optimal methods for decellularization in this study.

Laser Acupuncture Treatment on the Five Transport Points of the Spleen Meridian in Dextran-Sulfate-Sodium-Induced-Colitis in Rats (비경의 오수혈에 대한 830 nm 레이저침이 DSS로 유발된 흰쥐의 대장염에 미치는 영향)

  • Choi, Dong-Hee;Kim, Wang-In;Kim, Mi-Rea;Youn, Dae-Hwan;Na, Chang-Su
    • Korean Journal of Acupuncture
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    • v.31 no.2
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    • pp.56-65
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    • 2014
  • Objectives : The purpose of this study is to compare the effects of laser acupuncture to the 830 nm on the five transport points with the spleen meridian for treatment to intestinal disease in rat with dextran sulfate sodium(DSS)-induced colitis. Methods : Colitis was induced by DSS for 20 days. The laser therapy on the five transport points of spleen meridian (Laser Well Point-SP1(L-WE), Laser Brook Point-SP2(L-BR), Laser Stream Point-SP3(L-ST), Laser River Point-SP5(L-RI) and Laser Sea Point-SP9(L-SE) was practiced twice a week for 5 times. Colon length was measured using a measuring point. Histological evaluation of colitis was conducted by hematoxylin and eosin(H&E) staining. Reverse transcription polymerase chain reaction(RT-PCR) was determined using western blotting and quantitative reverse-transcriptase polymerase chain reaction, respectively. Results: Colon length increased significantly L-BR and L-ST points after 5 times of therapy. Damage to the colonic mucosa is an integral feature of the DSS model, so control colonic mucosa tissue was damaged in the areas of ulceration resulting in complete epithelial loss. However histological damage decreased on the epithelial lining at all points. Cyclooxygenase(COX)-2 concentrations decreased in all points groups and Interferon(IFN)-${\gamma}$ increased in L-WE, L-BR, L-RI and L-SE points but L-ST was decreased when compared with control. White blood cell(WBC) and neutrophils(NE) decreased after the fifth acupuncture on the all points. But hemoglobin(HGB) increased after the fifth acupuncture on the L-WE, L-BR, L-ST and L-RI points. Also Mean corpuscular hemoglobin(MCH) and Mean corpuscular hemoglobin concentration(MCHC) decreased after the fifth acupuncture on the all points. Conclusions: The present study indicated that five transport points of the spleen meridian can prevent the development of DSS-induced colitis in rat. Thereby suggesting that should be available for decreasing DSS-induced inflammation in a colonic mucosa of tissue.

Preparation and Characterization of Ipriflavone-Loaded Poly(L-lactide-co-glycolide) Scaffold for Tissue Engineered Bone (조직공학적 골을 위한 애프리플라본을 함유한 다공성 지지체의 제조 및 그 특성)

  • Jang, Ji-Wook;Lee, Bong;Han, Chang-Whan;Lee, Il-Woo;Lee, Hai-Bang;Khang, Gil-Son
    • Polymer(Korea)
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    • v.27 no.3
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    • pp.226-234
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    • 2003
  • Ipriflavone (IP), a non-hormonal isoflavone derivative, has been shown to interfere with bone remodeling by inhibiting bone resorption and stimulating bone formation. IP consistently increased the amount of Ca incorporated into the cell layer by mesenchymal stem cells (MSCs). In this study, we developed the novel IP loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds for the possibility of the application of the tissue engineered bone. IP/PLGA scaffo1ds were prepared by solvent casting/salt leaching method and were characterized by porosimeter, scanning electron microscopy, determination of residual salt amount, differential scanning calorimetry, and X-ray diffractometer, respectively. IP/PLGA scaffolds were implanted into the back of athymic nude mouse to observe the effect of IP on the osteoinduction compared with control PLGA scaffo1ds. Thin sections were cut from paraffin embedded tissues and histological sections were stained H&E, von Kossa, and immunohistochemical staining for Type I collagen and osteocalcin. It can be observed that the porosity was above 91.7% and the pore size was above 101 $\mu\textrm{m}$. Control scaffo1d and IP/PLGA scaffo1ds of 50% IP were implanted on the back of athymic nude mouse to observe the effect of IP on the induction of cells proliferation for 9 weeks. The evidence of calcification, osteoblast, and osteoid from the undifferentiated stem cells in the subcutaneous sites and other soft connective tissue sites having a preponderance of stem cells has been observed. From these results, it seems that IP plays an important role for bone induction in IP/PLCA scaffolds.

Effects of natural raw meal (NRM) on high-fat diet and dextran sulfate sodium (DSS)-induced ulcerative colitis in C57BL/6J mice

  • Shin, Sung-Ho;Song, Jia-Le;Park, Myoung-Gyu;Park, Mi-Hyun;Hwang, Sung-Joo;Park, Kun-Young
    • Nutrition Research and Practice
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    • v.9 no.6
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    • pp.619-627
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    • 2015
  • BACKGROUND/OBJECTIVES: Colitis is a serious health problem, and chronic obesity is associated with the progression of colitis. The aim of this study was to determine the effects of natural raw meal (NRM) on high-fat diet (HFD, 45%) and dextran sulfate sodium (DSS, 2% w/v)-induced colitis in C57BL/6J mice. MATERIALS/METHODS: Body weight, colon length, and colon weight-to-length ratio, were measured directly. Serum levels of obesity-related biomarkers, triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL), high density lipoprotein (HDL), insulin, leptin, and adiponectin were determined using commercial kits. Serum levels of pro-inflammatory cytokines including tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin (IL)-$1{\beta}$, and IL-6 were detected using a commercial ELISA kit. Histological study was performed using a hematoxylin and eosin (H&E) staining assay. Colonic mRNA expressions of TNF-${\alpha}$, IL-$1{\beta}$, IL-6, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) were determined by RT-PCR assay. RESULTS: Body weight and obesity-related biomarkers (TG, TC, LDL, HDL, insulin, leptin, and adiponectin) were regulated and obesity was prevented in NRM treated mice. NRM significantly suppressed colon shortening and reduced colon weight-to-length ratio in HFD+DSS induced colitis in C57BL/6J mice (P < 0.05). Histological observations suggested that NRM reduced edema, mucosal damage, and the loss of crypts induced by HFD and DSS. In addition, NRM decreased the serum levels of pro-inflammatory cytokines, TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 and inhibited the mRNA expressions of these cytokines, and iNOS and COX-2 in colon mucosa (P < 0.05). CONCLUSION: The results suggest that NRM has an anti-inflammatory effect against HFD and DSS-induced colitis in mice, and that these effects are due to the amelioration of HFD and/or DSS-induced inflammatory reactions.

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.

Id3 mRNA Expression on Folliculogenesis in Rat Ovary (쥐 난소에서 난포 발달에 따른 Id3 mRNA의 발현)

  • Hwang, Seong-Soo;Ko, Yeoung-Gyu;Lim, Hyun-Joo;Seong, Hwan-Hoo;Yoon, Jong-Taek;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.181-186
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    • 2007
  • Inhibitor of DNA binding protein or inhibitor of differentiation(Id) is largely considered as positive and/or negative regulators of proliferation, differentiation, angiogeneisis, and apoptosis. The four Id genes(Id1, Id2, Id3, and Id4) were known in mammals. However, little is known about the expression and function of these genes in reproductive physiology. Among them, this study was conducted to analyze the expression pattern of Id3 mRNA on folliculogenesis in rat ovary. After PMSG administration, the ovaries were obtained at 3, 6, 12, 24, 36, and 48hrs, fixed, dehydrated, and paraffin embedded. For in situ hybridization, anti-sense and sense Id3 cRNA probes were prepared and applied to the ovarian section. The ovarian sections were coated with NTB-2 emulsion. After that, the slides were developed and counterstained with hematoxylin and eosin staining. The hybridization signal was estimated on a scale of 1+ to 4+. In oocyte, the intensity of Id3 mRNA in primordial and primary follicles was scored at ${\geq}2+$, but the intensity was less than 1+ in secondary, dominant, and preovulatory follicles. In granulosa cells, the Id3 mRNA was strongly expressed(3+ or 4+) in dominant and preovulatory follicles. Taken together, Id3 mRNA was expressed specifically at follicle stages and follicular tissue and might be closely related with follicle development.

Protective effect of silk protein hydrolysates against tert-BHP induced liver damage (실크 단백질 가수분해물의 간 손상에 대한 보호효과)

  • Kim, Joo Hyoun;Suh, Hyung Joo;Choi, Hyeon-Son
    • Food Science and Preservation
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    • v.24 no.1
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    • pp.107-115
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    • 2017
  • The aim of this study was to investigate the hepatoprotecive effect of silk protein hydrolysates (SDH), which was prepared by acid hydrolysis, in rats. SDH itself did not exhibit any cytotoxic effect on hepatic tissues. SDH showed a protective effect on tert-butyl hydroperoxide (t-BHP)-induced hepatotoxicity and liver damage. SDH effectively reduced AST (aspartate aminotransferase) and ALT (alanine aminotransferase), which are biomarkers for liver damage, in a dose-dependent manner. Malondialdehyde (MDA), a lipid peroxidation product, was significantly reduced by SDH. A high dose of SDH (2 g/kg) reduced t-BHP-induced MDA production by 40%. Glutathione (GSH), which is an endogenous antioxidant molecule, was effectively increased by SDH treatment. GSH content was enhanced by around 2.5-fold, compared with t-BHP control, upon SDH (2 g/kg) treatment. Lactate dehydrogenase (LDH), which is an enzyme released by cell cytotoxicity, was greatly increased by t-BHP, but significantly decreased by SDH treatment. Furthermore, hematoxylin and eosin (H&E) staining showed that SDH suppressed t-BHP-induced lesions in liver tissue. Taken together, SDH might be used as a protective agent against liver damage.

Ginsenoside Rg1 alleviates Aβ deposition by inhibiting NADPH oxidase 2 activation in APP/PS1 mice

  • Zhang, Han;Su, Yong;Sun, Zhenghao;Chen, Ming;Han, Yuli;Li, Yan;Dong, Xianan;Ding, Shixin;Fang, Zhirui;Li, Weiping;Li, Weizu
    • Journal of Ginseng Research
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    • v.45 no.6
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    • pp.665-675
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    • 2021
  • Background: Ginsenoside Rg1 (Rg1), an active ingredient in ginseng, may be a potential agent for the treatment of Alzheimer's disease (AD). However, the protective effect of Rg1 on neurodegeneration in AD and its mechanism of action are still incompletely understood. Methods: Wild type (WT) and APP/PS1 AD mice, from 6 to 9 months old, were used in the experiment. The open field test (OFT) and Morris water maze (MWM) were used to detect behavioral changes. Neuronal damage was assessed by hematoxylin and eosin (H&E) and Nissl staining. Immunofluorescence, western blotting, and quantitative real-time polymerase chain reaction (q-PCR) were used to examine postsynaptic density 95 (PSD95) expression, amyloid beta (Aβ) deposition, Tau and phosphorylated Tau (p-Tau) expression, reactive oxygen species (ROS) production, and NAPDH oxidase 2 (NOX2) expression. Results: Rg1 treatment for 12 weeks significantly ameliorated cognitive impairments and neuronal damage and decreased the p-Tau level, amyloid precursor protein (APP) expression, and Aβ generation in APP/PS1 mice. Meanwhile, Rg1 treatment significantly decreased the ROS level and NOX2 expression in the hippocampus and cortex of APP/PS1 mice. Conclusions: Rg1 alleviates cognitive impairments, neuronal damage, and reduce Aβ deposition by inhibiting NOX2 activation in APP/PS1 mice.

Periodontal Tissue Response Following Different Types of Fixed Retainers in Young Adult Dogs (수종의 고정성 보정장치에 따른 유성견의 치주조직 반응)

  • Jo, Myung-Hun;Yoon, Young-Jooh;Kim, Kwang-Won
    • The korean journal of orthodontics
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    • v.31 no.1 s.84
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    • pp.85-95
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    • 2001
  • The purpose of this study was to evaluate the material for fixed type retainer, allowing physiologic tooth movement. and proper remodeling or periodontal tissue during retention period. The Present study was Performed to observe the histologic changes of periodontal tissue after application of different types of fixed type retainer after orthodontic tooth movement in young adult dogs. For this study, 4 young adult dogs were used as a experimental animal and experimental group was divided into three groups : experimental group 1 contained right side maxillayy third incisors and canines, experimental group 2 contained contralateral teeth of same animals, and control group contained mandibular premolars. And each dogs were applied the 4 different types of fixed type retainer to experimental group 1. The experimental teeth were ligated on the Sentalloy closed coil $spring^{\circledR}$(Tomy Co., Japan) from maxillary third incisors and canines and applied orthodontic force at initial 200gm-forced during 1 week. All the experimental animals were sacrificed on the 3rd week after the orthodontic teeth movement and then the specimens were taken, fixed in formalin, embeded in parafin, sectioned $6-8{\mu}m$ in thickness and stained with Hematoxylin-Eosin staining, and Masson's trichrome staining method. Examined under the light microscopy The following results were observed. 1. There were observed that decreased infiltration of giant tells in pressure side and increased the new bone forming in tension side on the specimen of 6-stranded 0.0195' $Respond^{\circledR}$(G&H Co., U.S.A.) group. Periodontal ligament fibers were much compressed or elongated in 3-stranded 0.018', 0.020' $Dentaflex^{\circledR}$(Dentarum Co., Germany), and Superbond $C&B^{\circledR}$(Sun Medical Co., Japan) groups. 2. In experimental group 1, necrotic bone inside the alveolar bone of pressure side, forming of the sharpey's fiber in osteoid tissue, and remodeling of the periodontal ligament were observed in all animals. 3. In experimental group 2, it was observed that the amount of bone resorption was equal or decreased in pressure side, and increased new bone forming and significantly decreased infiltration of giant cell than the experimental group 1. By this results, it considered that 6-stranded $Respond^{\circledR}$(G&H Co., U.S.A.) wire was the most useful material allowing early periodontal tissue remodeling.

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A Study on the changes of periodontal tissue after orthodontic tooth movement in young adult dogs (유성견에서 교정적 치아이동에 따른 치주조직 변화에 관한 연구)

  • Kang, Nam-Yong;Yoon, Young-John;Kim, Kwang-Won
    • The korean journal of orthodontics
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    • v.27 no.4 s.63
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    • pp.559-568
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    • 1997
  • For orthodontic tooth movement, optimal orthodontic force should be maintained without periodontal breakdown and alveolar bone should be remodeled physiologically Therefore, To obtain proper occlusion through tooth movement within alveolar bone, we should know the biomechanics of teeth and supporting 4issues. The present study was performed to observe histologic changes of periodontal tissue immediately after application of orthodontic force and during the retention period in growing young adult dogs. In this study, experimental group contained between mandibular left canine and 1st molar and control group contained contralateral teeth of same animal. The .018'x.022' stainless steel closed coil spring(Dentaurum Co.) was ligated on the experimental teeth at initial 200gm-force from mandibular canine to 1st molar The animals(4 to 6 months aged young adult dogs) were sacrificed on 0, 14, 28 days after the finish of appliance activation, and then tissue samples were divided into hematoxylin-eosin(HE) staining section, ground section, alkaline phosphatase(ALP) staining section, and tartrate-resistant acid phosphatase(TRAP) staining section. Thereafter, the preparations were examined under light microscopy The following results were obtained: 1. Immediately after the finish of appliance activation, the periodontal space was increased in tension side, but decreased in pressure side compared to that of control. The hyalinized zone was also observed in the periodontium. 2. After the 14-day retention, peridontal space was decreased in tension side and slightly increased in pressure side compared to that of immediately after the finish of appliance activation. The hyalinized zone was repaired and a few osteoblasts showing slightly new bone formation were seen. Osteoblasts were scarcely observed along the alveolar bone. 3. Aftter the 28-day retention, the periodontal fibers are normally repaired. A lot of TRAP(+) osteoclasts md increased alveolar bone resorption were observed in pressure side, and AP(+) osteoblast and increased new bone formation were observed in tension side.

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