• Title/Summary/Keyword: Cell membrane integrity

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Initial tissue response of biodegradable membrane in rat subcutaneous model (백서 피하층에서 흡수성 차단막의 초기 조직 반응)

  • Lim, Hyun-Chang;Chae, Gyung-Joon;Jung, Ui-Won;Kim, Chang-Sung;Lee, Yong-Keun;Cho, Kyoo-Sung;Kim, Chong-Kwan;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.37 no.4
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    • pp.839-848
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    • 2007
  • Purpose: Various kinds of biodegradable membranes are currently used in dental clinics. And the frequency and the necessity of their usage are increasing due to their numerous advantages. Therefore it is important to understand the difference of various membranes and histological reaction against implanted membranes. Materials and Methods: Biodegradable membranes of $Biogide^{(R)}$, $Resolute^{(R)}$, and $Tutodent^{(R)}$ were cut into small pieces by $1.0{\times}0.5cm$. The membranes were implanted 1.5cm apart from each other under the epithelium on the skull of 18 Sprague Dawley rats. The animals were sacrificed at 3, 7, and 14 days after surgical procedure. The specimens were examined by histological analysis. Results: 1. Early period after implantation of the membranes showed connective tissues surrounding membranes and there were a few inflammatory cells present. 2. In $Biogide^{(R)}$ and $Tutodent^{(R)}$ specimens, inflammatory cells and surrounding tissues were shown to infiltrate from outside with slight density difference inside. In $Resolute^{(R)}$ specimens, membranes were fragmented. Inflammatory cells and connective tissues were also observed inside. 3. In $Resolute^{(R)}$ specimen, giant cells were present which implicates that foreign body reaction has occurred. 4. $Biogide^{(R)}$ had lower integrity than other membranes and is not enough to be used alone in defect area. However, $Resolute^{(R)}$ had superior firmness than others. $Tutodent^{(R)}$ had middle level of integrity. Conclusion: This experimental model enabled to observe early inflammatory reactions and morphological changes of materials and can be used to develop and evaluate the efficacy of biodegradable membranes. Duplication of standardized human oral environment will be required in future experiments.

Osteogenic Potential of the Periosteum and Periosteal Augmentation for Bone-tunnel Healing

  • Youn Inchan;Suh J-K Francis;Choi Kuiwon
    • Journal of Biomedical Engineering Research
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    • v.26 no.2
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    • pp.101-110
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    • 2005
  • Periosteum and periosteum-derived progenitor cells have demonstrated the potential for stimulative applications in repairs of various musculoskeletal tissues. It has been found that the periosteum contains mesenchymal progenitor cells capable of differentiating into either osteoblasts or chondrocytes depending on the culture conditions. Anatomically, the periosteum is a heterogeneous multi-layered membrane, consisting of an inner cambium and an outer fibrous layer. The present study was designed to elucidate the cellular phenotypic characteristics of cambium and fibrous layer cells in vitro, and to assess whether structural integrity of the tendon in the bone tunnel can be improved by periosteal augmentation of the tendon­bone interface. It was found the cells from each layer showed distinct phenotypic characteristics in a primary monolayer culture system. Specifically, the cambium cells demonstrated higher osteogenic characteristics (higher alkaline phosphatase and osteocalcin levels), as compared to the fibrous cells. Also in vivo animal model showed that a periosteal augmentation of a tendon graft could enhance the structural integrity of the tendon-bone interface, when the periosteum is placed between the tendon and bone interface with the cambium layer facing toward the bone. These findings suggest that extra care needs to be taken in order to identify and maintain the intrinsic phenotypes of the heterogeneous cell types within the periosteum. This will improve our understanding of periosteum in applications for musculoskeletal tissue repairs and tissue engineering.

Effects of Tetracycline-loaded Poly(L-lactide) Barrier Membranes on Guided Bone Regeneration in Beagle Dog (테트라싸이클린 함유 차폐막을 이용한 골조직 유도 재생에 관한 연구)

  • Choi, Kwang-Soo;Kim, Tak;Yang, Dae-Seung;Kim, Eun-Cheol;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.31 no.2
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    • pp.299-315
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    • 2001
  • Application of membranes for guided tissue regeneration(GTR) have been confined to the subgingival barrier functions; however, many studies have provided evidence that some drugs, including tetracycline, initially can promote the growth of periodontal ligament or alveolar bone in peridontal therapy. Osseous regeneration in periodontal defects is increased by local administration of tetracycline due to its anti-collagenolytic effect, which enhances bone-forming ability via osteoblast cell chemotaxis and reduced bone resorption. The aim of this study was to evaluate effects of tetracycline loaded poly-L-lactide(PLLA) barrier membranes for guided bone regenerative potential. Tetracycline was incorporated into the PLLA membrane with the ratio 10% to PLLA by weight. Ability to guided bone regeneration of the membranes were tested by measuring new bone in the tibial defects($7{\times}10{\times}5\;mm^3$) of the beagle dog for 4,5, and 6 weeks. In control, drug-unloaded PLLA membranes were used in same size of defect. In histologic finding of the defect area, a few inflammatory cells were observed in both groups. These membrane were not perforated by connective tissue and maintained their mechanical integrity for the barrier function for 4-6 weeks. New bone formation was greater in defects covered by tetracycline-loaded membrane than in defects covered by drug- unloaded membranes. In bone regeneration guiding potential test, tetracycline-loaded membrane was more effective than drug- unloaded membranes(p<0.05). These results suggest that tetracycline-loaded PLLA membranes potentially enhance guided bone regenerative efficacy and might be a useful barrier for GTR in periodontal treatment.

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Magnetic-activated cell sorting improves high-quality spermatozoa in bovine semen

  • de Assumpcao, Teresinha Ines;Severo, Neimar Correa;Zandonaide, Joao Pedro Brandao;Macedo, Gustavo Guerino
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.2
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    • pp.91-98
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    • 2021
  • The objective of this study was to establish a selection process for high quality sperm in bovine semen using sperm separation by magnetic activation (MACS). For this, semen from 21 Nellore bulls was collected using an artificial vagina. To guarantee the presence of pathologies in the ejaculate, animals previously declassified in four consecutive spermiogram were used. Semen was analyzed in five statuses: (1) fresh semen (fresh); (2) density gradient centrifugation (DGC), percoll column; (3) non-apoptotic fraction after separation by MACS (MAC); (4) apoptotic fraction from the separation (MACPOOR); and (5) MAC followed by DGC (MACDGC). Using a computerized analysis system (CASA), motility was measured. The sperm morphology was evaluated by phase contrast, and the supravital test was completed with eosin/nigrosin staining. For DGC, 20 × 106 cells were used in a gradient of 90% and 45% percoll. MACS used 10 × 106 cells with 20 μL of nanoparticles attached to annexin V, and filtered through the MiniMACS magnetic separation column. Membrane integrity was assessed with SYBR-14/IP and mitochondrial potential with JC-1 by flow cytometry. Processing sperm by MACDGC, was more effective in obtaining samples with high quality sperm, verified by the total of abnormalities in the samples: 35.04 ± 2.29%, 21.50 ± 1.47%, 17.30 ± 1.10%, 30.68 ± 1.94% and 10.50 ± 1.46%, respectively for fresh, DGC, MAC, MACPOOR, and MACDGC. The subpopulation of non-apoptotic sperm had a high number of live cells (82.65%), membrane integrity (56.60%) and mitochondrial potential (83.98%) (p < 0.05). These findings suggest that this nanotechnological method, that uses nanoparticles, is efficient in the production of high-quality semen samples for assisted reproduction procedures in cattle.

The Functional Role of Lysosomes as Drug Resistance in Cancer (항암제 내성에 대한 라이소좀의 역할)

  • Woo, Seon Min;Kwon, Taeg Kyu
    • Journal of Life Science
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    • v.31 no.5
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    • pp.527-535
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    • 2021
  • Lysosomes are organelles surrounded by membranes that contain acid hydrolases; they degrade proteins, macromolecules, and lipids. According to nutrient conditions, lysosomes act as signaling hubs that regulate intracellular signaling pathways and are involved in the homeostasis of cells. Therefore, the lysosomal dysfunction occurs in various diseases, such as lysosomal storage disease, neurodegenerative diseases, and cancers. Multiple forms of stress can increase lysosomal membrane permeabilization (LMP), resulting in the induction of lysosome-mediated cell death through the release of lysosomal enzymes, including cathepsin, into the cytosol. Here we review the molecular mechanisms of LMP-mediated cell death and the enhancement of sensitivity to anticancer drugs. Induction of partial LMP increases apoptosis by releasing some cathepsins, whereas massive LMP and rupture induce non-apoptotic cell death through release of many cathepsins and generation of ROS and iron. Cancer cells have many drug-accumulating lysosomes that are more resistant to lysosome-sequestered drugs, suggesting a model of drug-induced lysosome-mediated chemoresistance. Lysosomal sequestration of hydrophobic weak base anticancer drugs can have a significant impact on their subcellular distribution. Lysosome membrane damage by LMP can overcome resistance to anticancer drugs by freeing captured hydrophobic weak base drugs from lysosomes. Therefore, LMP inducers or lysosomotropic agents can regulate lysosomal integrity and are novel strategies for cancer therapy.

Effects of Benzalkonium Chloride on the growth and survival of Human corneal epithelial cells (점안액 보존제 성분 Benzalkonium Chloride에 의해 유도된 각막상피세포의 세포고사 유도)

  • Kim, Jai-Min;Lee, Seok-Ju;Seo, Eun-Sun
    • Journal of Korean Ophthalmic Optics Society
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    • v.7 no.2
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    • pp.189-195
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    • 2002
  • The aim of this study was to investigate the action of benzalkonium chloride (BAC) used as a preservative in most ophthalmic topical solutions, on human corneal epithelial (HCE) cells in vitro. HCE cell line was exposed to BAC solutions at various concentrations (0.01%~0.0001%) for 15 minutes followed by 24 hours of cell recovery. Cell viability was assessed using MTT assay and chromatin condensation with a Hoechst 33342 test. The expression of membrane protein Fas and Fas ligand was examined by western blot and immunocytochemistry, and DNA fragmentation was studied by agarose gel electrophoresis. A significant decrease of membrane integrity with chromatin condensation was observed with BAC tested at concentrations of 0.005% and higher. BAC was cytotoxic preservatives in this study. An apoptotic mechanism appeared to be present at low concentrations of BAC, whereas a necrotic process appeared at higher concentrations. A functional Fas-mediated apoptotic pathway is present in cultured HCE cells and can be activated by upregulation of Fas expression with BAC.

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Novel Bacterial Surface Display System Based on the Escherichia coli Protein MipA

  • Han, Mee-Jung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.1097-1103
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    • 2020
  • Bacterial surface display systems have been developed for various applications in biotechnology and industry. Particularly, the discovery and design of anchoring motifs is highly important for the successful display of a target protein or peptide on the surface of bacteria. In this study, an efficient display system on Escherichia coli was developed using novel anchoring motifs designed from the E. coli mipA gene. Using the C-terminal fusion system of an industrial enzyme, Pseudomonas fluorescens lipase, six possible fusion sites, V140, V176, K179, V226, V232, and K234, which were truncated from the C-terminal end of the mipA gene (MV140, MV176, MV179, MV226, MV232, and MV234) were examined. The whole-cell lipase activities showed that MV140 was the best among the six anchoring motifs. Furthermore, the lipase activity obtained using MV140 as the anchoring motif was approximately 20-fold higher than that of the previous anchoring motifs FadL and OprF but slightly higher than that of YiaTR232. Western blotting and confocal microscopy further confirmed the localization of the fusion lipase displayed on the E. coli surface using the truncated MV140. Additionally the MV140 motif could be used for successfully displaying another industrial enzyme, α-amylase from Bacillus subtilis. These results showed that the fusion proteins using the MV140 motif had notably high enzyme activities and did not exert any adverse effects on either cell growth or outer membrane integrity. Thus, this study shows that MipA can be used as a novel anchoring motif for more efficient bacterial surface display in the biotechnological and industrial fields.

Study on Suitable Semen Additives Incorporation into the Extender Stored at Refrigerated Temperature

  • Bhakat, M.;Mohanty, T.K.;Raina, V.S.;Gupta, A.K.;Pankaj, P.K.;Mahapatra, R.K.;Sarkar, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.10
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    • pp.1348-1357
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    • 2011
  • The objective of this study was to compare the effect of Butylated Hydroxy Toluene (BHT), Pentoxifylline (PTX) and ${\alpha}$-tocopherol (Vit E) on semen quality parameters of Karan Fries bulls. The fortification of extender by various semen additives improves motility as well as fertility of spermatozoa. Split samples of 24 ejaculates of four Karan Fries bulls were extended in extender with or without various additives such as BHT, PTX and Vit E, and performance was evaluated at an interval of 0, 24, 48 and 72 h at refrigerated temperature (4-$7^{\circ}C$). Results of the present study revealed that addition of BHT, PTX and Vit E in extender improved sperm cell function, such as motility, viability, HOST, and acrosome integrity, as compared to the control during liquid storage up to 48 h of preservation at refrigerated temperature. There was no significant (p<0.05) difference between any of the additives up to 48 h of preservation. Overall, the results showed a significant (p<0.05) deterioration in motility after each storage interval. The results showed a significant deterioration in the acrosome integrity and plasma membrane integrity up to 48 h; subsequently, there was not much degradation of both the semen quality parameters. There was a significant increase in spermatozoal tail and total abnormality after each storage interval at refrigerator temperature (4 to $7^{\circ}C$); however, the head and mid-piece abnormalities were almost unaffected. Tail and total abnormality were least in extender fortified with BHT, PTX and Vit E at different hours of incubation as compared to the control. The addition of 1.5 mM BHT, 3.6 mM PTX and 1 mg/ml Vit E in the semen extender has more beneficial effect in terms of semen quality and preservability of spermatozoa.

Neuroprotective Effect of Scopoletin from Angelica dahurica on Oxygen and Glucose Deprivation-exposed Rat Organotypic Hippocampal Slice Culture

  • Son, Dong-Wook;Lee, Pyeong-Jae;Lee, Jong-Seok;Lee, Sang-Hyun;Choi, Sang-Yoon;Lee, Jong-Won;Kim, Sun-Yeou
    • Food Science and Biotechnology
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    • v.16 no.4
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    • pp.632-635
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    • 2007
  • This study examined the neuroprotective effect of scopoletin from Angelica dahurica against oxygen and glucose deprivation-induced neurotoxicity in a rat organotypic hippocampal slice culture. Scopoletin reduced the propidium iodide (PI) uptake, which is an indication of impaired cell membrane integrity. In addition, it inhibited the loss of NeuN, which represents the viability of neuronal cells. The results suggests that scopoletin from A. dahurica protects neuronal cells from the damage caused by oxygen and glucose deprivation.

Exploring Staphylococcus aureus Virulence Factors; Special Emphasis on Staphyloxanthin

  • Yehia, Fatma Al-zahraa A.;Yousef, Nehal;Askoura, Momen
    • Microbiology and Biotechnology Letters
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    • v.49 no.4
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    • pp.467-477
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    • 2021
  • Staphylococcus aureus is a well-known pathogen that can cause diseases in humans. It can cause both mild superficial skin infections and serious deep tissue infections, including pneumonia, osteomyelitis, and infective endocarditis. To establish host infection, S. aureus manages a complex regulatory network to control virulence factor production in both temporal and host locations. Among these virulence factors, staphyloxanthin, a carotenoid pigment, has been shown to play a leading role in S. aureus pathogenesis. In addition, staphyloxanthin provides integrity to the bacterial cell membrane and limits host oxidative defense mechanisms. The overwhelming rise of Staphylococcus resistance to routinely used antibiotics has necessitated the development of novel anti-virulence agents to overcome this resistance. This review presents an overview of the chief virulence determinants in S. aureus. More attention will be paid to staphyloxanthin, which could be a possible target for anti-virulence agents.