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      • KCI등재

        Nigella Sativa-Coated Hydroxyapatite Scaffolds: Synergetic Cues to Stimulate Myoblasts Differentiation and Offset Infections

        Amna Touseef,Alghamdi Abdullah A. A.,SHANG KE,Hassan M. Shamshi 한국조직공학과 재생의학회 2021 조직공학과 재생의학 Vol.18 No.5

        Background: At present osteoporosis has come into view as a major health concern. Skeletal diseases typified by weak and fragile bones have imposed threats of fissure. Hydroxyapatite (HAP) is known to induce osteoblast like differentiation and provide mechanical strength, hence, used in bone tissue engineering; whereas, Nigella sativa has also demonstrated potential to treat bone and muscle diseases. This study was aimed to develop potential orthopedic scaffold exploiting natural resources of Saudi Arabia which can be used as prospective tissue engineering implant. Methods: The bone scaffold was developed by grafting biogenic HAP with N. sativa essential oil. N. sativa was applied for boosting osteogenesis and to stimulate antimicrobial potential. Antimicrobial potential was investigated utilizing S. aureus bacteria. Spectroscopic and surface characters of N. sativa grafted HAP scaffolds were analyzed using Fourier-transform infrared spectroscopy, X-ray crystallography and Scanning electron microscopy. To ensure biocompatibility of scaffolds; we selected C2C12 cell-lines; best model to study mechanistic pathways related to osteoblasts and myoblasts differentiation. Results: Grafting of HAP with N. sativa did not affect typical spherical silhouette of nanoparticles. Characteristically; protein loaded polynucleated myotubes are result of in vitro myogenesis of C2C12 myoblasts in squat serum environment. Conclusion: It is first study of unique combination of N. sativa and HAP scaffold as a possible candidate of implantation for skeletal muscles regeneration. Outcome of this finding revealed N. sativa grafted HAP enhance differentiation significantly over that of HAP. The proposed scaffold will be an economical natural material for hard and soft tissue engineering and will aid in curing skeletal muscle diseases. Our findings have implications for treatment of muscular/bone diseases. Background: At present osteoporosis has come into view as a major health concern. Skeletal diseases typified by weak and fragile bones have imposed threats of fissure. Hydroxyapatite (HAP) is known to induce osteoblast like differentiation and provide mechanical strength, hence, used in bone tissue engineering; whereas, Nigella sativa has also demonstrated potential to treat bone and muscle diseases. This study was aimed to develop potential orthopedic scaffold exploiting natural resources of Saudi Arabia which can be used as prospective tissue engineering implant. Methods: The bone scaffold was developed by grafting biogenic HAP with N. sativa essential oil. N. sativa was applied for boosting osteogenesis and to stimulate antimicrobial potential. Antimicrobial potential was investigated utilizing S. aureus bacteria. Spectroscopic and surface characters of N. sativa grafted HAP scaffolds were analyzed using Fourier-transform infrared spectroscopy, X-ray crystallography and Scanning electron microscopy. To ensure biocompatibility of scaffolds; we selected C2C12 cell-lines; best model to study mechanistic pathways related to osteoblasts and myoblasts differentiation. Results: Grafting of HAP with N. sativa did not affect typical spherical silhouette of nanoparticles. Characteristically; protein loaded polynucleated myotubes are result of in vitro myogenesis of C2C12 myoblasts in squat serum environment. Conclusion: It is first study of unique combination of N. sativa and HAP scaffold as a possible candidate of implantation for skeletal muscles regeneration. Outcome of this finding revealed N. sativa grafted HAP enhance differentiation significantly over that of HAP. The proposed scaffold will be an economical natural material for hard and soft tissue engineering and will aid in curing skeletal muscle diseases. Our findings have implications for treatment of muscular/bone diseases.

      • KCI등재

        Electrospun Twin Fibers Encumbered with Intrinsic Antioxidant Activity as Prospective Bandage

        Touseef Amna,Fatehia N Gharsan,Ke Shang,M. Shamshi Hassan,길명섭,황인호 한국고분자학회 2019 Macromolecular Research Vol.27 No.7

        Our investigation for the first time explores the possibility of fabrication of a Capsaicin/polyurethane-based bioactive wound dressing. A micro/nanofibrous bandage was synthesized by electrospinning polyurethane supplemented with natural anti-inflammatory agent, Capsaicin. Herein, the best concentration (3%) of Capsaicin in polyurethane to get a consistent solution was standardized in order to acquire the composite micro/nanofibrous dressing. The as-spun bandage has been investigated by scanning electron microscopy and Fourier transform infrared spectroscopy, and biocompatibility of muscle cells on the bioactive bandage was also examined. Free radical scavenging activity of the fabricated micro/nanofiber bandage was estimated using 2,2-diphenyl-1-picrylhydrazyl assay. Inclusion of Capsaicin in polyurethane transformed the morphology as well as dimension of the fibers. Anti-oxidant wound bandage with diameters around 150-500 nm was fabricated by physical unification of polyurethane with natural Capsaicin. Here, polyurethane was used as a foundation polymer, which was blended with Capsaicin to attain desirable characteristics such as better anti-oxidant activity, hydrophilicity and excellent cell attachment. The polyurethane wound bandage possesses enlarged surface, proscribed evaporation, and fluid drainage ability. These results suggest the beneficial influence of antioxidant Capsaicin on wound repairing process. Therefore, a biologically active, natural compounds such as Capsaicin is material of choice for fabrication of future wound dressings.

      • Zinc oxide-doped poly(urethane) spider web nanofibrous scaffold via one-step electrospinning: a novel matrix for tissue engineering.

        Amna, Touseef,Hassan, M Shamshi,Sheikh, Faheem A,Lee, Hak Kyo,Seo, Kang-Seok,Yoon, Duhak,Hwang, I H Springer International 2013 Applied microbiology and biotechnology Vol.97 No.4

        <P>Zinc oxide (ZnO) nanostructures have been commonly studied for electronic purposes due to their unique piezoelectric and catalytic properties; however, recently, they have been also exploited for biomedical applications. The purpose of this study was to fabricate ZnO-doped poly(urethane) (PU) nanocomposite via one-step electrospinning technique. The utilized nanocomposite was prepared by using colloidal gel composed of ZnO and PU, and the obtained mats were vacuum dried at 60?C overnight. The physicochemical characterization of as-spun composite nanofibers was carried out by X-ray diffraction pattern, field emission scanning electron microscopy, energy-dispersive X-ray spectroscopy, electron probe microanalysis, and transmission electron microscopy, whereas the thermal behavior was analyzed by thermogravimetric analysis. The viability, attachment, and proliferation of NIH 3T3 mouse fibroblast cells on the ZnO/PU composite nanofibers were analyzed by in vitro cell compatibility test. The morphological features of the cells attached on nanofibers were examined by Bio-SEM. We conclude that the electrospun nanofibrous scaffolds with unique spider nets had good biocompatibility. Cytotoxicity experiments indicated that the mouse fibroblasts could attach to the nanocomposite after being cultured. Thus, the current work demonstrates that the as-synthesized ZnO/PU hybrid nanofibers represent a promising biomaterial to be exploited for various tissue engineering applications.</P>

      • SCIESCOPUSKCI등재

        Substantial Effect of Melanin Influencing Factors on In vitro Melanogenesis in Muzzle Melanocytes of Differently Colored Hanwoo

        Amna, Touseef,Park, Kyoung-Mi,Cho, In-Kyung,Choi, Tae-Jeong,Lee, Seung-Soo,Seo, Kang-Seok,Hwang, In-Ho Asian Australasian Association of Animal Productio 2012 Animal Bioscience Vol.25 No.7

        The present study was designed to investigate the effect of ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH), nitric oxide (NO) and L-cysteine on melanin production and expression of related genes MC1R, Tyr, Tyrp-1 and Tyrp-2 in muzzle melanocytes of differently colored three native Hanwoo cattle. Muzzle samples were taken from black, brindle and brown Hanwoo and purified melanocytes were cultured with ${\alpha}$-MSH, nitric oxide and L-cysteine at 100 nM, $50{\mu}M$ and 0.07 mg/ml of media respectively. The amounts of total melanin, eumelanin and mRNA expression at Tyr, Tyrp-1, Tyrp-2 and MC1R levels were quantified. ${\alpha}$-MSH and nitric oxide significantly increased (p<0.05) the amount of total melanin in black and brindle whereas eumelanin production in brown Hanwoo muzzle melanocytes. On the contrary, L-cysteine greatly (p<0.05) depressed the eumelanin production in black color but increased in brown. Simultaneously, up regulation of Tyr by nitric oxide and ${\alpha}$-MSH and down regulation of Tyr, Tyrp-2 and MC1R genes by L-cysteine were observed in muzzle melanocytes of all three phenotypes. The results of this study revealed nitric oxide and ${\alpha}$-MSH contribute hyper-pigmentation by enhancing eumelanogenesis whereas L-cysteine contributes to pheomelanin production in different colored Hanwoo muzzle melanocytes.

      • Virgin olive oil blended polyurethane micro/nanofibers ornamented with copper oxide nanocrystals for biomedical applications

        Amna, Touseef,Hassan, M Shamshi,Yang, Jieun,Khil, Myung-Seob,Song, Ki-Duk,Oh, Jae-Don,Hwang, Inho Dove Medical Press 2014 INTERNATIONAL JOURNAL OF NANOMEDICINE Vol.9 No.-

        <P>Recently, substantial interest has been generated in using electrospun biomimetic nanofibers of hybrids, particularly organic/inorganic, to engineer different tissues. The present work, for the first time, introduced a unique natural and synthetic hybrid micronanofiber wound dressing, composed of virgin olive oil/copper oxide nanocrystals and polyurethane (PU), developed via facile electrospinning. The as-spun organic/inorganic hybrid micronanofibers were characterized by scanning electron microscopy (SEM), energy dispersive X-ray analysis, X-ray diffraction, electron probe microanalysis, and transmission electron microscopy. The interaction of cells with scaffold was studied by culturing NIH 3T3 fibroblasts on an as-spun hybrid micronanofibrous mat, and viability, proliferation, and growth were assessed. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay results and SEM observation showed that the hybrid micronanofibrous scaffold was noncytotoxic to fibroblast cell culture and was found to benefit cell attachment and proliferation. Hence our results suggest the potential utilization of as-spun micronanoscaffolds for tissue engineering. Copper oxide–olive oil/PU wound dressing may exert its positive beneficial effects at every stage during wound-healing progression, and these micronanofibers may serve diverse biomedical applications, such as tissue regeneration, damaged skin treatment, wound healing applications, etc. Conclusively, the fabricated olive oil–copper oxide/PU micronanofibers combine the benefits of virgin olive oil and copper oxide, and therefore hold great promise for biomedical applications in the near future.</P>

      • SCIESCOPUS
      • Classy non-wovens based on animate L. gasseri-inanimate poly(vinyl alcohol): upstream application in food engineering

        Amna, Touseef,Hassan, M. Shamshi,Pandeya, Dipendra Raj,Khil, Myung-Seob,Hwang, I.H. Springer-Verlag 2013 Applied microbiology and biotechnology Vol.97 No.10

        <P>We explored electrospinning as a feasible and practicable mode for encapsulation and stabilization of Lactobacillus gasseri. The utilized nanocomposite was prepared using sol-gel composed of animate L. gasseri and inanimate PVA. The objective was to examine the ability of electrospinning method to protect functional properties of probiotic L. gasseri. The PVA was used as an encapsulation matrix as it is biocompatible and hydrophilic in nature thus facilitate an easy revival of bacteria. The characterization of as-spun bioproduct was done by energy-dispersive X-ray spectrometer, SEM, and TEM, whereas thermal behavior was analyzed by thermogravimetry. The viability was confirmed by traditional pour plate method and fluorescence microscopy. Furthermore, to test whether the functionality of L. gasseri was affected, the encapsulated L. gasseri were fed to mouse for colonization. Our results pointed out that encapsulated bacteria were viable for months, and their metabolism was not affected by immobilization; thus, they could be used in food engineering and trade.</P>

      • Impact on gene expression in response to silver-decorated titania nanomatrix using an in vitro satellite cell culture model

        Amna, Touseef,Hassan, M. Shamshi,Al-Deyab, Salem S.,Khil, Myung-Seob,Hwang, Inho Springer-Verlag 2016 Polymer bulletin Vol.73 No.7

        <P>Herein we report the synthesis of novel silver-ornamented titanium dioxide crossbreed nanofibers via electrospinning with improved cellular response for possible muscle tissue engineering applications. Titanium isopropoxide and silver nitrate served as precursors to get high aspect ratio silver-decorated titania nanofibers. For a model application in muscle tissue engineering, muscle satellite cells of local Korean Hanwoo were cultured on synthesized hybrid nanofibers. The standard microarray technique has been employed to screen the potential effects of synthesized nanofibers on genome after the culture of cells on hybrid nanofibers. EDX, EPMA, FT-IR, XRD and TEM analysis confirmed purity, distribution, interaction and crystalline nature of this material while the diameter of these nanofibers estimated from FE-SEM and TEM were between 200 and 300 nm. Cell counting with Kit-8 assay at regular time intervals and phase contrast microscopy data revealed that satellite cells proliferated well on nanofibers and cellular attachments were visible by SEM. Microarray analyses of gene expression response indicated clear changes in gene expression in a range of proliferation and apoptosis. Our results open-up new ways to develop and analyze gene expression biomarkers for satellite cell proliferation and finally muscle tissue engineering.</P>

      • Influence of Capsaicin on Inflammatory Cytokines Induced by Lipopolysaccharide in Myoblast Cells Under <i>In vitro</i> Environment

        Shang, Ke,Amna, Touseef,Amina, Musarat,Al-Musayeib, Nawal M.,Al-Deyab, Salem S.,Hwang, Inho Medknow PublicationsMedia Pvt Ltd 2017 Pharmacognosy magazine Vol.13 No.49

        <P><B>Background:</B></P><P>ellular damage initiated by reactive oxygen species (ROS) is the main cause of numerous severe diseases and therefore for this reason, the natural antioxidants have note worthy significance in human health. Capsaicin possesses noteworthy analgesic and anti-inflammatory properties. It also possesses healing effects for treatment of arthritis, diabetic neuropathy, gastric lesions, and cardiac excitability that is why it is incorporated in creams and gels.</P><P><B>Objective:</B></P><P>The present study was carried out to estimate the <I>in vitro</I> antioxidant and ROS scavenging activities of capsaicin against muscle precursor cells. Till date, no investigation has been carried out to study the effect of capsaicin on myoblasts.</P><P><B>Materials and methods:</B></P><P>Herein, the cytotoxicity was induced by endotoxin lipopolysaccharide (LPS) to analyze the effect of capsaicin on LPS induced inflammation and apoptosis on muscle cells. To find out the toxicity of endotoxin, myoblasts were exposed to different concentrations of LPS, viability and morphology was checkedby the means of CCK-8 test and microscopy, respectively. Apoptotic cell death was examined by fluorescence staining. Additionally, LPS-induced apoptosis was determined by mRNAexpression of calpain, caspase-3 and tumor necrosisfactor alpha (TNF-α), and were quantified by qRT-PCR.</P><P><B>Results:</B></P><P>The outcome of the presentstudy demonstrated that LPS stimulation generatestoxicity in dose-dependent manner. Pre-treatmentof myoblasts with capsaicin can considerably alleviate LPS-induced inflammation.</P><P><B>Conclusion:</B></P><P>In conclusion, this study indicates that dietetic supplementation of capsicum may help to alleviate/reduce the inflammatory effects and is therefore potent source of natural antioxidant agent which can be utilized to control muscle related diseases, such as myotube atrophy.</P><P><B>SUMMARY</B></P><P><P>In the present study cytotoxicity was induced by LPS to analyze the effect of capsaicin on LPS induced inflammation and apoptosis on muscle cells.</P><P>The results of this investigation demonstrated that LPS stimulation generates toxicity in dose dependent manner. Pre-treatment of myoblasts with capsaicin can considerably reduce LPS induced inflammation.</P><P>It has been concluded on the basis of results that the dietetic supplementation of capsicum may help to minimize inflammatory effects and are potent sources of natural antioxidants which can be utilized to control muscle related diseases such as atrophy.</P></P> >[FIG OMISSION]</BR><P><B>Abbreviation used:</B> AMP: Adenosine monophosphate, AO/EB: Acridine orange / Ethidium bromide, ATL: T-cell leukemi, CAP: Capsaicin, CCK-8: Cell counting Kit-8, CLSM: Laser Scanning Microscopy, DCF-DA: 2’, 7’-dichlorofluorescein diacetate, DMEM: Dulbecco’s modified Eagle’s medium, DPPH: α, α-diphenyl-β-picrylhydrazyl, FBS: Fetal bovine serum, KA: Kainic acid, LPS: Lipopolysaccharide, MDA: Malondialdehyde, NF-κB: Nuclear factor kgene binding, PBS: Phosphate buffer saline, pNA: p-nitroanilide, RNW: RNase free water, ROS: Reactive oxygen species, TNF-α: Tumor necrosis factor alpha, TRPV1: Transient receptor potential vanilloid 1</P>

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