• Title/Summary/Keyword: scaffold protein

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Radiolabeling of antibody-mimetic scaffold protein with 99mTc tricarbonyl precursor via hexahistidine (His6)-tag

  • Shim, Ha Eun;Kim, Do Hee;Lee, Chang Heon;Choi, Dae seong;Lee, Dong-Eun
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.5 no.1
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    • pp.11-17
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    • 2019
  • Recently, antibody-like scaffold proteins have received a great deal of interest in diagnosis and therapy applications because of their intrinsic features that are often required for tumor imaging and therapy. Intrinsic issues that are associated with therapeutic application of antibody-like scaffold proteins, particularly in cancer treatment, include an efficient and straightforward radiolabeling for understanding in vivo biodistribution and excretion route, and monitoring therapeutic responses. Herein, we report an efficient and straightforward method for radiolabeling of antibody-like scaffold proteins with the $[^{99m}Tc(OH_2)_3(CO)_3]^+$ ($^{99m}Tc$-tricarbonyl) by using a site-specific direct labeling method via hexahistidine-tag, which is a widely used for general purification of recombinant proteins with His-affinity chromatography. Repebody is a new class of antibody-like scaffold protein that consists of highly diverse leucine-rich repeat (LRR) modules. Although all possible biomedical applications with repebody are ongoing, it's in vivo biodistribution and excretion pathway has not yet been explored. In this study, hexahistidine ($His_6$)-tag bearing repebody (rEgH9) was labeled with [$^{99m}Tc$]-tricarbonyl. Repebody protein was radiolabeled with high radiolabeling efficiency (>90%) and radiolabeled compound was more than 99% pure after purification. These results clearly demonstrate that the present radiolabeling method will be useful molecular imaging study.

Ankyrin Repeat-Rich Membrane Spanning (ARMS)/Kidins220 Scaffold Protein Regulates Neuroblastoma Cell Proliferation through p21

  • Jung, Heekyung;Shin, Joo-Hyun;Park, Young-Seok;Chang, Mi-Sook
    • Molecules and Cells
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    • v.37 no.12
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    • pp.881-887
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    • 2014
  • Cell proliferation is tightly controlled by the cell-cycle regulatory proteins, primarily by cyclins and cyclin-dependent kinases (CDKs) in the $G_1$ phase. The ankyrin repeat-rich membrane spanning (ARMS) scaffold protein, also known as kinase D-interacting substrate of 220 kDa (Kidins 220), has been previously identified as a prominent downstream target of neurotrophin and ephrin receptors. Many studies have reported that ARMS/Kidins220 acts as a major signaling platform in organizing the signaling complex to regulate various cellular responses in the nervous and vascular systems. However, the role of ARMS/Kidins220 in cell proliferation and cell-cycle progression has never been investigated. Here we report that knockdown of ARMS/Kidins220 inhibits mouse neuroblastoma cell proliferation by inducing slowdown of cell cycle in the $G_1$ phase. This effect is mediated by the upregulation of a CDK inhibitor p21, which causes the decrease in cyclin D1 and CDK4 protein levels and subsequent reduction of pRb hyperphosphorylation. Our results suggest a new role of ARMS/Kidins220 as a signaling platform to regulate tumor cell proliferation in response to the extracellular stimuli.

Cross-Linked Collagen Scaffold from Fish Skin as an Ideal Biopolymer for Tissue Engineering

  • Biazar, Esmaeil;Kamalvand, Mahshad;Keshel, Saeed Heidari;Pourjabbar, Bahareh;Rezaei-Tavirani, Mustafa
    • Korean Journal of Materials Research
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    • v.32 no.4
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    • pp.186-192
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    • 2022
  • Collagen is one of the most widely used biological materials in medical design. Collagen extracted from marine organisms can be a good biomaterial for tissue engineering applications due to its suitable properties. In this study, collagen is extracted from fish skin of Ctenopharyngodon Idella; then, the freeze drying method is used to design a porous scaffold. The scaffolds are modified with the chemical crosslinker N-(3-Dimethylaminopropyl)-N'-ethyl carbodiimide hydrochloride (EDC) to improve some of the overall properties. The extracted collagen samples are evaluated by various analyzes including cytotoxicity test, SDS-PAGE, FTIR, DSC, SEM, biodegradability and cell culture. The results of the SDS-PAGE study demonstrate well the protein patterns of the extracted collagen. The results show that cross-linking of collagen scaffold increases denaturation temperature and degradation time. The results of cytotoxicity show that the modified scaffolds have no toxicity. The cell adhesion study also shows that epithelial cells adhere well to the scaffold. Therefore, this method of chemical modification of collagen scaffold can improve the physical and biological properties. Overall, the modified collagen scaffold can be a promising candidate for tissue engineering applications.

Sustained Release of Proteins Using Small Intestinal Submucosa Modified PLGA Scaffold (SIS로 개질된 PLGA 담체에서의 단백질의 서방화)

  • Ko, Youn-Kyung;Choi, Myung-Kyu;Kim, Soon-Hee;Kim, Geun-Ah;Lee, Hai-Bang;Rhee, John-M.;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.3
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    • pp.199-205
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    • 2008
  • In this study, we fabricated poly (lactide-co-glycolide) (PLGA) scaffold modified with small intestinal submucosa (SIS) as a drug delivery matrix of bioactive molecules. SIS derived from the submucosa layer of porcine intestine has been widely used as biomaterial because of low immune response. PLGA scaffold was prepared by the method of solvent casting/salt leaching. Novel composite scaffolds of SIS/PLGA were manufactured by simple immersion method of PLGA scaffold in SIS solution under vacuum. SEM observation shows that PLGA and SIS/PLGA scaffolds have interconnective and open pores. Especially, SIS/PLGA scaffold showed that micro-sponge of SIS with interconnected pore structures were formed in the pores of PLGA scaffold. In order to assay release profile of proteins, we manufactured FITC conjugated BSA loaded PLGA and SIS/PLGA scaffold. And the release amount was identified by fluorescence intensity using the fluorescence spectrophotometer. The initial burst of BSA containing SIS/PLGA scaffolds was lower than that of PLGA scaffolds resulting in constant release. And release of BSA in SIS/PLGA scaffold was fast and incremental because of the increased content of BSA. In conclusion, we confirmed that penetrated SIS solution prevented the initial burst of BSA and PLGA modified with SIS scaffold is useful as protein carriers with controlled release pattern.

Biomedical Application of Silk Sericin: Recent Research Trend

  • Seong-Gon Kim;Je-Yong Choi;HaeYong Kweon
    • International Journal of Industrial Entomology and Biomaterials
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    • v.48 no.1
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    • pp.1-12
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    • 2024
  • Silk sericin, a natural protein from silkworm cocoons, is emerging as a multifunctional biomaterial in biomedicine, particularly in tissue engineering and wound healing. Recent studies have highlighted its biocompatibility, biodegradability, and potential for chemical modification, which allows it to be incorporated into various scaffold architectures. This review article synthesizes current research, including the development of sericin-based hydrogel scaffolds for tissue engineering and sericin's role in enhancing wound healing. Key findings demonstrate sericin's ability to refine scaffold porosity and mechanical strength, expedite tissue healing, and reduce bacterial load in wounds. The integration of sericin into novel bioactive dressings and its use in peripheral nerve injury repair are also discussed, showcasing its adaptability and efficacy. The convergence of these studies illustrates the broad applications of sericin, from scaffold design to clinical interventions, making it a promising material in regenerative medicine and tissue engineering, with the potential to improve patient outcomes significantly.

Comparison of Soy and Pea Protein for Cultured Meat Scaffolds: Evaluating Gelation, Physical Properties, and Cell Adhesion

  • Do Hyun Kim;Seo Gu Han;Su Jin Lim;Seong Joon Hong;Hyuk Cheol Kwon;Hyun Su Jung;Sung Gu Han
    • Food Science of Animal Resources
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    • v.44 no.5
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    • pp.1108-1125
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    • 2024
  • Cultured meat is under investigation as an environmentally sustainable substitute for conventional animal-derived meat. Employing a scaffolding technique is one approach to developing cultured meat products. The objective of this research was to compare soy and pea protein in the production of hydrogel scaffolds intended for cultured meat. We examined the gelation process, physical characteristics, and the ability of scaffolds to facilitate cell adhesion using mesenchymal stem cells derived from porcine adipose tissue (ADSCs). The combination of soy and pea proteins with agarose and agar powders was found to generate solid hydrogels with a porous structure. Soy protein-based scaffolds exhibited a higher water absorption rate, whereas scaffolds containing agarose had a higher compressive strength. Based on Fourier transform infrared spectroscopy analysis, the number of hydrophobic interactions increased between proteins and polysaccharides in the scaffolds containing pea proteins. All scaffolds were nontoxic toward ADSCs, and soy protein-based scaffolds displayed higher cell adhesion and proliferation properties. Overall, the soy protein-agarose scaffold was found to be optimal for cultured meat production.

Successful strategy of treatment used to rhBMP-2 and LFA-collagen scaffold for BRONJ (임상가를 위한 특집 3 - rhBMP-2와 LFA-collagen scaffold를 이용한 BRONJ의 성공적인 치료 전략)

  • Kwon, Kyung-Hwan
    • The Journal of the Korean dental association
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    • v.52 no.4
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    • pp.218-233
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    • 2014
  • Bispbosphonates are a class of pharmaceutic agents, which induce apoptosis of osteoclast as well as impair osteoclastic activity to suppress bone resorption. Thus, bisphophonates are effectively used to treat osteoporosis, multiple myeloma and to prevent bone metastases of malignant cancer. However, recently dental disease have been reported associated with Bisphosphonates. Thus, there are a number of discussions about proper prevention and treatment of bisphosphonate-related osteonecrosis of jaw(BRONJ). Marshall R. Urist in 1965 made the seminal discovery that a specific protein, BMP(bone morphogenetic protein), found in the extracellular matrix of demineralized bone could induce bone formation newly when implanted in extraosseous tissues in a host. BMPs are multi-functional growth factors which are members of the transforming growth factor-beta super family and their ability is that plays a pivotal roll in inducing bone. About 18 BMP family members have been identified and characterized. Among of them, BMP-2 and BMP-7 have significant importance in bone development. In this study, patients of BRONJ were recieved who visited Department of oral and maxillofacial surgery, school of dentistry, Wonkwang university for past 3 years from 2011 to 2013. We focused on the results of the surgical intervention. We suggest that new strategy of treatment used to rhBMP-2 and LFA(Lidocaine-Fibrinogen-Aprotinin)-collagen scaffold for patients of BRONJ. The purpose of this paper is to give a brief overview of BMPs and to critically review the clinical data currently available on rhBMP-2 and LFA collage scaffold.

The Synergistic Effects of Agarose Scaffold Supplemented with Low-molecular-weight Silk Fibroin in Bone Tissue Regeneration

  • Park, Seung-Won;Goo, Tae-Won;Kim, Seong-Ryul;Kweon, Hae-Yong;Kang, Seok-Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.23 no.2
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    • pp.193-199
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    • 2011
  • Silk protein and agarose are widely known as biocompatible materials in the human body. A three-dimensional (3D) scaffold composed of agarose and low-molecular- weight silk fibroin (LSF) was fabricated and examined in terms of structural characteristics and cellular responses in bone tissue engineering. This study showed that mouse pluripotent precursor cells attached to and proliferated uniformly on and within the LSF-containing 3D scaffold. Interestingly, cell proliferation and attachment was shown to be higher in a 3D scaffold containing 0.02% LSF, as compared to other LSF concentrations. The results of this study suggest that agarose-LSF scaffolds may be useful materials for tissue engineering.

Efficient Solid Phase Library Synthesis of 7 -Alkoxy-1,3,4,5-tetrahydro-benzo [e][ 1.4] diazepin-2-one

  • Im, Isak;Kim, Yong-Chul
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.342.2-342.2
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    • 2002
  • The ${\beta}$-turn has been implicated as an important conformation for biological recognition of peptides or proteins. Benzodiazepine classes have been known as one of the non peptide ${\beta}$-turn mimic scaffolds. We have developed an efficient approach for the synthesis and derivatization of a scaffold of hydroxytetrahydrodizepinone class in order to screen compound library in various protein targets for new lead generations as well as for structure activity relationships of the scaffold. (omitted)

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