• Title/Summary/Keyword: mCPP

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Biological Activities of Calcium Polyphosphate (Calcium polyphosphate의 생물학적 활성도에 관한 연구)

  • Seol, Yang-Jo;Lee, Jae-Il;Lee, Yong-Moo;Lim, Yoon-Tak;Kim, Seok-Young;Ku, Young;Rhyu, In-Chul;Hahm, Byung-Do;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.213-231
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    • 2000
  • 이 연구의 목적은 다공성의 CPP 내부에 쥐의 장골의 골수에서 유래된 세포를 접종하고 3차 원적으로 배양하여 CPP가 골 형성을 위한 조직공학의 지지체로 적용가능한가를 연구하는 것과 Calcium PolyPhosphate(CPP)의 돌연변이 유발성을 검사하는 것이다. 무수 ($Ca(H_2PO_4)$)를 condensation하여 무결정의 ($Ca(PO_3)$)를 얻고 이를 용융하고 냉각시킨 후 분쇄하여 Calcium polyphosphate(CPP) powder를 얻었다. 다공성의 CPP는 5% $SiO_2$를 첨가하여 sponge 형태로 $450-550{\mu}m$ 소공의 크기를 가지는 것과(CPP-45ppi) $200-300{\mu}m$의 소공의 크기를 가지는 것(CCP-60ppi) 2가지로 제작하였다. 각각의 CPP matrices는 $5mm{\times}5mm{\times}1mm$의 블록 형태로 만들었다. 체중 100g 내외의 백서에서 장골(femur, tibia)을 채취하여 백서의 장골 골수 세포를 분리하여 배양한 후 24well에 CPP block을 넣고 CPP block 당 $10^5$개의 배양한 세포를 접종하였다. 배양 1, 7, 14, 및 21 일째에 각 well에서 trypsin EDTA를 이용하여 2회 반복하여 cell을 분리하였고, 원심분리한 후 hemacytometer로 측정하였다. 또, 45ppi와 60ppi, 그 리 고 Tissue Culture Polystyrene(control group)에 접종, 배양된 세포들의 염기성 인산분해효소활성도를 배양 7, 14, 및 21 일째에 각각 측정하였다. 각 기간별로 배양된 세포-CPP 혼합체내에서 세포의 부착 및 증식과 형성된 조직의 3차원적 형태를 관찰하기 위하여 주사전자현미경하에서의 관찰하였다. CPP의 돌연변이 유발성 검사 (mutagenicity test)를 위해 hypoxanthine-guanine phosphoribosyl transferase(HPRT) assay를 하였다. NIH3T3 cell line과 CHO-K1 cell line으로 각각 $1000{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$ 그리고 $1{\mu}g/m{\ell}$의 CPP 농도에서 측정하였다. 통계적 분석을 위해서 모든 측정은 각군당 4개체 이상 시험하였고, 각 측정값은 평균값${\pm}$표준편차로 나타내었다. 각 군간의 통계적 유의성 검정을 위해서 Analysis of variance(ANOVA)를 이용하였고 Tukey의 방법으로 사후분석을 실시하였다. 제작된 CPP matrices 소공들이 서로간에 연결이 잘 되어있는 형태였다. 두 가지로 제조된 CPP(45ppi와 60ppi) 모두에서 세포의 부착이 잘 일어났고, 부착된 세포의 분열도 잘 일어났다. 2 가지의 CPP 모두에서 7, 14, 21일째의 세포 수는 1일째에 비해 유의성 있게 증가하였다(P<0.01). 3차원적 구조인 Calcium PolyPhosphate에서 배양한 세포는 24well dish(tissue culture polystyrene)에서 평면적으로 배양한 대조군의 세포에서 보다 염기성 인산분해효소 (Alkaline Phosphatase)를 유의성 있게 높게 나타냈다. 주사전자현미경에서 세포-CPP 혼합체를 관찰한 결과, CPP block에 세포들이 잘부착되어 있었고, 시간이 지남에 따라 세포가 여러 층을 형성하면서 뭉치는 현상을 보였다. 또, HPRT assay 결과 , Calcium PolyPhosphate는 돌연변이 유발성을 보이지 않았다. 이상의 결과로 볼 때 CPP에는 세포부착이 잘 일어나고, 지지체 상에서 세포의 분열도 활발하게 일어나므로 골조직을 위한 조직공학의 우수한 지지체가 될 수 있을 것으로 사료된다.

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Formation and Characterization of Casein Phosphopeptide/Chitosan Oligosaccharide NanoComplex (케이신 포스포펩티드/키토올리고당 나노 복합체 형성과 특성 연구)

  • Baek, Yun-Seo;Ha, Ho-Kyung;Lee, Ji-Hong;Lee, Mee-Ryung;Lee, Won-Jae
    • Journal of Dairy Science and Biotechnology
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    • v.36 no.3
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    • pp.164-170
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    • 2018
  • The objectives of this study were to manufacture casein phosphopeptide (CPP)/chitosan oligosaccharide (CSO) nanocomplexes and to investigate the impacts of manufacturing variables, such as CPP concentration and pH, on their morphological and physicochemical characteristics. Transmission electron microscopy (TEM) and particle size analysis were used to assess the morphological and physicochemical properties of the CPP/CSO nano-complexes, respectively. Based on the images obtained by TEM, the spherical shapes of the CPP/CSO nanocomplexes ranged from 50 to 150 nm. As the concentration of CPP was increased and the pH was decreased, the average particle size of the nanocomplexes significantly (p<0.05) increased. The CPP/CSO nanocomplexes had a highly uniform distribution with a polydispersity index value of less than 0.3. In addition, they had a negative surface charge with a zeta-potential value between -17 and -26 mV. The CPP/CSO nanocomplexes showed good stability during the freeze-drying process. In conclusion, CPP/CSO nanocomplexes were successfully manufactured, and the CPP concentration and pH were found to be key factors that affected their morphological and physicochemical properties.

Effect of Dietary Caseinphosphopeptide Levels on Bone Metabolism in Ovariectomized Rats (카제인포스포펩티드의 수준별 섭취가 난소절제 흰쥐의 골대사에 미치는 영향)

  • Yang Daum;Lee Jin-Woo;Park Hong Joo;Kim Sun Hee;Chang Moon-Jeong
    • Journal of Nutrition and Health
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    • v.39 no.1
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    • pp.11-17
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    • 2006
  • The effect of the level of casein phosphopeptide (CPP) on mineral (Ca and P) bioavailabilties and bone biomarker of aged ovariectomized (OVX) Sprague-Dawley rats were studied as a model for postmenopausal bone loss. Forty five Spargue dawley rats, 220-230 g of body weight were fed a control diet (AIN 93M) or containing different level of CPP diet for 7 weeks: $0\%$ (sham control; SC, OVX control; OC), $1\%$ (OVX low CPP diet: OL), $2\%$ (OVX medium CPP diet; OM), $3\%$ (OVX high CPP diet; OH) Ca absorption was unaffected by increasing CPP content from 0 to $3\%$. Urinary Ca excretion was increased by OVX, and decreased by CPP significantly (p < 0.05) with no evident doserelationship. The urinary P excretion was increased by CPP intake in OVX rats. The fecal excretion of P given CPP decreased in OVX with dose dependent manner. Ca and P contents of femur significantly increased by adding 2 or $3\%$ of CPP when compared with OC group and OL group (p < 0.05). There were no significant differences in serum alkaline phosphatase activity and c-terminal telopeptide excretion in experimental groups. Although ovariectomy induced the increase in urinary c-terminal telopeptide excretion, 2 or $3\%$ of CPP in the diet decreased urinary c-terminal telopetide excretion significantly. These finding suggest the usefulness of CPP in the prevention of postmenopausal bone loss by decreasing urinary Ca excretion and bone resorption. Over 2 percent of CPP in the diet was effective to prevent postmenopausal bone loss.

Degradation rate of several types of Calcium Polyphosphate;Long term results (다양한 형태의 다공질 Calcium Polyphosphate의 생분해성에 관한 장기적인 연구)

  • Yang, S.M.;Seol, Y.J.;Kye, S.B.;Lee, I.K.;Lee, C.W.;Kim, S.Y.;Lee, Yong-Mu;Ku, Y.;Han, S.B.;Chung, C.P.;Choi, S.M.;Rhyu, I.C.
    • Journal of Periodontal and Implant Science
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    • v.33 no.2
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    • pp.301-310
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    • 2003
  • The purpose of this study is to evaluate the biocompatibility and the biorsorbability of several types of calcium polyphosphate made through change of manufacturing process for 12 month. To solve limitation of calcium phosphate, we developed a new ceramic, Calcium Polyphosphate(CPP), and report the biologic response to CPP in extraction sites of beagle dog. Porous CPP blocks were prepared by condensation of anhydrous $Ca(H_2PO_4)_2$ to form non-crystalline $Ca(PO_3)_2$ and then milled to produce CPP powder. CPP powder, CPP block, and CPP granules added with $Na_2O$ were implanted in extraction sockets and histologic observation were performed at 12 months later. Like 3 months results, histologic observation at 12 months revealed that CPP matrix were mingled with and directly apposed to new bone without any adverse tissue reaction, CPP powder show direct bony contact, but new bone formation and fibrous tissue encapsulation showed in CPP block. 10% $Na_2O$ CPP granules show more inflammatory cells infiltration around graft materials compared at 3 month, but 15% $Na_2O$ CPP granules show less. This result revealed that regardless of addition of $Na_2O$, CPP had a high affinity for bone and had been resorbed slowly. From this results, it was suggested that CPP is promising ceramic as a bone substitute and addition of $Na_2O$ help biodegradation but optimal concentration of $Na_2O$ and other additive component to increase degradation rate should be determined in further study.

Immuno-Enhancing Effects through Macrophages of Polysaccharides Isolated from Citrus Peels (진피로부터 분리한 다당의 대식세포를 통한 면역증진 효과)

  • Lee, Kyung-Ae;Park, Hye-Ryung
    • The Korean Journal of Food And Nutrition
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    • v.34 no.5
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    • pp.441-448
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    • 2021
  • This study was designed to investigate the intracellular signaling pathways and immunoenhancing effect of macrophage activation by crude polysaccharides (CPP) extracted from citrus peels. CPP did not affect the cytotoxicity of RAW264.7 cells, but showed dose-dependent effects on cell viability. Also, CPP showed high production of chemokine (nitric oxide (NO)) and cytokines (interleukin (IL)-6 and tumor necrosis factor (TNF)-α). CPP increased IL-6, TNF-α, and inducible nitric oxide synthase (iNOS) mRNA expression dose-dependently. CPP also strongly induced the phosphorylation of the ERK, p38, and IκBα pathways in RAW 264.7 cells. In anti-pattern recognition receptors (PRRs) experiments, the effect of CPP on NO production was strongly suppressed by neutralizing toll-like receptor (TLR)2, TLR4, and Dectin1 antibodies, whereas IL-6 and TNF-α production by CPP was mainly suppressed by mannose receptor (MR). Therefore, these results suggest that CPP treatment-induced NO production was regulated by the ERK, p38, and NF-κB pathways through TLR2, TLR4, and Dectin1 receptors, whereas IL-6 and TNF-α production was primarily regulated by the ERK, p38, and NF-κB pathways through MR receptors.

Regulation of Pluripotency-related Genes and Differentiation in Mouse Embryonic Stem Cells by Direct Delivery of Cell-penetrating Peptide-conjugated CARM1 Recombinant Protein

  • Choi, Sara;Jo, Junghyun;Seol, Dong-Won;Cha, Soo Kyung;Lee, Jeoung Eun;Lee, Dong Ryul
    • Development and Reproduction
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    • v.17 no.1
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    • pp.9-16
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    • 2013
  • Coactivator-associated arginine methyltransferase 1 (CARM1) is included in the protein arginine methyltransferase (PRMT) family, which methylates histone arginine residues through posttranslational modification. It has been proposed that CARM1 may up-regulate the expression of pluripotency-related genes through the alteration of the chromatin structure. Mouse embryonic stem cells (mESCs) are pluripotent and have the ability to self-renew. The cells are mainly used to study the genetic function of novel genes, because the cells facilitate the transmission of the manipulated genes into target mice. Since the up-regulated methylation levels of histone arginine residue lead to the maintenance of pluripotency in embryos and stem cells, it may be suggested that CARM1 overexpressing mESCs elevate the expression of pluripotency-related genes in reconstituted embryos for transgenic mice and may resist the differentiation into trophectoderm (TE). We constructed a fusion protein by connecting CARM1 and 7X-arginine (R7). As a cell-penetrating peptide (CPP), can translocate CARM1 protein into mESCs. CPP-CARM1 protein was detected in the nuclei of the mESCs after a treatment of 24 hours. Accordingly, the expression of pluripotency-related genes was up-regulated in CPP-CARM1-treated mESCs. In addition, CPP-CARM1-treated mESC-derived embryoid bodies (EBs) showed an elevated expression of pluripotency-related genes and delayed spontaneous differentiation. This result suggests that the treatment of recombinant CPP-CARM1 protein elevates the expression of pluripotency-related genes of mESCs by epigenetic modification, and this protein-delivery system could be used to modify embryonic fate in reconstituted embryos with mESCs.

CPP-ACP of artificially demineralized enamel surface and remineralization of material containing nano-sized carbonated apatite (인공 탈회된 법랑질 표면의 CPP-ACP와 nano-sized carbonated apatite 함유물질의 재광화효과)

  • Kim, Young-Sook
    • Journal of Korean society of Dental Hygiene
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    • v.8 no.1
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    • pp.13-22
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    • 2008
  • This study compared tooth's remineralization using enamel surface artificially demineralized with 0.1M lactate and HCL solution using Vicker's Hardness Number(VHN) to compare CPP-ACP and remineralization of nano-sized Carbonate Apatite's initial caries. Using pH circulation models divided into 0% nano-CA, 5% nano-CA, 10% nano-CA, 10% CPP-ACP and D.W. they were treated for 5 minutes, three times a day for 14 days to get the following results. 1. There were no significant differences among the initial surface hardness of samples demineralized surface of front tooth in 5 groups. and all 5 groups' surface hardness reduced significantly after demineralization of enamel. 2. When inquiring into hardness changes through pH circulation model, the highest hardness change was in 5% nano-CA group. Also. 10% nano-CA and 10% CPP-ACP groups increased significantly. but there was no significant difference statistically. In generalizing the above experiment results, nano-sized Carbonate Apatite showed remineralization, and compared to 10% CPP-ACP group, 5% nano-CA had remineralization to artificial caries. thus implies that when we develop method to contact with tooth of nano-CA in the future, it is expected to gain synergy effect on function of saliva, a natural remineralization material.

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Isolation and Characteristics of Lactic Acid Bacteria Producing Casein Phosphopeptides from Kimchi (Casein phosphopeptide를 생산하는 김치 유래 유산균의 분리 및 특성 연구)

  • Lee, Mi-Gyeong;Kwon, Hyo-Jeong;Byun, Ock-Hee;Bang, Bo-Yeon;Kim, Yu-Jin;Park, JungMin;Bai, Dong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.44 no.1
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    • pp.68-73
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    • 2016
  • Lactic acid bacteria showing both protease activity and the capacity to produce casein phosphopeptide (CPP) were isolated from Korean kimchi, a traditional food made from fermented vegetables. Among the 450 strains of isolated lactic acid bacteria, the strain MG-379 showed high protease activity and the highest ability to produce CPP. Characterization results showed that MG-379 was gram-positive and measured $0.6-0.8{\mu}m$ in diameter. DNA sequencing of MG-379 and comparison with other sequences using BLAST revealed a 100% identity with the sequence of Enterococcus faecalis. However, MG-379 showed a higher CPP-producing ability than E. faecalis KCCM 40450. Accordingly, MG-379 was newly named as E. faecalis MG-379. Amount of free calcium liberated by CPPs was 2227.5 and 1151.6 mg/kg for E. faecalis MG-379 and E. faecalis KCCM (control), respectively.

Investigation on the Heating Patterns Depending on the Packaging Materials During Microwave Cooking (포장 소재에 따른 전자레인지 가열 조리 패턴 조사)

  • Lee, Hwa Shin;Cho, Ah Reum;Moon, Sang Kwon;Yoon, Chan Suk;Lee, Keun Taik
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.21 no.1
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    • pp.27-34
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    • 2015
  • Heating patterns depending on the packaging materials were examined in order to investigate the causes of thermal deformation of packages used for ready-to-eat foods for microwave heating due to the non-uniformity of microwaves. Physical properties including tensile strength, heat-resistance and elongation of four different CPP grades were compared. High retortable CPP had higher sealing strength and heat resistance compared to the conventional CPPs. All CPP samples tested were proved to have melting temperatures around $160^{\circ}C$. However, they were all thermally deformed by microwave heating due to a limited penetration of microwave and non-uniform heating within the spicy sauce of high viscosity contained high salt, especially on the above the filling line and sealing edge of pouches. When the laminated stand-up pouches composed of G-PET/PET/PET/CPP and G-PET/PET/NY/CPP were retorted and microwaved, significant deformations were noticed in both samples after retorting. Besides, pouches contained titanium dioxide showed more intense thermal deformation than the control. When the $10{\mu}m$ aluminium foil was affixed on the pouch, small thermal deformation was observed only in the bottom layer. More studies are required to prevent the thermal deformation of packaging materials used for RTE foods during microwave heating by developing the technologies to increase the thermal stability of CPP layer and the modification of packaging design to modify the microwave access into the package.

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Tissue Response of Calcium Polyphosphate in Beagle Dog (비글견에서의 Calcium Polyphosphate의 조직반응에 관한 연구)

  • Yang, S.M.;Kim, W.H.;Lee, S.J.;Kim, S.Y.;Lim, Y.T.;Lee, J.Y.;Lee, Y.M.;K, Young;Chung, C.P.;Han, S.B.;Choi, S.M.;Rhyu, I.C.
    • Journal of Periodontal and Implant Science
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    • v.31 no.3
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    • pp.543-554
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    • 2001
  • 치주조직 재생을 도모하기 위한 전통적인 시술방법으로 여러가지 골이식재를 이용한 골이식술이 이용되고 있다. 이번 실험의 목적은 골형성을 위한 재료로 Calcium Polyphosphate(CPP)를 사용하여 비글견에서의 조직 반응과 골유도성을 보는 것이고 또한 다른 이식재들간에 신생골의 형성에 어느정도 영향을 주는 가를 알아보는 것이다. 이번 실험에 사용된 CPP는 무수 $Ca(H_2PO_4)$를 condensation하여 무결정의 $Ca(PO_3)_2$를 얻고 이를 용율하고 냉각시킨 후 분쇄하여 얻은 것으로 3세된 비글견에 이식하여 관찰하였다. 조직학적으로 CPP granule의 경우는 키토산이나 $Na_2O$를 넣은 경우 모두 bone 의 ingrowth가 관찰되었고 다른 섬유조직의 개재를 볼수 없었다 동결탈회건조골을 넣은 경우에는 주위로 골형성 보이지 않았고 단지 섬유성 조직이 관찰 되었다. 아무것도 넣지 않은 경우에 비해서 동결탈회건조골이나 키토산, $Na_2O$를 넣은 CPP granules 경우에 더 많은 비율로 신생 골의 양이 나타나는 것을 볼수 있었다. 아무것도 넣지 않은 대조군과 이식재를 넣은 군간에는 유의성이 있는 것으로 나타났고(p<0.05). 또한 CPP granules with chitosan과 CPP granules with $Na_2O$ 사이에는 신생골의 형성에 유의성이 없었다. 이것으로 보아 CPP granules with chitosan과 CPP granules with $Na_2O$는 모두 골유도성이 있고 신생골의 형성을 촉진하므로 치주질환으로 인한 골결손 부위에 사용할 수 있는 재료로 우수한 특성을 지닌다고 사료된다.

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