• Title/Summary/Keyword: Cellular Attachment

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Anti-metastatic Effects on B16F10 Melanoma Cells of Extracts and Two Prenylated Xanthones Isolated from Maclura amboinensis Bl. Roots

  • Siripong, Pongpun;Rassamee, Kitiya;Piyaviriyakul, Suratsawadee;Yahuafai, Jantana;Kanokmedhakul, Kwanjai
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3519-3528
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    • 2012
  • Inhibitory effects of Maclura amboinenesis Bl, one plant used traditionally for the treatment of cancers, on metastatic potential of highly metastatic B16F10 melanoma cells were investigated in vitro. Cell proliferation was assessed using the MTT colorimetric assay. Details of metastatic capabilities including invasion, migration and adhesion of B16F10 melanoma cells were examined by Boyden Chamber invasion and migration, scratch motility and cell attachment assays, respectively. The results demonstrated that n-hexane and chloroform extracts exhibited potent anti-proliferative effects (p<0.01), whereas the methanol and aqueous extracts had less pronounced effects after 24 h exposure. Bioactivity-guided chromatographic fractionation of both active n-hexane and chloroform extracts led to the isolation of two main prenylated xanthones and characterization as macluraxanthone and gerontoxanthone-I, respectively, their structures being identified by comparison with the spectral data. Interestingly, both exhibited potent effective effects. At non-toxic effective doses, n-hexane and chloroform extracts (10 and $30{\mu}g/ml$) as well as macluraxanthone and gerontoxanthone-I (3 and $10{\mu}M$) significantly inhibited B16F10 cell invasion, to a greater extent than $10{\mu}m$ doxorubicin, while reducing migration of cancer cells without cellular cytotoxicity. Moreover, exposure of B16F10 melanoma cells to high concentrations of chloroform ($30{\mu}g/ml$) and geratoxanthone-I ($20{\mu}M$) for 24 h resulted in delayed adhesion and retarded colonization. As insights into mechanisms of action, typical morphological changes of apoptotic cells e.g. membrane blebbing, chromatin condensation, nuclear fragmentation, apoptotic bodies and loss of adhesion as well as cell cycle arrest in the G1 phase with increase of sub-G1 cell proportions, detected by Hoechst 33342 staining and flow cytometry were observed, suggesting DNA damage and subsequent apoptotic cell death. Taken together, our findings indicate for the first time that active n-hexane and chloroform extracts as well as macluraxanthone and gerontoxanthone-I isolated from Maclura amboinensis Bl. roots affect multistep of cancer metastasis processes including proliferation, adhesion, invasion and migration, possibly through induction of apoptosis of highly metastatic B16F10 melanoma cells. Based on these data, M. amboinensis Bl. represents a potential candidate novel chemopreventive and/or chemotherapeutic agent. Additionally, they also support its ethno-medicinal usage for cancer prevention and/or chemotherapy.

Effects of SIS/PLGA Porous Scaffolds and Muscle-Derived Stem Cell on the Formation of Tissue Engineered Bone (SIS/PLGA 담체와 근육유래 줄기세포를 이용한 생체조직공학적 골재생)

  • Kim Soon Hee;Yun Sun Jung;Jang Ji Wook;Kim Moon Suk;Khang Gilson;Lee Hai Bang
    • Polymer(Korea)
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    • v.30 no.1
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    • pp.14-21
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    • 2006
  • Tissue engineering techniques require the use of a porous biodegradable/bioresorbable scaffold, which server as a three-dimensional template for initial cell attachment and subsequent tissue formation in both in vitro and in vivo. Small intestinal submucosa (SIS) has been investigated as a source of collagenous tissue with the potential to be used as biomaterials because of its inherent strength and biocompatibility. SIS-loaded poly(L-lactide-co-glicolide)(PLGA) scaffolds were prepared by solvent casting/particle leaching. Characterizations of SIS/PLGA scaffold were carried out by SEM, mercury porosimeter, and so on. Muscle-derived stem cells can be differentiated in culture into osteoblasts, chondrocytes, and even myoblasts by the controlling the culture environment. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide(MTT) test. Osteogenic differential cells were analyzed by alkaline phosphatase(ALP) activity. SIS/PLGA scaffolds were implanted into the back of athymic nude mouse to observe the effect of SIS on the osteoinduction compared with controlled PLGA scaffolds. Thin sections were cut from paraffin embedded tissues and histological sections were conducted hematoxylin and eosin (H&E), Trichrome, and von Kossa. We observed that bone formatioin of SIS/PLGA hybrid scaffold as natural/synthetic scaffold was better thean that of only PLGA scaffold. It canb be explained that SIS contains various kinds of bioactive molecules for osteoinduction.

Characteristics and Biocompatibility of Electrospun Nanofibers with Poly(L-lactide-co-ε-caprolactone)/Marine Collagen (전기 방사법을 통해 제조된 Poly(L-lactide-co-ε-caprolactone)/Marine Collagen 나노파이버의 특성 및 세포친화력 평가)

  • Kim, Woo-Jin;Shin, Young-Min;Park, Jong-Seok;Gwon, Hui-Jeong;Kim, Yong-Soo;Shin, Heung-Soo;Nho, Young-Chang;Lim, Youn-Mook;Chong, Moo-Sang
    • Polymer(Korea)
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    • v.36 no.2
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    • pp.124-130
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    • 2012
  • The uniform nanofibers of poly(L-lactide-$co$-${\varepsilon}$-caprolactone) (PLCL) with different contents of marine collagen (MC) were successfully prepared by electrospinning method. The effects of the major parameters in electrospinning process such as tip to target distance (TTD), voltage, nozzle size and flow rate on the average diameter of the electrospun nanofiber were investigated in generating composite nanofiber. The diameter and morphology of the nanofibers were confirmed by a scanning electron microscopy (SEM). Also, we measured a water contact angle to determine the surface wettability of the nanofibers. The average diameter of the nanofibers decreased as the value of TTD, MC contents, and voltages increased in comparison with that of pristine PLCL nanofiber. In contrast, the diameter of the nanofibers increased as the flow rate and inner diameter of nozzle increased in comparison with that of pristine PLCL. In addition, the hydrophilicity of the nanofiber and attachment of MG-63 cells on the sheets increased as incorporated collagen contents increased. Therefore, the marine collagen would be a potential material to enhance cellular interactivity of synthetic materials by mimicking the natural tissue.

Effects of Attachment and Proliferation of Retinal Pigment Epithelial Cells on Silk/PLGA Film (실크/PLGA 필름에서 실크 함량이 망막색소 상피세포의 부착 및 증식 거동에 미치는 영향)

  • Jo, Eun-Hye;Kim, Soo-Jin;Cho, Su-Jin;Lee, Ga-Young;Kim, On-You;Lee, Eun-Yong;Cho, Won-Hyung;Lee, Dong-Won;Khang, Gil-Son
    • Polymer(Korea)
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    • v.35 no.4
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    • pp.289-295
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    • 2011
  • Biomaterials for retinal tissue engineering must demonstrate several critical features for potential utility, including mechanical integrity, biocompatibility, and slow biodegradation. Silk film biomaterials were designed and characterized to meet these functional requirements. We prepared natural/synthetic hybrid silk/PLGA films using 0, 10, 20, 40, and 80 wt% of silk by a solvent evaporation method. MIT assay was used to confirm the number of cells attached on film at 1, 2, and 3 days, respectively. The morphology of cellular adhesion on films was also confirmed by scanning electron microscope (SEM). RT-PCR was conducted to confrrm mRNA expression of retinal pigment epithelitun (RPE) using RPE65 as a RPEs marker and the expression of cytokeratin were determined by immunofluorescence staining. We confirmed that the silk/PLGA film of 20~40 wt% silk was superior for the adhesion and proliferation of RPEs.

Inhibition of Cell Growth by Anoikis in Various Human Cancer Cell Lines Treated with an Extract of Smilax china L. (토복령 추출물이 처리된 여러 종류의 사람 암세포주에서 아노이키스 세포 사멸에 의한 세포 성장의 억제)

  • Kim, Min-Jae;Kim, Hyeon-Ji;Kim, Moo-Gyeong;Lee, Sung-Ho;Jeon, Byeong-Gyun
    • Journal of Life Science
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    • v.31 no.3
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    • pp.266-279
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    • 2021
  • The present study examined the cytotoxic effects of a Smilax china L. extract (SCLE) in human cancer (A-549, MCF-7, MDA-MB-231, U87-MG, AGS, MKN-74, and SNU-601) and normal MRC-5 fibroblasts, as well as in mesenchymal stem cells derived from dental tissue (DSC). The 50% inhibitory concentration (IC50) values for SCLE were significantly (p<0.05) lower in the cancer cell lines (A-549, MCF-7, MDA-MB-231, U87-MG, AGS, MKN-74 and SNU-601) than in the MRC-5 and DSC cells. Cell growth was significantly (p<0.05) more inhibited in the cancer cell lines treated with 200 ㎍/ml SCLE than in the normal MRC-5 and DSC, and anoikis-like floating cell morphology was observed in the SCLE-treated cancer cells. The cells detached by SCLE treatment were retrieved daily and assayed for viability and telomerase activity. Cells retrieved at 4 days showed significantly decreased viability and telomerase activity (p<0.05), as well as apoptosis-like abnormal morphology, when compared to cells retrieved in the previous 3 days. The ratio of apoptosis and cells in the G1 phase was significantly (p<0.05) increased in the A-549, AGS, and MCF-7 cancer cells treated with SCLE for 4 days compared to untreated controls. However, after SCLE treatment, cell adhesion was not increased by application of an inhibitor of the associated protein kinase (ROCK) that mainly contributes to the increase in cell attachment. This suggests that the cellular detachment by SCLE is probably controlled by a Rho-independent mechanism(s). These observations indicate that SCLE readily induces anoikis in cancer cells and could serve as a potent agent for cancer chemotherapy.

A study of the effects of PDGF-BB on the characteristics of bone stromal and periodontal ligament cells (혈소판유래성장인자-BB가 골간질세포와 치주인대세포의 성상에 미치는 영향)

  • Kwon, Young-Hyuk;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.949-965
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    • 1996
  • The main goal of periodontal therapy is to restore the lost periodontal tissue and establish the attachment appratus. Current acceptable therapeutic techniques are included : removal of diseased soft tissue, demineralization of exposed root surface, using the barrier membrane for preventing the downgrowth of gingival epithelial cell, insertion of graft materials as a scaffolding action, and biological mediators for promoting the cell activity. The latest concept one among them has been studied which based on the knowledge of cellular biology of destructed tissue. Platelet-derived growth factor(PDGF) is one of the polypeptide growth factor which have been reported as a biological mediator to regulate activities of wound healing progress including cell proliferation, migration, and metabolism. The purposes of this study is to evaluate the influences of the PDGF as biological mediator to periodontal ligament and bone marrow cell. Both right and left maxillary first molar were extracted from rat which had treated with 0.4% ${\beta}-Aminopropionitril$ for 5 days, and feeded until designed date to sacrifice under anesthesisa. Periodontal ligament were removed from the extracted socket of the rat, and cultured with Dulbecco's Modified Essential Medium(DMEM) contained with 10% Fetal Bovine Serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. Bone marrow cell were culture from bone marrow suspension with which washed out from femur with same medium. The study was performed to evaluate the effect of PDGF to periodontal ligament and bone cell, cell proliferation rate, total protein synthesis, and alkaline phosphatase activity of rat periodontal ligament(PDL) cell and bone stromal(RBS) cell in vitro. The effects of growth factors on both cells were measured at 3, 5th day after cell culture with (control group) or without growth factors(experimental group). The results were as follows: 1. The tendency of cell proliferation under the influence of PDGF showed more rapid proliferation pattern than control at 3 and 5 days after inoculation. 2. The activity of Alkaline phosphatase revealed 14, 80% increased respectively at 3, 5 days culture than control group. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(P<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P<0.05). 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group l(PDL 100%), 2(PDL 70%:GF 30%), and 3(PDL 50%:GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30%:GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05). From the above results, it is assumed that the PDGF on PDL cells and RMB cell culture. GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density. In conclusion, platelet-derived growth factor can promote rapid osteogenesis during early stage of periodontal tissue regeneration.

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The Effect of Price Discount Rate According to Brand Loyalty on Consumer's Acquisition Value and Transaction Value (브랜드애호도에 따른 가격할인율의 차이가 소비자의 획득가치와 거래가치에 미치는 영향)

  • Kim, Young-Ei;Kim, Jae-Yeong;Shin, Chang-Nag
    • Journal of Global Scholars of Marketing Science
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    • v.17 no.4
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    • pp.247-269
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    • 2007
  • In recent years, one of the major reasons for the fierce competition amongst firms is that they strive to increase their own market shares and customer acquisition rate in the same market with similar and apparently undifferentiated products in terms of quality and perceived benefit. Because of this change in recent marketing environment, the differentiated after-sales service and diversified promotion strategies have become more important to gain competitive advantage. Price promotion is the favorite strategy that most retailers use to achieve short-term sales increase, induce consumer's brand switch, in troduce new product into market, and so forth. However, if marketers apply or copy an identical price promotion strategy without considering the characteristic differences in product and consumer preference, it will cause serious problems because discounted price itself could make people skeptical about product quality, and the changes of perceived value might appear differently depending on other factors such as consumer involvement or brand attitude. Previous studies showed that price promotion would certainly increase sales, and the discounted price compared to regular price would enhance the consumer's perceived values. On the other hand, discounted price itself could make people depreciate or skeptical about product quality, and reduce the consumers' positivity bias because consumers might be unsure whether the current price promotion is the retailer's best price offer. Moreover, we cannot say that discounted price absolutely enhances the consumer's perceived values regardless of product category and purchase situations. That is, the factors that affect consumers' value perceptions and buying behavior are so diverse in reality that the results of studies on the same dependent variable come out differently depending on what variable was used or how experiment conditions were designed. Majority of previous researches on the effect of price-comparison advertising have used consumers' buying behavior as dependent variable. In order to figure out consumers' buying behavior theoretically, analysis of value perceptions which influence buying intentions is needed. In addition, they did not combined the independent variables such as brand loyalty and price discount rate together. For this reason, this paper tried to examine the moderating effect of brand loyalty on relationship between the different levels of discounting rate and buyers' value perception. And we provided with theoretical and managerial implications that marketers need to consider such variables as product attributes, brand loyalty, and consumer involvement at the same time, and then establish a differentiated pricing strategy case by case in order to enhance consumer's perceived values properl. Three research concepts were used in our study and each concept based on past researches was defined. The perceived acquisition value in this study was defined as the perceived net gains associated with the products or services acquired. That is, the perceived acquisition value of the product will be positively influenced by the benefits buyers believe they are getting by acquiring and using the product, and negatively influenced by the money given up to acquire the product. And the perceived transaction value was defined as the perception of psychological satisfaction or pleasure obtained from taking advantage of the financial terms of the price deal. Lastly, the brand loyalty was defined as favorable attitude towards a purchased product. Thus, a consumer loyal to a brand has an emotional attachment to the brand or firm. Repeat purchasers continue to buy the same brand even though they do not have an emotional attachment to it. We assumed that if the degree of brand loyalty is high, the perceived acquisition value and the perceived transaction value will increase when higher discount rate is provided. But we found that there are no significant differences in values between two different discount rates as a result of empirical analysis. It means that price reduction did not affect consumer's brand choice significantly because the perceived sacrifice decreased only a little, and customers are satisfied with product's benefits when brand loyalty is high. From the result, we confirmed that consumers with high degree of brand loyalty to a specific product are less sensitive to price change. Thus, using price promotion strategy to merely expect sale increase is not recommendable. Instead of discounting price, marketers need to strengthen consumers' brand loyalty and maintain the skimming strategy. On the contrary, when the degree of brand loyalty is low, the perceived acquisition value and the perceived transaction value decreased significantly when higher discount rate is provided. Generally brands that are considered inferior might be able to draw attention away from the quality of the product by making consumers focus more on the sacrifice component of price. But considering the fact that consumers with low degree of brand loyalty are known to be unsatisfied with product's benefits and have relatively negative brand attitude, bigger price reduction offered in experiment condition of this paper made consumers depreciate product's quality and benefit more and more, and consumer's psychological perceived sacrifice increased while perceived values decreased accordingly. We infer that, in the case of inferior brand, a drastic price-cut or frequent price promotion may increase consumers' uncertainty about overall components of product. Therefore, it appears that reinforcing the augmented product such as after-sale service, delivery and giving credit which is one of the levels consisting of product would be more effective in reality. This will be better rather than competing with product that holds high brand loyalty by reducing sale price. Although this study tried to examine the moderating effect of brand loyalty on relationship between the different levels of discounting rate and buyers' value perception, there are several limitations. This study was conducted in controlled conditions where the high involvement product and two different levels of discount rate were applied. Given the presence of low involvement product, when both pieces of information are available, it is likely that the results we have reported here may have been different. Thus, this research results explain only the specific situation. Second, the sample selected in this study was university students in their twenties, so we cannot say that the results are firmly effective to all generations. Future research that manipulates the level of discount along with the consumer involvement might lead to a more robust understanding of the effects various discount rate. And, we used a cellular phone as a product stimulus, so it would be very interesting to analyze the result when the product stimulus is an intangible product such as service. It could be also valuable to analyze whether the change of perceived value affects consumers' final buying behavior positively or negatively.

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A Study on an Effective Decellularization Technique for a Xenograft Cardiac Valve: the Effect of Osmotic Treatment with Hypotonic Solution (이종 심장 판막 이식편에서 효과적인 탈세포화 방법에 관한 연구; 저장성 용액(hypotonic solution)의 삼투압 처치법 효과)

  • Sung, Si-Chan;Kim, Yong-Jin;Choi, Sun-Young;Park, Ji-Eun;Kim, Kyung-Hwan;Kim, Woong-Han
    • Journal of Chest Surgery
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    • v.41 no.6
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    • pp.679-686
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    • 2008
  • Background: Cellular remnants in the bioprosthetic heart valve are known to be related to a host's immunologic response and they can form the nidus for calcification. The extracellular matrix of the decellularized valve tissue can also be used as a biological scaffold for cell attachment, endothelialization and tissue reconstitution. Thus, decellularization is the most important part in making a bioprosthetic valve and biological caffold. Many protocols and agents have been suggested for decellularization, yet there ave been few reports about the effect of a treatment with hypotonic solution prior to chemical or enzymatic treatment. This study investigated the effect of a treatment with hypotonic solution and the appropriate environments such as temperature, the treatment duration and the concentration of sodium dodecylsulfate (SDS) for achieving proper decellularization. Material and Method: Porcine aortic valves were decellularized with odium dodecylsulfate at various concentrations (0.25%, 0.5%), time durations (6, 12, 24 hours) and temperatures ($4^{\circ}C$, $20^{\circ}C$)(Group B). Same the number of porcine aortic valves (group A) was treated with hypotonic solution prior to SDS treatment at the same conditions. The duration of exposure to the hypotonic solution was 4, 7 and 14 hours and he temperature was $4^{\circ}C$ and $20^{\circ}C$, respectively. The degree of decellularization was analyzed by performing hematoxylin and eosin staining. Result: There were no differences in the degree of decellularization between the two concentrations (0.25% 0.5%) of SDS. Twenty four hours treatment with SDS revealed the best decellularization effect for both roups A and B at the temperature of $4^{\circ}C$, but there was no differences between the roups at $20^{\circ}C$. Treatment with hypotonic solution (group A) showed a better ecellularization effect at all the matched conditions. Fourteen hours treatment at $4^{\circ}C$ ith ypotonic solution prior to 80S treatment revealed the best decellularization effect. The treatment with hypotonic solution at $20^{\circ}C$ revealed a good decellularization effect, but his showed significant extracellular matrix destruction. Conclusion: The exposure of porcine heart valves to hypotonic solution prior to SDS treatment is highly effective for achieving decellularization. Osmotic treatment with hypotonic solution should be considered or achieving decellularization of porcine aortic valves. Further study should be carried out to see whether the treatment with hypotonic solution could reduce the exposure duration and concentration of chemical detergents, and also to evaluate how the structure of the extracellular matrix of the porcine valve is affected by the exposure to hypotonic solution.