Background: Metastasis and recurrence of primary cancer are the main causes of cancer mortality. Disseminated tumor cells refer to cancer cells that cause metastasis from primary cancer to other organs. Several recent studies have suggested that circulating tumor cells (CTCs) are associated with the clinical stage, cancer recurrence, cancer metastasis, and prognosis. There are several methods of isolating CTCs from whole blood; in particular, using a membrane filtration system is advantageous due to its cost-effectiveness and availability in clinical settings. In this study, an animal model of lung cancer was established in nude mice using the human large cell lung cancer cell line H460. Methods: Six-week-old nude mice were used. The H460 lung cancer cell line was injected subcutaneously into the nude mice. Blood samples were obtained from the orbital area before cell line injection, 2 weeks after injection, and 2 weeks after tumor excision. Blood samples were filtered using a polycarbonate 12-well Transwell membrane (Corning Inc., Corning, NY, USA). An indirect immunofluorescence assay was performed with the epithelial cell adhesion molecule antibody. The number of stained cells was counted using fluorescence microscopy. Results: The average size of the tumor masses was 35.83 mm. The stained cells were counted before inoculation, 2 weeks after inoculation, and 2 weeks after tumor excision. Cancer cells generally increased after inoculation and decreased after tumor resection. Conclusion: The CTC detection method using the commercial polycarbonate 12-well Transwell (Corning Inc.) membrane is advantageous in terms of cost-effectiveness and convenience.
Huang, Pingping;Su, Wen;Han, Rui;Lin, Hao;Yang, Jing;Xu, Libin;Ma, Lei
Journal of Microbiology and Biotechnology
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v.32
no.4
/
pp.522-530
/
2022
In this study we aimed to develop novel ZnO-NP/chitosan/β-glycerophosphate (ZnO-NP/CS/β-GP) antibacterial hydrogels for biomedical applications. According to the mass fraction ratio of ZnO-NPs to chitosan, mixtures of 1, 3, and 5% ZnO-NPs/CS/β-GP were prepared. Using the test-tube inversion method, scanning electron microscopy and Fourier-transform infrared spectroscopy, the influence of ZnO-NPs on gelation time, chemical composition, and cross-sectional microstructures were evaluated. Adding ZnO-NPs significantly improved the hydrogel's antibacterial activity as determined by bacteriostatic zone and colony counting. The hydrogel's bacteriostatic mechanism was investigated using live/dead fluorescent staining and scanning electron microscopy. In addition, crystal violet staining and MTT assay demonstrated that ZnO-NPs/CS/β-GP exhibited good antibacterial activity in inhibiting the formation of biofilms and eradicating existing biofilms. CCK-8 and live/dead cell staining methods revealed that the cell viability of gingival fibroblasts (L929) cocultured with hydrogel in each group was above 90% after 24, 48, and 72 h. These results suggest that ZnO-NPs improve the temperature sensitivity and bacteriostatic performance of chitosan/β-glycerophosphate (CS/β-GP), which could be injected into the periodontal pocket in solution form and quickly transformed into hydrogel adhesion on the gingiva, allowing for a straightforward and convenient procedure. In conclusion, ZnO-NP/CS/β-GP thermosensitive hydrogels could be expected to be utilized as adjuvant drugs for clinical prevention and treatment of peri-implant inflammation.
Journal of Dental Rehabilitation and Applied Science
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v.25
no.4
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pp.361-374
/
2009
For successful osteogenesis around the implants, interaction between implant surface and surrounding tissue is important. Biomechanical bonding and biochemical bonding are considered to influence the response of adherent cells. But the focus has shifted surface chemistry. The purpose of this study is to evaluate the MC3T3-E1 osteoblast like cell responses of magnesium (Mg) ion implanted titanium surface produced using a plasma source ion implantation method. Commercially pure titanium disc was used as substrates. The discs were prepared to produce four different surface, A: Machine turned surface, B: Mg implanted surface, C: sandblasted surface, D: sandblasted and Mg implanted surface. MC3T3 El osteoblastic like cells were cultured on the disc specimens. Cell adhesion, proliferation, differentiation, and synthesis of extracellular matrix were evaluated. The cell adhesion morphology was evaluated by SEM. RT PCR assay was used for assessment of cell adhesion, proliferation and differentiation. ALP activity was measured for cell differentiation. The results of this study were as follows: 1. SEM showed that cell on Mg ion groups was more proliferative than that of non Mg ion groups. On the machine turned surface, cell showed some degree of contact guidance in aligning with the machining grooves. 2. In RT PCR analysis, osteonectin and c-fos mRNA were more expressed on sandblasted and Mg ion implanted group. 3. ALP activity was not significantly different among all groups. Within the limitations of this study, the following conclusions were drawn: It might indicate Mg ion implanted titanium surface induce better bone response than non Mg ion groups.
Journal of the Korean Society of Food Science and Nutrition
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v.45
no.12
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pp.1717-1724
/
2016
Probiotic effects of Lactobacillus plantarum pF1 NITE-P1462 (Lp-pF1), L. plantarum KCCM 11352P (Lp-PNU), L. plantarum CBT LP3 KCTC 10782BP (Lp-CB), and L. plantarum KCTC 3099 (Lp-3099) isolated from kimchi and Lactococcus lactis KFCC 11510P (L-lactis) isolated from Doenjang were studied. Resistance to gastric and bile acid, adhesion to intestines in colon cells, thermal stability, and antioxidative and in vitro anticancer effects in HT-29 cancer cells were evaluated. L. plantarum strains showed improved tolerance of gastric and bile acids than L-lactis. Lp-pF1 had better adhesion ability in the intestine than Lp-PNU, Lp-3099, and L-lactis. Lp-pF1 also showed better heat resistance at $50^{\circ}C$, $70^{\circ}C$, and $80^{\circ}C$ than Lp-CB, Lp-3099, and L-lactis. In addition, Lp-pF1 exhibited greater antioxidant activity by scavenging DPPH radicals or hydroxyl radicals and anticancer effects in MTT assay than others. Taken together, these results suggest that L. plantarum isolated from kimchi showed higher probiotic activities with antioxidant and anticancer properties than Lac. lactis isolated from Doenjang. Lp-pF1 revealed the best probiotic activities among L. plantarum and could be used as a promising potential probiotics.
This study was designed to examine the suitable characteristics of potential probiotic bacteria. Possible probiotic bacteria, including Lactobacillus acidophilus DDS-1, Lb. acidophilus B-3208, Bifidobacterium bifidum KCTC 3357, Lb. plantarum, Leuconostoc mesenteroides ssp. mesenteroides ATCC 8293, and Lactococcus lactis ssp. lactis ATCC 7962 were selected. We then measured their acid and bile tolerances, adhesion properties in the gastrointestinal tract, antimicrobial activity against pathogenic bacteria, and immunomodulation activity. The acid tolerances of Lb. acidophilus DDS-1, Lb. acidophilus B-3208, Lb. plantarum, and Leu. mesenteroides ssp. mesenteroides ATCC 8293, in PBS (pH 2.5) for 2 hr, were high enough that 50% of the inocula survived. The bile tolerances of all bacteria, except Lc. lactis ssp. lactis ATCC 7962, were also observed at a 3% oxgall concentration in MRS broth. The results of the adhesion property assay showed that the total binding affinities of Lb. acidophilus DDS-1, Lb. acidophilus B-3208, and B. bifidum were about three times higher than those of the other bacteria. In testing their antimicrobial activities against pathogens, Lb. acidophilus B-3208, B. bifidum KCTC 3357, and Lb. plantarum inhibited the growth of pathogenic bacteria. For their immunomodulation activity, the cell wall fractions from Lb. acidophilus DDS-1 and Lb. acidophilus B-3208 showed the highest bone marrow cell proliferation activities. However, the cell wall fractions of Lb. acidophilus DDS-1 and B. bifidum, and the cytosol fraction of Lc. lactis ssp. lactis ATCC 7962 showed higher macrophage stimulation activities than those of the other bacteria. Since Lb. acidophilus DDS-1 and Lb. acidophilus B-3208 satisfy the requirements for probiotics, they can be considered suitable probiotic bacteria.
Purpose: The characteristics of oxidized titanium (Ti) surfaces varied according to treatment conditions such as duration time and temperature. Thermal oxidation can change Ti surface characteristics, which affect many cellular responses such as cell adhesion, proliferation, and differentiation. Thus, this study was conducted to evaluate the surface characteristics and cell response of thermally treated Ti surfaces. Methods: The samples were divided into 4 groups. Control: machined smooth titanium (Ti-S) was untreated. Group I: Ti-S was treated in a furnace at $300^{\circ}C$ for 30 minutes. Group II: Ti-S was treated at $500^{\circ}C$ for 30 minutes. Group III: Ti-S was treated at $750^{\circ}C$ for 30 minutes. A scanning electron microscope, atomic force microscope, and X-ray diffraction were used to assess surface characteristics and chemical composition. The water contact angle and surface energy were measured to assess physical properties. Results: The titanium dioxide ($TiO_2$) thickness increased as the treatment temperature increased. Additional peaks belonging to rutile $TiO_2$ were only found in group III. The contact angle in group III was significantly lower than any of the other groups. The surface energy significantly increased as the treatment temperature increased, especially in group III. In the 3-(4,5-Dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide assay, after 24 hours of incubation, the assessment of cell viability showed that the optical density of the control had a higher tendency than any other group, but there was no significant difference. However, the alkaline phosphatase activity increased as the temperature increased, especially in group III. Conclusions: Consequently, the surface characteristics and biocompatibility increased as the temperature increased. This indicates that surface modification by thermal treatment could be another useful method for medical and dental implants.
Cha, Se Rom;Jeong, Hyun Ki;Kim, Su Young;Kim, Eun Young;Song, Jeong Eun;Park, Chan Hum;Kwon, Soon Yong;Khang, Gilson
Polymer(Korea)
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v.39
no.3
/
pp.493-498
/
2015
For biomaterials for skin regeneration with minimized inflammatory response, high bioactivity and biocompatibility are highly required. Also, it should have a porous microstructure to improve cell adhesion and growth. In this study, we extracted a new collagen source from duck's feet which is by-product, and made the shape of sponges from duck's feet collagen (DC) to compare with DBP and SIS. To analyze physical and chemical property of the scaffold, SEM and FTIR were used. MTT assay was used to measure the attachment and proliferation of NIH/3T3 in the scaffolds. RTPCR was used to evaluate the expression of proinflammatory cytokine. Also, 1,1-diphenyl-2-picrylhydrazyl (DPPH) was used to measure the ability of antioxidant activity. Overall, this study shows that DC scaffold is biocompatible and has good physical property. Additionally, DC scaffold shows the potential as wound healing biomaterials.
Candida biofilms are organized microbial communities growing on the surfaces of host tissues or indwelling medical devices, and the biofilms show enhanced resistance against the conventional antifungal agents. The roots of Coptidis chinensis have been widely used for medicinal purposes in East Asia. The present study was aimed to assess the effect of C. chinensis aqueous extract upon preformed biofilms of 10 clinical Candida albicans isolates and the antifungal activities which contribute to inhibit the C. albicans biofilm formation. Its effect on preformed biofilms was judged using XTT [2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide)] reduction assay, and metabolic activity of all tested strains was reduced significantly ($57.3{\pm}14.7%$) at $98{\mu}g/ml$ of the C. chinensis extract. The extract damaged the cell membrane of C. albicans which was analyzed by fluorescein diacetate and propidium iodide staining. The anticandidal activity was fungicidal, and the extract obstructed the adhesion of C. albicans biofilms to polystyrene surfaces, arrested C. albicans cells at $G_o/G_1$ as well, and reduced the growth of biofilms or budding yeasts finally. The data suggest that C. chinensis has multiple antifungal effects on target fungi resulting in preventing the formation of biofilms. Therefore, C. chinensis holds great promise for exploring antifungal agents from natural products in treating and eliminating biofilm-associated Candida infection.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.31
no.5
/
pp.399-408
/
2005
In the progression of the Temporomandibular Joint Disorder(TMD), not only deformation and perforation of disc occur. But also fibrotic adhesion and inflammatory changes to the retrodiscal tissue can be seen in addition to the condylar degenerative change (e.g. osteoarthritis). However, the correct diagnosis,?planning for appropriate treatment, and prediction of prognosis are limited, because there are no means to stage the progression of the disorder. In this study relative signal intensity of retrodiscal tissue in MRI and the synovial fluid concentration of matrix metalloproteinase-2 (MMP-2), MMP-9, and Interleukin-6(IL-6) in the 23 temporomandibular joints(TMJ), from 17 patients with TMD were evaluated as a possible diagnostic marker. The relative signal intensity of retrodiscal tissue was referenced to brain gray matter with same region of interest(ROI) size. The concentrations of MMP-2, MMP-9, and IL-6 were evaluated by Enzyme Linked Immunosorbent Assay (ELISA). The collected data were compared with condylar degenerative change, joint effusion and disc position observed in MRI. The relative signal intensity of the retrodiscal tissue was increased significantly when degenerative changes were present. In addition, there was significantly high signal intensity in the presence of a disc displaced without reduction. The concentration of IL-6 was significantly increased when condylar degenerative change was no observed. And there were no changes in the levels of IL-6 according to disc position and joint effusion measurement. Moreover, there were no significant relevance between the concentration of total MMP-2 and active MMP-9 in synovial fluid, relative to degenerative changes in the mandibular condyle, to joint effusion, and to disc position observed on MRI images. In conclusion, the relative signal intensity of the retrodiscal tissue can be regarded as a mean of diagnosing the procession of TMD in a non-invasive manner. But more additional studies are required for the levels of MMP-2. MMP-9, and IL-6 to determine their potentials as a diagnostic marker for TMD.
A line of study reported that electroacupuncture(EA) modulate natural killer cell(NK Cell) activities. One report suggested that EA enhanced splenic interferon-gamma($IFN-{\gamma}$), interleukin-2(IL-2), and NK cell activity in Sprague-Dawley rats. Another study suggested that $IFN-{\gamma}$ mediates the up-regulation of NK cell activity, and endogenous ${\beta}$-endorphin secretion also play a role in the up-regulation of NK cell activity induced by EA stimulation. In order to better understand the molecular regulation underlying the activation of NK cell induced by EA, we have utilized cDNA microarray to elucidate how EA alters program of gene expression of spleen in rats. First, we divided three groups, group I was EA group treated with EA in restriction holder, group II was sham group with only holder stress, and last group III was control group with no treatment. We measured NK cell activity after EA stimulation three times for 2 days using $^{51}Cr$ release assay. Second, Biotin-labeled cDNA probes synthesized from EA group and sham group, were competitively hybridized to the microarray that contained variable genes. Such high-throughput screening has identified a number of EA-responsive gene candidates. Of these, we found that EA induced a subset of genes of genes that functionally could modulatory effects on NK cell activity. Genes(vascular cell adhesion molecule-1, protein-tyrosine kinase, CD94 mRNA) related to boost NK cell activity, were increased by EA And, genes(protein-tyrosine-phospatase mRNA, protein-tyrosine phosphatase(SHP-1) mRNA) related to inhibit NK cell activity, were decreased by EA. These EA-responsive genes may provide key insights from which to understand mechanisms of activation of NK cell induced by EA.
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