PURPOSE: This study was designed to present the effect of alendronate sodium on prevention of osteolysis after arthroplasty by suppression of macrophage. MATERIALS AND METHODS: Submicron-sized titanium particles were made from TiAlV cylinder and plate after milling, and by repeated washing, centrifugation and floating method. 12 dogs were given alendronate by oral administration for 3 months and blood was sampled at 3 weeks, 6 weeks and 3 months. Monocytes were separated from canine blood by isopycnic centrifugation with Percoll solution and cultured in media with LPS, L-NMA, TiAlV particle (x1 SAR and x5 SAR) for 24 hours. Nitric Oxide (NO) assay by Mendelow's method and MTT assay as a cell viability test were performed. RESULTS: Average diameter of TiAlV particles was 1.1+/-0.8 m (from 0.17 to 4.0 m). Particles less than 1.0 m were 55% and 3.0 m, 96%. Compared with control group, there was slight decrease of NO production in 3 weeks group with no statistic difference, but there was significant decrease in 6 weeks and 3 months group statistically. CONCLUSION: Administration of alendronate for more than 6 weeks may be effective in suppression of macrophage, prevention of osteolysis after arthroplasty.
Administration, Oral ; Alendronate* ; Animals ; Arthroplasty ; Cell Survival ; Centrifugation ; Centrifugation, Isopycnic ; Dogs ; Macrophages* ; Monocytes ; Nitric Oxide* ; Osteolysis ; Titanium

Background: In Vietnam, the proportion infertile couples was about 7-10% of reproductive couples. Assisted reproductive technologies appeared and expanded rapidly. In 1970, sperm preparation techniques were found out, artificial insemination became widespread. Sperm preparation stage is very important in assisted reproductive techniques and increasingly improved. Objectives: To compare the efficiency of two sperm preparation techniques: swim \ufffd?up and discontinuous density gradient. Subjects and method: This was a cross-sectional described study included 30 normal semen samples were chosen based on WHO 1999 criteria. We examined sperm vitality, concentration, motility, morphology before and after applied these techniques. Results: Both procedures resulted in a significant higher percentage of vitality, motility, morphology compared with the original semen sample (p<0.01), but the concentration reduced approximately 5 times from whole semen to sediment when swim \ufffd?up was used and 3 times with density gradient, the difference was statically significant (p< 0.01). Conclusion: Normal semen should be prepared by these method. But in oligozoospermic man, to obtain a higher concentration of sperm, semen should be prepared by density gradient method.
Spermatozoa ; Centrifugation ; Density Gradient ;

The influence of alpha-fetoprotein (AFP) on the bone marrow (BM) natural suppressor (NS) cells of intact Ehrlich carcinoma -bearing CBA mice was studied. Bone marrow NS cells were fractionated into three fractions by isopycnic centrifugation on percoll gradients: NS1 (rho=1.080 g/ml), NS2 (rho=1.090 g/ml) and NS3 (1.100>rho>1.090 g/ml). These fractions were highly different in their sensitivity to known NS cell inductors (interleukin (IL)-2, IL-3 or histamine). None of the NS fractions isolated from the intact mice spontaneously produced antiproliferative activity, however, they showed a high level of NS (antiproliferative and natural killer cell inhibitory) activity under the influence of AFP. A single injection of AFP to intact mice led to an increase of spontaneous NS activity and the inhibition of natural killer cell activity. NS activity, especially NS2, was increased in when tumor cells were subcutaneously inoculated three days after AFP injection. In the AFP-treated mice, the tumor mass at 14 days was 60% larger than that in the untreated mice. Our data confirmed that AFP is a tumor marker that can inhibit cancer immunity and plays a role in cancer pathogenesis.
alpha-Fetoproteins ; Animals ; Bone Marrow ; Centrifugation, Isopycnic ; Interleukin-3 ; Killer Cells, Natural ; Mice ; Mice, Inbred CBA ; Povidone ; Silicon Dioxide

No abstract available.
Centrifugation, Density Gradient* ; Humans* ; Spermatozoa*

No abstract available.
Centrifugation ; Weightlessness

OBJECTIVE: This study was carried out to investigate the correlations of the sperm DNA fragmentation index (DFI) with semen parameters and apoptosis, and to investigate the effects of density-gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) on reducing the proportion of sperm with DNA fragmentation and protamine deficiency. METHODS: Semen analysis and a sperm DNA fragmentation assay were performed to assess the correlations between semen parameters and the DFI in 458 semen samples. Sperm with progressive motility or non-apoptosis were isolated by DGC or MACS, respectively, in 29 normozoospermic semen samples. The effects of DGC or MACS alone and of DGC and MACS combined on reducing the amount of sperm in the sample with DNA fragmentation and protamine deficiency were investigated. RESULTS: The sperm DFI showed a significant correlation (r=–0.347, p<0.001) with sperm motility and morphology (r=–0.114, p<0.05) but not with other semen parameters. The DFI (11.5%±2.0%) of semen samples was significantly reduced by DGC (8.1%±4.1%) or MACS alone (7.4%±3.9%) (p<0.05). The DFI was significantly further reduced by a combination of DGC and MACS (4.1%±1.3%, p<0.05). Moreover, the combination of DGC and MACS (1.6%±1.1%, p<0.05) significantly reduced the protamine deficiency rate of semen samples compared to DGC (4.4%±3.2%) or MACS alone (3.4%±2.2%). CONCLUSION: The combination of DGC and MACS may be an effective method to isolate high-quality sperm with progressive motility, non-apoptosis, high DNA integrity, and low protamine deficiency in clinical use.
Apoptosis ; Centrifugation* ; Centrifugation, Density Gradient ; Chromatin* ; DNA Fragmentation ; DNA* ; Methods ; Product Packaging* ; Semen ; Semen Analysis ; Sperm Motility ; Spermatozoa*

There are several different methods of purifying Treponema pallidum(TP) from rabbit testicular tissue. Among them, we compared the use of differential centrifugation, which has been most widely used, to Percoll density gradient centrifugatian, a newly applied method, in purifying TP from rabbit testicular tissue by checking the protein concentration of the TP suspension, hemagglutination assay using sheep erythrocytes sensitized by TP, IgM-TP-enzyme-linked immunosorhent, assay(IgM-TP-ELISA) and eJect,ron microscopic observation. The protein concent,ration af TP antigen suspension (2x10(8)TP/ml) purified by Percoll density gradient centrifugation (lower band) was the lowest (129.0pg/ml) when compared to those purified by differential centrifugation (324.0pg/ml) and Percoll density gradient centrifugatian (upper band) (560.2pg/ml). Sheep erythrocytes sensitized by TP purified by Percoll density gradient centrifugation(lower band) showed the same resiilts as those using a commercii1 TPHA kit when tested with positive and negative control sera. The sensitivity and specificity of the IgM-TP-ELISA were 88.5%(23/26') and 86.4%(19/2Z) respectively using TP as an antigen purified by differential centrifugation. The rates were 96.29% (25/26) and 95.5%(2l/22) using TP purified by Percoll density gradient centrifugation. As shown by the electron microscopy, T. pizllida purified by clifferential centritugation and Percoll density gradient centrifugatiori were structurally unaltered, and Percoll-purified TP contained much less tissue debris than TP prepared by differ ential centrifugation. Therefor e, Percoll density gradient centrifugation is considered to be a better method of purifying TP from rabbit testicular tissue when compared to differential centrifugatian, as a matter of fact, Perrol1 density gradient centrifugation has been applieci successfully in the study of the physiology, recombinant DNA techniques, and antigenic structure of TP and to the preparation of the antigen for the FTA-ARS and TP-ELISA
Centrifugation ; Centrifugation, Density Gradient ; DNA, Recombinant ; Erythrocytes ; Hemagglutination ; Microscopy, Electron ; Physiology ; Sensitivity and Specificity ; Sheep ; Treponema pallidum* ; Treponema*

BACKGROUND: Adipose tissues include multipotent cells, the same as bone marrow-derived mesenchymal stem cells. The stromal vascular fractions (SVFs) from adipose tissues represent a heterogeneous cell population. The purpose of this study was to isolate and purify adipose-derived stem cells (ASCs) in SVFs by the density gradient method. METHODS: SVFs were extracted from the subcutaneous, epididymal, mesenteric and retroperitoneal adipose tissue of 8 weeks old male Sprague-Dawley rats (n = 15) and these were separated into 4 layers according to a Nycodenz gradient (Fx-1: < 11%, Fx-2: 11-13%, Fx-3: 13-19% and Fx-4: 19-30%). The post-confluent SVFs were cultured in adipogenic medium for 2 days, in insulin medium for 2 days and in 10% fetal bovine serum medium for 5 days. To observe lipid droplets in SVFs, we performed Oil Red O staining. RESLTS: The SVFs' cellular fractions (Fx-1, Fx-2, Fx-3 and Fx-4) were isolated by density gradient centrifugation from the adipose tissues of rats. The SVFs extracted to fraction 3 (Fx-3) had the most abundant cells compared to that of the other fractions. However fraction 1 (Fx-1) or 2 (Fx-2) had a superior ability to make lipid droplets. The adipogenic differentiation of Fx-1 or 2 was higher than that of the unfractionated cells. The SVFs extracted from retroperitoneal adipose tissue had the highest efficiency for adipogenic differentiation, whereas the SVFs from mesenteric adipose tissue did not differentiate. CONCLUSION: This density gradient fractionated method leads to efficient isolation and purification of cells with the characteristics of ASCs.
Adipose Tissue ; Animals ; Azo Compounds ; Centrifugation ; Centrifugation, Density Gradient ; Humans ; Insulin ; Intra-Abdominal Fat ; Iohexol ; Male ; Mesenchymal Stromal Cells ; Rats ; Rats, Sprague-Dawley ; Stem Cells

Separation of Langerhans cells in epidermis of 16 healthy Korean individuals were performcd. Separation of Langerhans cells by density gradient centrifugation on a colloidal sillica(percoll) polyvinilpyrrolidone gradient. And autologous, allogeneic mixed skin cell leukocyte culture reaction was done with each fractionatcd cpidermal cell suspensions. Also lymphocytes, epidermal cells was cultured in media alone, respectively. The results was quantitated by the incorporation of H-thymidine by p-liquid scintillation counter. The densities of I angerhans cells within the epidermal cells, fraction-2 was most higher concentration (22.0+2.8%) and fraction-5 was most lower concentration (3.4+ l.9%). 2. In the comparison of the results of Langehans cells enriched and depleted population in autologous mixed skin cell leukocyte culture reaction, the former was higher than the latter on lymphocyte stimulatory capacity. There was significant differences(p<0.005) And also same as result in allogeneic mixed skin cell leukocyte culture reaction. 3. Langerhans cells enriched fraction in this study was more lymphocyte stimulatory capacity than depleted fraction in allogeneic mixed skin cell leukocyte culture(p<0.01~0.05). Ailogeneic mixed skin cell leukocyte culture reaction was more lymphocyte stimulatory capacity than the autologous(p<0.005~0.05).
Centrifugation, Density Gradient* ; Colloids* ; Epidermis ; Humans* ; Langerhans Cells* ; Leukocytes* ; Lymphocytes ; Scintillation Counting ; Silicon Dioxide* ; Skin* ; Suspensions

OBJECTIVETo study the effect of Percoll selection technique on normal morphology and acrosin activity of human spermatoza.

METHODSThe sperm morphology and sperm acrosin activity were analyzed by automated sperm morphology analyzer(ASMA) and spectrocolorimetry.

RESULTSThe normal morphology sperm rate and acrosin activity were significantly increased after Percoll selection technique (P < 0.001).

CONCLUSIONPercoll selection technique could affect normal morphology sperm ratio and acrosin activity.

Acrosin ; metabolism ; Adult ; Centrifugation, Density Gradient ; Fertilization in Vitro ; Humans ; Male ; Spermatozoa ; cytology ; enzymology