• Title/Summary/Keyword: In-vitro techniques

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Evaluation of the efficacy of unipolar and bipolar spinal dorsal root ganglion radiofrequency thermocoagulation in the treatment of postherpetic neuralgia

  • Zhu, Jianjun;Luo, Ge;He, Qiuli;Yao, Ming
    • The Korean Journal of Pain
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    • v.35 no.1
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    • pp.114-123
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    • 2022
  • Background: Different views have been proposed on the radiofrequency treatment modes and parameters of radiofrequency thermocoagulation of the spinal dorsal root ganglion for the treatment of postherpetic neuralgia (PHN). It is urgent to identify a more effective therapy for patients with PHN. Methods: Patients who underwent radiofrequency thermocoagulation therapy for PHN were retrospectively reviewed and were divided into a radiofrequency thermocoagulation (CRF) and double neddles radiofrequency thermocoagulation (DCRF). The pain scores (numerical rating scale, NRS) were evaluated at the following time points: before the operation, 1 day, 3 months, 6 months, 1 year, and 2 years after operation. The incidence of complications and the degree of pain relief were evaluated. The in vitro ovalbumin experiment was used to indicate the effects of radiofrequency thermocoagulation. Results: Compared with the preoperative NRS scores, the postoperative NRS scores decreased significantly; the NRS scores of the DCRF group was lower than that of the CRF group at all time points from 6 months to 2 years following the operation. The total effective rate of the DCRF group was significantly higher than that of the CRF group at 2 years following the operation. The incidence of numbness in the DCRF group was higher than that noted in the CRF group. The ovalbumin experiments in vitro indicated that the effects of radiofrequency thermocoagulation were optimal when the distance between the two needles was 5 mm. Conclusions: DCRF with a 5 mm spacing exhibits a longer duration and higher effective rate in the treatment of PHN and is worth promoting.

The efficacy of different implant surface decontamination methods using spectrophotometric analysis: an in vitro study

  • Roberto Giffi;Davide Pietropaoli;Leonardo Mancini;Francesco Tarallo;Philipp Sahrmann;Enrico Marchetti
    • Journal of Periodontal and Implant Science
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    • v.53 no.4
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    • pp.295-305
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    • 2023
  • Purpose: Various methods have been proposed to achieve the nearly complete decontamination of the surface of implants affected by peri-implantitis. We investigated the in vitro debridement efficiency of multiple decontamination methods (Gracey curettes [GC], glycine air-polishing [G-Air], erythritol air-polishing [E-Air] and titanium brushes [TiB]) using a novel spectrophotometric ink-model in 3 different bone defect settings (30°, 60°, and 90°). Methods: Forty-five dental implants were stained with indelible ink and mounted in resin models, which simulated standardised peri-implantitis defects with different bone defect angulations (30°, 60°, and 90°). After each run of instrumentation, the implants were removed from the resin model, and the ink was dissolved in ethanol (97%). A spectrophotometric analysis was performed to detect colour remnants in order to measure the cumulative uncleaned surface area of the implants. Scanning electron microscopy images were taken to assess micromorphological surface changes. Results: Generally, the 60° bone defects were the easiest to debride, and the 30° defects were the most difficult (ink absorption peak: 0.26±0.04 for 60° defects; 0.32±0.06 for 30° defects; 0.27±0.04 for 90° defects). The most effective debridement method was TiB, independently of the bone defect type (TiB vs. GC: P<0.0001; TiB vs. G-Air: P=0.0017; TiB vs. GE-Air: P=0.0007). GE-Air appeared to be the least efficient method for biofilm debridement. Conclusions: T-brushes seem to be a promising decontamination method compared to the other techniques, whereas G-Air was less aggressive on the implant surface. The use of a spectrophotometric model was shown to be a novel but promising assessment method for in vitro ink studies.

In-Vitro Study of Urokinase Thrombolysis Following Stereotactic Aspiration of Intracerebral Hematoma

  • Son, Wonsoo;Park, Jaechan;Kang, Dong-Hun;Han, Young-Min;Choi, Yeon-Ju;Ohk, Boram
    • Journal of Korean Neurosurgical Society
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    • v.63 no.3
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    • pp.380-385
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    • 2020
  • Objective : A consensus regarding the ideal regimen for urokinase (UK) thrombolysis subsequent to stereotactic spontaneous intracerebral hemorrhage aspiration has yet to be established. The purpose of this study is to evaluate the efficacy of UK thrombolysis relative to when the regimen is changed. Methods : Venous blood from 30 heathy volunteers was obtained for this in-vitro study. Various concentrations of UK solution were added to microcentrifuge tubes containing the clotted blood. The efficacy of UK thrombolysis was identified by checking the weight of lysed hematoma following various time intervals with different concentrations of UK solution. Group one, the "3×4" group involved four administrations every 3 hours over 12 hours, and group two, the "6×2" group involved two administrations every 6 hours over 12 hours. Results : More hematoma was lysed in the 3×4 group than the 6×2 group across all concentration levels (however, the differences were only significant between groups at the 500 and 1000 IU concentration levels, p<0.05). There were no significant differences of lysed hematoma among the various UK solution concentrations within groups. Conclusion : This study suggests that frequent administrations of UK thrombolysis may result in a greater degree of lysed hematoma in comparison to a higher concentration of UK.

The effects of chitosan on the human periodontal ligament fibroblasts in vitro (키토산이 치주인대 섬유아세포에 미치는 영향)

  • Paik, Jeong-Won;Lee, Hyun-jung;Yoo, Yun-Jung;Cho, Kyoo-Sung;Kim, Chong-Kwan;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.31 no.4
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    • pp.823-832
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    • 2001
  • Periodontal therapy has dealt primarily with attempts at arresting progression of disease, however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. The effect of chitosan(poly-N-acetyl glucosaminoglycan), a carbohydrate biopolymer extracted from chitin, on periodontal ligament regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on the human periodontal ligament fibroblasts(hPDLFs) in vitro, with special focus on their proliferative properties by M'IT assay, the synthesis of type I collagen by reverse transcription-polymerase chain reaction(RT-PCR) and the activity of alkaline phosphatase(ALP). Fibroblast populations were obtained from individuals with a healthy periodontium and cultured with ${\alpha}MEM$ as the control group. The experimental groups were cultured with chitosan in concentration of 0.01,0.1, 1,2mg/ml. The results are as follows; 1. Chitosan-induced proliferative responses of hPDLFs reached a plateau at the concentration of O.lmg/ml(p<0.05). 2. When hPDLFs were stimulated with 0.lmg/ml chitosan, mRNA expression of type I collagen was up-regulated. 3. When hPDLFs were stimulated with 0.lmg/ml chitosan, ALP activity was significantly up-regulated(p<0.05). In summary, chitosan(0.lmg/ml) enhanced the type I collagen synthesis in the early stage, and afterwards, facilitated differentiation into osteogenic cells. The results of this in vitro experiment suggest that chitosan potentiates the differentiation of osteoprogenitor cells and may facilitate the formation of bone.

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Production of Chimera by Embryos Aggregation Techniques in Bovine - Review-

  • Suzuki, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.8
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    • pp.1188-1195
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    • 2001
  • A tetraparental chimeric bull was successfully produced by aggregating bovine IVF embryos of F1 (female Holstein${\times}$male Japanese Black) and F1(female Japanese Brown${\times}$male Limousin) and culturing in vitro without the zona pellucida at Yamaguchi Research Station in Japan. In the microsatellite genotyping, 12% (28/228) microsatellite primer sets ware potentially useful for this parentage analysis in the chimeric bull, 78.6% (22/28) of microsatellite present in the chimeric bull were uniquely contributed from the Japanese Black and 21.4% (6/28) from Limousin. This chimeric bull semen was used in producing IVF embryos. The chromosome preparations were made from peripheral lymphocytes. Based on chromosome analysis the Chimera had apparently normal chromosomes (29 acrocentric pairs, one large sub metacentric X chromosome and one small sub metacentric Y chromosome). The proportion of acrosome reacted spermatozoa after 1 h of incubation was higher (p<0.01) with the Chimera than with the Holstein and in Japanese Brown bulls. But did not differ from Japanese Black and Limousin bull sperm. Fertilization rates observed after 5 h of sperm-oocyte incubation with Chimera sperm were higher (p<0.05) than with Japanese Brown and (p<0.01) than with Holstein sperm, but did not differ from Japanese Black and Limousin sperm. The cleavage rates of IVF oocytes inseminated with Chimera sperm were also higher (p<0.001) compared with Holstein, (p<0.01) Japanese Brown and (p<0.05) Limousin, but did not differ from Japanese Black sperm. The blastocyst rates of IVM oocytes inseminated with sperm were higher (p<0.05) than in Limousin, Japanese Brown and Holstein, but did not differ from Japanese Black. Chimeric cattles were produced by aggregation of parthenogenetic (Japanese Brown) and in vitro fertilized (Holstein) bovine embryos at the Yamaguchi Research Station in Japan and by aggregation of parthenogenetic (Red Angus) and in vitro fertilized (Holstein) embryos at the St. Gabriel Research Station in Louisiana. The aggregation rate of the reconstructed demi-embryos cultured in vitro without agar embedding was significantly lower than with agar embedding. The aggregation was also lower when the aggregation resulted from a whole parthenogenetic and IVF-derieved embryos cultured without agar than when cultured with agar. The development rate to blastocysts, however, was not different among the treatment. To verify parthenogenetic and the cells derieved from the male IVF embryos in blastocyst formation, 51 embryos were karyotyped, resulting in 27 embryos having both XX and XY chromosome plates in the same sample, 14 embryos with XY and 10 embryos with XX. The viability and the percentage of zonafree chimeric embryos at 24 h following cryopreservation in EG plus T with 10% PVP were significantly greater than those cryopreserved without PVP. Pregnancies were diagnosed in both stations after the transfer of chimeric blastocysts. Twin male and single chimeric calves were delivered at the Yamaguchi station, with each having both XX and XY chromosomes detected. Three pregnancies resulted from the transfer of 40 chimeric embryos at the Louisiana station. Two pregnancies were Jost prior to 4 months and one phenotypically chimeric viable male born.

Factors Affecting the Isolation of Mesophyll Protoplasts from Populus euramericana cv. I-214 (이태리포푸라 I-214 엽육조직(葉肉組織)에서 원형질체(原形質體) 분리(分離)에 미치는 몇가지 요인(要因))

  • Park, Young Goo;Son, Sung Ho
    • Journal of Korean Society of Forest Science
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    • v.74 no.1
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    • pp.29-36
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    • 1986
  • A method isolating Populus euramericana cv. I-214 mesophyll protoplasts was developed to facilitate application of genetic engineering techniques to this species. The suitable medium for shoot multiplication in vitro was MS basal medium with $0.1mg/{\ell}$ BAP. The effects of several factors influencing protoplast isolation could be evaluated quickly by using leaf in vitro and known volumes of maceration and washing media. The best yields of mesophyll protoplasts were obtained using leaves in vitro in 2.0% Cellulase R-10, 0.8% Macerozyme R-10, 1.2% Hemicellulase, 2.0% Driselase, 0.05% Pectolyase Y-23, and O.6M Mannitol in addition to DTT and MES buffer adjusted to pH 5.6. Over $2.4{\times}10^6$ protoplasts per gram of leaf were produced using these conditions. For protoplast purification, the most favorable sucrose concentration of floating solution was 0.6M after washing them with CPW solution. This method of screening factors affecting protoplast isolation could be applicable to other species.

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Recent advances in intravital microscopy for investigation of dynamic cellular behavior in vivo

  • Choo, Yeon Woong;Jeong, Juhee;Jung, Keehoon
    • BMB Reports
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    • v.53 no.7
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    • pp.357-366
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    • 2020
  • Currently, most biological research relies on conventional experimental techniques that allow only static analyses at certain time points in vitro or ex vivo. However, if one could visualize cellular dynamics in living organisms, that would provide a unique opportunity to study key biological phenomena in vivo. Intravital microscopy (IVM) encompasses diverse optical systems for direct viewing of objects, including biological structures and individual cells in live animals. With the current development of devices and techniques, IVM addresses important questions in various fields of biological and biomedical sciences. In this mini-review, we provide a general introduction to IVM and examples of recent applications in the field of immunology, oncology, and vascular biology. We also introduce an advanced type of IVM, dubbed real-time IVM, equipped with video-rate resonant scanning. Since the realt-ime IVM can render cellular dynamics with high temporal resolution in vivo, it allows visualization and analysis of rapid biological processes.

Design and Optimization of Solid Dispersed Osmotic Pump Tablets of Aceclofenac, A Better Approach to Treat Arthritis

  • Edavalath, Sudeesh;Rao, B. Prakash
    • Journal of Pharmaceutical Investigation
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    • v.41 no.4
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    • pp.217-225
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    • 2011
  • The aim of this work was to prepare porous osmotic pump tablets for controlled delivery of Aceclofenac. Aceclofenac solid dispersion was prepared to improve the solubility by using the drug - carrier (Mannitol) ratio of 1:1. The osmotic pump tablets were prepared using the solid dispersed product of Aceclofenac. The formulation contains potassium chloride as osmotic agent, cellulose acetate as semipermeable membrane, poly ethylene glycol (PEG 4000) as pore former and sodium lauryl sulphate (SLS) as solubility enhancer. The formulations were designed by the general factors such as osmotic agent and pore former. All formulations were evaluated for various physical parameters and, the in vitro release studies were conducted as per USP. The drug release kinetic studies such as zero order, first order, and Higuchi and Korsmeyer peppas were determined and compared. All the formulations gave more controlled release compared to the marketed tablet studied. Numerical optimization techniques were applied to found out the best formulation by considering the parameter of in vitro drug release kinetics and dissolution profile standards. It was concluded that the porous osmotic pump tablets (F7) composed of Aceclofenac solid dispersion/Potassium chloride/Lactose/Sodium lauryl sulphate/Magnesium Stearate (400/40/95/10/5, mg/tab) and coating composition with Cellulose acetate/ PEG 4000 (60/40 %w/w) is the most satisfactory formulation. The porous osmotic pump tablets provide prolonged, controlled, and gastrointestinal environment-independent drug release.

Triterpenoids from Schisandra henryi with Cytotoxic Effect on Leukemia and Hela Cells In Vitro

  • Chen, Ye-Gao;Wu, Zheng-Cai;Lv, Yu-Ping;Gui, Shi-Hong;Wen, Jin;Liao, Xin-Rong;Yuan, Li-Ming;Halaweish, Fathi
    • Archives of Pharmacal Research
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    • v.26 no.11
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    • pp.912-916
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    • 2003
  • Four known lanostane triterpenoids, schiprolactone A (1), schisanlactone B (2), nigranoic acid (3) and schisandronic acid (4) Were isolated from the stems of Schisandra henryi for the first time. Their structures were characterized by IR, MS and NMR techniques. Compounds 1, 2 and 4 showed moderate cytotoxic activity against Leukemia cells in vitro. Cytotoxic activity of compounds 1-4 showed $IC_{50}$ of 0.0097, 0.01, 0.097 and 0.0099 $\mu$ mol/mL respectively toward Leukemia cells and $IC_{50}$ of 0.097, 0.1, 0.097 and 0.099 $\mu$mol/mL toward Hela cells respectively. It is the first report that these compounds possess cytotoxic activity on Leukemia and Hela cells.

Micropropagation of a rare plant species, Astragalus membranaceus Bunge var. alpinus N. (희귀식물 제주황기의 미세번식)

  • Han, Mu Seok;Noh, Seol Ah;Kwak, Myung Cheol;Moon, Heung Kyu
    • Journal of Plant Biotechnology
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    • v.41 no.2
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    • pp.100-106
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    • 2014
  • In order to develop an efficient in vitro micropropagation technique for a rare plant species, Astragalus membranaceus Bunge var. alpinus N., shoot proliferation and in vitro or in vivo rootings were conducted and hyperhydrated leaf generated from cultures was histologically observed. During shoot induction, no distinct effect on multiple shoot induction was found between BA and kinetin treatment. BA enhanced the number of internodes, whereas kinetin stimulated shoot elongation. Hyperhydrated leaf composed of bigger cells and retarded palisade parenchyma and showed irregular cell arrangement compared to normal leaf. Especially starch content in hyperhydrated leaf was significantly reduced. The best rooting rate was achieved by B5 medium among three different medium (B5, MS and WPM) and 0.1mg/L IBA treatment induced the highest rooting ratio (80%). No statistical difference was induced by explant types (apical bud or axillary bud) in terms of rooting ratio. In vivo cutting induced rooting rate up to 65% by 0.5% IBA/Talc powder treatment. Although in vivo rooting rate was less efficient compared to in vitro rooting, better survival rate was observed after soil acclimatization. Present study suggested that above micropropagation techniques can be used for rapid multiplication as well as in vitro or in vivo conservation of the species.