• Proceedings of the Korean Vacuum Society Conference
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• 2011.02a
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• pp.241-241
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• 2011

최근들어 저온플라즈마를 이용한 생물학적 응용분야가 각광을 받고 있다. 특히 전기전도도를 가진 전해질 내에서 형성된 액상 플라즈마는 열손상없이 암, 세균 및 비정상 장기조직의 제거가 가능하다는 점에서 기존 시술들이 가지는 문제를 해결할 수 있다. 허리통증을 유발하는 탈출 수핵을 대용량으로 제거하기위한 플라즈마발생 전극에 관한 연구가 수행되었다. 수핵 분해량을 늘리기 위해서는 플라즈마를 통하여 다량의 수산화기 라디컬을 형성, 수핵표면에 조사해야 한다. 이를 위하여 6개의 텅스텐 전극표면에서 기포를 발생시켜 플라즈마 발생면적을 넓힐 수 있었다. 텅스텐 전극들은 캡톤코딩과 세라믹 스페이서를 통하여 분리되었고, 전극의 후방에는 SUS 재질의 환형 접지전극을 배치하여 6개의 텅스텐 전극표면에서 모두 기포가 발생할 수 있도록 하였다. 시술적용시 플라즈마 및 전극이 가지는 제한 조건은 단백질 변성을 막기위한 섭씨 45도 이하의 온도 상승과 조직에 대한 기계적인 손상 방지를 위한 2.5 mm 이하의 전체 전극 굵기이다. 이를 만족하는 가운데 수산화기 라디컬 형성을 증대할 수 있는 전극의 구조를 결정하기 위하여 1-D 전기 열유체 모델 도입하였다. 모델에서 도출된 기포의 두께를 바탕으로 다중전극간의 거리 조절을 통하여 플라즈마 방전구조를 전극 - 전극 (기포두께${\times}2$ > 전극간 거리)과 전극 - 기포표면 (기포두께${\times}2$ < 전극간 거리)으로 통제하였다. 형성된 플라즈마의 소모전력, 전자 밀도및 수산화기 라디컬의 회전온도를 분석하기 위하여 0.9% 염화나트륨 수용액, 1.6 S/m, 전해질에서 플라즈마 형성를 형성하고 전기신호 및 광학신호를 관측하였다. 전극에 인가된 전압은 340 VRMS이며 운전주파수는 380 kHz이다. 실험 결과, 전극 - 기포표면 방전구조는 전극 -전극 방전구조에 비하여 전해질의 저항역할로 인하여 방전전류가 3.4 Ipp에서 1.6 Ipp로 감소하였으나, 기포표면에서의 물분자의 분해로 인하여 수산화기 라디컬에서의 발광세기는 약 4배 증가하였다. 또한 수산화기의 회전온도 분포상에서도 전극 - 기포표면 방전은 주변 물분자의 열교환으로 인하여 전극 -전극간 방전의 1500K 에 비하여 낮은 400K를 보였다. 이는 전극-기포표면 방전구조의 전극이 낮은 온도의 수산화기를 다량으로 형성할 수 있음을 시사하며, 카데바를 이용한 실험에서 220초에 걸쳐 약 87%의 수핵을 기계적 손상 및 단백질 변형없이 효과적으로 제거함을 확인하였다.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2012.05a
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• pp.741-742
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• 2012

• The Korean Journal of Pain
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• v.18 no.1
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• pp.34-38
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• 2005

Background: Over the years, disc surgery has progressively evolved in the direction of decreasing trauma and its invasiveness. Conventional open surgery has many complications, such as scarring, instability, bleeding and a relative high mortality rate. Minimally invasive spinal surgery is now an alternative to a traditional discectomy. Herein, we present an operative technique, and the early results, for a percutaneous endoscopic lumbar discectomy in herniated lumbar disc disease. Methods: 43 patients, including 27 men and 16 women, with ages ranging from 18 to 66 years, were enrolled in this study. All the patients showed a protruded or extruded soft disc herniation at the lumbar level on magnetic resonance imaging and computed tomography. A percutaneous endoscopic lumbar discectomy was applied to the patients, and clinical responses evaluated using MacNab's criteria. Results: 40 patients were regarded as showing successful responses (93.1%), and there were no severe complications, such as a hematoma, nerve injury, postoperative dysesthesia or death. One patient underwent fusion surgery for remnant back pain six month later. Conclusions: We conclude that, in properly selected patients, a percutaneous endoscopic lumbar discectomy is a safe, noninvasive and effective treatment modality for herniated lumbar intervertebral disc disease.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.171-178
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• 2003

This study was conducted to investigate the effect of electric fusion methods on cell fusion rate and embryo development by somatic cell nuclear transfer in Korean Native Cattle. The KNC ear cell was cultured in vitro for confluence in serum starvation condition(DMEM＋0.05％ FBS) for cell confluence. The zona pellucida of IVM oocytes were partially dissection using micro pipette. Ear cells were transferred into an enucleated oocyte. The reconstructed embryos were electrically fused with Zimmermann Cell Fusion Medium(ZCFM). Nuclear transfer embryos were activated with a combination of 10${\mu}{\textrm}{m}$ calcium ionophore(5 min) and 2.0mM 6-DMAP(3 hr). The activated embryos were cultured in CR1 -aa medium contains 0.3％ BSA or 10％ FBS at 37$^{\circ}C$, 90％ $N_2$, and 5％ $CO_2$in incubator for 6 days. The fusion rates were 51.6％(chamber) and 68.9％(needle), respectively and there were significantly difference between the fusion method(P<0.05). But, lysis rates were not significantly different(10.7％, 11.5％), respectively. The cleavage rates were significantly different between the chamber method(73.2％) and needle method(80.3％), respectively(P<0.05). The rates of early embryos(2∼4cells) and blastocysts of chamber and needle methods were 54.1％, 61.1％ and 18.4％, 26.3％ respectively, and needle method was significantly higher than chamber method(P<0.05). But, morulae formation rate were not significantly differences between the chamber(6.7％) and needle(6.2) method(P <0.05). These result suggest that electric fusion of needle method was to be profitable for nuclear transfer embryo fusion rate, blastocyst formation rate and reduce of oocyte lysis.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.137-146
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• 2006

The objective of this study was to examine the effect of donor cell types, the source of recipient oocytes and estrous synchronization on pregnancy and delivery rates of somatic cell nuclear transfer (SCNT) embryos in Korean native goats. Recipient oocytes were surgically collected after superovulation. Ear cells and fetal fibroblasts were collected and cultured in serum-starvation condition (TCM-199 + 0.5% FBS) for cell confluence. The zonae pellucidae of in vivo- and in vitro-matured oocytes were partially drilled using a laser system. Single somatic cell was transferred into the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3 M mannitol. After the fusion, embryos were activated by Ionomycin+6-DMAP. NT embryos were cultured in mSOF medium supplemented with 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $O_2$, 90% $N_2$ for 12 to 20 hr. One hundred and two SCNT embryos were transferred into 20 recipients and pregnancy rate at days 30 was 20.0%. Of them, one developed to term and delivered 1 kid. Ear cells showed significantly higher fusion (63.8 vs. 26.5%) and pregnancy rates (20.0 vs. 0.0%) than those of fetal fibroblast (p<0.05). The recipients synchronized by CIDR showed significantly lower pregnancy rates compared to that of recipient in natural estrus ($0.0{\sim}25.0%$ vs. 100%) (p<0.05). Cloned kid was born from the recipient in natural estrus. For the synchronization of estrus between recipient and donor, there was no difference between treatments (${\pm}0$ vs. +12 hr) in pregnancy rate. The first healthy cloned kid (Jinsoonny) was produced by transfer of SCNT embryos derived from in vivo oocytes and ear cells into a recipient goat whose estrus was synchronized with the donor. These results imply that donor cells for nuclear transfer may affect the success rate, and the estrus synchronization between donor and recipient animals can also be important.

• Journal of the Korean Orthopaedic Association
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• v.54 no.3
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• pp.211-218
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• 2019

Herniation of the intervertebral disc is a medical disease manifesting as a bulging out of the nucleus pulposus or annulus fibrosis beyond the normal position. Most lumbar disc herniation cases have a favorable natural course. On the other hand, surgical intervention is reserved for patients with severe neurological symptoms or signs, progressive neurological symptoms, cauda equina syndrome, and those who are non-responsive to conservative treatment. Numerous surgical methods have been introduced, ranging from conventional open, microscope assisted, tubular retractor assisted, and endoscopic surgery. Among them, microscopic discectomy is currently the standard method. Biportal endoscopic spinal surgery (BESS) has several merits over other surgical techniques, including separate and free handling of endoscopy and surgical instruments, wide view of the surgical field with small skin incisions, absence of the procedure of removing fog from the endoscope, and lower infection rate by continuous saline irrigation. In addition, existing arthroscopic instruments for the extremities and conventional spinal instruments can be used for this technique and surgery for recurred disc herniation is applicable because delicate surgical procedures are performed under a brightness of 2,700 to 6,700 lux and a magnification of 28 to 35 times. Therefore, due to such advantages, BESS is a novel technique for the surgical treatment of lumbar disc herniation.

• Reproductive and Developmental Biology
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• v.28 no.1
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• pp.21-27
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• 2004

This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Recipient bovine and porcine oocytes were obtained from slaughterhouse and were matured in vitro according to established protocols. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS and primary fibroblast cultures were established in TCM-199 with 10% FBS. The matured oocytes were dipped in D-PBS plus 10% FBS ＋ 7.5 $\mu$ g/ml cytochalasin B and 0.05M sucrose. Enucleation were accomplished by aspirating the first polar body and partial cytoplasm which containing metaphase II chromosomes using a micropipette with an out diameter of 20∼30 $\mu$m. A Single donor cell was individually transferred into the perivitelline space of each enucleated oocyte. The reconstructed oocytes were electric fusion with 0.3M mannitol fusion medium. After the electrofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. And porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6 ∼8 day at $39^{\circ}C, 5% CO_2 $in air. Interspecies nuclear transfer by recipient bovine oocytes were fused with electric length 1.95 kv/cm and 2.10 kv/cm. There was no significant difference between two electric length in fusion rate(47.7 and 44.6%) and in cleavage rate(41.9 and 54.5%). Using electric length 1.95 kv/cm and 2.10 kv/cm in caprine-porcine NT oocytes, there was also no significant difference between two treatments in fusion rate(51.3 and 46.1%) and in cleavage rate(75.0 and 84.9%). The caprine-bovine NT oocytes fusion rate was lower(P＜0.05) in 1 pulse for 60 $\mu$sec(19.3%), than those from 1 pulse for 30 $\mu$sec(50.8%) and 2 pulse for 30 $\mu$sec(31.0%). The cleavage rate was higher(P＜0.05) in 1 pulse for 30 $\mu$sec(53.3%) and 2 pulse for 30 $\mu$sec(50.0%), than in 1 pulse for 60 $\mu$sec(18.2%). The caprine-porcine NT oocytes fusion rate was 48.1% in 1 pulse for 30 $\mu$sec, 45.2% in 2 pulse for 30 $\mu$sec and 48.6% in 1 pulse for 60 $\mu$sec. The cleavage rate was higher(P＜0.05) in 1 pulse for 30 $\mu$sec(78.4%) and 1 pulse for 60 $\mu$sec(79.4%), than in 2 pulse for 30 $\mu$sec(53.6%). In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer and 30.6% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P＜0.05) in interspecies nuclear transfer(5.1%) than parthenotes(37.4%).

• Journal of Yeungnam Medical Science
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• v.15 no.1
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• pp.164-172
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• 1998

The Graf stabilization has been introduced in treating lumbar spinal disorder associated with posterior instability. This study reviewed some problems of the Graf instrumentation as a soft stabilizer. The purpose of this study is to analyse the problems of the soft stabilization in spinal instability. We reviewed 145 cases which were operative treatment using the Graf instrument for lumbar spinal disorder associated with posterior instability at our department from May, 1991 to Dec, 1995. The mean follow up periods was 29 months ranging from 24 months to 6 years 8 months. Of the 145 cases, 22 cases were showed the problem. The diagnostic method were simple x-ray, flexion-extension lateral stress view and CT scan. Results were as follows: Adjacent segmental instability was 10 cases(6.9%), disc space narrowing was 8 cases(5.5%), screw loosening was 3 cases(2.1%) and breakage of the Graf band was 1 case(0.6%). The problems of the soft stabilization were adjacent segmental instability, disc space narrowing, screw loosening, and breakage of the Graf band. But the rate of adjacent segmental instability and disc space narrowing was lower than other lumbar spinal instrumentation.

• Polymer(Korea)
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• v.37 no.6
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• pp.669-676
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• 2013

Demineralized bone particle (DBP) is a biomaterial used widely in the field of tissue engineering. In this study, in order to study the effect of DBP/poly(lactic-co-glycolic acid) (PLGA) scaffold on disc regeneration in vivo environment, we prepared the porous DBP/PLGA hybrid scaffold. Disc defect was induced by removing the nucleus pulposus tissue after incision the annulus fibrosus tissue in half and scaffolds were transplanted. After 1, 2 and 3 months later, the extracted discs were confirmed by collagen synthesis and glycosaminoglycan (sGAG). We conducted histology (H&E, Safranin-O, Alcian blue, Type I Collagen, Type II Collagen). From the results, it was confirmed that collagen and sGAG content were high in DBP/PLGA scaffold, and the regeneration of intervertebral disc was possible.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.213-221
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• 2001

This study was conducted to examine in vitro developmental ability of porcine embryos after somatic cell nuclear transfer. The porcine ear fell was cultured in vitro for confluency in serum-starvation condition(TCM-199 + 0.5% FBS) far 3~6 days of cell confluency. The zona pellucida of IVM oocytes were partially drilled using laser system. Single somatic cell was individually transferred into enucleated oocytes. And the reconstructed embryos were electrically fused(single DC 1.9kv/cm, 30$\mu$ sec) with 0.3M mannitol. After electrofusion, embryos were activated(single AC 5v/mm, 5sec) and cultured in HCSU-23 medium containing 10% FBS at 39$^{\circ}C$, 5% $CO_2$ in air for 6 to 8 days. The fusion rate of donor cells was 45.6, 36.8 and 46.1% in 3~4, 5~6 days of serum starvation and non serum starvation(N-S), and were 52.7. 53.0 and 51.7% in 1~2. 5~6 and 13~14 passages of donor cell culture, respectively. No significant difference was found in the fusion rate of donor cells by the duration of serum starvation treatment or the number of donor cell passages. By the size of donor cells, however, the fusion rate was significantly higher(P<0.05) for reconstructed embryos derived from 25r $\mu$m $\geq$ site of donor cells (65.3%) than that of 25~30$\mu$ m(42.5%) or 30$\mu$ m(45.5%)$\leq$ cells. The cleavage rate was significantly (P<0.05) higher in 3~4 darts of serum starvation treatment(67.1%) than that in N-S (50.7%) or 5~6 days of starvation(57.1%). The activation rate by the size of donor cells in fused oocytes was 56.5, 68.8 and 58.5%, respectively, and was not significant.