• 제목/요약/키워드: Intervertebral disc(IVD)

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3D 바이오프린팅과 무지지체 조직공학 기술 기반 추간판 복합 조직 제작 (Fabrication of Tissue Engineered Intervertebral Disc Using Enable 3D bio-printing and Scaffod-Free technologies)

  • 김병국;박진호;박상혁
    • 대한의용생체공학회:의공학회지
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    • 제39권1호
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    • pp.22-29
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    • 2018
  • Intervertebral disc(IVD) mainly consists of Annulus fibrosus(AF) and Nucleus pulposus(NP), playing a role of distributing a mechanical load on vertebral body. IVD tissue engineering has been developed the methods to achieve anatomic morphology and restoration of biological function. The goal of present study is to identify the possibilities for creating a substitute of IVD the morphology and biological functions are the same as undamaged complete IVD. To fabricate the AF and NP combine biphasic IVD tissue, AF tissue scaffolds have been printed by 3D bio-printing system with natural biomaterials and NP tissues have been prepared by scaffold-free culture system. We evaluated whether the combined structure of 3D printed AF scaffold and scaffold-free NP tissue construct could support the architecture and cell functions as IVD tissue. 3D printed AF scaffolds were printed with 60 degree angle stripe patterned lamella structure(the inner-diameter is 5mm, outer-diameter is 10 mm and height is 3 mm). In the cytotoxicity test, the 3D printed AF scaffold showed good cell compatibility. The results of histological and immunohistochemical staining also showed the newly synthesized collagens and glycosaminoglycans, which are specific makers of AF tissue. And scaffold-free NP tissue actively synthesized glycosaminoglycans and type 2 collagen, which are the major components of NP tissue. When we combined two engineered tissues to realize the IVD, combined biphasic tissues showed a good integration between the two tissues. In conclusion, this study describes the fabrication of Engineered biphasic IVD tissue by using enable techniques of tissue engineering. This fabricated biphasic tissue would be used as a model system for the study of the native IVD tissue. In the future, it may have the potential to replace the damaged IVD in the future.

The Effects of TWEAK, Fn14, and TGF-$\beta1$ on Degeneration of Human Intervertebral Disc

  • Huh, Hoon;Lee, Yong-Jik;Kim, Jung-Hee;Kong, Min-Ho;Song, Kwan-Young;Choi, Gun
    • Journal of Korean Neurosurgical Society
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    • 제47권1호
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    • pp.30-35
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    • 2010
  • Objective: The purpose of this study is to explain the effect and reciprocal action among tumor necrosis factor (TNF) like weak inducer of apoptosis (TWEAK), fibroblast growth factor-inducible 14 (Fn14), and transforming growth factor-$\beta1$ (TGF-$\beta1$) on degeneration of human intervertebral disc (IVD). Methods: Human intervertebral disc tissues and cells were cultured with Dulbecco's Modified Eagle's Medium/Nutrient F-12 Ham (DMEM/F-12) media in $37^{\circ}C$, 5% $CO_2$ incubator. When IVD tissues were cultured with TWEAK, Fn14 that is an antagonistic receptor for TWEAK and TGF-$\beta1$, the level of sulfated glycosaminoglycan (sGAG) was estimated by dimethyl methyleneblue (DMMB) assay and sex determining region Y (SRY)-box 9 (Sox9) and versican messenger ribonucleic acid (mRNA) levels were estimated by reverse transcriptase polymerase chain reaction (RT-PCR). Results: When human IVD tissue was cultured for nine days, the sGAG content was elevated in proportion to culture duration. The sGAG was decreased significantly by TWEAK 100 ng/mL, however, Fn14 500 ng/mL did not change the sGAG production of IVD tissue. The Fn14 increased versican and Sox9 mRNA levels decreased with TWEAK in IVD tissue TGF-$\beta1$ 20 ng/mL elevated the sGAG concentration 40% more than control. The sGAG amount decreased with TWEAK was increased with Fn14 or TGF-$\beta1$ but the result was insignificant statistically. TGF-$\beta1$ increased the Sox9 mRNA expression to 180% compared to control group in IVD tissue. Sox9 and versican mRNA levels decreased by TWEAK were increased with TGF-$\beta1$ in primary cultured IVD cells, however, Fn14 did not show increasing effect on Sox9 and versican. Conclusion: This study suggests that TWEAK would act a role in intervertebral disc degeneration through decreasing sGAG and the mRNA level of versican and Sox9.

Rabbit Model for in vivo Study of Intervertebral Disc Degeneration and Regeneration

  • Kong, Min-Ho;Do, Duc-H.;Miyazaki, Masashi;Wei, Feng;Yoon, Sung-Hwan;Wang, Jeffrey C.
    • Journal of Korean Neurosurgical Society
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    • 제44권5호
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    • pp.327-333
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    • 2008
  • Objective: The purpose of this study is to verify the usefulness of the rabbit model for disc degeneration study. Materials: The L1-L2, L2-L3, L3-L4. or L4-L5 lumbar intervertebral disc (IVD) of 9 mature male New Zealand White rabbits were injured by inserting a 16-gauge needle to a depth of 5 mm in the left anterolateral annulus fibrosus while leaving L5-L6 IVD uninjured. Three other rabbits also received intradiscal injections of rabbit disc cells transfected with adenovirus and bone morphogenetic protein-2 (ad-BMP-2) at L4-L5 in addition to injury by 16-gauge needle at the L1-L2 level. Using digitized radiographs, measurements of IVD height were made and analyzed by using the disc height index (DHI). Magnetic resonance imaging (MRI) scans of the injured discs, injected discs, and uninjured L5-L6 discs were performed at 15 weeks post surgery and compared with preoperative MRI scans. Results: All twelve rabbits showed consistent results of disc degeneration within 15 weeks following annular puncture. DHIs of injured discs were significantly lower than that of the uninjured L5-L6 discs (p<0.05). The mean value of disc degeneration grade of injured discs was significantly higher than that of uninjured discs (p<0.05). The injection of disc cell transfected with ad-BMP-2 did not induce disc regeneration at 15 weeks after injection. Conclusion: This study showed that the injured disc had a significant change in DHI on simple lateral radiograph and disc degeneration grade on MRI scans within 15 weeks in all rabbits. Rabbit annular puncture model can be useful as a disc degeneration model in vivo.

Analysis of Molecular Expression in Adipose Tissue-Derived Mesenchymal Stem Cells : Prospects for Use in the Treatment of Intervertebral Disc Degeneration

  • Jin, Eun-Sun;Min, Joongkee;Jeon, Sang Ryong;Choi, Kyoung Hyo;Jeong, Je Hoon
    • Journal of Korean Neurosurgical Society
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    • 제53권4호
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    • pp.207-212
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    • 2013
  • Objective : Recent studies have shown encouraging progress toward the use of autogenic and allogenic mesenchymal stem cells (MSCs) to arrest, or even lead to partial regeneration in, intervertebral disc (IVD) degeneration. However, this technology is still in its infancy, and further development is required. The aim of this study was to analyze whether rat adipose-derived mesenchymal stem cells (ADMSC) can differentiate towards IVD-like cells after treatment with transforming growth factor ${\beta}3$ (TGF-${\beta}3$) in vitro. We also performed quantitative analysis of gene expression for ADMSC only, ADMSCs treated with TGF-${\beta}3$, and co-cultured ADMSCs treated with TGF-${\beta}3$. Methods : ADMSCs were sub-cultured to homogeneity and used in fluorocytometry assays for CD11, CD45, and CD90/Thy1. ADMSCs were differentiated in spheroid culture towards the chondrogenic lineage by the presence of TGF-${\beta}3$, dexamethasone, and ascorbate. We also co-cultured pure ADMSCs and nucleus pulposus cells in 24-well plates, and performed immunohistochemical staining, western blotting, and RT-PCR for quantitative analysis of gene expression. Results : Results of fluorocytometry were positive for CD90/Thy1 and negative for CD11 and CD45. TGF-${\beta}3$-mediated induction of ADMSCs led to the expression of the differentiation markers of intervertebral disc-like cells, such as aggrecan, collagen II, and sox-9. Co-cultured ADMSCs treated with TGF-${\beta}3$ showed higher expression of differentiation markers and greater extracellular matrix production compared with ADMSCs treated with TGF-${\beta}3$ alone. Conclusion : ADMSC treated with TGF-${\beta}3$ may be an attractive source for regeneration therapy in degenerative IVD. These findings may also help elucidate the pathologic mechanism of MSC therapy in the degeneration of IVD in vivo.

DBP 스폰지와 DBP/PLGA 지지체에서의 인간 디스크세포 거동분석 비교 (The Comparison of Sponges and PLGA Scaffolds Impregnated with DBP on Growth Behaviors of Human Intervertebral Disc Cells)

  • 이선경;홍희경;김수진;김용기;송이슬;하윤;이동원;강길선
    • 폴리머
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    • 제34권5호
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    • pp.398-404
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    • 2010
  • 본 연구팀은 DBP를 함침시킨 물성이 서로 다른 스폰지와 PLGA 지지체를 제작한 후 세포 부착, 증식 및 형태 유지를 알아보기 위한 실험을 수행하였다. WST 분석법과 SEM 관찰을 통하여 스폰지에 비해서 PLGA 지지체에서의 세포의 증식이 활발한 것을 확인하였고, RT-PCR을 통해 디스크세포에서 특이적으로 발현하는 제 2형 콜라겐과 어그리칸의 발현을 확인하였다. WST 결과, 세포 증식률은 DBP/PLGA 지지체가 DBP를 함침시킨 스폰지보다 세포 증식률이 높음을 확인하였다. 본 연구팀은 스폰지보다 PLGA 지지체가 인간디스크의 표현형 유지 및 증식에 있어서 긍정적인 영향을 미치는 것을 확인하였다.

방사선 일반 정면검사에서 허리뼈 추간판 계측 값에 따른 입사각 적용 (Application of Incidence Angle on Lumbar Spine Anteroposterior General Radiography Image according to Measured Intervertebral Disc Angle)

  • 문슬지아;김경립;조희정;성순기;곽종혁
    • 한국콘텐츠학회논문지
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    • 제19권6호
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    • pp.471-480
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    • 2019
  • 정상인을 대상으로 체질량지수, 전만각도에 따른 L-spine 3~4, L-spine 4~5, L-spine 5~Sacrum 1의 각각의 추간판각도를 비교 분석하여 입사각을 제시하였다. 체질량지수의 L-spine 3~4, L-spine 4~5, L-spine 5~Sacrum 1의 정면 입사각도는 머리 쪽 방향으로 5.66도, 13.23도, 29.13도였으며, 전만각도의 L-spine 3~4, L-spine 4~5, L-spine 5~Sacrum 1의 정면 입사각도는 머리 쪽 방향으로 6.32도, 16.09도, 35.36도였다. 체질량지수, 전만각도에 따른 정면 입사각도에 따라 적용한 팬텀의 일반 전후방향 영상의 왜곡정도를 면적비율로 비교한 결과 L-spine 4~5, L-spine 5~Sacrum 1의 추간판각도에서 유의미한 차이가 있었고(p<0.05), 전만각도와 추간판각도는 양의 상관관계를 보였다(p<0.05). 팬텀의 전만각도에 따른 추간판각도 계측 값을 머리 쪽 방향으로 입사각을 L4는 11도, L5는 26도로 적용하여 영상의 유용성을 평가한 결과, 왜곡비율면적은 L4에서 14.90%에서 12.11%로 줄어들었고, L5에서는 15.25%에서 13.72%로 줄어들었다. 계측된 추간판 각도에 따른 입사각을 적용한 허리뼈 일반 전후방향 영상에서 허리뼈 4번, 5번을 목적으로 하는 허리뼈 정면 영상에서 왜곡을 줄여, 정확한 구조의 정면 추체상과 인접하는 관절 간 영상을 얻을 수 있었고, 목적하는 부위의 영상의 질과 진단적 정보를 향상시킬 수 있었다.

교통사고로 유발된 요통 환자의 요천각, 전만각 및 IVD각에 대한 임상적 고찰 (The Clinical Study of the Ferguson's Angle, Lumbar Lordotic Angle, Lumbar IVD Angle of Low Bcak Pain Patients Induced in Traffic Accident)

  • 이길재;박경무;임제연;송윤경;임형호
    • 한방재활의학과학회지
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    • 제19권2호
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    • pp.227-239
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    • 2009
  • Objectives : The purpose of this study is to investigate the relation of Ferguson's angle, Lumbar lordotic angle, Lumbar IVD angle in traffic accident patients. Methods : Total patients were classified into TA(traffic accident) inpatients and non-TA(lumbago) inpatients. We analyzed the characteristics of each group Ferguson's angle, Lumbar lordotic angle, L4-5 IVD angle, L5-S1 IVD angle on X-ray film. Results : 1. Ferguson's angle and Lumbar lordotic angle were smaller than normal range. Non-TA group's angles were smaller than TA group's angles. 2. L4-5 IVD angle of female showed smaller than that of male in TA groups. The older age was, the smaller L4-5 IVD angle was at non-TA groups. 3. L5-S1 IVD angle of female showed smaller than that of male in total groups. The older age was, the smaller L5-S1 IVD angle was at non-TA groups. 4. Ferguson's angle, Lumbar lordotic angle, Lumbar IVD angle were related with direct proportion in total groups. 5. Non-TA groups showed more effective VAS variation than TA groups. Conclusions : Lumbar angles were intimately related with each others. And lumbar angles have an effect on curative value.