• Journal of Biomedical Engineering Research
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• v.39 no.1
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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.

• Journal of Korean Neurosurgical Society
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• v.47 no.1
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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.

• Journal of Korean Neurosurgical Society
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• v.44 no.5
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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.

• Journal of Korean Neurosurgical Society
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• v.53 no.4
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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.

• Polymer(Korea)
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• v.34 no.5
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• pp.398-404
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• 2010

We fabricated sponge and poly(lactide-co-glycolide)(PLGA) scaffolds impregnated demineralized bone particle(DBP)(DBP/PLGA) and investigated proper condition to proliferation and phenotype maintenance of intervertebral disc(IVD) cells by comparison between DBP/PLGA scaffold and DBP sponge. DBP/PLGA scaffolds were prepared by solvent casting/salt leaching. Human IVD cells were seeded in scaffolds of two types. Cell viability and proliferation according to scaffolds were analyzed by WST assay and SEM. RT-PCR was assessed to measure mRNA expression of aggrecan and type II collagen of human IVD cells. In WST assay results, cell viability in scaffolds impregnated DBP/PLGA scaffold were higher than DBP sponge. We could observe that disc cell mRNA expressed better in DBP/PLGA scaffold than DBP sponge. We concluded that the using of DBP/PLGA in terms of scaffold fabrication for bio-disc with human IVD cells is helpful growth of disc cells maintenance of phenotypes.

• The Journal of the Korea Contents Association
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• v.19 no.6
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• pp.471-480
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• 2019

L-spine 3~4, L-spine 4~5, and L-spine 5~Sacrum 1 intervertebral disc(IVD) angle according to gender, age, body mass index(BMI), lumbar lordosis angle(LLA) were compared and analyzed. The anteriorposterior incidence angle of L-spine 3 ~ 4, L-spine 4 ~ 5 and L-spine 5~Sacrum 1 in body mass index were 5.66, 13.23 and 29.13 degrees in the head direction and L-spine 3 4, L-spine 4 ~ 5, L-spine 5~Sacrum 1 had 6.32 degrees, 16.09 degrees and 35.36 degrees in the head direction. The distortion area ratio comparison was performed with the phantom image using the proposed incidence angle. There was a significant difference in L-spine 4~5 and L-spine 5~Sacrum 1 IVD angle relative to body mass index and LLA(p<0.05), IVD angle and LLA were positively correlated(p<0.05).As a result of evaluating the usefulness of the image by applying the incidence angles of the disc angles according to the phantom angle of deviation to the head direction as 11 degrees for L4 and 26 degrees for L5, the distortion ratio area decreased from 14.90% to 12.11% in L4, And from 15.25% to 13.72% in L5. In anteriorposterior image of the Lumbar spine applying the incidence angle according to the measured disc angle, it is possible to reduce the distortion to purpose L4, L5. And improved the quality and diagnostic information of the target site.

• Journal of Korean Medicine Rehabilitation
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• v.19 no.2
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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.