Tissue Engineering of the Intervertebral Disc with Cultured Nucleus Pulposus Cells Using Atelocollagen Scaffold and Gene Therapy

STUDY DESIGN: This is an in-vitro experiment using rabbit intervertebral disc (IVD) cells and growth factors. OBJECTIVES: We wanted to determine the effect of types I, and II atelocollagen and growth factor gene therapy for matrix regeneration of rabbit IVD cells. SUMMARY OF THE LITERATURE REVIEW: Adenovirus-medicated growth factor gene therapy is efficient for matrix regeneration of the IVD. Atellocollagen has provided a favorable environment for matrix synthesis. However, a combined approach using gene and cell therapy in an atelocollagen scaffold has not yet been attempted. MATERIALS AND METHODS: Rabbit IVD cells were transduced with Ad/TGF-beta1 and Ad/BMP-2. The cells were then implanted to the atelocollagen scaffold. The [methyl-3H]thymidine incorporation for DNA synthesis and the [35S]sulfur incorporation for proteoglycan synthesis were measured. RT-PCR was performed for assessing the aggrecan, collagen type I, collagen type II and osteocalcin mRNA expressions. RESULTS: The rabbit IVD cells with Ad/TGF-beta1 and that were cultured in type I atelocollagen showed a 130% increase in new proteoglycan synthesis, while the rabbit IVD cells with Ad/TGF-beta1 and that were cultured in type II atelocollagen showed a 180% increase in new proteoglycan synthesis (p<0.05). The rabbit IVD cells with Ad/BMP-2 and that were cultured in type I atelocollagen showed a 70% increase in new proteoglycan synthesis, while the rabbit IVD cells with Ad/BMP-2 and that were cultured in type II atelocollagen showed a 95% increase (p<0.05). Rabbit IVD cells with Ad/TGF-beta1 and Ad/BMP-2 and that were cultured in type I and II atelocollagen demonstrated increased collagen type I and II mRNA expressions without an osteocalcin mRNA expression (p<0.05). CONCLUSION: Cell and gene therapy in an atelocollagen scaffold provided a efficient mechanism for chondrogenic matrix regeneration of rabbit IVD cells.ope

아데노바이러스를 이용한 유전자치료법과 아텔로콜라겐 지지체를 매개로 한 인간 추간판의 조직공학적 재생

Dept. of Medical Science/석사[한글] [영문] Tissue engineering is regarded as a new experimental technique for the biological treatment about degenerative intervertebral disc (IVD) diseases. Among artificial scaffolds, atelocollagen has an advantage in safety over others. And growth factors were known to stimulate matrix synthesis of IVD cells, among of them, transforming growth factor-β1 and bone morphogenetic protein-2 are the best candidates for IVD regeneration. Gene transfer to IVD cells provided sustained mechanism for matrix synthesis. Hence, the object of this study was to examine the cellular proliferation, proteoglycan (PG) synthesis, and phenotypical expression of IVD cells transduced by adenovirus transforming growth factor beta-1(Ad/TGF-β1) construct and adenovirus bone morphogenetic protein-2(Ad/BMP-2) construct and seeded on type I atelocollagen scaffolds.The IVD cells were isolated by sequential enzymatic digestion. Atelocollagen scaffold were prepared by lypophilization and cross-linking method. Ad/TGF-β1 or Ad/BMP-2 treated IVD cells were cultured for 7days. At 3 and 7 days, cellular proliferation, newly synthesized proteoglycan, mRNA expressions and the inner morphology of cell seeded scaffolds was determined by scanning electron microscopy (SEM).As result, transduced IVD cultures in atelocollagen type I with Ad/TGF-β1 and mixture group demonstrated increase in proteoglycan synthesis, but did not show significant change in mRNA expressions, compared to control. On the other hand, the IVD of transduced with Ad/ TGF-β1 in types I atelocollagen scaffolds decresed in cellular proliferation compared with others. And there were not showed osteocalcin mRNA expression in any groups.Human IVD cells were viable in type I atelocollagen scaffolds. Type I atelocollagen scaffold was suitable for cell proliferation, extracellular matrix synthesis. And Ad/TGF-β1 and Ad/BMP-2 combination was potential for IVD regeneration. Taken together, IVD cells in atelocollagen scaffolds, with therapeutic gene, provide a mechanism for tissue engineering of IVD.ope

The Effects of a Story-Structure Learning Strategy on Comprehension and Reading Attitude of Students of Poor Reading Performance

본 연구는 초등학교 4학년 읽기학습부진학생을 대상으로 이야기구조 학습전략이 독해력과 읽기 태도에 미치 는 효과를 검증하고자 하였다. 연구 대상은 00광역시에 소재한 M초등학교 4학년 읽기학습부진학생으로 실험집 단 10명, 통제집단 10명이다. 실험 처지는 주 2회 8주간 총 16회였으며 매회 40분이 소요되었다. 실험집단에게 는 1단계 이야기 분석 단계, 2단계 전략 중심의 지도, 3단계 독해력 표현의 과정으로 이야기구조 학습전략을 실시하였고 통제집단에게는 전통적인 읽기지도를 하였다. 주요 연구 결과는 다음과 같았다. 첫째, 집단 간에는 유의한 차이가 없었지만, 이야기구조 학습전략 프로그램은 실험집단의 읽기학습부진학생의 독해력 향상에 유의 한 영향을 미쳤다. 둘째, 이야기구조 학습전략 프로그램은 읽기학습부진학생의 읽기 태도에 긍정적인 영향을 미 쳤다. 이상의 연구결과로 미루어볼 때 이야기구조 활용 전략을 지도할 경우 읽기학습부진학생의 읽기 태도와 독해력에 긍정적인 영향을 미칠 것으로 기대된다. The purpose of this study was to investigate the effects of a story-structure learning strategy on the comprehension and reading attitude of fourth-graders of poor reading performance. A total of 20 fourth-graders of poor reading performance at M Elementary School in I City participated to this study. The treatment group was taught to use story-structure learning strategies when comprehend stories, while the control group was taught with traditional reading comprehension practices. The main findings of this study were as follow: First, the story-structure learning strategy program had significantly improved comprehension of the treatment group's students with poor reading achievements. Second, the story-structure learning strategy program had positive effects on the reading attitude of the students of poor reading performance. Thus, it seemed that the story-structure learning strategy might help poor readers comprehend text material with multiple paragraphs