조직공학적 지지체와 성체줄기세포를 이용한 골 재생 및 신경 재생에 관한 연구

Abstract

In chapter 2, Bone marrow stromal cells (BMSCs) were harvested from the femurs and tibias of adult female Fischer rat. The ability of the PLGA with BMSCs to be integrated and to promote nerve regeneration in the transected rat spinal cord was investigated. Fischer rat received an implant consisting of the BMSCs suspended in the PLGA in the gap (T8 ~ T9) created by the spinal cord resection. For histological evaluation, the implants were removed after 4 and 8 weeks. Thin sections were cut from paraffin embedded tissue and histological sections were stained H&E, NSE and NF staining. Motor functional outcome measurements using the Basso-Beattie-Bresnehan (BBB) score performed weekly to 8 weeks post-injury. It was observed that the effects of the PLGA with BMSCs on neuroinduction (Group Ⅲ, scaffold, cell, cytokine) are stronger that PLGA without BMSCs (Group Ⅱ, scaffold) and blank model (Group Ⅰ). In chapter 3, Temperature sensitive MPEG-PCL diblock copolymer in this work could utilize as a potential carrier of BDNF for nerve regeneration. Methoxy poly(ethylene glycol) (MPEG)-b-poly(ε-caprolactone) (PCL) diblock copolymer was synthesized by ring-opening of ε-caprolactone (ε-CL) in the presence of a monomer activator with the terminal alcohol of MPEG as an initiator. The temperature sensitive behavior of the prepared MPEG-PCL diblock copolymer solution was examined. The polymer solution formed translucent sol at the room temperature. As the temperature increased from room temperature, the sol became gel. Brain-derived neurotrophic factor (BDNF) loaded MPEG-PCL diblock copolymer solution was prepared to examine the release behavior of BDNF. The release of BDNF in MPEG-PCL gel showed the prolonged release profile for 21 days. In chapter 4, To screen for small molecules inducers of neuronal differentiation, rat bone marrow mesenchymal stem cells isolated from adult female Fischer rat were used. Neuronal differentiation were analyzed with immunohistochemistry staining of Tuj1, immunofluorescence staining of NSE and CNPase. In addition, we confirmed that gene expression of neuronal specific marker such as Tuj1 and NSE by RT-PCR. It has been shown that rBMSCs can differentiate into neuron and oligodendrocyte in vitro by small molecules as KR63240 and KR63244. The last chapter, we describe the manufacture and characterization of in situ-forming chitosan gels containing chitosan and GP, and their ability to offer a suitable scaffold for rBMSCs in vitro and in vivo. Finally, we injected rBMSC-containing chitosan gel and evaluated its incorporation into surrounding tissues without the addition of exogenous biological factors.