CAJ | 학술논문

背景：骨髓间充质干细胞发挥成骨作用需要支架材料的辅助，一方面支架材料不仅可将细胞运载至骨缺损区域，另一方面还可作为新骨生长的框架结构。胶原-壳聚糖复合材料是骨组织工程较为理想的支架材料之一，同时其具有骨诱导性，比常规支架材料更优越的成骨能力。骨搬移技术在临床上在修复长段骨缺损方面已得到广泛应用，但也存在成骨慢、外固定时间长、骨不连等缺憾。如何进一步加快骨形成速度，减少并发症发生，已成当前亟待解决的问题。实验假设：骨髓间充质干细胞复合胶原-壳聚糖支架移植能提高胫骨缺损骨搬移修复效果。 　　方法/设计：随机对照动物实验。分为体外和体内实验两部分。体外实验中取月龄一两个月的新西兰大白兔股骨骨髓，提取骨髓间充质干细胞，培养至第3代，将细胞悬液滴于胶原-壳聚糖支架材料，构建骨髓间充质干细胞复合胶原-壳聚糖支架。体内实验选用24只三四月龄新西兰大白兔，被随机分配接受如下干预：骨搬移、支架植入、骨搬移联合支架植入。研究的主要观察指标为植入材料与骨缺损界面的生长情况、X 射线检测的缺损区骨修复情况、苏木精-伊红染色及扫描电镜观察缺损区成骨情况、免疫组织化学染色检测成骨区Ⅰ型胶原蛋白的表达情况、扫描电子显微镜观察移植材料与宿主骨的界面键合情况、超微结构及新骨的生成。 　　讨论：实验结果将有助于确定对骨缺损进行骨搬移治疗过程中，应用骨髓间充质干细胞复合胶原-壳聚糖支架移植促进骨缺损再生修复效果的可行性。
배경：골수간충질간세포발휘성골작용수요지가재료적보조，일방면지가재료불부가장세포운재지골결손구역，령일방면환가작위신골생장적광가결구。효원-각취당복합재료시골조직공정교위이상적지가재료지일，동시기구유골유도성，비상규지가재료경우월적성골능력。골반이기술재림상상재수복장단골결손방면이득도엄범응용，단야존재성골만、외고정시간장、골불련등결감。여하진일보가쾌골형성속도，감소병발증발생，이성당전극대해결적문제。실험가설：골수간충질간세포복합효원-각취당지가이식능제고경골결손골반이수복효과。 　　방법/설계：수궤대조동물실험。분위체외화체내실험량부분。체외실험중취월령일량개월적신서란대백토고골골수，제취골수간충질간세포，배양지제3대，장세포현액적우효원-각취당지가재료，구건골수간충질간세포복합효원-각취당지가。체내실험선용24지삼사월령신서란대백토，피수궤분배접수여하간예：골반이、지가식입、골반이연합지가식입。연구적주요관찰지표위식입재료여골결손계면적생장정황、X 사선검측적결손구골수복정황、소목정-이홍염색급소묘전경관찰결손구성골정황、면역조직화학염색검측성골구Ⅰ형효원단백적표체정황、소묘전자현미경관찰이식재료여숙주골적계면건합정황、초미결구급신골적생성。 　　토론：실험결과장유조우학정대골결손진행골반이치료과정중，응용골수간충질간세포복합효원-각취당지가이식촉진골결손재생수복효과적가행성。
BACKGROUND:Bone marrow mesenchymal stem cels play an osteogenic role under the assistance of scaffold materials. The scaffold cannot only deliver the cels to the bone defect area, but also act as a new bone growth framework. Colagen-chitosan composite is one of ideal scaffold materials in bone tissue engineering, which has osteoinductive ability and better osteogenic ability than conventional scaffolds. Bone transport technology has been widely used in the clinical repair of long bone defects, but it has some deficiencies, such as slow osteogenesis, long time for external fixation and nonunion. How to further accelerate bone formation and reduce complications has become the current problem to be solved. Here, it is hypothesized that bone marrow mesenchymal stem cels/ colagen/chitosan composite scaffold can increase the therapeutic effect of bone transport in the repair of tibial bone defects. METHODS/DESIGN:This study is a randomized controled animal experiment, includingin vitro andin vivo tests.In vitro test: Bone marrow mesenchymal stem cels are isolated from the bone marrow of New Zealand rabbits aged 1-2 months, and passaged to the third generation. Then, cel suspension is added onto the colagen-chitosan scaffold to construct the bone marrow mesenchymal stem cels/colagen/chitosan composite scaffold.In vivo test: Twenty-four New Zealand rabbits at 3-4 months are selected and randomly assigned to receive bone transport, scaffold implantation, bone transport+scaffold implantation, respectively. The primary outcome measures are the growth of implant materials and bone defect interface, X-ray detection of bone defect repair, hematoxylin-eosin staining and scanning electron microscope observation of bone formation in the bone defect region, immunohistochemical detection of type I colage expression in the osteogenic region, scanning electron microscope observation of interface bonding between implant materials and host bone, ultrastructure and bone formation. DISCUSSION:The results from this animal experiment wil help to determine the feasibility of bone marrow mesenchymal stem cels/colagen/chitosan composite scaffold to accelerate bone repair during bone defect repair using bone transport technology.