CAJ | 학술논문

目的制备骨形态发生蛋白-2( BMP-2)/胶原/掺锶羟基磷灰石材料并探讨其修复大鼠颅骨缺损的可行性和有效性.方法扫描电镜观察Ⅰ型胶原制备单纯胶原、胶原/羟基磷灰石、胶原/掺锶羟基磷灰石、BMP-2/胶原/掺锶羟基磷灰石4组骨修复材料表面结构.用BMP-2/胶原/掺锶羟基磷灰石材料浸提液进行细胞毒性试验和体外溶血试验评价其生物相容性.在大鼠头颅制备颅骨极限骨缺损模型,分别植入4种骨修复材料.术后12周CT扫描观察骨缺损修复影像学.苏木素-伊红(HE)和Masson染色观察骨缺损组织学变化,并在骨缺损及其周围新生骨部位行骨桥蛋白( OPN)和β-连环蛋白(β-catenin)免疫组织化学染色.结果在扫描电镜下观察发现单纯的胶原材料为交织样物质结构,胶原/羟基磷灰石材料为交织晶体板状结构胶原/掺锶羟基磷灰石和BMP-2/胶原/掺锶羟基磷灰石材料晶体结构为单晶体交织状.BMP-2/胶原/掺锶羟基磷灰石材料浸提液对细胞相对增殖率(RGR)无显著影响(P＞0.05),材料的细胞毒性为1级.骨缺损CT扫描平均CT值分别为(98.5±10.2)、(208.4±19.5)、(418.4±27.1)、(476.8±30.5)hu,BMP-2/胶原/掺锶羟基磷灰石材料缺损部位CT值最高.HE和Masson染色见BMP-2/胶原/掺锶羟基磷灰石组骨质愈合完全,原骨缺损处多为红色成熟骨.胶原/掺锶羟基磷灰石组植入区内蓝色的新生骨较多.胶原/羟基磷灰石材料组,植入区在植入材料边缘新生骨形成,界限仍然清晰.单纯胶原组骨质未愈合,骨缺损处为淡蓝色条索状结构,中间未见骨形成.对比其他3组,BMP-2/胶原/掺锶羟基磷灰石组存在大量棕色的OPN和β-catenin染色阳性新生骨组织,差异有统计学意义(P＜0.05).结论 BMP-2/胶原/掺锶羟基磷灰石材料促进骨修复能力强于单纯胶原、胶原/羟基磷灰石、胶原/掺锶羟基磷灰石材料.
목적 제비골형태발생단백-2( BMP-2)/효원/참송간기린회석재료병탐토기수복대서로골결손적가행성화유효성.방법 소묘전경관찰Ⅰ형효원제비단순효원、효원/간기린회석、효원/참송간기린회석、BMP-2/효원/참송간기린회석4조골수복재료표면결구.용BMP-2/효원/참송간기린회석재료침제액진행세포독성시험화체외용혈시험평개기생물상용성.재대서두로제비로골겁한골결손모형,분별식입4충골수복재료.술후12주CT소묘관찰골결손수복영상학.소목소-이홍(HE)화Masson염색관찰골결손조직학변화,병재골결손급기주위신생골부위행골교단백( OPN)화β-련배단백(β-catenin)면역조직화학염색.결과 재소묘전경하관찰발현단순적효원재료위교직양물질결구,효원/간기린회석재료위교직정체판상결구효원/참송간기린회석화BMP-2/효원/참송간기린회석재료정체결구위단정체교직상.BMP-2/효원/참송간기린회석재료침제액대세포상대증식솔(RGR)무현저영향(P＞0.05),재료적세포독성위1급.골결손CT소묘평균CT치분별위(98.5±10.2)、(208.4±19.5)、(418.4±27.1)、(476.8±30.5)hu,BMP-2/효원/참송간기린회석재료결손부위CT치최고.HE화Masson염색견BMP-2/효원/참송간기린회석조골질유합완전,원골결손처다위홍색성숙골.효원/참송간기린회석조식입구내람색적신생골교다.효원/간기린회석재료조,식입구재식입재료변연신생골형성,계한잉연청석.단순효원조골질미유합,골결손처위담람색조색상결구,중간미견골형성.대비기타3조,BMP-2/효원/참송간기린회석조존재대량종색적OPN화β-catenin염색양성신생골조직,차이유통계학의의(P＜0.05).결론 BMP-2/효원/참송간기린회석재료촉진골수복능력강우단순효원、효원/간기린회석、효원/참송간기린회석재료.
Objective To investigate the osteogenetic capacticy of bone morphogenetic protein-2 ( BMP-2 )/collagen/sr-HA scafffold. Methods Preparation of collagen,collagen/HA,collagen/sr-HA,BMP-2/collagen/sr-HA scaffolds.And the scaffolds were tested by scan electron microscope.To evaluate BMP-2/collagen/sr-HA scaffolds biocompatibility,the extract of the composite material was used to carry out a cytotoxicity test and in vitro hemolytic test.Cranial defects were created on 24 Sprague-Dawley rats.The Cranial defects were implanted the 4 scaffohls. Up to the 12 weeks,the rats were scanned hy CT and the whole calvaria were scanned with 0.5 mm in thickness.The rat calvaria from four groups were observed by histological staining and Immunohistochemistry. Results The toxicity of BMP-2/collagen/sr-HA to L929 cells was one degree.Three-dimensional reconstruction of computed tomography indicated that the cranial defects of BMP-2/collagen/sr-HA group were almost repair completely. The CT scores of these gourp were (98.5±10.2),(208.4±19.5),(418.4±27.1),(476.8±30.5) hu.And the density of radiodense areas of BMP-2/collagen/sr-HA group were highest among all groups.Histological observation showed a large amount of new and mature bone had been observed in the BMP-2/collagen/sr-HA groups,but in the collagen/HA and collagen/sr-HA the number of new bone were less.Immunohistochemistry in collagen/HA and collagen/sr-HA group the β-catenin andosteopontin (OPN) signals appeared on some cells at the boundary of the formed bone,in BMP-2/collagen/sr-HA group more intense and concentrated β-catenin and OPN signals were observed on the cells surrounding the newly formed extracellular matrix.Conclsion The osteogenic capability of BMP-2/collager/sr-HA scaffold is obviously superior to col/HA and col/sr-HA.BMP-2/collagen/sr-HA scaffold biomimetic scaffold material can be a potential repairing material for bone defects.