CAJ | 학술논문

目的评价重组人肝细胞生长因子(hHGF)基因修饰的成骨细胞经髓芯减压移植促进早期股骨头缺血坏死的血管新生及局部骨修复的效果.方法二步酶消化法和差速贴壁法分离培养胎兔成骨细胞,利用脂质体介导hHGF基因转染成骨细胞,然后通过髓芯减压移植于兔缺血坏死的股骨头内,同时设置单纯细胞移植组和髓芯减压组,于术后第2、4、8周通过CT、组织学和微血管墨汁灌注等观察血管形成及成骨情况.结果分离培养的胎兔成骨细胞经Ⅰ型胶原和碱性磷酸酶鉴定纯度较高.hHGF基因转染成骨细胞体外及体内检测均有hHGF蛋白的表达.通过髓芯减压移植细胞8周后转基因组和单纯成骨细胞组的新生骨小梁与单纯髓芯减压组比较有差异性,而术后2、4周转基因组的新生血管数(29.5±1.6)和(34.0±1.7)较对照组(20.6±1.9)和(25.6±2.2)差异有统计学意义(P＜0.01).结论转染hHGF基因的成骨细胞移植于缺血坏死的股骨头后,早期可促进血管新生,并增加骨形成,显著促进坏死骨修复.
목적 평개중조인간세포생장인자(hHGF)기인수식적성골세포경수심감압이식촉진조기고골두결혈배사적혈관신생급국부골수복적효과.방법 이보매소화법화차속첩벽법분리배양태토성골세포,이용지질체개도hHGF기인전염성골세포,연후통과수심감압이식우토결혈배사적고골두내,동시설치단순세포이식조화수심감압조,우술후제2、4、8주통과CT、조직학화미혈관묵즙관주등관찰혈관형성급성골정황.결과 분리배양적태토성골세포경Ⅰ형효원화감성린산매감정순도교고.hHGF기인전염성골세포체외급체내검측균유hHGF단백적표체.통과수심감압이식세포8주후전기인조화단순성골세포조적신생골소량여단순수심감압조비교유차이성,이술후2、4주전기인조적신생혈관수(29.5±1.6)화(34.0±1.7)교대조조(20.6±1.9)화(25.6±2.2)차이유통계학의의(P＜0.01).결론 전염hHGF기인적성골세포이식우결혈배사적고골두후,조기가촉진혈관신생,병증가골형성,현저촉진배사골수복.
Objective To evaluate the effects of transplantation of human hepatocyte growth factor (hHGF)gene-modified osteoblasts combined with core decompression in treatment of avascular necrosis of femoral head(ANFH).Methods The plasmid pcDNA3.1(+)-hHGF containing hHGF gene was constructed.Osteoblasts were isolated from fetal rabbits,cultured,and transfect3d with the plasmid pcDNA3.1(+)-hHGF or blank plasmid pcDNA3.1(+),or used as controls.Thirty-six adult New Zealand rabbits were made into ANFH models,underwent core decompression,and were randomly divided into 3 groups.Group A,transplanted with osteoblasts transfected with pcDNA3.1(+)-hHGF plasmid,Group B,transplanted with osteoblasts not transfected with pcDNA3.1(+)-hHGF plasmid,and Group C,injected with PBS medium.2,4,and 8 weeks later samples of femoral head were obtained to undergo CT,histological examination,and capillary ink infusion so as to observe the angiogenesis and osteogenesis.Results The pcDNA3.1(+)-hHGF transfected osteoblasts showed stable expression of hHGF.The numbers of newly formed vessels of the femoral heads of the group transfected with pcDNA3.1(+)-hHGF-transfected osteoblasts 2 and 4 weeks later were(29.47±1.64)and(34.02±1.72)/cm2 respectively,both significantly higher than those of the group transfected with blank plasmid-trsansfected osteoblasts[(20.61±1.91)and(25.57±2.20)/cm2 respectively,both P＜0.01].Eight weeks later the numbers of mature trabeeular bone and bone marrow of Groups A and B were significantly higher than those of Group C.Conclusion Core decompresson combined with transplantation of HGF gene-modified osteoblasts promotes angiogenesis,enhances bone formation,and improves the restoration of avascular necrosis of femoral head.