干细胞移植改善股骨头坏死缺血状态的实验及临床效应:20只模型兔及188例患者资料分析
간세포이식개선고골두배사결혈상태적실험급림상효응:20지모형토급188례환자자료분석
Stem cell transplantation for ischemic femoral head necrosis Analysis in 20 model rabbits and 188 patients
  • 万方数据
    中国组织工程研究与临床康复 중국조직공정연구여림상강복
    JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
    2008年 8期 1558-1562 ,共5页

    杨晓凤%王红梅%许忆峰%张轶斌%吴雁翔%吕欣%吕乃武%单鸿%马淑彦

    양효봉%왕홍매%허억봉%장질빈%오안상%려흔%려내무%단홍%마숙언

    骨髓间充质干细胞%动脉内干细胞移植%股骨头坏死%血管新生 骨髓間充質榦細胞%動脈內榦細胞移植%股骨頭壞死%血管新生
    골수간충질간세포%동맥내간세포이식%고골두배사%혈관신생
    背景:缺血性股骨头坏死由于股骨头局部滋养血管损伤、供血不足致病,迄今尚无最佳治疗方案,改善股骨头血液供应成为治疗本病的关键.目的:通过动物实验和临床观察,验证自体骨髓间充质干细胞移植血管再生技术改善股骨头坏死缺血状态的效果.设计:对比动物实验和自身对照临床观察.单位:解放军第四六三医院细胞治疗中心.资料:①动物:选用日本大耳白兔20只,雌雄不拘,体质量3.0～4.0 kg,购于沈阳军区总医院动物实验中心.此动物实验符合动物伦理学要求.②对象:选择2004-07/2007-07在解放军第四六三医院细胞治疗中心住院的具有完整随访资料的缺血性股骨头坏死患者188例(335髋),男113例,女75 例,年龄18～72岁.均经髋关节X线摄片、核素扫描、MRI、CT检查确诊;对治疗和实验知情同意.实验经医院伦理委员会批准.方法:①动物实验:实验于2004-01/2004-06在沈阳军区总医院动物实验中心完成.采用液氮冷冻股骨头缺血性坏死模型建立方法,双后肢股骨头均制作缺血模型,右侧为移植组,左侧为对照组.抽取骨髓,将所获单个核细胞经右股动脉注入;同法将生理盐水注入左侧股动脉内.②临床观察:密度梯度离心缺血性股骨头坏死患者自体骨髓,分离单个核细胞.行旋股内、旋股外及闭孔动脉干细胞移植术.主要观察指标:①4周后行DSA动脉造影观察双侧血管新生情况.4,12周后取双侧股骨头标本,行病理切片观察新骨再生及股骨头修复程度.②于移植后第3,6,12,24个月观察患者髋关节疼痛、行走间距及步态、髋关节外展与内旋功能变化.于干细胞移植后6个月行股骨头供血动脉造影术,观察血管新生及股骨头供血动脉充盈情况.于干细胞移植后6,12,24个月行CT,X射线,MRI检查,观察股骨头区形态学及缺血状态变化.于干细胞移植前,移植后3,6,12,24个月进行髋关节Harris评分评价髋关节功能.结果:动物实验结果:①20只兔均进入结果分析.②DSA动脉造影结果:移植后4周移植组兔右后肢股骨头区供血动脉较对照组股动脉明显增多.③病理结果:移植后12周右侧股骨头软骨、板层骨及骨小梁明显修复,左侧股骨头坏死无改善.临床观察:188例患者均进入结果分析.①症状改善:188例患者中髋关节疼痛缓解164例(87.3%),关节功能改善147例(78.4%),行走间距延长150例(80.0%).②影像学改变:移植后6个月12例患者DSA动脉造影显示,股骨头供血动脉较移植前明显增多、增粗,血流速度增快;12～24个月后X射线平片,CT,MRI检查24例患者坏死的股骨头区可见骨质病变获得改善.③髋关节Harris评分:干细胞移植后6,12,24个月高于移植前,差异有非常显著性意义(t=-3.423,-6.714,-9.039,P﹤0.01).结论:经动脉自体骨髓间充质干细胞移植可改善股骨头坏死缺血状态,是治疗缺血性股骨头坏死的有效手段.
    배경:결혈성고골두배사유우고골두국부자양혈관손상、공혈불족치병,흘금상무최가치료방안,개선고골두혈액공응성위치료본병적관건.목적:통과동물실험화림상관찰,험증자체골수간충질간세포이식혈관재생기술개선고골두배사결혈상태적효과.설계:대비동물실험화자신대조림상관찰.단위:해방군제사륙삼의원세포치료중심.자료:①동물:선용일본대이백토20지,자웅불구,체질량3.0～4.0 kg,구우침양군구총의원동물실험중심.차동물실험부합동물윤리학요구.②대상:선택2004-07/2007-07재해방군제사륙삼의원세포치료중심주원적구유완정수방자료적결혈성고골두배사환자188례(335관),남113례,녀75 례,년령18～72세.균경관관절X선섭편、핵소소묘、MRI、CT검사학진;대치료화실험지정동의.실험경의원윤리위원회비준.방법:①동물실험:실험우2004-01/2004-06재침양군구총의원동물실험중심완성.채용액담냉동고골두결혈성배사모형건립방법,쌍후지고골두균제작결혈모형,우측위이식조,좌측위대조조.추취골수,장소획단개핵세포경우고동맥주입;동법장생리염수주입좌측고동맥내.②림상관찰:밀도제도리심결혈성고골두배사환자자체골수,분리단개핵세포.행선고내、선고외급폐공동맥간세포이식술.주요관찰지표:①4주후행DSA동맥조영관찰쌍측혈관신생정황.4,12주후취쌍측고골두표본,행병리절편관찰신골재생급고골두수복정도.②우이식후제3,6,12,24개월관찰환자관관절동통、행주간거급보태、관관절외전여내선공능변화.우간세포이식후6개월행고골두공혈동맥조영술,관찰혈관신생급고골두공혈동맥충영정황.우간세포이식후6,12,24개월행CT,X사선,MRI검사,관찰고골두구형태학급결혈상태변화.우간세포이식전,이식후3,6,12,24개월진행관관절Harris평분평개관관절공능.결과:동물실험결과:①20지토균진입결과분석.②DSA동맥조영결과:이식후4주이식조토우후지고골두구공혈동맥교대조조고동맥명현증다.③병리결과:이식후12주우측고골두연골、판층골급골소량명현수복,좌측고골두배사무개선.림상관찰:188례환자균진입결과분석.①증상개선:188례환자중관관절동통완해164례(87.3%),관절공능개선147례(78.4%),행주간거연장150례(80.0%).②영상학개변:이식후6개월12례환자DSA동맥조영현시,고골두공혈동맥교이식전명현증다、증조,혈류속도증쾌;12～24개월후X사선평편,CT,MRI검사24례환자배사적고골두구가견골질병변획득개선.③관관절Harris평분:간세포이식후6,12,24개월고우이식전,차이유비상현저성의의(t=-3.423,-6.714,-9.039,P﹤0.01).결론:경동맥자체골수간충질간세포이식가개선고골두배사결혈상태,시치료결혈성고골두배사적유효수단.
    BACKGROUND: Ischemic femoral head necrosis is caused by local vascular injury and blood-supply insufficiency. There exists no optimal treatment for the ischemic femoral head necrosis. Thus, the improvement of the blood supply to the femoral head seems to be a key point for the treatment. OBJECTIVE: To verify the curative effects of autologous bone marrow mesenchymal stem cell transplantation induced vascular regeneration on the improvement of ischemic femoral head necrosis via animal experiments and clinical observations. DESIGN: Contrast animal experiment and self-controlled clinical observation.SETTING: Cell Therapy Center, the 463 Hospital of Chinese PLA.MATERIALS AND PARTICIPANTS: ① Animals: Twenty Japanese white rabbits in either gender and weighing 3.0-4.0 kg were purchased from Animal Experimental Center, General Hospital of Shenyang Military Area Command of Chinese PLA. The animal experiments were coincident with the ethical standards. ② Participants: 188 patients with ischemic femoral head necrosis (335 hips) having whole following-up data were selected from Cell Therapy Center, the 463 Hospital of Chinese PLA from July 2004 to July 2007. There were 113 males and 75 females, and their ages ranged from 18 to 72 years. Diagnosis was done by using X-ray photographs, nuclide scanning, MRI and CT examinations. All patients provided the informed consent, and the study was approved by the local research ethics committee. METHODS: ① Animal experiments: The experiment was carried out at the Animal Experiment Center, General Hospital of Shenyang Military Area Command of Chinese PLA from January to June 2004. Ischemic femoral head necrosis models were established at both hindlimbs by using liquid-nitrogen refrigeration. The right side was regarded as transplantation group and the left one as control group. Mononuclear cells extracted from bone marrow were poured in the right femoral artery, while saline was poured in the left femoral artery. ②Clinical observations: Mononuclear cells were separated from autologous bone marrow of patients with ischemic femoral head necrosis after density gradient centrifugation. MAIN OUTCOME MEASURES: ①Four weeks later, angiogenesis at both femoral arteries was observed by arteriography by using digital subtraction anglography (DSA). Moreover, bilateral femoral head samples underwent pathological sections to observe bone regeneration and repair of femoral head 4 and 12 weeks later. ②Items including hip pain, walking distance and gait, abduction and internal rotation function changes of hip joint were observed in 3, 6, 12 and 24 months after stem cell transplantation in media femoral circum flex artery, lateral femoral circum flex artery and obturator artery. In 6 months after stem cell transplantation, angiogenesis and blood supply of femoral head were observed by using arteriography. In 6, 12 and 24 months after stem cell transplantation, morphological and ischemic changes of femoral head were observed by using CT, X-ray and MRI examinations. Harris scores were used to evaluate function of hip joint before and in 3, 6, 12 and 24 months after stem cell transplantation.RESULTS: Animal experiment: Twenty rabbits were involved in the final analysis. ① DSA-arteriography results: In 4 weeks after transplantation, blood-supply arteries in femoral head of right hindlimb in the transplantation group were more than those in the control group. ② Pathological results: In 12 weeks after transplantation, cartilage, lamellar bone and bone trabecula in the left femoral head were repaired remarkably, but left femoral head necrosis was not improved. Clinical observations: 188 patients were involved in the final analysis. ① Improvement of symptoms: Among 188 patients, 164 (87.3%) had remission of hip pain, 147 (78.4%) had function improvement, and 150 (80.0%) had elongation of walking distance. ② Imaging changes: At 6 months after transplantation, DSA-arteriography in 12 patients demonstrated that blood-supply arteries in femoral head were increased and thickened remarkably as compared with those before transplantation, and the blood flow was rapid. At 12-24 months after transplantation, lesion of bone matrix in 24 patients was improved under the X-ray, CT and MRI examinations. ③ Harris scores of hip joint: The scores at 6, 12 and 24 months after transplantation were significantly higher than those before transplantation (t= -3.423, -6.714, -9.039, P < 0.01).CONCLUSION: Autologous bone marrow mesenchymal stem cell transplantation can effectively improve and treat ischemic femoral head necrosis.
    원문보기 원문다운로드 사이트로이동
저자의 최근 논문