体外冲击波疗法促进成骨的基础及相关临床研究
|
摘要
目的:本研究将体外冲击波(Extracorporeal Shock Wave, ESW)分别作用于体外培养的骨膜组织和骨髓间充质干细胞(human mesenchymal stem cells, hMSCs),了解ESW对骨膜成骨细胞及骨髓间充质干细胞代谢的影响,为揭示体外冲击波疗法(Extracorporeal Shock Wave Therapy, ESWT)治疗成骨障碍性疾病机理提供依据。同时应用ESWT治疗股骨头坏死及肩部软组织慢性损伤疾病,研究ESWT治疗成人非创伤性股骨头缺血性坏死的疗效及治疗方法,同时还探讨了彩色超声定位ESWT治疗肩部疾患中的应用价值,比较超声定位与痛点定位ESWT的疗效差异。
方法:ESW干预体外培养的骨膜组织及成体骨髓间充质干细胞后,分别用骨膜组织H~3-TdR掺入放射性自显影、透射电镜及相差显微镜观察,并用MTT、流式细胞计数及免疫组化染色等方法检测ESW干预前后骨膜成骨细胞H~3-TdR标记率、细胞超微结构的变化,以及ESW对hMSCs增殖周期和分泌碱性磷酸酶、骨钙素、Ⅰ型胶原等的影响。本研究还将能量密度为0.16-0.20mJ/mm~2的体外冲击波用于治疗69例Ⅰ～Ⅲ期股骨头缺血性坏死患者,然后分别于治疗前、治疗后3月、6月及12月行双侧髋关节X线、MRI检查及髋关节Harris评分。此外,采取随机分组的方法比较超声定位ESWT与痛点定位ESWT治疗肩峰下滑囊炎(65例)及肱二头肌长头腱炎(86例)的疗效差异。对上述各部分实验数据采用相应合理的统计学分析。
结果:(1) ESW干预后1、2周实验侧骨膜成骨细胞H~3-TdR标
Objective: In order to reveal the mechanism of Extracorporeal Shock Wave therapy (ESWT) in accelerating osteogenesis, DNA metabolish change in periosteal osteaoblast and human mesenchymal stem cells (hMSCs) was studied with multi-approach. Meanwhile, ESWT was applied to the treatment of avascular necrosis of the femoral head (ANFH) in adults. Its curative effect and application was discussed again. The effect of ESWT of ultrasonic orientation was also compared with the ESWT of pain-point orientation for subacromial bursitis (SB) and inflammation of long-tendon of biceps muscle (ILBM) in this study. Methods: After the periosteal osteaoblast and hMSCs was stimulated by Extracorporeal Shock Wave (ESW) in vitro, their mitotic activity, cellular ultrastructue, metabolish change, proliferation and differentiation was observed and evaluated by autoradiography, transmission electron micrograph, flow cytometer, cytochemical staining for osteocalcin and collagen I expression, the Alkaline phosphatese activity. ESWT with the energy density of 0.16~0.20mJ/mm~2 was applied to the treatment to ANFH of 69 patients, and its curative effects were evaluated by the MRI, X-ray, and the Harris hip score, before ESWT, and after ESWT for 3, 6, 12 months. Another group of patients, including 65 cases of SB and 86 cases of ILBM also accepted ESWT with the same density, but they were
randomly divided into two groups, ESWT by ultrasonic orientation andESWT resorted to tenderness point and local anatomy symbol (control group). The energy density, times and the interval stage of ESWT were same in each group. The statistic of every part of this study was made, precisely and felicitously.Results: First, the mitotic activity of periosteal osteoblasts increased significantly, compared with a control group (P<0.05) after 1 week, 2 weeks and 4 week when experemental group being performed to ESW, then the periosteal ossification effect was intensified. ESW accelerated cellular proliferation and differentiation obviously, and the form and structure of the periosteal osteoblast and hMSCs changed, cytochemical staining results show that the expression of alkaline phosphatese, osteocalcin and collagen I improved with the interval stage of ESW, andthe difference is obvious. Second, 69 cases of ANFH were followed up for 1 year after ESWT. The necrosis range occupied were measured by MRI, and averaged from 49.9%(before ESWT), to 19.6%(3 months), to 6.8%(6 months), and to 5.6%(12 months) respectively. The Harris hip score average from 47.2 points before ESWT, to 77.9 after ESWT for 3 months, to 90.2 points after ESWT for 6 months, and to 92.2 points after ESWT for 12 months respectively, the difference was significant between ESWT group and self-control group(P<0.01). No collapse of the femoral
方法:ESW干预体外培养的骨膜组织及成体骨髓间充质干细胞后,分别用骨膜组织H~3-TdR掺入放射性自显影、透射电镜及相差显微镜观察,并用MTT、流式细胞计数及免疫组化染色等方法检测ESW干预前后骨膜成骨细胞H~3-TdR标记率、细胞超微结构的变化,以及ESW对hMSCs增殖周期和分泌碱性磷酸酶、骨钙素、Ⅰ型胶原等的影响。本研究还将能量密度为0.16-0.20mJ/mm~2的体外冲击波用于治疗69例Ⅰ～Ⅲ期股骨头缺血性坏死患者,然后分别于治疗前、治疗后3月、6月及12月行双侧髋关节X线、MRI检查及髋关节Harris评分。此外,采取随机分组的方法比较超声定位ESWT与痛点定位ESWT治疗肩峰下滑囊炎(65例)及肱二头肌长头腱炎(86例)的疗效差异。对上述各部分实验数据采用相应合理的统计学分析。
结果:(1) ESW干预后1、2周实验侧骨膜成骨细胞H~3-TdR标
Objective: In order to reveal the mechanism of Extracorporeal Shock Wave therapy (ESWT) in accelerating osteogenesis, DNA metabolish change in periosteal osteaoblast and human mesenchymal stem cells (hMSCs) was studied with multi-approach. Meanwhile, ESWT was applied to the treatment of avascular necrosis of the femoral head (ANFH) in adults. Its curative effect and application was discussed again. The effect of ESWT of ultrasonic orientation was also compared with the ESWT of pain-point orientation for subacromial bursitis (SB) and inflammation of long-tendon of biceps muscle (ILBM) in this study. Methods: After the periosteal osteaoblast and hMSCs was stimulated by Extracorporeal Shock Wave (ESW) in vitro, their mitotic activity, cellular ultrastructue, metabolish change, proliferation and differentiation was observed and evaluated by autoradiography, transmission electron micrograph, flow cytometer, cytochemical staining for osteocalcin and collagen I expression, the Alkaline phosphatese activity. ESWT with the energy density of 0.16~0.20mJ/mm~2 was applied to the treatment to ANFH of 69 patients, and its curative effects were evaluated by the MRI, X-ray, and the Harris hip score, before ESWT, and after ESWT for 3, 6, 12 months. Another group of patients, including 65 cases of SB and 86 cases of ILBM also accepted ESWT with the same density, but they were
randomly divided into two groups, ESWT by ultrasonic orientation andESWT resorted to tenderness point and local anatomy symbol (control group). The energy density, times and the interval stage of ESWT were same in each group. The statistic of every part of this study was made, precisely and felicitously.Results: First, the mitotic activity of periosteal osteoblasts increased significantly, compared with a control group (P<0.05) after 1 week, 2 weeks and 4 week when experemental group being performed to ESW, then the periosteal ossification effect was intensified. ESW accelerated cellular proliferation and differentiation obviously, and the form and structure of the periosteal osteoblast and hMSCs changed, cytochemical staining results show that the expression of alkaline phosphatese, osteocalcin and collagen I improved with the interval stage of ESW, andthe difference is obvious. Second, 69 cases of ANFH were followed up for 1 year after ESWT. The necrosis range occupied were measured by MRI, and averaged from 49.9%(before ESWT), to 19.6%(3 months), to 6.8%(6 months), and to 5.6%(12 months) respectively. The Harris hip score average from 47.2 points before ESWT, to 77.9 after ESWT for 3 months, to 90.2 points after ESWT for 6 months, and to 92.2 points after ESWT for 12 months respectively, the difference was significant between ESWT group and self-control group(P<0.01). No collapse of the femoral
引文
1.邢更彦,刘玉祥,姚建祥,等.应用体外冲击波技术治疗肩部软组织慢性损伤疾病.中国疗养医学杂志,1996,5(3):18.
2.邢更彦,姚建祥,刘玉祥,等.应用体外冲击波碎石技术治疗肱骨外上髁炎.中华理疗杂志,1995,18(4):235.
3.邢更彦.骨膜环切促进长骨纵向生长的放射性自显影研究.武警医学,1996,7(4):192.
4. Haupt G. Use of extracorporeal shock waves in the treatment of pseudarthrosis, tendinopathy and other orthopaedic diseases. J Urology, 1997, 158: 4～11.
5.邢更彦,井茹芳,刘树茂,等.体外冲击波对骨膜组织骨不连及骨折延迟愈合的影响.中华理疗杂志,1998,21(6):331—333.
6. Rompe JD, Zoellnen J, Nafe B. Shock wave therapy versus conventional surgery in the shoulder. Clin Orthop, 2001, 387: 72～82.
7. Ludwig J, Lauber S, Lauer HJ, et al. High energy Extracorporeal shock wave therapy of femoral head necrosis in adults. Clin Orthop, 2001, 387:119～126.
8.邢更彦,杨传铎,武化云,等.体外冲击波治疗成人股骨头缺血性坏死的初步研究.中国临床康复杂志,2002,6:3009～3010.
9.邢更彦,白晓东,杜明奎,等.体外冲击波治疗成人股骨头缺血性坏死的疗效观察.中华物理医学与康复杂志,2003,25:472～474.
10. Uslu MM, Bozdogan O, Guney S, et al. The effect of extracorporeal shock wave treatment (ESWT) on bone defects. An experimental study. Bull Hosp Jt Dis, 1999, 58(2): 114-118.
11. Schaden W, Fischer A, Sailer A. Extracorporeal shock wave therapy of nonunion or delayed osseous union. Clin Orthop, 2001, 387: 90～94.
12. Wang FS, Yang KD, Chen RF, et al. Extracorporeal shock wave promotes growth and differentiation of bone-marrow stromal cells towards osteoprogenitors associated with induction of TGF-betal. J bone Joint Surg Br, 2002, 84: 457-461.
13. Chen Y J, Wurtz T, Wang C J, et al. Recruitment of mesenchymal stem cells and expression of TGF-betal and VEGF in the early stage of shock wave-promoted bone regeneration of segmental defect in rats. J Orthop Res, 2004, 22: 526-534.
14. Wang FS, Wang CJ, Sheen-Chen SM, et al. Superoxide mediates shock wave induction of ERK-dependent osteogenic transcription factor(CBFA1)and mesenehymal cell differentiation toward osteoprogenitors. J Biol Chem, 2002, 277:10931-10937.
15. Wang FS, Wang C J, Chen Y J, et al. Ras induction of superoxide activates ERK dependent angiogenic transcription factor HIF-1 alpha and VEGF-A expression in shock wave-stimulated osteoblasts. J Biol Chem, 2004, 279: 10331-10337.
16. Wang FS, Wang C J, Huang H J, et al. Physical shock wave mediates membrane
2.邢更彦,姚建祥,刘玉祥,等.应用体外冲击波碎石技术治疗肱骨外上髁炎.中华理疗杂志,1995,18(4):235.
3.邢更彦.骨膜环切促进长骨纵向生长的放射性自显影研究.武警医学,1996,7(4):192.
4. Haupt G. Use of extracorporeal shock waves in the treatment of pseudarthrosis, tendinopathy and other orthopaedic diseases. J Urology, 1997, 158: 4～11.
5.邢更彦,井茹芳,刘树茂,等.体外冲击波对骨膜组织骨不连及骨折延迟愈合的影响.中华理疗杂志,1998,21(6):331—333.
6. Rompe JD, Zoellnen J, Nafe B. Shock wave therapy versus conventional surgery in the shoulder. Clin Orthop, 2001, 387: 72～82.
7. Ludwig J, Lauber S, Lauer HJ, et al. High energy Extracorporeal shock wave therapy of femoral head necrosis in adults. Clin Orthop, 2001, 387:119～126.
8.邢更彦,杨传铎,武化云,等.体外冲击波治疗成人股骨头缺血性坏死的初步研究.中国临床康复杂志,2002,6:3009～3010.
9.邢更彦,白晓东,杜明奎,等.体外冲击波治疗成人股骨头缺血性坏死的疗效观察.中华物理医学与康复杂志,2003,25:472～474.
10. Uslu MM, Bozdogan O, Guney S, et al. The effect of extracorporeal shock wave treatment (ESWT) on bone defects. An experimental study. Bull Hosp Jt Dis, 1999, 58(2): 114-118.
11. Schaden W, Fischer A, Sailer A. Extracorporeal shock wave therapy of nonunion or delayed osseous union. Clin Orthop, 2001, 387: 90～94.
12. Wang FS, Yang KD, Chen RF, et al. Extracorporeal shock wave promotes growth and differentiation of bone-marrow stromal cells towards osteoprogenitors associated with induction of TGF-betal. J bone Joint Surg Br, 2002, 84: 457-461.
13. Chen Y J, Wurtz T, Wang C J, et al. Recruitment of mesenchymal stem cells and expression of TGF-betal and VEGF in the early stage of shock wave-promoted bone regeneration of segmental defect in rats. J Orthop Res, 2004, 22: 526-534.
14. Wang FS, Wang CJ, Sheen-Chen SM, et al. Superoxide mediates shock wave induction of ERK-dependent osteogenic transcription factor(CBFA1)and mesenehymal cell differentiation toward osteoprogenitors. J Biol Chem, 2002, 277:10931-10937.
15. Wang FS, Wang C J, Chen Y J, et al. Ras induction of superoxide activates ERK dependent angiogenic transcription factor HIF-1 alpha and VEGF-A expression in shock wave-stimulated osteoblasts. J Biol Chem, 2004, 279: 10331-10337.
16. Wang FS, Wang C J, Huang H J, et al. Physical shock wave mediates membrane