人骨骼肌活检手术使用修改伯格斯坦技术
Summary
The purpose of this video is to present the modified Bergström skeletal muscle biopsy technique on human subjects.
Abstract
经皮活检技术使研究人员和临床医生收集的骨骼肌肉组织样本。该技术是安全的和高度有效的。这部影片描述了使用修改后的伯格斯坦针，以获得人类受试者的股外侧肌肉组织样本经皮活检技术。该BERGSTROM针包括一个外套管具有在前端侧的小开口（“窗口”）和一个内套管针带切削刃的前端。在局部麻醉和无菌条件下，将针通过皮肤，皮下组织和筋膜切开前进到骨骼肌。接着，抽吸施加到内套管针，所述外套管针被拉回，骨骼肌组织被吸入到所述外套管被抽吸的窗口，并且所述内套管针迅速闭合，从而切断或削波的骨骼肌组织样品。针旋转90°60，另有裁切。这个过程可以重复3次以上。此多刀切割技术通常产生100-200毫克或更多的健康受试者的样品与可以做立即之前，期间和运动或其他介入的回合之后。随着切口部位的后活检敷料，受试者通常恢复日常活动的时候了，并能充分参与身体活动中的48-72小时。受试者应避免重阻力锻炼48小时通过切口在筋膜，减少肌肉症的风险。
Introduction
经皮或“半开放”活检技术被用来获得从人患者和研究受试者的骨骼肌组织标本。假肥大型（1806年至1875年），相信作为第一个构建针头套管针经皮活检1以获取活体骨骼肌。在20世纪60年代，伯格斯坦推出了经皮活检针类似由杜氏2-4描述。二十年后，Evans 等人的 5通过经由BERGSTROM针的切割套管针施加抽吸改性的技术。这种修饰可以增强组织3的产率，以5倍6,7和被用在临床和生物医学研究的设置。这种技术已经并将继续培养肌病的诊断和我们的骨骼肌的结构和功能的理解。
经皮肌活检技术是straightforward。如果做得正确，严格遵守无菌技术操作，相关的风险是最小的。肌肉活检程序被视为一种研究工具，用于研究项目。一些学术机构允许经过培训的教师与研究人员博士，与医生的直接监督，以获得肌肉活检等人要求委员会认证的医师来执行的技术。在过去的13年8月15日在亚利桑那州立大学活检团队已经成功完成超过1,600肌肉活检。该视频的目的是描述了改性BERGSTROM针经皮肌活检技术5获得骨骼肌组织样品从人类受试者的股外侧。
Protocol
所描述的骨骼肌过程遵循阿巴拉契亚州立大学伦理审查委员会的指导方针。 注：该小组由操作者（博士或MD训练的活检技术），和至少一个，但理想的是，三个或更多个技术员。操作员负责进行及监督骨骼肌活检过程的各个方面。一技术员（技术员＃1）是立即用于辅助操作者肌肉提取的所有方面负责。这包括在“非无菌手”和施加抽吸，用无菌注射器。第二技师（技师＃2）?…
Representative Results
如上文所描述的肌肉活检程序允许研究者快速一致收集骨骼肌组织样品。典型的产率在健康，运动受试者是200毫克或更多的单次通过以3-4的剪辑。该过程需要15-20分钟，其中大部分是花在准备在切口。在运动为基础的研究，肌肉样品往往采取前后运动，以恢复过程中收集的一个或两个样本。利用这一设计，在运动后的毛发（对侧）大腿被限幅和切口位点是预先标有一个细点永久性标记，而运动前?…
Discussion
这部影片提供了在亚利桑那州立大学人类表现实验室使用的骨骼肌活检过程的步骤一步的总结。此过程中，小的修改，已被用于在过去的13年，收集约1600肌肉活检。肌肉活检样本中的运动营养基础研究提供了重要的数据，从而导致重要的研究发现。
有需要注意进行改性BERGSTROM骨骼肌活检技术时的许多重要的技术。首先，活检队必须在程序的各个阶段保持严格无菌操作。当定?…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
没有资金申报。
Materials
| Name of Material / Equipment | Company | Catalog Number | Comments / Description |
| Bergström biopsy needle, 5 x 100mm | Stille Surgical Inc. | 119-29187-50 | 1 ea |
| Durasorb blue pad | Fisher | 22-031-340 | case of 300 |
| Prep Razor Disposable | Moore Medical | 89760 | 1 ea |
| Shave Cream | Moore Medical | 92869 | 1 can |
| Alcohol Swab | Moore Medical | 98721 | box/200 |
| Povidone-iodine Swabsticks (3 pack) | Moore Medical | 90691 | pack of 50 |
| Face masks | Fisher | 19-039-690 | case of 50 |
| Sterile Gloves; sz 6.5 | Moore Medical | 68128 | box of 50 pairs |
| Sterile Gloves; sz 7.0 | Moore Medical | 68129 | box of 50 pairs |
| Sterile Gloves; sz 8.5 | Moore Medical | 68132 | box of 50 pairs |
| Fenestrated Towel Drape | Moore Medical | 92712 | case of 50 |
| Lidocaine (1% HCl, w/o Epi | Dealmed | 427902 | 30cc vial |
| Ethyl Chloride spray | Dealmed | 386020 | 3.5 fl. Oz. bottle |
| 5 mL syringe w/ 21G needle | Fisher | 14-827-48 | case of 100 |
| 25G 5/8" needle | Fisher | 14-826AA | case of 100 |
| 25G 1 1/2" needle | Fisher | 14-826-49 | case of 100 |
| Single-Use Scalpels w/ #11 blade | Fisher | 0-8927-5B | box of 20 |
| 60 mL Syringe | Fisher | 13-689-8 | case of 40 |
| ~30 inch extension tubing | Fisher | 14-169-7A | roll of 50ft |
| Metal stopcock | Fisher | 01-290-38 | each |
| Polypropylene tubing connector w/ tapered ends, 3.2 to 5.5 mm | Fisher | 15-315-28A | case of 100 |
| Yellow pipette tips (1-200 µL) | Fisher | 02-681-2 | cut off ~15-18 mm of the tip |
| Topical surgical adhesive | Medline | MSC091076 | case of 36 tubes |
| Topical surgical adhesive | Medline | MSC091076Z | box of 6 tubes |
| Butterfly stitches (adhesive strips) 1/2" x 4" (6 per pack) | Moore Medical | 93050 | box/300 |
| Bandadge, Strip, 3/4 x 3 in. | Fisher | 19-027-202 | pack of 100 |
| Adhesive bandage (5m rolls) | Fisher | 19-027-768 | case of 36 |
| 1" paper surgical tape | Fisher | 19-027-761 | case of 12 |
| Sutures (4-0 reverse cutting, FS2) | Moore Medical | 82926 | box of 36 |
| 4 x 4 sterile gauze pads | Moore Medical | 66949 | pkg/50 |
| 2 x 2 non-sterile gauze pads | Moore Medical | 37336 | pkg/200 |
| 3 x 4 Non-Adherent Pads | Moore Medical | 74389 | bx/100 |
| Autoclave bags 3.5 x 9 | Moore Medical | 40390 | box of 200 |
| Autoclave bags 5.25 x 10 | Moore Medical | 40391 | box of 200 |
| Autoclave indicator tape | Moore Medical | 31608 | box of 250 |
| Lab Coats—small | Fisher | 23-900-520A | 10/pk |
| Lab Coats—medium | Fisher | 23-900-520B | 10/pk |
| Lab Coats—large | Fisher | 23-900-520C | 10/pk |
| Germicidal Disposable Cloth Wipes | Moore Medical | 62879 | Can/160 |
Riferimenti
- Duchene, G. B. A. Recherches sur la paralysie musculaire pseudohypertrophique ou paralysie myo-sclerosique. Arch Gen Med. 11 (30), (1868).
- Bergstrom, J. Muscle electrolytes in man. Determined by neutron activation analysis on needle biopsy specimens. Scand J Clin Lab Invest (England). 14, (1962).
- Bergstrom, J. Percutaneous needle biopsy of skeletal muscle in physiological and clinical research). Scand J Clin Lab Invest. 35, 609-616 (1975).
- Bergstrom, J., Hultman, E. A study of the glycogen metabolism during exercise in man. Scand J Clin Lab Invest. 19, 218-228 (1967).
- Evans, W. J., Phinney, S. D., Young, V. R. Suction applied to a muscle biopsy maximizes sample size. Med Sci Sports Exerc. 14, 101-102 (1982).
- Hennessey, J. V., Chromiak, J. A., Della Ventura, S., Guertin, J., MacLean, D. B. Increase in percutaneous muscle biopsy yield with a suction-enhancement technique. J Appl Phys. 82, 1739-1742 (1997).
- Tarnopolsky, M. A., Pearce, E., Smith, K., Lach, B. Suction-modified Bergstrom muscle biopsy technique: experience with 13,500 procedures. Muscl., & Nerve. 43, 717-725 (2011).
- Nieman, D. C., et al. Carbohydrate ingestion influences skeletal muscle cytokine mRNA and plasma cytokine levels after a 3-h run. J Appl Physiol. 94, 1917-1925 (2003).
- Nieman, D. C., et al. Influence of carbohydrate ingestion on immune changes after 2 h of intensive resistance training. J Appl Physiol. 96, 1292-1298 (2004).
- Utter, A. C., et al. Carbohydrate supplementation and perceived exertion during prolonged running. Med Sci Sports Exerc. 36, 1036-1041 (2004).
- Nieman, D. C., et al. Muscle cytokine mRNA changes after 2.5 h of cycling: influence of carbohydrate. Med Sci Sports Exerc. 37, 1283-1290 (2005).
- Nieman, D. C., et al. Effects of quercetin and EGCG on mitochondrial biogenesis and immunity. Med Sci Sports Exerc. 41, 1467-1475 (2009).
- Nieman, D. C., et al. Quercetin’s Influence on Exercise Performance and Muscle Mitochondrial Biogenesis. Med Sci Sports Exerc. 42, 338-345 (2010).
- Dumke, C. L., et al. Quercetin’s effect on cycling efficiency and substrate utilization. Appl Physiol Nut Metab. 34, 993-1000 (2009).
- Nieman, D. C., et al. Quercetin’s influence on exercise-induced changes in plasma cytokines and muscle and leukocyte cytokine mRNA. J Appl Physiol. 103, 1728-1735 (2007).
- . Guideline for Disinfection and Sterilization in Healthcare Facilities. http://www.cdc.gov/hicpac/Disinfection_Sterilization/toc.html. , (2008).
- Faria, F. E., et al. The onset and duration of mobilization affect the regeneration in the rat muscle. Histol Histopath. 23, 565-571 (2008).
- Carlson, B. M., Shepard, B., Komorowski, T. E. A histological study of local anesthetic-induced muscle degeneration and regeneration in the monkey. J Ortho Res. 8, 485-494 (1990).
- Yagiela, J. A., Benoit, P. W., Buoncristiani, R. D., Peters, M. P., Fort, N. F. Comparison of myotoxic effects of lidocaine with epinephrine in rats and humans. Anesth Analg. 60, 471-480 (1981).
- Coyle, E. F., Coggan, A. R., Hemmert, M. K., Ivy, J. L. Muscle glycogen utilization during prolonged strenuous exercise when fed carbohydrate. J Appl Physiol. 61, 165-172 (1986).
- Gibala, M. J., et al. Brief intense interval exercise activates AMPK and p38 MAPK signaling and increases the expression of PGC-1alpha in human skeletal muscle. J Appl Physiol. 106, 929-934 (1985).
- Phillips, S. M., Tipton, K. D., Ferrando, A. A., Wolfe, R. R. Resistance training reduces the acute exercise-induced increase in muscle protein turnover. Am J Physiol. 276, 118-124 (1999).
- Davis, J. M., Murphy, E. A., Carmichael, M. D., Davis, B. Quercetin increases brain and muscle mitochondrial biogenesis and exercise tolerance. Am J Physiol Regul Integr Comp Physiol. 296, 1071-1077 (2009).
- Vissing, K., Andersen, J. L., Schjerling, P. Are exercise-induced genes induced by exercise. FASEB J. 19, 94-96 (2005).
- Lundby, C., et al. Gene expression in human skeletal muscle: alternative normalization method and effect of repeated biopsies. Eur J Appl Physiol. 95, 351-360 (2005).
- Coyle, E. F., Coggan, A. R., Hemmert, M. K., Ivy, J. L. Muscle glycogen utilization during prolonged strenuous exercise when fed carbohydrate. J Appl Physiol. 61, 165-172 (1985).
Citazione di questo articolo
Shanely, R. A., Zwetsloot, K. A., Triplett, N. T., Meaney, M. P., Farris, G. E., Nieman, D. C. Human Skeletal Muscle Biopsy Procedures Using the Modified Bergström Technique. J. Vis. Exp. (91), e51812, doi:10.3791/51812 (2014).