早期膝骨性关节炎模型大鼠的建立与验证
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摘要
背景:木瓜蛋白酶诱导膝关节骨性关节炎与其他化学诱导试剂相比优势在于对胶原蛋白和软骨细胞没有直接影响。目的:通过软骨的表面形态、放射学和组织病理学进一步验证木瓜蛋白酶诱导大鼠早期膝关节骨性关节炎模型的有效性。方法:20只成年雄性Wistar大鼠,由南京市江宁区青龙山动物繁殖场提供,实验方案经南京市六合区人民医院医学伦理委员会批准。将大鼠随机分成对照组和实验组:分别于实验开始后第1,4,7天,给予对照组右膝关节腔内注射4%生理盐水;实验组右膝关节腔内注射4%木瓜蛋白酶。实验开始后第10天观察关节软骨形态,行X射线、三维CT检查;之后麻醉下处死大鼠即刻进行组织病理学检查,并使用Mankins和OARSI评分验证结果。结果与结论:①与对照组相比,实验组大鼠明显跛行,注射侧膝关节肿胀明显,后肢不愿着地;放射学检查可见明显关节间隙变窄,三维CT关节间隙值变小[对照组(0.680 0±0.016 3) mm,实验组(0.558 0±0.033 9) mm,P<0.05];②实验组右膝关节组织显微镜下观察可见早期的炎症特征;2组病理学Mankins和OARSI评分差异均有显著性意义(P <0.05);③结果说明,大鼠右膝关节腔内注射木瓜蛋白酶诱发的早期膝骨性关节炎可用于相关实验研究,因为它提供了关节运动、触觉异常性疼痛、进行性放射性变性和关节软骨微观炎症的可测量变化,代表了骨关节炎评估,为研究早期膝关节骨性关节炎的发病机制提供依据。
BACKGROUND: Papain inducing knee osteoarthritis has no direct effect on collagen and chondrocyte compared with other chemical inducers.OBJECTIVE: To evaluate the effectiveness of early osteoarthritis model of knee joint in rats by the surface morphology, radiology and histopathology of cartilage.METHODS: Twenty adult male Wistar rats were provided by Qinglongshan Animal Breeding Farm of Jiangning District, Nanjing, and the study was approved by the Ethics Committee of Liuhe Hospital Affiliated to Yangzhou University Medical College. The rats were randomly divided into control and experimental groups. At 1, 4 and 7 days, 4% normal saline and 4% papain were injected into the rat right knee joint cavity in the control and experimental groups, respectively. Rats were killed at 10 days. The articular cartilage morphology,X-ray and three-dimensional CT images were observed before the execution. Histopathological examination was performed immediately after the execution, and the results were verified by Mankins and OARSI scores.RESULTS AND CONCLUSION:(1) Compared with the control group, in the experimental group, the limp was obvious, the knee joint swelling on the injection side was obvious, and the hind limbs were unwilling to land. Radiological examination showed that the joint space narrowed obviously, and the value of three-dimensional CT joint space was significantly decreased [(0.680 0±0.016 3)vs.(0.558 0±0.033 9) mm, P < 0.05].(2) Microscopic examination showed early stage inflammation characteristics at the right knee joint in the experimental group. There were significant differences in the Mankins and OARSI scores(P < 0.05).(3) In summary, early knee osteoarthritis induced by intra-articular injection of papain into the right knee joint of rats can be used in relevant clinical studies, because it provides measurable changes in joint motion, tactile abnormal pain, progressive radiodegeneration and micro-inflammation of articular cartilage, representing the evaluation of osteoarthritis, and providing a basis for studying the pathogenesis of early knee osteoarthritis.
BACKGROUND: Papain inducing knee osteoarthritis has no direct effect on collagen and chondrocyte compared with other chemical inducers.OBJECTIVE: To evaluate the effectiveness of early osteoarthritis model of knee joint in rats by the surface morphology, radiology and histopathology of cartilage.METHODS: Twenty adult male Wistar rats were provided by Qinglongshan Animal Breeding Farm of Jiangning District, Nanjing, and the study was approved by the Ethics Committee of Liuhe Hospital Affiliated to Yangzhou University Medical College. The rats were randomly divided into control and experimental groups. At 1, 4 and 7 days, 4% normal saline and 4% papain were injected into the rat right knee joint cavity in the control and experimental groups, respectively. Rats were killed at 10 days. The articular cartilage morphology,X-ray and three-dimensional CT images were observed before the execution. Histopathological examination was performed immediately after the execution, and the results were verified by Mankins and OARSI scores.RESULTS AND CONCLUSION:(1) Compared with the control group, in the experimental group, the limp was obvious, the knee joint swelling on the injection side was obvious, and the hind limbs were unwilling to land. Radiological examination showed that the joint space narrowed obviously, and the value of three-dimensional CT joint space was significantly decreased [(0.680 0±0.016 3)vs.(0.558 0±0.033 9) mm, P < 0.05].(2) Microscopic examination showed early stage inflammation characteristics at the right knee joint in the experimental group. There were significant differences in the Mankins and OARSI scores(P < 0.05).(3) In summary, early knee osteoarthritis induced by intra-articular injection of papain into the right knee joint of rats can be used in relevant clinical studies, because it provides measurable changes in joint motion, tactile abnormal pain, progressive radiodegeneration and micro-inflammation of articular cartilage, representing the evaluation of osteoarthritis, and providing a basis for studying the pathogenesis of early knee osteoarthritis.
引文
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[15]Bendele AM.Animal models of osteoarthritis.J Musculoskelet Neuronal Interact.2001;1(4):363-376.
[16]Poole R,Blake S,Buschmann M,et al.Recommendations for the use of preclinical models in the study and treatment of osteoarthritis.Osteoarthritis Cartilage.2010;18,Supplement3:S10-16.
[17]McCoy AM.Animal models of osteoarthritis:comparisons and key considerations.Vet Pathol.2015;52(5):803-818.
[18]Jimenez PA,Glasson SS,Trubetskoy OV,et al.Spontaneous osteoarthritis in Dunkin Hartley guinea pigs:histologic,radiologic,and biochemical changes.Lab Anim Sci.1997;47(6):598-601.
[19]Glyn-Jones S,Palmer AJR,Agricola R,et al.Osteoarthritis.Lancet.2015;386(9991):376-387.
[20]Havdrup T,Henricson A,Telhag H.Papain-induced mitosis of chondrocytes in adult joint cartilage:an experimental study in full-grown rabbits.Acta Orthop Scand.1982;53:119-124.
[21]Pritzker KP.Animal models for osteoarthritis:processes,problems and prospects.Ann Rheum Dis.1994;53(6):406-420.
[22]Kikuchi T,Sakuta T,Yamaguchi T.Intra-articular injection of collagenase induces experimental osteoarthritis in mature rabbits.Osteoarthr Cartil.1998;6:177-186.
[23]Khan HM,Ashraf M,Hashmi AS,et al.Papain induced progressive degenerative changes in articular cartilage of rat femorotibial joint and its histopathological grading.J Anim Plant Sci.2013;23(2):350-358.
[24]Braun HJ,Gold GE.Diagnosis of osteoarthritis:imaging.Bone.2012;51(2):278-288.
[25]Kraus VB,Blanco FJ,Englund M,et al.Call for standardized definitions of osteoarthritis and risk stratification for clinical trials and clinical use.Osteoarthritis Cartilage.2015;23(8):1233-1241.
[26]Ferrazzo KL,Osório LB,Ferrazzo VA.CT images of a severe TMJ osteoarthritis and differential diagnosis with other joint disorders.Case Rep Dentistry.2013;2013:5.
[27]Piscaer TM,Waarsing JH,Kops N,et al.In vivo imaging of cartilage degeneration usingμCT-arthrography.Osteoarthritis Cartilage.2008;16(9):1011-1017.
[28]谢希,高洁生.骨关节炎动物模型研究进展[J].医学综述,2005,11(1):67-69.
[29]Park J,Lee J,Kim KI,A Pathophysiological Validation of Collagenase II-Induced Biochemical Osteoarthritis Animal Model in Rabbit.Tissue Eng Regen Med.2018;15(4):437-444.
[30]Kuyinu EL,Narayanan G,Nair LS,et al.Animal models of osteoarthritis:classification,update,and measurement of outcomes.J Orthop Surg Res.2016;11:19.
[31]Murakami K,Nakagawa H,Nishimura K,et al.Changes in peptidergic fiber density in the synovium of mice with collagenase-induced acute arthritis.Can J Physiol Pharmacol.2015;93(6):435-441.
[32]Kumagai K,Suzuki S,Kanri Y,et al.Spontaneously developed osteoarthritis in the temporomandibular joint in STR/ort mice.Biomed Rep.2015;3(4):453-456.
[33]Staines KA,Poulet B,Wentworth DN,et al.The STR/ort mouse model of spontaneous osteoarthritis-an update.Osteoarthritis Cartilage.2017;25(6):802-808.
[34]Motomura H,Seki S,Shiozawa S,et al.A selective c-Fos/AP-1inhibitor prevents cartilage destruction and subsequent osteophyte formation.Biochem Biophys Res Commun.2018;497(2):756-761.
[35]Morse A,McDonald MM,Kelly NH,et al.Mechanical load increases in bone formation via a sclerostin-independent pathway.J Bone Miner Res.2014;29(11):2456-2467.
[36]Christiansen BA,Guilak F,Lockwood KA,et al.Non-invasive mouse models of post-traumatic osteoarthritis.Osteoarthritis Cartilage.2015;23(10):1627-1638.
[2]Chen N,Wang J,Mucelli A,et al.Electroacupuncture is beneficial for knee osteoarthritis:the evidence from meta-analysis of randomized controlled trials.Am J Chin Med.2017;45:965-985.
[3]吴清洪,杨朝湘,邹华章,等.猪膝关节早期骨性关节炎模型的建立[J].动物医学进展,2018,39(6):33-37.
[4]Mankin HJ,Dorfman H,Lippillo L,et al.Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips.J Bone joint Surg(Am).1971;53(3):523-537.
[5]Pritzker KP,Gay S,Jimenez SA,et al.Osteoarthritis cartilage histopathology:grading and staging.Osteoarthritis Cartilage.2006;14(10):13-29.
[6]Rutgers M,van Pelt MJP,Dhert WJA,et al.Evaluation of histological scoring systems for tissue-engineered,repaired and osteoarthritic cartilage.Osteoarthritis Cartilage.2010;18(1):12-23.
[7]Aigner T,Cook JL,Gerwin N,et al.Histopathology atlas of animal model systems-overview of guiding principles.Osteoarthritis Cartilage.2010;18,Supplement 3:S2-6.
[8]苏睿.膝关节骨性关节炎治疗进展研究[J].中医临床研究,2016,8(2):145-146.
[9]Karsdal MA,Christiansen C,Ladel C,et al.Osteoarthritis-a case for personalized health care?Osteoarthritis Cartilage.2014;22(1):7-16.
[10]Matthews GL.Disease modification:promising targets and impediments to success.Rheum Dis Clin N Am.2013;39(1):177-187.
[11]许蓓,许进.骨关节炎发病机制及治疗进展[J].浙江医学,2017,39(21):1833-1835.
[12]尹宏,俞宁,钱卫庆.奇正消痛贴治疗膝骨性关节炎临床疗效观察[J].白求恩军医学院学报,2010,12(6):8.
[13]中华医学会骨科学分会关节外科学组.骨关节炎诊疗指南(2018年版)[J].中华骨科杂志,2018,38(12):705-715.
[14]Lampropoulou-Adamidou K,Lelovas P,Karadimas EV,et al.Useful animal models for the research of osteoarthritis.Eur JOrthop Surg Traumatol.2014;24(3):263-271.
[15]Bendele AM.Animal models of osteoarthritis.J Musculoskelet Neuronal Interact.2001;1(4):363-376.
[16]Poole R,Blake S,Buschmann M,et al.Recommendations for the use of preclinical models in the study and treatment of osteoarthritis.Osteoarthritis Cartilage.2010;18,Supplement3:S10-16.
[17]McCoy AM.Animal models of osteoarthritis:comparisons and key considerations.Vet Pathol.2015;52(5):803-818.
[18]Jimenez PA,Glasson SS,Trubetskoy OV,et al.Spontaneous osteoarthritis in Dunkin Hartley guinea pigs:histologic,radiologic,and biochemical changes.Lab Anim Sci.1997;47(6):598-601.
[19]Glyn-Jones S,Palmer AJR,Agricola R,et al.Osteoarthritis.Lancet.2015;386(9991):376-387.
[20]Havdrup T,Henricson A,Telhag H.Papain-induced mitosis of chondrocytes in adult joint cartilage:an experimental study in full-grown rabbits.Acta Orthop Scand.1982;53:119-124.
[21]Pritzker KP.Animal models for osteoarthritis:processes,problems and prospects.Ann Rheum Dis.1994;53(6):406-420.
[22]Kikuchi T,Sakuta T,Yamaguchi T.Intra-articular injection of collagenase induces experimental osteoarthritis in mature rabbits.Osteoarthr Cartil.1998;6:177-186.
[23]Khan HM,Ashraf M,Hashmi AS,et al.Papain induced progressive degenerative changes in articular cartilage of rat femorotibial joint and its histopathological grading.J Anim Plant Sci.2013;23(2):350-358.
[24]Braun HJ,Gold GE.Diagnosis of osteoarthritis:imaging.Bone.2012;51(2):278-288.
[25]Kraus VB,Blanco FJ,Englund M,et al.Call for standardized definitions of osteoarthritis and risk stratification for clinical trials and clinical use.Osteoarthritis Cartilage.2015;23(8):1233-1241.
[26]Ferrazzo KL,Osório LB,Ferrazzo VA.CT images of a severe TMJ osteoarthritis and differential diagnosis with other joint disorders.Case Rep Dentistry.2013;2013:5.
[27]Piscaer TM,Waarsing JH,Kops N,et al.In vivo imaging of cartilage degeneration usingμCT-arthrography.Osteoarthritis Cartilage.2008;16(9):1011-1017.
[28]谢希,高洁生.骨关节炎动物模型研究进展[J].医学综述,2005,11(1):67-69.
[29]Park J,Lee J,Kim KI,A Pathophysiological Validation of Collagenase II-Induced Biochemical Osteoarthritis Animal Model in Rabbit.Tissue Eng Regen Med.2018;15(4):437-444.
[30]Kuyinu EL,Narayanan G,Nair LS,et al.Animal models of osteoarthritis:classification,update,and measurement of outcomes.J Orthop Surg Res.2016;11:19.
[31]Murakami K,Nakagawa H,Nishimura K,et al.Changes in peptidergic fiber density in the synovium of mice with collagenase-induced acute arthritis.Can J Physiol Pharmacol.2015;93(6):435-441.
[32]Kumagai K,Suzuki S,Kanri Y,et al.Spontaneously developed osteoarthritis in the temporomandibular joint in STR/ort mice.Biomed Rep.2015;3(4):453-456.
[33]Staines KA,Poulet B,Wentworth DN,et al.The STR/ort mouse model of spontaneous osteoarthritis-an update.Osteoarthritis Cartilage.2017;25(6):802-808.
[34]Motomura H,Seki S,Shiozawa S,et al.A selective c-Fos/AP-1inhibitor prevents cartilage destruction and subsequent osteophyte formation.Biochem Biophys Res Commun.2018;497(2):756-761.
[35]Morse A,McDonald MM,Kelly NH,et al.Mechanical load increases in bone formation via a sclerostin-independent pathway.J Bone Miner Res.2014;29(11):2456-2467.
[36]Christiansen BA,Guilak F,Lockwood KA,et al.Non-invasive mouse models of post-traumatic osteoarthritis.Osteoarthritis Cartilage.2015;23(10):1627-1638.