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J Zhejiang Univ (Med Sci)  2019, Vol. 48 Issue (5): 517-525    DOI: 10.3785/j.issn.1008-9292.2019.10.09
Mechanical stress promotes cartilage repair in inflammatory environment
YAO Wangxiang1,2(),DAI Hanghao1(),GUI Jianchao1,*()
1. Department of Sports Medicine and Joint Surgery, the Affiliated Nanjing Hospital of Nanjing Medical University, Nanjing 210006, China
2. Department of Orthopedics, Hangzhou First People's Hospital, Hangzhou 310006, China
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Objective: To investigate the effect and mechanism of mechanical stress on cartilage repair in inflammatory environment. Methods: The chondrogenic progenitor cells (CPCs) were isolated from the knee joint cartilage of patients with osteoarthritis (OA) undergoing total knee arthroplasty. The CPCs were cultured and expanded in a 3-D scaffold constructed with alginate. Intermittent hydrostatic pressure (IHP) was applied in a inflammatory environment induced by IL-1β, and Western blot was used to detect the expression of MAPK signaling pathway proteins. Cell proliferation was detected by CCK-8 method, and the expression of related genes like matrix metallo-proteinases 13 (MMP-13) and a disintegrins and metalloproteinase with thrombospondin motif 5 (ADAMTS-5) was detected by real-time RT-PCR. The anterior cruciate ligament of the rats was cut to construct the knee joint OA model, and the appropriate mechanical stress was constructed with external fixation to distract the knee joint in order to observe the repair of the cartilage and to explore its mechanism. Results: Adding 0.01 ng/ml IL-1β in cell culture inhibited the proliferation of CPCs. After IHP application, the expression of MAPK pathway protein was decreased, the mRNA expression of MMP-13 and ADAMTS-5 was reduced. The inhibition of IL-1β on CPCs was counteracted by IHP. Four weeks after the anterior cruciate ligament resected, the articular cartilage degeneration was observed in rats. The Mankin score in the OA treatment (joint distraction) group was lower, and the cartilage repair was better than that of the control group (P < 0.01). Animal experiments found that the suitable mechanical stress reduced the expression of P-p38, MMP-13 and COLL-X, inhibited cartilage cells apoptosis and promoted the repair of OA cartilage. Conclusion: Mechanical stress can promote the proliferation of CPCs, reduce the expression of matrix degrading enzymes, and promote the repair of OA cartilage by inhibiting MAPK signaling pathway.

Key wordsOsteoarthritis/pathology      Pressure      Chondrocytes      Interleukin-1beta      Matrix metalloproteinase 13      Mitogen-activated protein kinases      Cells, cultured      Disease models, animal     
Received: 03 July 2019      Published: 04 January 2020
CLC:  R684.3  
Corresponding Authors: GUI Jianchao     E-mail:;;
Cite this article:

YAO Wangxiang,DAI Hanghao,GUI Jianchao. Mechanical stress promotes cartilage repair in inflammatory environment. J Zhejiang Univ (Med Sci), 2019, 48(5): 517-525.

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目的: 探索在炎性环境中施加机械应力对软骨修复的影响及机制。方法: 提取骨关节炎(OA)患者膝关节软骨中的软骨前体细胞(CPC),培养扩增后在海藻酸构建的支架上三维培养,在IL-1β塑造的炎症环境中施加周期性静水压(IHP),蛋白免疫印迹法检测MAPK信号通路的变化,CCK-8法检测细胞增殖,实时荧光定量PCR检测基质金属蛋白酶13(MMP-13)和解聚素与金属蛋白酶5(ADAMTS-5)基因表达的变化。切断大鼠的前交叉韧带构建膝关节OA模型,通过外固定支架牵开关节腔构建合适的机械应力,观察关节牵引能否促进软骨的修复及其机制。结果: 0.01 ng/mL IL-1β可以抑制CPC增殖,施加IHP后可以减少MAPK信号通路蛋白的表达,减少MMP-13ADAMTS-5基因表达,促进炎性环境下的CPC增殖。切断前交叉韧带4周后,膝关节OA模型成功构建。OA大鼠经关节牵引后软骨的Mankin评分更低(P < 0.01),软骨修复较对照组更佳。软骨标本的免疫组织化学染色发现关节牵引可减少P-p38、MMP-13及X型胶原的表达,减少细胞凋亡,促进OA软骨的修复。结论: 机械应力可以通过抑制MAPK信号通路,促进CPC增殖,减少基质降解酶表达,促进OA软骨修复。

关键词: 骨关节炎/病理学,  压力,  软骨细胞,  白细胞介素1β,  基质金属蛋白酶13,  丝裂原激活蛋白激酶类,  细胞, 培养的,  疾病模型, 动物 
Tab 1 Primers sequences for real-time PCR
Fig 1 Morphology of chondrogenic progenitor cells
Fig 2 Effects of IL-1β and intermittent hydrostatic pressure on proliferation of chondrogenic progenitor cells
Fig 3 Mechanical stress applied to cell-alginate microspheres during one week of pre-culture inhibits IL-1β activation of MAPK signaling pathway
Fig 4 Mechanical stress in IL-1β can reduce the expression of MMP-13 and ADAMTS-5 mRNA in chondrogenic progenitor cells
Fig 5 Gross observation, radiological evaluation, HE and MASSON staining of femur cartilage in osteoarthritis treatment and control groups
Fig 6 The apoptosis of chondrocytes observed by TUNEL staining
Fig 7 Differential expression of P-p38, MMP-13 and COLL-X in the cartilage of osteoarthritis treatment and control groups
[1]   GOLDRING S R , GOLDRING M B . Changes in the osteochondral unit during osteoarthritis: structure, function and cartilage-bone crosstalk[J]. Nat Rev Rheumatol, 2016, 12 (11): 632- 644
doi: 10.1038/nrrheum.2016.148
[2]   ZHEN G , WEN C , JIA X et al. Inhibition of TGF–β signaling in subchondral bone mesenchymal stem cells attenuates osteoarthritis[J]. Nat Med, 2013, 19 (6): 704- 712
doi: 10.1038/nm.3143
[3]   BANNURU R R , OSANI M C , VAYSBROT E E et al. OARSI guidelines for the non-surgical management of knee, hip, and polyarticular osteoarthritis[J]. Osteoarthritis Cartilage, 2019, 27 (11): 1578- 1589
doi: 10.1016/j.joca.2019.06.011
[4]   COLLINS J A , ARBEEVA L , CHUBINSKAYA S et al. Articular chondrocytes isolated from the knee and ankle joints of human tissue donors demonstrate similar redox-regulated MAP kinase and Akt signaling[J]. Osteoarthritis Cartilage, 2019, 27 (4): 703- 711
doi: 10.1016/j.joca.2018.12.010
[5]   PELLETIER J P , FERNANDES J C , BRUNET J et al. In vivo selective inhibition of mitogen-activated protein kinase kinase 1/2 in rabbit experimental osteoarthritis is associated with a reduction in the development of structural changes[J]. Arthritis Rheum, 2003, 48 (6): 1582- 1593
doi: 10.1002/art.11014
[6]   ROBINSON W H , LEPUS C M , WANG Q et al. Low-grade inflammation as a key mediator of the pathogenesis of osteoarthritis[J]. Nat Rev Rheumatol, 2016, 12 (10): 580- 592
doi: 10.1038/nrrheum.2016.136
[7]   KOELLING S , KRUEGEL J , IRMER M et al. Migratory chondrogenic progenitor cells from repair tissue during the later stages of human osteoarthritis[J]. Cell Stem Cell, 2009, 4 (4): 324- 335
doi: 10.1016/j.stem.2009.01.015
[8]   JOOS H , WILDNER A , HOGREFE C et al. Interleukin-1 beta and tumor necrosis factor alpha inhibit migration activity of chondrogenic progenitor cells from non-fibrillated osteoarthritic cartilage[J]. Arthritis Res Ther, 2013, 15 (5): R119
doi: 10.1186/ar4299
[9]   LI Y , ZHOU J , YANG X et al. Intermittent hydrostatic pressure maintains and enhances the chondrogenic differentiation of cartilage progenitor cells cultivated in alginate beads[J]. Dev Growth Differ, 2016, 58 (2): 180- 193
doi: 10.1111/dgd.12261
[10]   SCHIPHOF D , DE KLERK B M , KOESB W et al. Good reliability, questionable validity of 25 different classification criteria of knee osteoarthritis: a systematic appraisal[J]. J Clin Epidemiol, 2008, 61 (12): 1205- 1215
doi: 10.1016/j.jclinepi.2008.04.003
[11]   MCNULTY A L , ROTHFUSZ N E , LEDDY H A et al. Synovial fluid concentrations and relative potency of interleukin-1 alpha and beta in cartilage and meniscus degradation[J]. J Orthop Res, 2013, 31 (7): 1039- 1045
doi: 10.1002/jor.22334
[12]   NATENSTEDT J , KOK A C , DANKELMAN J et al. What quantitative mechanical loading stimulates in vitro cultivation best?[J]. J Exp Orthop, 2015, 2 (1): 15
doi: 10.1186/s40634-015-0029-x
[13]   CHEN Y , SUN Y , PAN X et al. Joint distraction attenuates osteoarthritis by reducing secondary inflammation, cartilage degeneration and subchondral bone aberrant change[J]. Osteoarthritis Cartilage, 2015, 23 (10): 1728- 1735
doi: 10.1016/j.joca.2015.05.018
[14]   FU S , THOMPSON C L , ALI A et al. Mechanical loading inhibits cartilage inflammatory signalling via an HDAC6 and IFT-dependent mechanism regulating primary cilia elongation[J]. Osteoarthritis Cartilage, 2019, 27 (7): 1064- 1074
doi: 10.1016/j.joca.2019.03.003
[15]   VAN VALBURGA A , VAN ROERMUND P M , LAMMENS J et al. Can Ilizarov joint distraction delay the need for an arthrodesis of the ankle? A preliminary report[J]. J Bone Joint Surg Br, 1995, 77 (5): 720- 725
[16]   TABEIAN H, BETTI B F, DOS SANTOS CIRQUEIRA C, et al. IL-1β damages fibrocartilage and upregulates MMP-13 expression infibrochon-drocytes in the condyle of the temporomandibular joint[J]. Int J Mol Sci, 2019, 20(9). pii: E2260.
[17]   ROSENZWEIG D H , QUINN T M , HAGLUND L . Low-frequency high-magnitude mechanical strain of articular chondrocytes activates p38 MAPK and induces phenotypic changes associated with osteoarthritis and pain[J]. Int J Mol Sci, 2014, 15 (8): 14427- 14441
doi: 10.3390/ijms150814427
[18]   CHEN J , YUAN Z , LIU Y et al. Improvement of in vitro three-dimensional cartilage regeneration by a novel hydrostatic pressurebioreactor[J]. Stem Cells Transl Med, 2017, 6 (3): 982- 991
doi: 10.5966/sctm.2016-0118
[19]   HEINEG?RD D , SAXNE T . The role of the cartilage matrix in osteoarthritis[J]. Nat Rev Rheumatol, 2011, 7 (1): 50- 56
[20]   ZHANG Y , PIZZUTE T , LI J et al. sb203580 preconditioning recharges matrix-expanded human adult stem cells forchondrogenesis in an inflammatory environment-a feasible approach for autologous stem cell based osteoarthritic cartilage repair[J]. Biomaterials, 2015, 64 88- 97
doi: 10.1016/j.biomaterials.2015.06.038
[21]   JI B , MA Y , WANG H et al. Activation of the P38/CREB/MMP13 axis is associated with osteoarthritis[J]. Drug Des Devel Ther, 2019, 13 2195- 2204
doi: 10.2147/DDDT.S209626
[22]   WANG Z W , CHEN L , HAO X R et al. Elevated levels of interleukin-1β, interleukin-6, tumor necrosis factor-α and vascular endothelial growth factor in patients with knee articular cartilageinjury[J]. World J Clin Cases, 2019, 7 (11): 1262- 1269
doi: 10.12998/wjcc.v7.i11.1262
[23]   TETSUNAGA T , NISHIDA K , FURUMATSU T et al. Regulation of mechanical stress-induced MMP-13 and ADAMTS-5 expression by RUNX-2 transcriptional factor in SW1353 chondrocyte-like cells[J]. Osteoarthritis Cartilage, 2011, 19 (2): 222- 232
doi: 10.1016/j.joca.2010.11.004
[24]   WANG M , SAMPSON E R , JIN H et al. MMP13 is a critical target gene during the progression of osteoar-thritis[J]. Arthritis Res Ther, 2013, 15 (1): R5
doi: 10.1186/ar4133
[25]   ZHOU B , CHEN D , XU H et al. Proliferation of rabbit chondrocyte and inhibition of IL-1β-induced apoptosis through MEK/ERK signaling by statins[J]. In Vitro Cell Dev Biol Anim, 2017, 53 (2): 124- 131
doi: 10.1007/s11626-016-0086-1
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