Overexpressing Runx2 of BMSCs Improves the Repairment of knee Cartilage Defects

Author(s): Jing Hu, Wen-Zhong Zou, Ling Li, Zheng-shuai Shi, Xiang-Zhong Liu, Han-tao Cai, Ao-fei Yang, Da-ming Sun, Liang-liang Xu, Yi Yang, Zhang-Hua Li*

Journal Name: Current Gene Therapy

Volume 20 , Issue 5 , 2020


  Journal Home
Translate in Chinese
Become EABM
Become Reviewer
Call for Editor

Graphical Abstract:


Abstract:

Background: Recruitment of gene modifying bone marrow mesenchymal stem cells (BMSCs) has been considered an alternative to single-cell injection in articular cartilage repair.

Purpose: This study aimed to investigate whether the effect of runt-related transcription factor 2(Runx2) overexpression bone marrow mesenchymal stem cells in vivo could improve the quality of repaired tissue of a knee cartilage defect in a rabbit model.

Methods: Thirty-two New Zealand rabbits were randomly divided into four groups. The blank group (Con) did not receive anything, the model group (Mo) was administered saline, the simple stem cell group (MSCs) received MSCs injection, and the Runx2 transfection group (R-MSCs) received Runx2 overexpression MSCs injection. After adapting to the environment for a week, a 5 mm diameter cylindrical osteochondral defect was created in the center of the medial femoral condyle. Cell and saline injections were performed in the first and third weeks after surgery. The cartilage repair was evaluated by macroscopically and microscopically at 4 and 8 weeks.

Results: Macroscopically, defects were filled and surfaces were smoother in the MSCs groups than in the Mo group at 4th week. Microscopically, the R-MSCs group showed coloration similar to surrounding normal articular cartilage tissue at 8 weeks in masson trichrome staining. The COL-II, SOX9, and Aggrecan mRNA expressions of MSCs were enhanced at 4 weeks compared with R-MSCs, then the expression reduced at 8 weeks, but was still higher than Mo group level (P<0.05). The western blot examination revealed that the COL-IIand SOX9 expression of MSCs was higher than R-MSCs at 4 weeks, then the expression reduced at 8 weeks, but was still higher than the Mo level (P<0.05). The IL-1β content in the joint fluid also revealed that cartilage repair with R-MSCs was better than that with MSCs at 8 weeks (P<0.05).

Conclusion: The R-MSCs group showed cellular morphology and arrangement similar to surrounding normal articular cartilage tissue, and Runx2 overexpression of MSCs resulted in overall superior cartilage repair as compared with MSCs at 8 weeks.

Keywords: Runx2, knee cartilage repair, bone marrow mesenchymal stem cells, cartilage defects, chondral defects, animal model.

[1]
Rahim S, Rahim F, Shirbandi K, Haghighi BB, Arjmand B. Sports injuries: diagnosis, prevention, stem cell therapy, and medical sport strategy. Adv Exp Med Biol 2019; 1084: 129-44.
[http://dx.doi.org/10.1007/5584_2018_298] [PMID: 30539427]
[2]
Curl WW, Krome J, Gordon ES, Rushing J, Smith BP, Poehling GG. Cartilage injuries: a review of 31,516 knee arthroscopies. Arthroscopy 1997; 13(4): 456-60.
[http://dx.doi.org/10.1016/S0749-8063(97)90124-9] [PMID: 9276052]
[3]
Kuo AC, Rodrigo JJ, Reddi AH, Curtiss S, Grotkopp E, Chiu M. Microfracture and bone morphogenetic protein 7 (BMP-7) synergistically stimulate articular cartilage repair. Osteoarthritis Cartilage 2006; 14(11): 1126-35.
[http://dx.doi.org/10.1016/j.joca.2006.04.004] [PMID: 16765606]
[4]
Guettler JH, Demetropoulos CK, Yang KH, Jurist KA. Osteochondral defects in the human knee: influence of defect size on cartilage rim stress and load redistribution to surrounding cartilage. Am J Sports Med 2004; 32(6): 1451-8.
[http://dx.doi.org/10.1177/0363546504263234] [PMID: 15310570]
[5]
Theodoropoulos J, Dwyer T, Whelan D, Marks P, Hurtig M, Sharma P. Microfracture for knee chondral defects: a survey of surgical practice among Canadian orthopedic surgeons. Knee Surg Sports Traumatol Arthrosc 2012; 20(12): 2430-7.
[http://dx.doi.org/10.1007/s00167-012-1925-6] [PMID: 22362098]
[6]
Matsusue Y, Yamamuro T, Hama H. Arthroscopic multiple osteochondral transplantation to the chondral defect in the knee associated with anterior cruciate ligament disruption. Arthroscopy 1993; 9(3): 318-21.
[http://dx.doi.org/10.1016/S0749-8063(05)80428-1] [PMID: 8323618]
[7]
Zellner J, Grechenig S, Pfeifer CG, et al. Clinical and radiological regeneration of large and deep osteochondral defects of the knee by bone augmentation combined with matrix-guided autologous chondrocyte transplantation. Am J Sports Med 2017; 45(13): 3069-80.
[http://dx.doi.org/10.1177/0363546517717679] [PMID: 28777662]
[8]
Koga H, Shimaya M, Muneta T, et al. Local adherent technique for transplanting mesenchymal stem cells as a potential treatment of cartilage defect. Arthritis Res Ther 2008; 10(4): R84.
[http://dx.doi.org/10.1186/ar2460] [PMID: 18664254]
[9]
Jiang S, Guo W, Tian G, et al. Clinical application status of articular cartilage regeneration techniques: tissue-engineered cartilage brings new hope. Stem Cells Int 2020; 2020: 5690252.
[http://dx.doi.org/10.1155/2020/5690252] [PMID: 32676118]
[10]
Wang J, Zhang W, He GH, Wu B, Chen S. Transfection with CXCR4 potentiates homing of mesenchymal stem cells in vitro and therapy of diabetic retinopathy in vivo. Int J Ophthalmol 2018; 11(5): 766-72.
[PMID: 29862173]
[11]
Wolf D, Wolf AM. Mesenchymal stem cells as cellular immunosuppressants. Lancet 2008; 371(9624): 1553-4.
[http://dx.doi.org/10.1016/S0140-6736(08)60666-2] [PMID: 18468526]
[12]
Chamberlain G, Fox J, Ashton B, Middleton J. Concise review: mesenchymal stem cells: their phenotype, differentiation capacity, immunological features, and potential for homing. Stem Cells 2007; 25(11): 2739-49.
[http://dx.doi.org/10.1634/stemcells.2007-0197] [PMID: 17656645]
[13]
Fernandes TL, Shimomura K, Asperti A, et al. Development of a novel large animal model to evaluate human dental pulp stem cells for articular cartilage treatment. Stem Cell Rev Rep 2018; 14(5): 734-43.
[http://dx.doi.org/10.1007/s12015-018-9820-2] [PMID: 29728886]
[14]
Bianco P, Gehron Robey P. Marrow stromal stem cells. J Clin Invest 2000; 105(12): 1663-8.
[http://dx.doi.org/10.1172/JCI10413] [PMID: 10862779]
[15]
Song SY, Hong J, Go S, et al. Interleukin-4 gene transfection and spheroid formation potentiate therapeutic efficacy of mesenchymal stem cells for osteoarthritis. Adv Healthc Mater 2020; 9(5): e1901612.
[http://dx.doi.org/10.1002/adhm.201901612] [PMID: 31977158]
[16]
Xu J, Li Z, Hou Y, Fang W. Potential mechanisms underlying the Runx2 induced osteogenesis of bone marrow mesenchymal stem cells. Am J Transl Res 2015; 7(12): 2527-35.
[PMID: 26885254]
[17]
Enomoto H, Enomoto-Iwamoto M, Iwamoto M, et al. Cbfa1 is a positive regulatory factor in chondrocyte maturation. J Biol Chem 2000; 275(12): 8695-702.
[http://dx.doi.org/10.1074/jbc.275.12.8695] [PMID: 10722711]
[18]
Chen CG, Thuillier D, Chin EN, Alliston T. Chondrocyte-intrinsic Smad3 represses Runx2-inducible matrix metalloproteinase 13 expression to maintain articular cartilage and prevent osteoarthritis. Arthritis Rheum 2012; 64(10): 3278-89.
[http://dx.doi.org/10.1002/art.34566] [PMID: 22674505]
[19]
Sasaki T, Akagi R, Akatsu Y, et al. The effect of systemic administration of G-CSF on a full-thickness cartilage defect in a rabbit model MSC proliferation as presumed mechanism: G-CSF for cartilage repair. Bone Joint Res 2017; 6(3): 123-31.
[http://dx.doi.org/10.1302/2046-3758.63.BJR-2016-0083] [PMID: 28258115]
[20]
Li ZH, Liao W, Cui XL, et al. Intravenous transplantation of allogeneic bone marrow mesenchymal stem cells and its directional migration to the necrotic femoral head. Int J Med Sci 2011; 8(1): 74-83.
[http://dx.doi.org/10.7150/ijms.8.74] [PMID: 21234272]
[21]
Fathi E, Farahzadi R, Valipour B, Sanaat Z. Cytokines secreted from bone marrow derived mesenchymal stem cells promote apoptosis and change cell cycle distribution of K562 cell line as clinical agent in cell transplantation. PLoS One 2019; 14(4): e0215678.
[http://dx.doi.org/10.1371/journal.pone.0215678] [PMID: 31009502]
[22]
Fathi E, Farahzadi R, Sheikhzadeh N. Immunophenotypic characterization, multi-lineage differentiation and aging of zebrafish heart and liver tissue-derived mesenchymal stem cells as a novel approach in stem cell-based therapy. Tissue Cell 2019; 57: 15-21.
[http://dx.doi.org/10.1016/j.tice.2019.01.006] [PMID: 30947959]
[23]
Li ZH, Liao W, Zhao Q, et al. Effect of Cbfa1 on osteogenic differentiation of mesenchymal stem cells under hypoxia condition. Int J Clin Exp Med 2014; 7(3): 540-8.
[PMID: 24753746]
[24]
Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006; 8(4): 315-7.
[http://dx.doi.org/10.1080/14653240600855905] [PMID: 16923606]
[25]
Rajabzadeh N, Fathi E, Farahzadi R. Stem cell-based regenerative medicine. Stem Cell Investig 2019; 6: 19.
[http://dx.doi.org/10.21037/sci.2019.06.04] [PMID: 31463312]
[26]
Caplan AI. Review: mesenchymal stem cells: cell-based reconstructive therapy in orthopedics. Tissue Eng 2005; 11(7-8): 1198-211.
[http://dx.doi.org/10.1089/ten.2005.11.1198] [PMID: 16144456]
[27]
Gao J, Zhang G, Xu K, et al. Bone marrow mesenchymal stem cells improve bone erosion in collagen-induced arthritis by inhibiting osteoclasia-related factors and differentiating into chondrocytes. Stem Cell Res Ther 2020; 11(1): 171.
[http://dx.doi.org/10.1186/s13287-020-01684-w] [PMID: 32381074]
[28]
Stricker S, Fundele R, Vortkamp A, Mundlos S. Role of Runx genes in chondrocyte differentiation. Dev Biol 2002; 245(1): 95-108.
[http://dx.doi.org/10.1006/dbio.2002.0640] [PMID: 11969258]
[29]
Kuboki T, Kanyama M, Nakanishi T, et al. Cbfa1/Runx2 gene expression in articular chondrocytes of the mice temporomandibular and knee joints in vivo. Arch Oral Biol 2003; 48(7): 519-25.
[http://dx.doi.org/10.1016/S0003-9969(03)00088-8] [PMID: 12798155]
[30]
Chen J, Wang Y, Chen C, et al. Exogenous heparan sulfate enhances the TGF-β3-induced chondrogenesis in human mesenchymal stem cells by activating TGF-β/Smad signaling Stem Cells Int 2016; 2016: 1520136.
[http://dx.doi.org/10.1155/2016/1520136] [PMID: 26783399]
[31]
Fernandes TL, Gomoll AH, Lattermann C, Hernandez AJ, Bueno DF, Amano MT. Macrophage: A Potential Target on Cartilage Regeneration. Front Immunol 2020; 11(111): 111.
[http://dx.doi.org/10.3389/fimmu.2020.00111] [PMID: 32117263]
[32]
Ying J, Wang P, Zhang S, et al. Transforming growth factor-beta1 promotes articular cartilage repair through canonical Smad and Hippo pathways in bone mesenchymal stem cells. Life Sci 2018; 192: 84-90.
[http://dx.doi.org/10.1016/j.lfs.2017.11.028] [PMID: 29158053]
[33]
Taylor DW, Ahmed N, Parreno J, et al. Collagen type XII and versican are present in the early stages of cartilage tissue formation by both redifferentating passaged and primary chondrocytes. Tissue Eng Part A 2015; 21(3-4): 683-93.
[http://dx.doi.org/10.1089/ten.tea.2014.0103] [PMID: 25315796]
[34]
Hino K, Saito A, Kido M, et al. Master regulator for chondrogenesis, Sox9, regulates transcriptional activation of the endoplasmic reticulum stress transducer BBF2H7/CREB3L2 in chondrocytes. J Biol Chem 2014; 289(20): 13810-20.
[http://dx.doi.org/10.1074/jbc.M113.543322] [PMID: 24711445]
[35]
Tsukamoto I, Inoue S, Teramura T, Takehara T, Ohtani K, Akagi M. Activating types 1 and 2 angiotensin II receptors modulate the hypertrophic differentiation of chondrocytes. FEBS Open Bio 2013; 3: 279-84.
[http://dx.doi.org/10.1016/j.fob.2013.07.001] [PMID: 23905010]
[36]
Kraskiewicz H, Paprocka M, Bielawska-Pohl A, et al. Can supernatant from immortalized adipose tissue MSC replace cell therapy? An in vitro study in chronic wounds model. Stem Cell Res Ther 2020; 11(1): 29-38.
[http://dx.doi.org/10.1186/s13287-020-1558-5] [PMID: 31964417]
[37]
Wang X, Song X, Li T, et al. Aptamer-functionalized bioscaffold enhances cartilage repair by improving stem cell recruitment in osteochondral defects of rabbit knees. Am J Sports Med 2019; 47(10): 2316-26.
[http://dx.doi.org/10.1177/0363546519856355] [PMID: 31233332]
[38]
Tao SC, Yuan T, Zhang YL, Yin WJ, Guo SC, Zhang CQ. Exosomes derived from miR-140-5p-overexpressing human synovial mesenchymal stem cells enhance cartilage tissue regeneration and prevent osteoarthritis of the knee in a rat model. Theranostics 2017; 7(1): 180-95.
[http://dx.doi.org/10.7150/thno.17133] [PMID: 28042326]


Rights & PermissionsPrintExport Cite as

Article Details

VOLUME: 20
ISSUE: 5
Year: 2020
Published on: 11 December, 2020
Page: [395 - 404]
Pages: 10
DOI: 10.2174/1566523220666201005110339
Price: $65

Article Metrics

PDF: 22