PROTECTION EFFECTS OF BONE MARROW MESENCHYMAL STEM CELLS PARACRINE ON CHONDROCYTES INJURED BY INTERLEUKIN 1β_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

XULizhang ¹ , DENGGuoying ¹ , WANGWeiheng ¹ , HUANGXiaodong ¹ , YANGXiangqun ² ,  YEXiaojian ¹

1. Department of Orthopeadics, Changzheng Hospital, the Second Military Medical University, Shanghai, 200003, P. R. China;
2. Department of Anatomy, Second Military Medicine University, Shanghai, 200003, P. R. China;

Corresponding author：

YEXiaojian, Email: xjye2012@126.com

Keywords：

Bone marrow mesenchymal stem cells; Indirect co-culture; Paracrine; Anti-inflammation; Anti-apoptosis; Tissue engineering; Seed cells

DOI：

10.7507/1002-1892.20150214

Video：

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Objective To study the potential protective effects of bone marrow mesenchymal stem cells (BMSCs) on chondrocytes injured by interleukin 1β (IL-1β), and the resistant capacity of chondrocytes when co-cultured indirectly with BMSCs against IL-1β. Methods Six Sprague Dawley (SD) rats were randomly divided into experimental group (articular cartilage defects) and control group. The content and gene expression of IL-1β were detected at 6 hours after surgical intervention by quantitative real time RCR (qRT-PCR) and ELISA. BMSCs repairing function test: the 18-holes cultured chondrocytes were randomly divided into 3 groups (n=6): cells of blank group were not treated;cells of injured group and co-cultured group were intervened by IL-1β, and Transwell chamber was used to establish co-culture system of BMSCs with chondrocyte in co-cultured group. The mRNA relative expressions of cysteinyl aspartate specific proteinase 3 (Caspase 3), a disintegrin and metalloprotease with Thrombospondin motifs 4 (ADAMTS-4), and ADAMTS-5 were measured via qRT-PCR in chondrocytes, meanwhile Caspase-3 content was detected via ELISA, and the cell apoptosis rate was detected via flow cytometry. BMSCs protecting function test: the 12-holes cultured chondrocytes were randomly divided into 2 groups (n=6), Transwell chamber was used to establish co-culture system of BMSCs with chondrocyte in co-cultured group before the 2 groups were both intervened by IL-1β, then the same detected indexes were taken as the BMSCs repairing function test. Results Animal in vivo studies showed that relative expression of IL-1β mRNA and IL-1β contents were significantly higher in experimental group than control group (P<0.05). BMSCs repair tests showed that mRNA relative expressions of Caspase-3, ADAMTS-4, and ADAMTS-5, Caspase-3 content, and cell apoptosis rate were significantly higher in injured group and co-cultured group than blank group, and in injured group than co-cultured group (P<0.05). BMSCs protect tests showed that mRNA relative expressions of Caspase-3, ADAMTS-4, and ADAMTS-5, Caspase-3 content, and cell apoptosis rate in co-cultured group were significantly lower than those in control group (P<0.05). Conclusion BMSCs, as seed cells for tissue engineering, have potential for applications to anti-inflammation and anti-apoptosis.

Citation： XULizhang, DENGGuoying, WANGWeiheng, HUANGXiaodong, YANGXiangqun, YEXiaojian. PROTECTION EFFECTS OF BONE MARROW MESENCHYMAL STEM CELLS PARACRINE ON CHONDROCYTES INJURED BY INTERLEUKIN 1β. Chinese Journal of Reparative and Reconstructive Surgery, 2015, 29(8): 996-1002. doi: 10.7507/1002-1892.20150214 Copy

1.	Xie A,Nie L,Shen G,et al.The application of autologous plateletrich plasma gel in cartilage regeneration.Mol Med Rep,2014,10(3):1642-1648.
2.	Hu R,Ling W,Xu W,et al.Fibroblast-like cells differentiated from adipose-derived mesenchymal stem cells for vocal fold wound healing.PLoS One,2014,9(3):e92676.
3.	Toth A,Fodor K,Blazsó P,et al.Generation of induced pluripotent stem cells by using a mammalian artificial chromosome expression system.Acta Biol Hung,2014,65(3):331-345.
4.	Liss MA,Skarecky D,Morales B,et al.The application of regional hypothermia using transrectal cooling during radical prostatectomy:mitigation of surgical inflammatory damage to preserve continence.J Endourol,2012,26(12):1553-1557.
5.	Wang F,Wu L,Li L,et al.Monotropein exerts protective effects against Il-1β-induced apoptosis and catabolic responses on osteoarthritis chondrocytes.Int Immunopharmacol,2014,23(2):575-580.
6.	李成,周海斌.骨髓间充质干细胞的旁分泌能影响成骨细胞生物学功能.中国组织工程研究,2014,18(10):1477-1483.
7.	Li B,Zhang H,Zeng M,et al.Bone marrow mesenchymal stem cells protect alveolar macrophages from lipopolysaccharide-induced apoptosis partially by inhibiting the Wnt/β-catenin pathway.Cell Biol Int,2014,39(2):192-200.
8.	王锋,吴小涛,王运涛,等.非接触共培养条件下人BMSCs对氧化应激环境中退变髓核细胞凋亡的保护研究.中国修复重建外科杂志,2010,24(4):391-398.
9.	Tsuzaki M,Guyton G,Garrett W,et al.IL-1 beta induces COX2,MMP-1,-3 and-13,ADAMTS-4,IL-1 beta and IL-6 in human tendon cells.J Orthop Res,2003,21(2):256-264.
10.	王正辉,吴宝俊,Kamal Mustafa.RNA干扰技术构建组织工程软骨的实验研究.中国修复重建外科杂志,2012,26(1):106-111.
11.	Chen S,Huang Y,Zhou ZJ,et al.Upregulation of tumor necrosis factor α and ADAMTS-5,but not ADAMTS-4,in human intervertebral cartilage endplate with modic changes.Spine (Phila Pa 1976),2014,39(14):E817-825.
12.	Zhai FX,Liu XF,Fan RF,et al.Runx3 is involved in caspase-3-dependent apoptosis induced by a combination of 5-aza-CdR and TSA in leukaemia cell lines.J Cancer Res Clin Oncol,2012,138(3):439-449.
13.	Leonardi R,Crimi S,Almeida LE,et al.ADAMTS-4 and ADAMTS-5 expression in human temporomandibular joint discs with internal derangement,correlates with degeneration.J Oral Pathol Med,2014.[Epub ahead of print].
14.	Li W,Du C,Wang H,et al.Increased serum ADAMTS-4 in knee osteoarthritis:apotential indicator for the diagnosis of osteoarthritis in early stages.Genet Mol Res,2014,13(4):9642-9649.
15.	Yagi H,Soto-Gutierrez A,Navarro-Alvarez N,et al.Reactive bone marrow stromal cells attenuate systemic inflammation via sTNFR1.Mol Ther,2010,18(10):1857-1864.
16.	Danchuk S,Ylostalo JH,Hossain F,et al.Human multipotent stromal cells attenuate lipopolysaccharide-induced acute lung injury in mice via secretion of tumor necrosis factor-alpha-induced protein 6.Stem Cell Res Ther,2011,2(3):27.
17.	Yagi H,Soto-Gutierrez A,KitagawaY,et al.Bone marrow mesenchymal stromal cells attenuate organ injury induced by LPS and Burn.Cell Transplant,2010,19(6):823-830.
18.	Haider HKh,Jiang S,Idris NM,et al.IGF-1-overexpressing mesenchymal stem cells accelerate bone marrow stem cell mobilization via paracrine activation of SDF-1alpha/CXCR4 signaling to promote myocardial repair.Circ Res,2008,103(11):1300-1308.
19.	Aggarwal S,Pittenger MF.Human mesenchymal stem cells modulate allogeneic immune cell responses.Blood,2005,105(4):1815-1822.
20.	Ulivi V,Tasso R,Cancedda R,et al.Mesenchymal stem cell paracrine activity is modulated by platelet lysate:induction of an inflammatory response and secretion of factors maintaining macrophages in a proinflammatory phenotype.Stem Cells Dev,2014,23(16):1858-1869.
21.	Eliopoulos N,Zhao J,Bouchentouf M,et al.Human marrow-derived mesenchymal stromal cells decrease cisplatin renotoxicity in vitro and in vivo and enhance survival of mice post-intraperitoneal injection.Am J Physiol Renal Physiol,2010,299(6):F1288-1298.
22.	Kuwana H,Terada Y,Kobayashi T,et al.The phosphoinositide-3 kinase gamma-Akt pathway mediates renal tubular injury in cisplatin nephrotoxicity.Kidney Int,2008,73(4):430-445.
23.	Morigi M,Rota C,Montemurro T,et al.Life-sparing effect of human cord blood-mesenchymal stem cells in experimental acute kidney injury.Stem Cells,2010,28(3):513-522.

1. Xie A,Nie L,Shen G,et al.The application of autologous plateletrich plasma gel in cartilage regeneration.Mol Med Rep,2014,10(3):1642-1648.
2. Hu R,Ling W,Xu W,et al.Fibroblast-like cells differentiated from adipose-derived mesenchymal stem cells for vocal fold wound healing.PLoS One,2014,9(3):e92676.
3. Toth A,Fodor K,Blazsó P,et al.Generation of induced pluripotent stem cells by using a mammalian artificial chromosome expression system.Acta Biol Hung,2014,65(3):331-345.
4. Liss MA,Skarecky D,Morales B,et al.The application of regional hypothermia using transrectal cooling during radical prostatectomy:mitigation of surgical inflammatory damage to preserve continence.J Endourol,2012,26(12):1553-1557.
5. Wang F,Wu L,Li L,et al.Monotropein exerts protective effects against Il-1β-induced apoptosis and catabolic responses on osteoarthritis chondrocytes.Int Immunopharmacol,2014,23(2):575-580.
6. 李成,周海斌.骨髓间充质干细胞的旁分泌能影响成骨细胞生物学功能.中国组织工程研究,2014,18(10):1477-1483.
7. Li B,Zhang H,Zeng M,et al.Bone marrow mesenchymal stem cells protect alveolar macrophages from lipopolysaccharide-induced apoptosis partially by inhibiting the Wnt/β-catenin pathway.Cell Biol Int,2014,39(2):192-200.
8. 王锋,吴小涛,王运涛,等.非接触共培养条件下人BMSCs对氧化应激环境中退变髓核细胞凋亡的保护研究.中国修复重建外科杂志,2010,24(4):391-398.
9. Tsuzaki M,Guyton G,Garrett W,et al.IL-1 beta induces COX2,MMP-1,-3 and-13,ADAMTS-4,IL-1 beta and IL-6 in human tendon cells.J Orthop Res,2003,21(2):256-264.
10. 王正辉,吴宝俊,Kamal Mustafa.RNA干扰技术构建组织工程软骨的实验研究.中国修复重建外科杂志,2012,26(1):106-111.
11. Chen S,Huang Y,Zhou ZJ,et al.Upregulation of tumor necrosis factor α and ADAMTS-5,but not ADAMTS-4,in human intervertebral cartilage endplate with modic changes.Spine (Phila Pa 1976),2014,39(14):E817-825.
12. Zhai FX,Liu XF,Fan RF,et al.Runx3 is involved in caspase-3-dependent apoptosis induced by a combination of 5-aza-CdR and TSA in leukaemia cell lines.J Cancer Res Clin Oncol,2012,138(3):439-449.
13. Leonardi R,Crimi S,Almeida LE,et al.ADAMTS-4 and ADAMTS-5 expression in human temporomandibular joint discs with internal derangement,correlates with degeneration.J Oral Pathol Med,2014.[Epub ahead of print].
14. Li W,Du C,Wang H,et al.Increased serum ADAMTS-4 in knee osteoarthritis:apotential indicator for the diagnosis of osteoarthritis in early stages.Genet Mol Res,2014,13(4):9642-9649.
15. Yagi H,Soto-Gutierrez A,Navarro-Alvarez N,et al.Reactive bone marrow stromal cells attenuate systemic inflammation via sTNFR1.Mol Ther,2010,18(10):1857-1864.
16. Danchuk S,Ylostalo JH,Hossain F,et al.Human multipotent stromal cells attenuate lipopolysaccharide-induced acute lung injury in mice via secretion of tumor necrosis factor-alpha-induced protein 6.Stem Cell Res Ther,2011,2(3):27.
17. Yagi H,Soto-Gutierrez A,KitagawaY,et al.Bone marrow mesenchymal stromal cells attenuate organ injury induced by LPS and Burn.Cell Transplant,2010,19(6):823-830.
18. Haider HKh,Jiang S,Idris NM,et al.IGF-1-overexpressing mesenchymal stem cells accelerate bone marrow stem cell mobilization via paracrine activation of SDF-1alpha/CXCR4 signaling to promote myocardial repair.Circ Res,2008,103(11):1300-1308.
19. Aggarwal S,Pittenger MF.Human mesenchymal stem cells modulate allogeneic immune cell responses.Blood,2005,105(4):1815-1822.
20. Ulivi V,Tasso R,Cancedda R,et al.Mesenchymal stem cell paracrine activity is modulated by platelet lysate:induction of an inflammatory response and secretion of factors maintaining macrophages in a proinflammatory phenotype.Stem Cells Dev,2014,23(16):1858-1869.
21. Eliopoulos N,Zhao J,Bouchentouf M,et al.Human marrow-derived mesenchymal stromal cells decrease cisplatin renotoxicity in vitro and in vivo and enhance survival of mice post-intraperitoneal injection.Am J Physiol Renal Physiol,2010,299(6):F1288-1298.
22. Kuwana H,Terada Y,Kobayashi T,et al.The phosphoinositide-3 kinase gamma-Akt pathway mediates renal tubular injury in cisplatin nephrotoxicity.Kidney Int,2008,73(4):430-445.
23. Morigi M,Rota C,Montemurro T,et al.Life-sparing effect of human cord blood-mesenchymal stem cells in experimental acute kidney injury.Stem Cells,2010,28(3):513-522.

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PROTECTION EFFECTS OF BONE MARROW MESENCHYMAL STEM CELLS PARACRINE ON CHONDROCYTES INJURED BY INTERLEUKIN 1β

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