EFFECTS OF Schwann CELLS PROMOTING NITRIC OXIDE SECRETION OF BONE MARROW MESENCHYMAL STEM CELLS DERIVED ENDOTHELIAL CELLS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

CAIXiyu ^1,2 , ZHANGXinxin ^2,3 , JIANGXiaorui ^2,4 , JIANGShan ² ,  PEIGuoxian ⁵

1. Department of Orthopaedics, the First Affiliated Hospital of Zhengzhou University, Zhengzhou Henan, 450052, P. R. China;
2. Department of Orthopaedic Traumatology, Nanfang Hospital, Southern Medical University;
3. Department of Orthopaedics, Cancer Hospital and Institute, Chinese Academy of Medical Sciences, Peking Union Medical College;
4. Department of Hand and Foot Orthopaedics, Yantai Yuhuangding Hospital, Qingdao University Medical College;
5. Institute of Orthopaedics of Chinese PLA, Xijing Hospital, the Forth Military Medical University;

Corresponding author：

PEIGuoxian, Email: nfperry@163.com

Keywords：

Bone tissue engineering; Bone marrow mesenchymal stem cells; Angiogenesis; Neuralization; Nitric oxide; Schwann cells; Rat

DOI：

10.7507/1002-1892.20140220

Video：

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Objective To study the effect of Schwann cells (SCs) promoting the function of nitric oxide (NO) secretion of bone marrow mesenchymal stem cells (BMSCs) derived endothelial cells so as to lay the experimental foundation for research of the effect of nerves on vessels during the process of tissue engineering bone formation. Methods SCs were collected from 1-day-old Sprague Dawley (SD) rats,and identified through S100 immunohistochemistry (IHC).BMSCs were collected from 2-week-old SD rats and induced into endothelial cells (IECs),which were identified through von Willebrand factor (vWF) and CD31 immunofluorescence (IF).Transwell system was used for co-culture of SCs and IECs without contact as the experimental group,and simple culture of IECs served as the control group.The NO concentration in the medium was measured at 1,3,5,and 7 days after culture; the mRNA expressions of nitric oxide synthetase 2 (NOS2) and NOS3 were detected by real-time fluorescence quantitative PCR (RT-qPCR) at 1,3,7,and 10 days. Results SCs and IECs were identified through morphology and immunology indexes of S100 IHC,vWF and CD31 IF.Significant differences were found in the NO concentration among different time points in 2 groups (P<0.05); the NO concentration of the experimental group was significantly higher than that of the control group at the other time points (P<0.05) except at 3 days.NOS2 mRNA expression of the experimental group was significantly higher than that of the control group (P<0.05); difference was significant in the NOS2 mRNA expression among different time points in 2 groups (P<0.05).NOS3 mRNA expression of the experimental group was significantly higher than that of the control group at the other time points (P<0.05) except at 10 days.No significant difference was found in NOS3 mRNA expression among different time points in the experimental group (F=6.673,P=0.062),but it showed significant differences in the control group (F=36.581,P=0.000). Conclusion SCs can promote NO secretion of BMSCs derived endothelial cells,which is due to promoting the activity of NOS.

Citation： CAIXiyu, ZHANGXinxin, JIANGXiaorui, JIANGShan, PEIGuoxian. EFFECTS OF Schwann CELLS PROMOTING NITRIC OXIDE SECRETION OF BONE MARROW MESENCHYMAL STEM CELLS DERIVED ENDOTHELIAL CELLS. Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(8): 998-1003. doi: 10.7507/1002-1892.20140220 Copy

1.	Johnson EO,Troupis T,Soucacos PN.Tissue-engineered vascularized bone grafts:basic science and clinical relevance to trauma and reconstructive microsurgery.Microsurgery,2011,31(3):176-182.
2.	Griffith LG,Naughton G.Tissue engineering-current challenges and expanding opportunities.Science,2002,295(5557):1009-1014.
3.	Pick M,Perry C,Lapidot T,et al.Stress-induced cholinergic signaling promotes inflammation-associated thrombopoiesis.Blood,2006,107(8):3397-3406.
4.	Fukuda T,Takeda S.Secondary osteoporosis or secondary contributors to bone loss in fracture.Regulation of bone homeostasis by nerve system.Clin Calcium,2013,23(9):1279-1283.
5.	Ohlsson C,Engdahl C,Borjesson AE,et al.Estrogen receptor-alpha expression in neuronal cells affects bone mass.Proc Natl Acad Sci U S A,2012,109(3):983-988.
6.	Bhandari V,Choo-Wing R,Chapoval SP,et al.Essential role of nitric oxide in VEGF-induced,asthma-like angiogenic,inflammatory,mucus,and physiologic responses in the lung.Proc Natl Acad Sci U S A,2006,103(29):11021-11026.
7.	Lei J,Vodovotz Y,Tzeng E,et al.Nitric oxide,a protective molecule in the cardiovascular system.Nitric Oxide,2013,35:175-185.
8.	Elefteriou F.Regulation of bone remodeling by the central and peripheral nervous system.Arch Biochem Biophys,2008,473(2):231-236.
9.	Haug SR,Heyeraas KJ.Modulation of dental inflammation by the sympathetic nervous system.J Dent Res,2006,85(6):488-495.
10.	Lee NJ,Herzog H.NPY regulation of bone remodelling.Neuropeptides,2009,43(6):457-463.
11.	Park MJ,Kwak HJ,Lee HC,et al.Nerve growth factor induces endothelial cell invasion and cord formation by promoting matrix metalloproteinase-2 expression through the phosphatidylinositol 3-kinase/Akt signaling pathway and AP-2 transcription factor.J Biol Chem,2007,282(42):30485-30496.
12.	孙春艳,胡豫,吴涛,等.脑源性神经营养因子对血管新生的作用.中华病理学杂志,2006,35(4):238-239.
13.	Tunyogi-Csapo M,Koreny T,Vermes C,et al.Role of fibroblasts and fibroblast-derived growth factors in periprosthetic angiogenesis.J Orthop Res,2007,25(10):1378-1388.
14.	Knott AB,Bossy-Wetzel E.Nitric oxide in health and disease of the nervous system.Antioxid Redox Signal,2009,11(3):541-554.
15.	Ricciardolo FL,Sterk PJ,Gaston B,et al.Nitric oxide in health and disease of the respiratory system.Physiol Rev,2004,84(3):731-765.
16.	Cirino G,Fiorucci S,Sessa WC.Endothelial nitric oxide synthase:the Cinderella of inflammation? Trends Pharmacol Sci,2003,24(2):91-95.
17.	Wang WZ,Fang XH,Stepheson LL,et al.Acute microvascular action of vascular endothelial growth factor in skeletal muscle ischemia/reperfusion injury.Plast Reconstr Surg,2005,115(5):1355-1365.
18.	Altavilla D,Galeano M,Bitto A,et al.Lipid peroxidation inhibition by raxofelast improves angiogenesis and wound healing in experimental burn wounds.Shock,2005,24(1):85-91.
19.	Tao J,Tu YT,Li JW,et al.Endogenous production of nitric oxide contributes to proliferation effect of vascular endothelial growth factor-induced malignant melanoma cell.Clin Exp Dermatol,2006,31(1):94-99.

1. Johnson EO,Troupis T,Soucacos PN.Tissue-engineered vascularized bone grafts:basic science and clinical relevance to trauma and reconstructive microsurgery.Microsurgery,2011,31(3):176-182.
2. Griffith LG,Naughton G.Tissue engineering-current challenges and expanding opportunities.Science,2002,295(5557):1009-1014.
3. Pick M,Perry C,Lapidot T,et al.Stress-induced cholinergic signaling promotes inflammation-associated thrombopoiesis.Blood,2006,107(8):3397-3406.
4. Fukuda T,Takeda S.Secondary osteoporosis or secondary contributors to bone loss in fracture.Regulation of bone homeostasis by nerve system.Clin Calcium,2013,23(9):1279-1283.
5. Ohlsson C,Engdahl C,Borjesson AE,et al.Estrogen receptor-alpha expression in neuronal cells affects bone mass.Proc Natl Acad Sci U S A,2012,109(3):983-988.
6. Bhandari V,Choo-Wing R,Chapoval SP,et al.Essential role of nitric oxide in VEGF-induced,asthma-like angiogenic,inflammatory,mucus,and physiologic responses in the lung.Proc Natl Acad Sci U S A,2006,103(29):11021-11026.
7. Lei J,Vodovotz Y,Tzeng E,et al.Nitric oxide,a protective molecule in the cardiovascular system.Nitric Oxide,2013,35:175-185.
8. Elefteriou F.Regulation of bone remodeling by the central and peripheral nervous system.Arch Biochem Biophys,2008,473(2):231-236.
9. Haug SR,Heyeraas KJ.Modulation of dental inflammation by the sympathetic nervous system.J Dent Res,2006,85(6):488-495.
10. Lee NJ,Herzog H.NPY regulation of bone remodelling.Neuropeptides,2009,43(6):457-463.
11. Park MJ,Kwak HJ,Lee HC,et al.Nerve growth factor induces endothelial cell invasion and cord formation by promoting matrix metalloproteinase-2 expression through the phosphatidylinositol 3-kinase/Akt signaling pathway and AP-2 transcription factor.J Biol Chem,2007,282(42):30485-30496.
12. 孙春艳,胡豫,吴涛,等.脑源性神经营养因子对血管新生的作用.中华病理学杂志,2006,35(4):238-239.
13. Tunyogi-Csapo M,Koreny T,Vermes C,et al.Role of fibroblasts and fibroblast-derived growth factors in periprosthetic angiogenesis.J Orthop Res,2007,25(10):1378-1388.
14. Knott AB,Bossy-Wetzel E.Nitric oxide in health and disease of the nervous system.Antioxid Redox Signal,2009,11(3):541-554.
15. Ricciardolo FL,Sterk PJ,Gaston B,et al.Nitric oxide in health and disease of the respiratory system.Physiol Rev,2004,84(3):731-765.
16. Cirino G,Fiorucci S,Sessa WC.Endothelial nitric oxide synthase:the Cinderella of inflammation? Trends Pharmacol Sci,2003,24(2):91-95.
17. Wang WZ,Fang XH,Stepheson LL,et al.Acute microvascular action of vascular endothelial growth factor in skeletal muscle ischemia/reperfusion injury.Plast Reconstr Surg,2005,115(5):1355-1365.
18. Altavilla D,Galeano M,Bitto A,et al.Lipid peroxidation inhibition by raxofelast improves angiogenesis and wound healing in experimental burn wounds.Shock,2005,24(1):85-91.
19. Tao J,Tu YT,Li JW,et al.Endogenous production of nitric oxide contributes to proliferation effect of vascular endothelial growth factor-induced malignant melanoma cell.Clin Exp Dermatol,2006,31(1):94-99.

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Chinese Journal of Reparative and Reconstructive Surgery

EFFECTS OF Schwann CELLS PROMOTING NITRIC OXIDE SECRETION OF BONE MARROW MESENCHYMAL STEM CELLS DERIVED ENDOTHELIAL CELLS

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