Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
JCR Impact Factor (IF) – 1.736
5-Year Impact Factor – 2.135
Index Copernicus  – 168.52
MEiN – 70 pts

ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
Periodicity – monthly

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Advances in Clinical and Experimental Medicine

2020, vol. 29, nr 3, March, p. 345–353

doi: 10.17219/acem/112602

Publication type: original article

Language: English

License: Creative Commons Attribution 3.0 Unported (CC BY 3.0)

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Effect of glucocorticoids on the function of microvascular endothelial cells in the human femoral head bone

Yufeng Lu1,B,C,D,F, Qingsheng Yu2,C, Wanshou Guo3,A, Yangquan Hao1,E,F, Wei Sun3,C, Liming Cheng3,B

1 Department of Joint Surgery, Osteonecrosis and Joint Reconstruction Ward, Honghui Hospital, Xi’an Jiaotong University, China

2 Beijing Key Laboratory of Arthritic and Rheumatic Diseases, China-Japan Friendship Hospital, China

3 Department of Orthopedic Surgery, Centre for Osteonecrosis and Joint-Preserving & Reconstruction, Beijing Key Laboratory of Arthritic and Rheumatic Diseases, China-Japan Friendship Hospital, China

Abstract

Background. The pathogenesis of glucocorticoid (GC)-induced osteonecrosis (ON) of the femoral head remains unclear. Recent research has suggested that it is closely associated with injured bone microvascular endothelial cells (BMECs). However, few studies have used BMECs to perform research pertaining ON of the femoral head.
Objectives. The objective of this study was to investigate the functional changes of BMECs treated with a GC and to detect the changes in related genes using microarrays.
Material and Methods. Cells were isolated using an enzymatic method and identified with EC markers, such as von Willebrand factor (vWF), CD31 and vascular endothelial cadherin (VE-cadherin). Bone microvascular endothelial cells were treated with 0.1 mg/mL and 0.3 mg/mL of hydrocortisone to establish a GC-damaged model of BMECs. The mRNA microarrays were used to detect the differential expression profiles between BMECs with and without GC damage.
Results. Primary cells appeared as having a cobblestone-like morphology. Immunofluorescence staining revealed that the cells were 100% positive for vWF and CD31, and near 100% positive for VE-cadherin. It also confirmed that the cells were BMECs. Bone microvascular endothelial cells treated with 0.1 mg/mL of hydrocortisone showed shrinkage, and those treated with 0.3 mg/mL of hydrocortisone mostly showed apoptosis. The mRNA microarray showed that genes associated with endothelial cells, such as endothelin 1 (ET-1) receptor, angiotensin II (AII) receptor, intercellular adhesion molecule 1 (ICAM-1), and plasminogen activator inhibitor 1 (PAI-1), were upregulated, and genes associated with endothelial nitric oxide synthase (eNOS), endothelin 1 (ET-1), prostaglandin I2 (PGI2) synthase, PGI2 receptor, vascular endothelial growth factor (VEGF), prostaglandin E (PGE) synthase, and PGE receptor were downregulated. The results of quantitative polymerase chain reaction (qPCR) validation were consistent with the findings of mRNA microarrays.
Conclusion. Glucocorticoids promoted BMECs to express vasoconstrictors and procoagulant factors and related receptors, and decreased the expression of vasodilators and their receptors.

Key words

glucocorticoids, microarray analysis, cell culture techniques, microvascular endothelial cells, real-time polymerase chain reaction

References (40)

  1. Mont M, Pivec R, Banerjee S, Issa K, Elmallah RK, Jones LC. High-dose corticosteroid use and risk of hip osteonecrosis: Meta-analysis and systematic literature review. J Arthroplasty. 2015;30(9):1506–1512.
  2. Wang Y, Li Y, Mao K, Li J, Cui Q, Wang GJ. Alcohol-induced adipogenesis in bone and marrow: A possible mechanism for osteonecrosis. Clin Orthop Relat Res. 2003;410:213–224.
  3. Sharareh B, Schwarzkopf R. Dysbaric osteonecrosis: A literature review of pathophysiology, clinical presentation, and management. Clin J Sport Med. 2015;25(2):153–161.
  4. Milner PF, Kraus AP, Sebes JI, et al. Sickle cell disease as a cause of osteonecrosis of the femoral head. N Engl J Med. 1991;325(21):1476–1481.
  5. Nakamura J, Konno K, Orita S, et al. Distribution of hip pain in patients with idiopathic osteonecrosis of the femoral head. Mod Rheumatol. 2017;27(3):503–507.
  6. Kobayashi S, Kubo T, Iwamoto Y, Fukushima W, Sugano N. Nationwide multicenter follow-up cohort study of hip arthroplasties performed for osteonecrosis of the femoral head. Int Orthop. 2018;42(7):1661–1668.
  7. Ikeuchi K, Hasegawa Y, Seki T, Takegami Y, Amano T, Ishiguro N. Epidemiology of nontraumatic osteonecrosis of the femoral head in Japan. Mod Rheumatol. 2015;25(2):278–281.
  8. Fukushima W, Fujioka M, Kubo T, Tamakoshi A, Nagai M, Hirota Y. Nationwide epidemiologic survey of idiopathic osteonecrosis of the femoral head. Clin Orthop Relat Res. 2010;468(10):2715–2724.
  9. Kang JS, Park S, Song JH, Jung YY, Cho MR, Rhyu KH. Prevalence of osteonecrosis of the femoral head: A nationwide epidemiologic analysis in Korea. J Arthroplasty. 2009;24(8):1178–1183.
  10. Youm YS, Lee SY, Lee SH. Apoptosis in the osteonecrosis of the femoral head. Clin Orthop Surg. 2010;2(4):250–255.
  11. Mutijima E, De Maertelaer V, Deprez M, Malaise M, Hauzeur JP. The apoptosis of osteoblasts and osteocytes in femoral head osteonecrosis: Its specificity and its distribution. Clin Rheumatol. 2014;33(12):1791–1795.
  12. Ren X, Fan W, Shao Z, Chen K, Yu XX, Liang Q. A metabolomic study on early detection of steroid-induced avascular necrosis of the femoral head. Oncotarget. 2018;9(8):7984–7995.
  13. Zhang Q, L V J, Jin L. Role of coagulopathy in glucocorticoid-induced osteonecrosis of the femoral head. J Int Med Res. 2018;46(6):2141–2148.
  14. Akaike M, Matsumoto T. Glucocorticoid-induced reduction in NO bioavailability and vascular endothelial dysfunction. Clin Calcium. 2007;17(6):864–870.
  15. Kerachian MA, Séguin C, Harvey EJ. Glucocorticoids in osteonecrosis of the femoral head: A new understanding of the mechanisms of action. J Steroid Biochem Mol Biol. 2009;114(3–5):121–128.
  16. Vogt CJ, Schmid-Schönbein GW. Microvascular endothelial cell death and rarefaction in the glucocorticoid-induced hypertensive rat. Microcirculation. 2001;8(2):129–139.
  17. Li Y, Chen J, Zhang Z, Wang K, Tong Z, Yan H. The experimental study on treatment of glucocorticoid-induced ischemic necrosis of femoral head by Gu Fu Sheng capsule. J Tradit Chin Med. 2004;24(4):303–307.
  18. Yang Y, Lou J, Li Z, Sun W, Wang B, Jia Y. Effect of glucocorticoid on production of reactive oxygen species in bone microvascular endothelial cells [in Chinese]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2011;25(5):533–537.
  19. Wnuczko K, Szczepański M. Endothelium: Characteristics and functions [in Polish]. Pol Merkur Lekarski. 2007;23(133):60–65.
  20. Van Veldhuizen PJ, Neff J, Murphey MD, Bodensteiner D, Skikne BS. Decreased fibrinolytic potential in patients with idiopathic avascular necrosis and transient osteoporosis of the hip. Am J Hematol. 1993;44(4):243–248.
  21. Glueck CJ, Glueck HI, Mieczkowski L, Tracy T, Speirs J, Stroop D. Familial high plasminogen activator inhibitor with hypofibrinolysis: A new pathophysiologic cause of osteonecrosis? Thromb Haemost. 1993;69(5):460–465.
  22. Asano T, Takahashi KA, Fujioka M, et al. Relationship between postrenal transplant osteonecrosis of the femoral head and gene polymorphisms related to the coagulation and fibrinolytic systems in Japanese subjects. Transplantation. 2004;77(2):220–225.
  23. Yu Q, Guo W, Cheng L, Lu Y, Li P. Preliminary study of impact of steroids on expression profile and transcriptome of bone microvascular endothelial cells [in Chinese]. Zhonghua Yi Xue Za Zhi. 2014;94(48):3817–3820.
  24. Lu YF, Yu QS, Guo WS, Cheng LM, Zhang Y. A method for isolated culture of bone microvascular endothelial cells of human femoral head [in Chinese]. Zhongguo Gu Shang. 2014;27(10):843–847.
  25. Microsurgery Department of the Orthopedics Branch of the Chinese Medical Doctor Association; Group from the Osteonecrosis and Bone Defect Branch of the Chinese Association of Reparative and Reconstructive Surgery; Microsurgery and Reconstructive Surgery Group of the Orthopedics Branch of the Chinese Medical Association; et al. Chinese Guideline for the Diagnosis and Treatment of Osteonecrosis of the Femoral Head in Adults. Orthop Surg. 2017;9(1):3–12.
  26. Youm YS, Lee SY, Lee SH. Apoptosis in the osteonecrosis of the femoral head. Clin Orthop Surg. 2010;2(4):250–255.
  27. Chotanaphuti T, Heebthamai D, Chuwong M, Kanchanaroek K. The prevalence of thrombophilia in idiopathic osteonecrosis of the hip. J Med Assoc Thai. 2009;92(Suppl 6):S141–146.
  28. Zalavras CG, Vartholomatos G, Dokou E, Malizos KN. Genetic background of osteonecrosis: Associated with thrombophilic mutations? Clin Orthop Relat Res. 2004;422:251–255.
  29. De Bruyn PP. Structural substrates of bone marrow function. Semin Hematol. 1981;18(3):179–193.
  30. Tavassoli M. The marrow–blood barrier. Br J Haematol. 1979;41(3):297–302.
  31. Drescher W, Li H, Lundgaard A, Bünger C, Hansen ES. Endothelin-1-induced femoral head epiphyseal artery constriction is enhanced by long-term corticosteroid treatment. J Bone Joint Surg Am. 2006;88(Suppl 3):173–179.
  32. Shatanawi A, Lemtalsi T, Yao L, Patel C, Caldwell RB, Caldwell RW. Angiotensin II limits NO production by upregulating arginase through a p38 MAPK-ATF-2 pathway. Eur J Pharmacol. 2015;746:106–114.
  33. Montezano AC, Nguyen Dinh Cat A, Rios FJ, Touyz RM. Angiotensin II and vascular injury. Curr Hypertens Rep. 2014;16(6):431.
  34. He W, Xu C, Fan Y, et al. Effects of the Chinese drugs for activating blood circulation on plasma TXB2 and 6-keto-PGF1alpha contents in rabbits with glucocorticoid-induced femoral head necrosis. J Tradit Chin Med. 2004;24(3):233–237.
  35. Fang WT, Li HJ, Zhou LS. Protective effects of prostaglandin E1 on human umbilical vein endothelial cell injury induced by hydrogen peroxide. Acta Pharmacol Sin. 2010;31(4):485–492.
  36. Zeng Z, Wang B, Pan H. Relation between osteonecrosis of the femoral head and PAI-1 4G/5G gene polymorphism: A meta-analysis. A Int J Clin Exp Med. 2015;8(11):20337–20342.
  37. Yamamoto Y, Ishizu A, Ikeda H, Otsuka N, Yoshiki T. Dexamethasone increased plasminogen activator inhibitor-1 expression on human umbilical vein endothelial cells: An additive effect to tumor necrosis factor-alpha. Pathobiology. 2004;71(6):295–301.
  38. Kim H, Cho C, Cho Y, Cho S, Yoon K, Kim K. Significant associations of PAI-1 genetic polymorphisms with osteonecrosis of the femoral head. BMC Musculoskelet Disord. 2011;12:160.
  39. Lawson C, Wolf S. ICAM-1 signaling in endothelial cells. Pharmacol Rep. 2009;61(1):22–32.
  40. Li X, Jin L, Cui Q, Wang GJ, Balian G. Steroid effects on osteogenesis through mesenchymal cell gene expression. Osteoporos Int. 2005;16(1):101–108.