Changes in serum calcitonin gene-related peptide levels in animal models of femoral head osteonecrosis

doi:10.3969/j.issn.2095-4344.2015.05.003

Abstract

Abstract:

BACKGROUND: Calcitonin gene-related peptide is a peptide secreted from neuropeptides, has multiple important physiological functions, and plays a key role in normal metabolism of bone tissue, bone defect repair and reconstruction.
OBJECTIVE: To observe the changes in the levels of serum calcitonin gene-related peptide of animal models of osteonecrosis of the femoral head.
METHODS: A total of 45 New Zealand white rabbits were randomly divided into three groups: hormone modeling group, liquid nitrogen modeling group and normal control group. At 4, 6, 8, 9 and 10 weeks after model establishment, serum calcitonin gene-related peptide levels were determined. At 9 and 10 weeks, pathological detection was performed.
RESULTS AND CONCLUSION: Clear bone necrosis was visible at 8 weeks. Simultaneously, serum calcitonin gene-related peptide levels were significantly higher in the hormone modeling and liquid nitrogen modeling groups than in the normal control group. These data suggested that osteonecrosis of the femoral head was possibly strongly associated with serum calcitonin gene-related peptide levels.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

全文链接：

Key words: Femur Head Necrosis, Models, Animal, Calcitonin Gene-Related Peptide

CLC Number:

R318

Huang Tao, Kaisaierjiang Aihemaiti, Cui Yong. Changes in serum calcitonin gene-related peptide levels in animal models of femoral head osteonecrosis[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(5): 667-673.

Figures/Tables 3

2.1 实验动物数量分析液氮建模组实验动物术后因感染死亡1只，正常对照组实验动物因腹泻死亡2只，余下实验动物正常存活。 2.2 模型质量 MRI观察结果：激素建模组实验动物在造模第4周时行MRI检查，结果显示双侧股骨头出现斑片状低信号区域(图1A)。造模第8周时MRI图像显示在T2WI图像上股骨头出现混杂性低信号，提示股骨头已经出现坏死；在T1WI和T2WI上呈明显的低信号区域，提示增生反应区存在(图1B)。液氮建模组在术后第4周时MRI检查图像显示双侧股骨头及头下信号不均匀，股骨头表面软骨退变?变薄，髋臼内少量积液(图1C)。至第8周时MRI图像显示股骨头出现明显坏死区，呈低信号，未见股骨头塌陷，提示股骨头已经坏死(图1D)。 X射线观察结果：激素建模组动物在造模第4周时行X射线检查，结果提示股骨头密度明显增高，关节间隙基本正常；造模第8周时股骨头轮廓呈现出不规则改变，周边出现不规则透光带，骺板较清晰，关节间隙稍微增宽(图2A，B)。液氮建模组动物在术后第4周时行X射检查，结果显示股骨头大体轮廓基本正常，但股骨头局部呈现密度不均匀影像，骺板尚清晰，未观察到股骨头出现明显塌陷，髋关节间隙无明显变化；第8周时股骨头体积缩小、股骨头表面毛躁、不光滑、局部出现塌陷、骺板比较模糊、关节间隙增宽(图2C，D)。病理检测结果：激素建模组于造模第8，9，10周时分别处死5只实验动物行病理学检查。光镜观察结果提示，第8周时，动物骨小梁内骨陷窝增加，骨细胞核固缩，骨小梁之间脂肪细胞也增多、增大，骨小梁明显稀疏(图3A)；造模第9周时，髓内脂肪细胞增大，增多，造血组织(骨小梁间充质细胞)下降(图3B)；造模第10周时，脂肪细胞增多极为明显，髓腔几乎全部被脂肪细胞填满，正常骨髓组织明显减少(图3C)。液氮建模组于造模第8，9周时分别处死5只实验动物，第10周时处死剩余4只实验动物行病理学检查。造模第8周时观察结果提示，股骨头冷冻区域软骨下出现出血、凝血；冷冻区周边骨小梁分布紊乱，发生断裂，且空骨陷窝数量明显增加；髓腔内出现肉芽组织增生、出血、充血，可见由脂肪细胞相互融合而形成的小囊(图4A)；造模第9周时观察股骨头冷冻区域内骨小梁结构、走形极其紊乱，有匍行附着，空骨陷窝出现在周边大部分骨小梁内；软骨下小血管数量减少，周围软骨出现脱落(图4B)；造模第10周时，出现较大面积的软骨剥脱现象，软骨覆盖下的骨质因软骨的剥脱而暴露；死骨出现在骨髓腔周围，并可见沉积性生长的新生骨，骺板细胞堆积、变形(图4C)。正常对照组病理检查结果显示，软骨层均匀，软骨细胞无坏死，软骨陷窝无空虚(图5A)；骨小梁走形规则，有少量的空骨陷窝(图5B，C)；骨髓内脂肪细胞大小均匀，无增大及增多现象，与造血组织比例均匀(图5D)。"

References

[1] 张鹤山,李子荣.股骨头坏死诊断与治疗的专家建议[J].中华骨科杂志,2007,27(2):146-148.
[2] Poyner D,Marshall I,Brain SD.The CGRP Family. Lands, Bioscience. USA:Texas, 1999:173-252.
[3] 李瑞琦,张国平,李宜炯,等.兔激素性股骨头坏死模型的研究[J].现代中西医结合杂志,2013,22(28):3087-3089,3170.
[4] 周正丽,张潜,彭笳宸,等.液氮冷冻兔股骨头坏死模型制备新方法及可靠性评价[J].解剖学报,2012,43(2):284-288.
[5] Mont MA, Jones LC, Einhorn TA,et al.Osteonecrosis of the femoral head. Potential treatment with growth and differentiation factors.Clin Orthop Relat Res.1998;(355 Suppl): S314-335.
[6] Jones JP Jr.Alcoholism, hypercortisonism, fat embolism and osseous avascular necrosis. 1971.Clin Orthop Relat Res. 2001;(393):4-12.
[7] Chao YC, Wang SJ, Chu HC,et al.Investigation of alcohol metabolizing enzyme genes in Chinese alcoholics with avascular necrosis of hip joint, pancreatitis and cirrhosis of the liver.Alcohol Alcohol. 2003;38(5):431-436.
[8] Kim HK, Herring JA.Pathophysiology, classifications, and natural history of Perthes disease.Orthop Clin North Am. 2011;42(3):285-295.
[9] 陈演.股骨头坏死病因学研究的某些进展[J].华西医学,2009, 24(7):1899-1901.
[10] Aaron RK,Gray RL.Osteonecrosis:etiology,natural history, pathophysiology and diagnosid//Cailaghan JJ, Rosenberg AG,Rubash JE. Theaduhhip.2nd ed. Philadelphia: Lippincott Williams&Wilkins,2007:463-476.
[11] Jones LC, Hungerford DS.Osteonecrosis: etiology, diagnosis, and treatment.Curr Opin Rheumatol. 2004;16(4):443-449.
[12] Suh KT, Kim SW, Roh HL, et al.Decreased osteogenic differentiation of mesenchymal stem cells in alcohol-induced osteonecrosis.Clin Orthop Relat Res. 2005;(431):220-225.
[13] Bjurholm A, Kreicbergs A, Schultzberg M,et al. Neuroendocrine regulation of cyclic AMP formation in osteoblastic cell lines (UMR-106-01, ROS 17/2.8, MC3T3-E1, and Saos-2) and primary bone cells.J Bone Miner Res. 1992; 7(9):1011-1019.
[14] Michelangeli VP, Fletcher AE, Allan EH,et al.Effects of calcitonin gene-related peptide on cyclic AMP formation in chicken, rat, and mouse bone cells.J Bone Miner Res. 1989; 4(2):269-272.
[15] Bernard GW, Shih C.The osteogenic stimulating effect of neuroactive calcitonin gene-related peptide.Peptides. 1990; 11(4):625-632.
[16] Vignery A, McCarthy TL.The neuropeptide calcitonin gene-related peptide stimulates insulin-like growth factor I production by primary fetal rat osteoblasts.Bone. 1996;18(4): 331-335.
[17] Calland JW, Harris SE, Carnes DL Jr.Human pulp cells respond to calcitonin gene-related peptide in vitro.J Endod. 1997;23(8):485-489.
[18] Shih C, Bernard GW.Calcitonin gene related peptide enhances bone colony development in vitro.Clin Orthop Relat Res. 1997;(334):335-344.
[19] Villa I, Dal Fiume C, Maestroni A,et al.Human osteoblast-like cell proliferation induced by calcitonin-related peptides involves PKC activity.Am J Physiol Endocrinol Metab. 2003; 284(3):E627-633.
[20] Villa I, Mrak E, Rubinacci A,et al.CGRP inhibits osteoprotegerin production in human osteoblast-like cells via cAMP/PKA-dependent pathway.Am J Physiol Cell Physiol. 2006;291(3):C529-537.
[21] Imai S, Matsusue Y.Neuronal regulation of bone metabolism and anabolism: calcitonin gene-related peptide-, substance P-, and tyrosine hydroxylase-containing nerves and the bone. Microsc Res Tech. 2002;58(2):61-69.
[22] Tippins JR, Morris HR, Panico M,et al.The myotropic and plasma-calcium modulating effects of calcitonin gene-related peptide (CGRP).Neuropeptides. 1984;4(5):425-434.
[23] Chambers TJ, Magnus CJ.Calcitonin alters behaviour of isolated osteoclasts.J Pathol. 1982;136(1):27-39.
[24] Valentijn K, Gutow AP, Troiano N,et al.Effects of calcitonin gene-related peptide on bone turnover in ovariectomized rats. Bone. 1997;21(3):269-274.
[25] Cornish J, Callon KE, Bava U,et al.Effects of calcitonin, amylin, and calcitonin gene-related peptide on osteoclast development.Bone. 2001;29(2):162-168.
[26] Ishizuka K, Hirukawa K, Nakamura H,et al.Inhibitory effect of CGRP on osteoclast formation by mouse bone marrow cells treated with isoproterenol.Neurosci Lett. 2005;379(1):47-51.
[27] 孙应明,罗颂椒,赵昱辉,等.降钙素基因相关肽对体外大鼠破骨细胞形成的影响[J].临床口腔医学杂志,2005,21(1):3-5.
[28] 徐琳,谭颖徽,王建华,等. 失神经支配兔下颌骨骨折愈合过程中CGRP对OPG/RANKL表达的影响[J].第三军医大学学报,2005, 27(10):988-990.
[29] Hukkanen M, Konttinen YT, Santavirta S,et al.Effect of sciatic nerve section on neural ingrowth into the rat tibial fracture callus. Clin Orthop Relat Res. 1995;(311):247-257.
[30] Figini M, Emanueli C, Grady EF,et al.Substance P and bradykinin stimulate plasma extravasation in the mouse gastrointestinal tract and pancreas.Am J Physiol. 1997;272 (4 Pt 1):G785-793.
[31] Elefteriou F.Regulation of bone remodeling by the central and peripheral nervous system.Arch Biochem Biophys. 2008; 473(2): 231-236.
[32] Jones KB, Mollano AV, Morcuende JA,et al.Bone and brain: a review of neural, hormonal, and musculoskeletal connections. Iowa Orthop J. 2004;24:123-132.
[33] Elefteriou F.Neuronal signaling and the regulation of bone remodeling.Cell Mol Life Sci. 2005;62(19-20):2339-2349.
[34] Huebner AK, Keller J, Catala-Lehnen P,et al.The role of calcitonin and alpha-calcitonin gene-related peptide in bone formation.Arch Biochem Biophys. 2008;473(2):210-217.
[35] Gajda M, Litwin JA, Cichocki T,et al.Development of sensory innervation in rat tibia: co-localization of CGRP and substance P with growth-associated protein 43 (GAP-43).J Anat. 2005; 207(2):135-144.
[36] Hukkanen M, Konttinen YT, Santavirta S,et al.Rapid proliferation of calcitonin gene-related peptide- immunoreactive nerves during healing of rat tibial fracture suggests neural involvement in bone growth and remodelling. Neuroscience. 1993 Jun;54(4):969-979.
[37] Schlomer JJ, Storey BB, Ciornei RT,et al.Calcitonin gene-related peptide inhibits early B cell development in vivo.J Leukoc Biol. 2007;81(3):802-808.
[38] Coderre TJ, Basbaum AI, Levine JD.Neural control of vascular permeability: interactions between primary afferents, mast cells, and sympathetic efferents.J Neurophysiol. 1989; 62(1):48-58.
[39] Wang X, Han CD, Fiscus RR,et al.Hypotension- and endotoxin-induced alterations in calcitonin gene-related peptide: modulation by dexamethasone.Circ Shock. 1991; 34(2): 217-223.