Serum N‑terminal telopeptide of type I collagen as an early marker of fracture nonunion in rabbits

Lin,Jian‑Ping; Shi,Zhan‑Jun; Shen,Ning‑Jiang; Wang,Jian; Li,Zao‑Min; Xiao,Jun

doi:10.3892/etm.2016.3839

Serum N‑terminal telopeptide of type I collagen as an early marker of fracture nonunion in rabbits

Authors:
- Jian‑Ping Lin
- Zhan‑Jun Shi
- Ning‑Jiang Shen
- Jian Wang
- Zao‑Min Li
- Jun Xiao
View Affiliations

Affiliations: Department of Orthopedic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of Orthopedic Surgery, Hainan Provincial People's Hospital, Haikou, Hainan 570311, P.R. China
Published online on: October 26, 2016 https://doi.org/10.3892/etm.2016.3839
Pages: 3595-3601
Copyright: © Lin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of the present study was to establish an experimental animal model of fracture nonunion, and to investigate the changes in serum biomarker concentrations in fracture nonunion. A total of 20 purebred New Zealand rabbits were divided into two group: A bone defect group and a bone fracture group. In the bone defect group, a 15‑mm section of bone (including the periosteum) was removed from the mid‑radius, and the medullary cavities were closed with bone wax. In the bone fracture group, the mid‑radius was fractured. X‑rays were taken and blood samples were collected preoperatively and at 2, 3, 4, 5, 6, 7, 8, 10 and 12 weeks after the surgical procedure. The serum concentrations of osteocalcin (OC) and bone‑specific alkaline phosphatase (BSAP) served as markers of bone formation, and those of C‑terminal telopeptide of type I collagen (CTX), N‑terminal telopeptide of type I collagen (NTX) and tartrate‑resistant acid phosphatase 5b (TRACP 5b) served as markers of bone resorption. The concentration levels of the markers were measured using a biotin double‑antibody sandwich enzyme‑linked immunosorbent assay. In the bone defect group, bone callus was observed on X‑ray at 2 weeks in three rabbits and the bone calluses stabilized at 5 weeks; however, none of the bones had healed at 8 weeks. In the bone fracture group, the fracture line was distorted at 2 weeks and bone calluses formed at 6‑8 weeks. In the bone defect group, the serum BSAP and TRACP 5b concentrations increased following the surgical procedure, peaked at 4 weeks, began to decrease at 5 weeks and stabilized after 6 weeks. The serum OC concentrations did not change significantly following the surgical procedure. The serum CTX concentrations fluctuated during the first 4 weeks, peaked at 5 weeks, then decreased and stabilized after 6 weeks. The serum NTX concentrations fluctuated during the first 4 weeks, were significantly lower at 5 weeks compared with the other time points and stabilized after 6 weeks. These results suggested that a bone nonunion model can be established in New Zealand rabbits by resecting a 15‑mm section of bone from the mid‑radius prior to bone wax blocking. Measurement of the serum BSAP, CTX, NTX, and TRACP 5b concentrations may be useful for the early detection of bone nonunion. The serum NTX concentrations changed significantly in rabbits with bone nonunion. Further studies are required in order to determine the feasibility of using serum NTX concentrations for the early diagnosis of bone nonunion.

View Figures

View References

1	Court-Brown CM and McQueen MM: Nonunions of the proximal humerus: their prevalence and functional outcome. J Trauma. 64:1517–1521. 2008. View Article : Google Scholar : PubMed/NCBI
2	Blokhuis TJ, de Bruine JH, Bramer JA, den Boer FC, Bakker FC, Patka P, Haarman HJ and Manoliu RA: The reliability of plain radiography in experimental fracture healing. Skeletal Radio. 30:151–156. 2001. View Article : Google Scholar
3	Lotz J, Gaertner T, Hahn M and Prellwitz W: Collagen type I metabolism after bone surgery. Arch Orthop Trauma Surg. 119:212–216. 1999. View Article : Google Scholar : PubMed/NCBI
4	Sambrook P and Cooper C: Osteoporosis. Lancet. 367:2010–2018. 2006. View Article : Google Scholar : PubMed/NCBI
5	Lin JP, Song SF and Yao LL: Research progress of fracture healing and it's early diagnosis. Ortho J China. 24:1876–1878. 2009.
6	Gamero P and Somay-Rendu E: Biochemical markers of bone loss rate and prevalence of osteoporosis at multiple skeletal sites in Chinese women. Osteoporos Int. 13:669–676. 2002. View Article : Google Scholar : PubMed/NCBI
7	Laboratory animal welfare, . Public Health Service policy on humane care and use of laboratory animals by awardee institutions; notice. Fed Regist. 50:19584–19585. 1985.PubMed/NCBI
8	Kokubu T, Hak DJ, Hazelwood SJ and Reddi AH: Development of an atrophic nonunion model and comparison to a closed healing fracture in rat femur. J Orthop Res. 21:503–510. 2003. View Article : Google Scholar : PubMed/NCBI
9	Reed AA, Joyner CJ, Brownlow HC and Simpson AH: Human atrophic fracture nonunion are not avascular. J Orthop Res. 20:593–599. 2002. View Article : Google Scholar : PubMed/NCBI
10	Mumaghan M, Li G and Mash DR: Nonsteroidal anti-inflammatory drug induced fracture nonunion: An inhibition of angiognensis? JBJS Am. 88:141–147. 2006.
11	Nunamaker DM: Experimental models of fracture repair. Clin Orthop. (Suppl 355). 561998. View Article : Google Scholar : PubMed/NCBI
12	Peter CP and Cook WO: Effect of alendronate on fracture healing and bone remodeling in dogs. J Orthop Res. 14:741996. View Article : Google Scholar : PubMed/NCBI
13	Korkmaz M, Oztürk H, Bulut O, Unsaldi T and Kaloğlu C: The effect of definitive continuous distraction employed with the Ilizarov type external fixation system on fracture healing: An experimental rabbit model. Acta Orthop Traumatol Turc. 39:247–257. 2005.(In Turkish). PubMed/NCBI
14	Johnson EE, Urist MR, Schmalzried TP, Chotivichit A, Huang HK and Finerman GA: Autogeneic cancellous bone grafts in extensive segmental ulnar defects in dogs. Effects of xenogeneic bovine bone morphogenetic protein without and with interposition of soft tissues and interruption of blood supply. Clin Orthop Relat Res. 254–265. 1989.PubMed/NCBI
15	Cook SD, Wolfe MW, Salkeld SL and Rueger DC: Effect of recombinant human osteogenic protein-1 on healing of segmental defects in non-human primate. J Bone Joint Surg Am. 77:734–750. 1995. View Article : Google Scholar : PubMed/NCBI
16	Kasten P, Vogel J, Geiger F, Niemeyer P, Luginbühl R and Szalay K: The effect of platelet-rich plasma on healing in critical-size long-bone defects. Biomaterials. 29:3983–3992. 2008. View Article : Google Scholar : PubMed/NCBI
17	Niemeyer P, Szalay K, Luginbühl R, Südkamp NP and Kasten P: Transplantation of human mesenchymal stem cells in a non-autogenous setting for bone regeneration in a rabbit critical-size defect model. Acta Biomater. 6:900–908. 2010. View Article : Google Scholar : PubMed/NCBI
18	Felix R, Herrmann W and Fleisch H: Stimulation of precipitation of calcium phosphate by matrix vesicles. Biochem J. 170:681–691. 1978. View Article : Google Scholar : PubMed/NCBI
19	Anderson HC, Stechschulte DJ Jr, Collins DE, Jacobs DH, Morris DC, Hsu HH, Redford PA and Zeiger S: Matrix vesicle biogenesis in vitro by rachitic and normal rat chondrocytes. Am J Pathol. 136:391–398. 1990.PubMed/NCBI
20	Parker MJ, Raghavan R and Gurusamy K: Incidence of fracture-healing complications after femoral neck fractures. Clin Orthop Relat Res. 458:175–179. 2007.PubMed/NCBI
21	Gothlin G and Ericsson JL: Fine structural localization of alkaline phosphatase in the fracture callus of the rat. Histochemie. 36:225–236. 1973. View Article : Google Scholar : PubMed/NCBI
22	Pedersen BJ, Schlemmer A, Hassager C and Christiansen C: Changes in the carboxyl-terminal propeptide of type I procollagen and other markers of bone formation upon five days of bed rest. Bone. 17:91–95. 1995. View Article : Google Scholar : PubMed/NCBI
23	Lian J, Stewart C, Puchacz E, Mackowiak S, Shalhoub V, Collart D, Zambetti G and Stein G: Structure of the rat osteocalcin gene and regulation of vitamin D-dependent expression. Proc Natl Acad Sci USA. 86:1143–1147. 1989. View Article : Google Scholar : PubMed/NCBI
24	Noite PA, Krans A Vander, Patka P, Janssen IM, Ryaby JP and Albers GH: Low-intensity pulsed ultrasound device for the noninvasive treatment of nonunions. J Trauma. 51:693–702. 2001. View Article : Google Scholar : PubMed/NCBI
25	Geiger F, Lorenz H, Xu W, Szalay K, Kasten P, Claes L, Augat P and Richter W: VEGF producing bone marrow stromal cells (BMSC) enhance vascularization and resorption of a natural coral bone substitute. Bone. 41:516–522. 2007. View Article : Google Scholar : PubMed/NCBI
26	García-Pérez MA, Moreno-Mercer J, Tarín JJ and Cano A: Similar efficacy of low and standard doses of transdermal estradiol in controlling bone turnover in postmenopausal women. Gynecol Endocrinol. 22:179–184. 2006. View Article : Google Scholar : PubMed/NCBI
27	Kyro A, Usenius JP, Aarnio M, Kunnamo I and Avikainen V: Are smokers a risk group for delayed healing of tibial shaft fractures? Ann Chir Gynaecol. 82:254–262. 1993.PubMed/NCBI
28	Adams CI, Keating JF and Court-Brown CM: Cigarette smoking and open tibial fractures. Injury. 32:61–65. 2001. View Article : Google Scholar : PubMed/NCBI
29	Kurdy NM: Serology of abnormal fracture healing: The role of PINP, PICP, and BsALP. J Orthop Trauma. 14:48–53. 2000. View Article : Google Scholar : PubMed/NCBI
30	Southwood LL, Frisbie DD, Kawcak CE and McIlwraith CW: Evaluation of serum biochemical markers of bone metabolism for early diagnosis of nonunion and infected nonunion fractures in rabbits. Am J Vet Res. 64:727–735. 2003. View Article : Google Scholar : PubMed/NCBI
31	Moghaddam A, Müller U, Roth HJ, Wentzensen A, Grützner PA and Zimmermann G: TRACP 5b and CTX as osteological markers of delayed fracture healing. Injury. 42:758–764. 2011. View Article : Google Scholar : PubMed/NCBI
32	Kaplan FS, Hayes WC, Keaveny TM, Boskey AL, Einhorn TA and Iannotti JP: Form and function of boneOrthopaedic basic science. Simon SS: Amercan Academy of Orthopaedic Surgeons; Rosemont, IL: pp. 127–184. 1994
33	Linkhart SG, Linkhart TA, Taylor AK, Wergedal JE, Bettica P and Baylink DJ: Synthetic peptidebased immunoassay for amino-terminal propeptide of type I procollagen: application for evaluation of bone formation. Clin Chem. 39:2254–2258. 1993.PubMed/NCBI
34	Eastell R, Yergey AL, Vieira NE, Cedel SL, Kumar R and Riggs BL: Interrelationship among vitamin D metabolism, true calcium absorption, parathyroid function, and age in women: Evidence of an age-related intestinal resistance to 1,25-dihydroxyvitamin D action. J Bone Miner Res. 6:125–132. 1991. View Article : Google Scholar : PubMed/NCBI
35	Sato Y, Kaji M, Higuchi F, Yanagida I, Oishi K and Oizumi K: Changes in bone and calcium metabolism following hip fracture in elderly patients. Osteoporos Int. 12:445–449. 2001. View Article : Google Scholar : PubMed/NCBI
36	Delmas PD: Biochemical markers of bone turnover for the clinical assessment of metabolic bone disease. Endocrinol Metab Clin North Am. 19:1–18. 1990.PubMed/NCBI
37	Seibel MJ: Molecular markers of bone turnover:Biochemical technical and analytical aspects. Osteoporos Int. 11:18–29. 2000. View Article : Google Scholar
38	Lin JP, Song SF and Yao LL: Feasibility of predicting fracture risk with bone turnover markers and bone mineral density. Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 14:317–320. 2010.
39	Szulc P and Delmas PD: Biochemical markers of bone turnover in men. Calcif Tissue Int. 69:229–234. 2001. View Article : Google Scholar : PubMed/NCBI
40	Laurer HL, Hagenbourger O, Quast S, Herrmann W and Marzi I: Sequential changes and pattern of bone specific alkaline phosphtase after trauma. Eur J Trauma. 1:33–38. 2000. View Article : Google Scholar
41	Dahabreh Z, Calori GM, Kanakaris NK, Nikolaou VS and Giannoudis PV: A cost analysis of treatment of tibial fracture nonunion by bone grafting or bone morphogenetic protein-7. Int Orthop. 33:1407–1414. 2009. View Article : Google Scholar : PubMed/NCBI
42	Borys J, Grabowska SZ, Antonowicz B, Dryl D, Citko A and Rogowski F: The concentuation of C-terminal propeptide of type I procollagen in blood serum in the course of mandibular fracture healing(preliminary report). Rocz Akad Med Bialmyst. 46:251–262. 2001.
43	Ingle BM, Hay SM, Bottjer HM and Eastell R: Changes in bone mass and bone turnover following ankle fracture. Osteporos Int. 10:408–415. 1999. View Article : Google Scholar
44	Kon T, Cho TJ, Aizawa T, Yamazaki M, Nooh N, Graves D, Gerstenfeld LC and Einhorn TA: Expression of osteoprotegerin, receptor activator of NF-kappaB ligand (osteoprotegerin ligand) and related proinflammatory cytokines during fracture healing. J Bone Miner Res. 16:1004–1014. 2001. View Article : Google Scholar : PubMed/NCBI
45	Hanson DA, Weis MA, Bollen AM, Maslan SL, Singer FR and Eyre DR: A specific immunoassay for monitoring human bone resorption: Quantitation of type I collagen cross-linked N-telopeptides in urine. J Bone Miner Res. 7:1251–1258. 1992. View Article : Google Scholar : PubMed/NCBI
46	Rosen HN, Moses AC, Garber J, Ross DS, Lee SL and Greenspan SL: Utility of biochemical markers of bone turnover in the follow-up of patients treatment with biphosphonates. Calcif Tissue Int. 63:363–368. 1998. View Article : Google Scholar : PubMed/NCBI