Investigation of the effects of inflammatory and metabolic factors on fracture union in head trauma and long bone fractures : Effect of head trauma and long bone fracture on biomediator levels

Abdülkadir  Sarı; Berna  Erdal; Aliye  Çelikkol; Mehmet Ümit  Çetin

doi:10.28982/josam.1084466

Authors

Abdülkadir Sarı Tekirdag Namık Kemal University Faculty of Medicine, Department of Orthopedics and Traumatology, Tekirdag, Turkey https://orcid.org/0000-0003-3416-5666
Berna Erdal Tekirdag Namık Kemal University Faculty of Medicine, Department of Medical Microbiology, Tekirdag, Turkey https://orcid.org/0000-0003-3375-7926
Aliye Çelikkol Tekirdag Namık Kemal University Faculty of Medicine, Department of Medical Biochemistry, Tekirdag, Turkey https://orcid.org/0000-0002-3799-4470
Mehmet Ümit Çetin Tekirdag Namık Kemal University Faculty of Medicine, Department of Orthopedics and Traumatology, Tekirdag, Turkey https://orcid.org/0000-0001-9827-8892

DOI:

https://doi.org/10.28982/josam.1084466

Keywords:

biomediator, bone metabolism, fracture healing, interleukin, traumatic brain injury

Abstract

Background/Aim: Fractures are the most common form of trauma in current orthopedic practice. Although studies have shed light on the relationship between the factors affecting the healing process after fracture, this process is still not fully understood. In this study, we aimed to investigate the changes in serum biomediator levels and fracture healing in different trauma patterns, such as head trauma (HT), long bone fracture (LBF), a combination of HT + LBF injury (CI), and in different time points of the healing period.

Methods: Forty Wistar rats were included in the study and divided into five groups. Group 1, the donor group, included rats with HT; Group 2 included rats with LBFs who were administered the serum taken from rats in Group 1; Group 3 included the rats with isolated LBFs; and Group 4 the rats with CI. Group 5 comprised the control rats. An experimental closed HT and fracture model was applied to rats. The rats in Groups 2, 3 and 4 were sacrificed on the 10^th, 20^th, and 30^thdays. The biomediator levels in the serum taken after sacrification were studied, while closed femoral fracture models were examined radiologically.

Results: Statistically significant differences were found among the groups regarding radiological scores on the 10^th, 20^th, and 30^th days. On Day 10, Group 2a had significantly higher scores than Group 3a (P=0.03), and Group 3a had lower scores than Group 4a (P=0.01). On Day 20, Group 2b had significantly higher scores than Group 3b (P=0.004) but lower than Group 4b (P=0.03). On Day 30, Group 2c had significantly higher scores than Group 3c but lower than Group 4c (P=0.001). The mean Ca, TGF beta 1, beta-catenin, IL-10, IL-17A, TNF alpha, CRP, Wnt-16, ALP, GH, PTH, IL-1 beta, IL-6, and IL-22 levels were significantly different among the groups (P<0.05). No significant difference was observed in the biomediator levels among the groups at different time points of the healing period.

Conclusion: We concluded that inflammatory cytokines (IL-1 beta, IL-6, IL-17A, IL-17F, IL-23, and TNF alpha) were elevated in the early period in individuals with isolated head trauma and that this effect could be transferred to other individuals by serum transfer. On the other hand, the negative relationship between the IL-10 level, which is a negative modulator in fracture union, and callus thickness was significant. Our study contributes by providing a molecular description of the positive union effect transferred between individuals by serum. We believe our findings will play a significant role in developing new therapeutic agents for fracture healing.

Downloads

Download data is not yet available.

References

Yu MD, Su BH, Zhang XX. Morphologic and molecular alteration during tibia fracture healing in rat. Eur Rev Med Pharmacol Sci. 2018 Mar;22(5):1233-40. doi: 10.26355/eurrev_201803_14463.

Zhang T, Yao Y. Effects of inflammatory cytokines on bone/cartilage repair. J Cell Biochem. 2019 May;120(5):6841-6850. doi: 10.1002/jcb.27953. DOI: https://doi.org/10.1002/jcb.27953

Echeverri LF, Herrero MA, Lopez JM, Oleaga G. Early stages of bone fracture healing: formation of a fibrin-collagen scaffold in the fracture hematoma. Bull Math Biol. 2015 Jan;77(1):156-83. doi: 10.1007/s11538-014-0055-3. DOI: https://doi.org/10.1007/s11538-014-0055-3

Copuroglu C, Calori GM, Giannoudis PV. Fracture non-union: who is at risk? Injury. 2013 Nov;44(11):1379-82. doi: 10.1016/j.injury.2013.08.003. DOI: https://doi.org/10.1016/j.injury.2013.08.003

Oryan A, Monazzah S, Bigham-Sadegh A. Bone injury and fracture healing biology. Biomed Environ Sci. 2015 Jan;28(1):57-71. doi: 10.3967/bes2015.006.

Morioka K, Marmor Y, Sacramento JA, Lin A, Shao T, Miclau KR, et al. Differential fracture response to traumatic brain injury suggests dominance of neuroinflammatory response in polytrauma. Sci Rep. 2019 Aug 21;9(1):12199. doi: 10.1038/s41598-019-48126-z. DOI: https://doi.org/10.1038/s41598-019-48126-z

Doblare M, Garcia JM, Gomez MJ. Modelling bone tissue fracture and healing: a review. Eng Fract Mech. 2004;71:1809-40. DOI: https://doi.org/10.1016/j.engfracmech.2003.08.003

Marmarou A, Foda MA, van den Brink W, Campbell J, Kita H, Demetriadou K. A new model of diffuse brain injury in rats. Part I: Pathophysiology and biomechanics. J Neurosurg. 1994 Feb;80(2):291-300. doi: 10.3171/jns.1994.80.2.0291. DOI: https://doi.org/10.3171/jns.1994.80.2.0291

Silva Ddos S, Brito JN, Ibiapina JO, Lima MF, Medeiros AR, Queiroz BH, et al. Traumatic brain injury: clinical and pathological parameters in an experimental weightdrop model. Acta Cir Bras. 2011 Apr;26(2):94-100. doi: 10.1590/s0102-86502011000200004. DOI: https://doi.org/10.1590/S0102-86502011000200004

Bonnarens F, Einhorn TA. Production of a standard closed fracture in laboratory animal bone. J Orthop Res. 1984;2(1):97-101. doi: 10.1002/jor.1100020115. DOI: https://doi.org/10.1002/jor.1100020115

Spencer RF. The effect of head injury on fracture healing. A quantitative assessment. J Bone Joint Surg Br. 1987 Aug;69(4):525-8. doi: 10.1302/0301-620X.69B4.3611151. DOI: https://doi.org/10.1302/0301-620X.69B4.3611151

Brandi ML. How innovations are changing our management of osteoporosis. Medicographia. 2010;32:1-6.

Schindeler A, McDonald MM, Bokko P, Little DG. Bone remodeling during fracture repair: The cellular picture. Semin Cell Dev Biol. 2008 Oct;19(5):459-66. doi: 10.1016/j.semcdb.2008.07.004. DOI: https://doi.org/10.1016/j.semcdb.2008.07.004

Mountziaris PM, Mikos AG. Modulation of the inflammatory response for enhanced bone tissue regeneration. Tissue Eng Part B Rev. 2008 Jun;14(2):179-86. doi: 10.1089/ten.teb.2008.0038. DOI: https://doi.org/10.1089/ten.teb.2008.0038

Smith R. Head injury, fracture healing and callus. J Bone Joint Surg Br. 1987 Aug;69(4):518-20. doi: 10.1302/0301-620X.69B4.3611149. DOI: https://doi.org/10.1302/0301-620X.69B4.3611149

Giannoudis PV, Mushtaq S, Harwood P, Kambhampati S, Dimoutsos M, Stavrou Z, et al. Accelerated bone healing and excessive callus formation in patients with femoral fracture and head injury. Injury. 2006 Sep;37 Suppl 3:S18-24. doi: 10.1016/j.injury.2006.08.020. Erratum in: Injury. 2007 Oct;38(10):1224. DOI: https://doi.org/10.1016/j.injury.2006.08.020

Gautschi OP, Cadosch D, Frey SP, Skirving AP, Filgueira L, Zellweger R. Serum-mediated osteogenic effect in traumatic brain-injured patients. ANZ J Surg. 2009 Jun;79(6):449-55. doi: 10.1111/j.1445-2197.2008.04803.x. DOI: https://doi.org/10.1111/j.1445-2197.2008.04803.x

Wang YH, Liu Y, Rowe DW. Effects of transient PTH on early proliferation, apoptosis, and subsequent differentiation of osteoblast in primary osteoblast cultures. Am J Physiol Endocrinol Metab. 2007 Feb;292(2):E594-603. doi: 10.1152/ajpendo.00216.2006. DOI: https://doi.org/10.1152/ajpendo.00216.2006

Khallaf FG, Kehinde EO, Hussein S. Bone Healing and Hormonal Bioassay in Patients with Long-Bone Fractures and Concomitant Head Injury. Med Princ Pract. 2016;25(4):336-42. doi: 10.1159/000445250. DOI: https://doi.org/10.1159/000445250

Estrores IM, Harrington A, Banovac K. C-reactive protein and erythrocyte sedimentation rate in patients with heterotopic ossification after spinal cord injury. J Spinal Cord Med. 2004;27(5):434-7. doi: 10.1080/10790268.2004.11752233. DOI: https://doi.org/10.1080/10790268.2004.11752233

Nam D, Mau E, Wang Y, Wright D, Silkstone D, Whetstone H, et al. T-lymphocytes enable osteoblast maturation via IL-17F during the early phase of fracture repair. PLoS One. 2012;7(6):e40044. doi: 10.1371/journal.pone.0040044. DOI: https://doi.org/10.1371/journal.pone.0040044

Wildemann B, Schmidmaier G, Ordel S, Stange R, Haas NP, Raschke M. Cell proliferation and differentiation during fracture healing are influenced by locally applied IGF-I and TGF-beta1: comparison of two proliferation markers, PCNA and BrdU. J Biomed Mater Res B Appl Biomater. 2003 Apr 15;65(1):150-6. doi: 10.1002/jbm.b.10512. DOI: https://doi.org/10.1002/jbm.b.10512

Alam I, Alkhouli M, Gerard-O'Riley RL, Wright WB, Acton D, Gray AK, et al. Osteoblast-Specific Overexpression of Human WNT16 Increases Both Cortical and Trabecular Bone Mass and Structure in Mice. Endocrinology. 2016 Feb;157(2):722-36. doi: 10.1210/en.2015-1281. DOI: https://doi.org/10.1210/en.2015-1281

Salehi A, Jullienne A, Baghchechi M, Hamer M, Walsworth M, Donovan V, et al. Up-regulation of Wnt/β-catenin expression is accompanied with vascular repair after traumatic brain injury. J Cereb Blood Flow Metab. 2018 Feb;38(2):274-89. doi: 10.1177/0271678X17744124. DOI: https://doi.org/10.1177/0271678X17744124

Bao Q, Chen S, Qin H, Feng J, Liu H, Liu D, et al. An appropriate Wnt/β-catenin expression level during the remodeling phase is required for improved bone fracture healing in mice. Sci Rep. 2017 Jun 2;7(1):2695. doi: 10.1038/s41598-017-02705-0. DOI: https://doi.org/10.1038/s41598-017-02705-0

Huang Y, Zhang X, Du K, Yang F, Shi Y, Huang J, et al. inhibition of β-catenin signaling in chondrocytes induces delayed fracture healing in mice. J Orthop Res. 2012 Feb;30(2):304-10. doi: 10.1002/jor.21505. DOI: https://doi.org/10.1002/jor.21505

Zhao C, Yu T, Dou Q, Guo Y, Yang X, Chen Y. Knockout of TLR4 promotes fracture healing by activating Wnt/β-catenin signaling pathway. Pathol Res Pract. 2020 Feb;216(2):152766. doi: 10.1016/j.prp.2019.152766. DOI: https://doi.org/10.1016/j.prp.2019.152766

Chan JK, Glass GE, Ersek A, Freidin A, Williams GA, Gowers K, et al. Low-dose TNF augments fracture healing in normal and osteoporotic bone by up-regulating the innate immune response. EMBO Mol Med. 2015 May;7(5):547-61. doi: 10.15252/emmm.201404487. DOI: https://doi.org/10.15252/emmm.201404487

Kon T, Cho TJ, Aizawa T, Yamazaki M, Nooh N, Graves D, et al. Expression of osteoprotegerin, receptor activator of NF-kappaB ligand (osteoprotegerin ligand) and related pro-inflammatory cytokines during fracture healing. J Bone Miner Res. 2001 Jun;16(6):1004-14. doi: 10.1359/jbmr.2001.16.6.1004. DOI: https://doi.org/10.1359/jbmr.2001.16.6.1004

Hengartner NE, Fiedler J, Ignatius A, Brenner RE. IL-1β inhibits human osteoblast migration. Mol Med. 2013 Apr 30;19(1):36-42. doi: 10.2119/molmed.2012.00058. DOI: https://doi.org/10.2119/molmed.2012.00058

Ozeki N, Mogi M, Hase N, Hiyama T, Yamaguchi H, Kawai R, et al. Wnt16 Signaling Is Required for IL-1β-Induced Matrix Metalloproteinase-13-Regulated Proliferation of Human Stem Cell-Derived Osteoblastic Cells. Int J Mol Sci. 2016 Feb 6;17(2):221. doi: 10.3390/ijms17020221. DOI: https://doi.org/10.3390/ijms17020221

Feyen JH, Elford P, Di Padova FE, Trechsel U. Interleukin-6 is produced by bone and modulated by parathyroid hormone. J Bone Miner Res. 1989 Aug;4(4):633-8. doi: 10.1002/jbmr.5650040422. DOI: https://doi.org/10.1002/jbmr.5650040422

Beeton CA, Chatfield D, Brooks RA, Rushton N. Circulating levels of interleukin-6 and its soluble receptor in patients with head injury and fracture. J Bone Joint Surg Br. 2004 Aug;86(6):912-7. doi: 10.1302/0301-620x.86b6.14176. DOI: https://doi.org/10.1302/0301-620X.86B6.14176

Thomas MV, Puleo DA. Infection, inflammation, and bone regeneration: a paradoxical relationship. J Dent Res. 2011 Sep;90(9):1052-61. doi: 10.1177/0022034510393967. DOI: https://doi.org/10.1177/0022034510393967

Kitami S, Tanaka H, Kawato T, Tanabe N, Katono-Tani T, Zhang F, et al. IL-17A suppresses the expression of bone resorption-related proteinases and osteoclast differentiation via IL-17RA or IL-17RC receptors in RAW264.7 cells. Biochimie. 2010 Apr;92(4):398-404. doi: 10.1016/j.biochi.2009.12.011. DOI: https://doi.org/10.1016/j.biochi.2009.12.011

Xu J, Wang Y, Li J, Zhang X, Geng Y, Huang Y, et al. IL-12p40 impairs mesenchymal stem cell-mediated bone regeneration via CD4+ T cells. Cell Death Differ. 2016 Dec;23(12):1941-51. doi: 10.1038/cdd.2016.72. DOI: https://doi.org/10.1038/cdd.2016.72

Li T, Zhang YM, Han D, Hua R, Guo BN, Hu SQ, et al. Involvement of IL-17 in Secondary Brain Injury After a Traumatic Brain Injury in Rats. Neuromolecular Med. 2017 Dec;19(4):541-54. doi: 10.1007/s12017-017-8468-4. DOI: https://doi.org/10.1007/s12017-017-8468-4

Grütz G. New insights into the molecular mechanism of interleukin-10-mediated immunosuppression. J Leukoc Biol. 2005 Jan;77(1):3-15. doi: 10.1189/jlb.0904484. DOI: https://doi.org/10.1189/jlb.0904484

Yang S, Ding W, Feng D, Gong H, Zhu D, Chen B, et al. Loss of B cell regulatory function is associated with delayed healing in patients with tibia fracture. APMIS. 2015 Nov;123(11):975-85. doi: 10.1111/apm.12439. DOI: https://doi.org/10.1111/apm.12439

Sarı A, Dinçel YM, Çetin MÜ, İnan S. Can fracture healing be accelerated by serum transfer in head trauma cases? An experimental head trauma model in rats. Jt Dis Relat Surg. 2021;32(2):306-12. doi: 10.52312/jdrs.2021.8. DOI: https://doi.org/10.52312/jdrs.2021.8