The influence of bone metabolism gene polymorphism on the level of encoded proteins in patients with long bone fractures

Introduction. Polymorphism of bone metabolism genes makes an important contribution to the course of reparative processes in fractures and can contribute to the disruption of consolidation.

The aim of the study. To identify the level of encoded proteins (OPG, IL6, TGFβ1, EGF) in patients with limb bone fractures depending on the carriage of bone metabolism gene polymorphisms (TNFRSF11B-1181G>C, IL6-174C>G, TGFβ1-25Arg>Pro, EGFR-2073A>T).

Materials and methods. The case-control study was performed in 108 patients: Group 1 (n = 64) with uncomplicated course; Group 2 (n = 46) – delayed consolidation. The analyzed groups of patients are comparable in clinical and epidemiological parameters and treatment. The control group was represented by 92 practically healthy individuals of the same sex and age. Two months after the injury, the levels of OPG, IL-6, TGFβ1, EGF were determined by ELISA, and gene polymorphism (TNFRSF11B-1181G>C, IL6174C>G, TGFβ1-25Arg>Pro, EGFR-2073A>T) was determined using standard primer sets “Litech” (Moscow). Statistical processing was performed by the IBM SPSS Statistics Version 25.0 program.

Results. The highest level of OPG was recorded in carriers of the CC genotype of the TN-FRSF11BG1181C gene in all studied groups, including the control group. In the group of patients with bone tissue repair disorders, the OPG content increased 1.1-fold relative to carriers of the GC genotype and 1.7-fold compared to carriers of the GG genotype. Analysis of the effects of SNP of the IL6C174G gene, the TGFb1Arg25Pro gene, and the EGFRA2073T gene showed opposite results: carriers of the GG, ProPro, and TT genotypes, respectively, had significantly lower concentrations of encoded proteins (IL-6, TGFb1, and EGF).

Conclusion. The content of OPG, IL-6, TGFβ1, EGF decreases in the carriage of the geno-types: -1181G/Gofthe TNFRSF11Bgene,-174G/Gofthe IL6 gene, -25Pro/Pro of the TGFβ1 gene, -2073T/T of the EGFR gene, respectively. The SNPs in question can be used to predict delayed consolidation in patients with limb bone fractures.

1. Castillo IA, Heiner JA, Meremikwu RI, Kellam J, Warner SJ. Where are we in 2022? A summary of 11,000 open tibia fractures over four decades. Journal of orthopaedic trauma. 2023; 37(8): e326-e334. doi: 10.1097/BOT.0000000000002602

2. Ekegren CL, Edwards ER, de Steiger R, Gabbe BJ. Incidence, costs and predictors of non-union, delayed union and mal-union following long bone fracture. International journal of environmental research and public health. 2018; 15(12): 2845. doi: 10.3390/ijerph15122845

3. Rupp M, Biehl C, Budak M, Thormann U, Heiss C, Alt V. Diaphyseal long bone nonunions – types, aetiology, economics, and treatment recommendations. International orthopaedics. 2018; 42(2): 247-258. doi: 10.1007/s00264-017-3734-5

4. Bowers KM, Anderson DE. Delayed Union and Nonunion: Current Concepts, Prevention, and Correction: A Review. Bioengineering (Basel, Switzerland). 2024; 11(6): 525. doi: 10.3390/bioengineering11060525

5. Yang N, Liu Y. The role of the immune microenvironment in bone regeneration. International journal of medical sciences. 2021; 18(16): 3697-3707. doi: 10.7150/ijms.61080

6. Katsimbri P. The biology of normal bone remodelling. European journal of cancer care. 2017; 26(6). doi: 10.1111/ecc.12740

7. Staroselnikov AN. Some pathogenetic mechanisms of delayed consolidation of fractures of long bones of the lower extremities / dissertation ... candidate of medical sciences: 3.3.3 / Staroselnikov Artem Nikolaevich. Chita, 2024; 145. (In Russ.).

8. Azevedo Filho FA, Cotias RB, Azi ML, Teixeira AA. Reliability of the radiographic union scale in tibial fractures (RUST). Revista brasileira de ortopedia. 2016; 52(1): 35-39. doi: 10.1016/j.rboe.2016.05.006

9. Mudrov VA. Statistical analysis algorithms of quantitative features in biomedical researchusing the SPSS software package. Transbaikal Medical Bulletin. 2020; 1: 140-150. (In Russ.). doi: 10.52485/19986173_2020_1_140

10. Mudrov VA. Statistical analysis algorithms of qualitativefeatures in biomedical researchusing the SPSS software package. Transbaikal Medical Bulletin. 2020; 1: 151-163. (In Russ.). doi: 10.52485/19986173_2020_1_151

11. Ono T, Takayanagi H. Osteoimmunology in bone fracture healing. Current osteoporosis report. 2017; 15(4): 367-375. doi: 10.1007/s11914-017-0381-0

12. Huang Z, Pei X, Graves DT. The interrelationship between diabetes, IL-17 and bone loss. Current osteoporosis reports. 2020; 18(1): 23-31. doi: 10.1007/s11914-020-00559-6

13. Graves DT, Ding Z, Yang Y. The impact of diabetes on periodontal diseases. Periodontology 2000. 2020; 82(1): 214-224. doi: 10.1111/prd.12318

14. Naot D, Wilson LC, Allgrove J, Adviento E, Piec I, Musson DS, et al. Juvenile Paget’s disease with compound heterozygous mutations in TNFRSF11B presenting with recurrent clavicular fractures and a mild skeletal phenotype. Bone. 2020; 130: 115098. doi: 10.1016/j.bone.2019.115098

15. Graves DT, Alshabab A, Albiero ML, Mattos M, Corrêa JD, Chen S, et al. Osteocytes play an important role in experimental periodontitis in healthy and diabetic mice through expression of RANKL. Journal of clinical periodontology. 2018; 45(3): 285-292. doi: 10.1111/jcpe.12851

16. Könnecke I, Serra A, El Khassawna T, Schlundt C, Schell H, Hauser A, et al. T and B cells participate in bone repair by infiltrating the fracture callus in a two-wave fashion. Bone. 2014; 64: 155-65. doi: 10.1016/j.bone.2014.03.052

17. Liu ZW, Zhang YM, Zhang LY, Zhou T, Li YY, Zhou GC, et al. Duality of Interactions Between TGF-β and TNF-α During Tumor Formation. Frontiers in immunology. 2022; 12: 810286. doi: 10.3389/fimmu.2021.810286

18. Zuo T, Liu Y, Li C, Tang J, Guo K. Correlations of IL-6 and TGF-β Gene Polymorphisms and Expressions With Osteoporotic, Thoracolumbar, Vertebral Compression Fracture. Alternative therapies in health and medicine. 2023; 29(3): 120-126.