Predictive genetic markers of coagulation, inflammation and apoptosis in Perthes disease—Serbian experience

Abstract

Perthes disease is one of the most common forms of pediatric femoral head osteonecrosis with an unknown etiology. Coagulation factors were the first genetic factors suspected to have a role in the pathogenesis of this disease, but studies showed inconsistent results. It is described that inflammation is present during early stages of Perthes disease, but its genetic aspect has not been studied extensively. Little is known regarding the status of apoptotic factors during the repair process that leads to the occurrence of hip deformity in patients. Therefore, the aim of this study was to analyze major mediators involved in coagulation, inflammation, and apoptotic processes as possible causative factors of Perthes disease. The study cohort consisted of 37 patients. Gene variants of TNF-α, FV, FII, and MTHFR genes were determined by PCR-RFLP, while IL-3 and PAI-1 were genotyped by direct sequencing. The expression level of Bax, Bcl-2, Bcl2L12, Fas and FasL was analyzed by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) technique. Our results showed a significantly increased level of expression of pro-apoptotic factor Bax along with significantly higher Bax/Bcl-2 ratio in the patient group.

Conclusion: The results presented indicate that apoptosis could be one of the factors contributing to the lack of balanced bone remodeling process in Perthes patients.

What is Known:
The etiology of Perthes disease is unknown. The role of genetic factors involved in the coagulation process has been studied, showing inconsistent results so far.
Genetic factors involved in inflammation and apoptotic processes that could contribute to development of hip deformity have not been studied extensively.
What is New:
Our results show significantly increased level of expression of the pro-apoptotic factor Bax as well as significantly higher Bax/Bcl-2 ratios in patient group, indicating that apoptosis could be one of the factors contributing to the lack of a balanced bone remodeling process in Perthes patients.

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Fig. 1

Abbreviations

BAX:

Bcl-2-associated X protein

BCL-2:

B cell lymphoma 2

BCL2L12:

BCL2-like 12

FAS:

Fas receptor

FASL:

Fas ligand

FV:

Factor V Leiden

FII:

Factor II

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

IL-3:

Interleukin-3

LCPD:

Legg–Calve–Perthes’ disease

MTHFR:

Methylenetetrahydrofolate reductase

PAI-1:

Plasminogen activator inhibitor-1

PBMNC:

Peripheral blood mononuclear cells

TNF-α:

Tumor necrosis factor alpha

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Acknowledgments

This work was supported by the Ministry of Education, Science and Technological Development, Republic of Serbia (Grant No. III41004). This study is in memoriam of Prof. Zoran Vukasinovic, who was the research coordinator for the subproject: “Orthopedic rare diseases: molecular basis of Perthes disease.” His enthusiasm and willpower contributed most to the opening of this new research topic in Serbia.

Ethical statements

The study was approved by the Ethics Committee of the Institute for Orthopedic Surgery “Banjica,” Belgrade, Serbia (I-86/34; 7-2, 12. 12. 2012.). Written informed consent was obtained for all patients.

Conflict of interest

The authors declare that they have no conflict of interest.

Author’s Contributions

DS, ZB and ZŽ performed the case and control sample collection and clinical management of patients. ZTŠ and DM participated in clinical management of patients and participated in studz design. VDJ provided control sample for coagulation study and participated in data analyzes. SS and VS performed the laboratory work and statistical analysis. SS and GN analyzed data and wrote first draft of the paper. VS and SP designed the study and wrote the final version of the paper. VS take the primary responsibility for the paper.

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Correspondence to Vesna Spasovski.

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Sanja Srzentić and Gordana Nikčević contributed equally to this work.

Communicated by Beat Steinmann

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Srzentić, S., Nikčević, G., Spasovski, D. et al. Predictive genetic markers of coagulation, inflammation and apoptosis in Perthes disease—Serbian experience. Eur J Pediatr 174, 1085–1092 (2015). https://doi.org/10.1007/s00431-015-2510-z

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Keywords

  • Blood coagulation factors
  • Bone remodeling
  • Inflammation
  • Apoptosis
  • Perthes disease