Morphometric Features of Growth Plate Cells in the Proximal Tibia in Blount's Disease

The aim of the study was to study the morphometric parameters of growth plate cells in the proximal tibia in Blount's disease.

Material and methods. The study was conducted on cells isolated from the medial growth plates of the proximal tibia in Blount's disease grade III–IV. The material was obtained from 12 children (4 boys and 8 girls) (n=12) aged 3–9 years at the clinic of pediatric orthopedics of the Ya.L. Tsivyan Novosibirsk Research Institute of Traumatology and Orthopedics. The cells were cultured from passages 1 to 4, fixed, and stained for key chondrogenic (collagen type II and SOX₉) and neural (SOX₂, SOX₁₀, PAX₃, PAX₆, MSI₁, Nestin, HNK₁, TUBB₃, NF₂₀₀) markers. Morphometric analysis of immunopositive cells was performed using ImageJ software. The obtained data were processed using standard statistical methods with the application of the Shapiro–Wilk test and Student's t-test.

Results. Fluorescence staining of most growth plate cells revealed a positive reaction for the studied chondrogenic markers. Among these immunopositive cells, bipolar and multipolar cells with two or more processes expressing early (SOX₂, SOX₁₀, PAX₃, PAX₆, MSI₁, Nestin, HNK₁) and late (TUBB₃, NF₂₀₀) neural markers were identified. During cultivation, an increase in the proportion of positively stained cells for the majority of the studied neural markers was observed, along with an increase in fluorescence intensity, an increase in the number of processes, and an increase in the mean process length. Statistically significant differences in morphometric parameters (nuclear area, cytoplasmic area, and cell area) combined with differences in the nuclear-to-cytoplasmic ratio were found between cells expressing chondrogenic and neuronal markers.

Conclusion. The results indicate the presence of cells of neural origin in the growth plate of the proximal tibia in Blount's disease. Positive staining for the transcription factors PAX₃, PAX₆, SOX₂, SOX₁₀, as well as for the markers HNK₁, MSI₁, Nestin, and TUBB₃, supports the hypothesis regarding the role of neural crest cells in the development of this pathology.

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