Effect of recombinant Sox9 protein on the expression of cartilage-specific genes in human dermal fibroblasts cell culture

Authors

DOI:

https://doi.org/10.3897/rrpharmacology.8.90447

Abstract

Introduction: Damage to the hyaline layer of large joints resulting from injuries or age-related changes restricts their mobility. The repair of these disorders is an actual issue in medicine. One of the promising therapies is the usage of cell engineering constructs based on a biodegradable scaffold and a modified cell culture. A frequently used method to modify the proliferation of cell culture for tissue engineering of hyaline cartilage, which makes it possible to introduce an experimental technique into clinical practice, is the application of recombinant proteins that affect chondrogenesis and lead to increase synthesis of extracellular matrix proteins. The goal of this work was to elucidate the effect of the key transcription factor in the chondrogenesis process – Sox9 protein – on the expression of genes responsible for chondrogenesis (Tgfβ3, Sox9, Acan, Comp, Col2a1).

Materials and methods: Human dermal fibroblasts were used as a cell culture; recombinant Sox9 was added at each change of medium; the modification was carried out for 21 days, and difference in gene expression was determined by real-time PCR and -ΔΔCt method.

Results and discussion: To assess the effectiveness of fibroblast modification, we analyzed the changing of expression of genes responsible for chondrogenesis (Tgfß3, Sox9, Col2a1, Acan, Comp). We studied the direct effect of different concentrations of the recombinant Sox9 protein on the proliferation of dermal fibroblasts in the chondrogenic direction. We showed that the addition of the recombinant Sox9 protein in various concentration did not significantly change the expression of both the genes encoding proteins of the extracellular matrix of hyaline cartilage (Acan, Col2a1, Comp) and the genes encoding chondrogenesis inducers (Tgfß3, Sox9).

Conclusion: As a result of the experiments, it was shown that the recombinant Sox9 protein has practically no effect on chondrogenic differentiation and does not significantly change the expression of chondrogenesis genes.

Keywords:

Sox9, chondrogenesis, fibroblasts, gene expression, recombinant proteinroloquinolones

Author Contribution

Mikhail S. Bozhokin, Institute of Cytology of the Russian Academy of Science; Vreden National Medical Research Centre of Traumatology and Orthopedics

Assistant researcher, Vreden National Medical Research Center for Traumatology and Orthopedics.

Svetlana A. Bozhkova, Vreden National Medical Research Centre of Traumatology and Orthopedics

Dr. Habil. Sci. (Med.), Head of the Research Department of Prevention and Treatment of Wound Infection and Department of Clinical Pharmacology.

Julia V. Sopova, St. Petersburg State University; Saint-Petersburg branch of Vavilov Institute of General Genetics

PhD in Biology, leading researcher, Center of Tansgenesic and Genome Editing, Institute of Translational Biomedicine.

Elena R. Mikhailova, Institute of Cytology of the Russian Academy of Science

PhD in Biology, researcher, Laboratory of Cell Protection Mechanisms.

Daria V. Marchenko, Institute of Cytology of the Russian Academy of Science

Researcher, Laboratory of Cell Protection Mechanisms, Institute of Cytology of the Russian Academy of Sciences.

Mikhail G. Khotin, Institute of Cytology of the Russian Academy of Science

PhD in Biology, Head of the Laboratory of Biomedical Technologies and Testing with Pilot Production, Cell Technologies Center.

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Published

15-12-2022

How to Cite

Bozhokin MS, Bozhkova SA, Sopova JV, Mikhailova ER, Marchenko DV, Khotin MG (2022) Effect of recombinant Sox9 protein on the expression of cartilage-specific genes in human dermal fibroblasts cell culture. Research Results in Pharmacology 8(4): 109–114. https://doi.org/10.3897/rrpharmacology.8.90447

Issue

Section

Review article