Effect of recombinant Sox9 protein on the expression of cartilage-specific genes in human dermal fibroblasts cell culture
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 scaffol...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Belgorod National Research University
2022-12-01
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| Series: | Research Results in Pharmacology |
| Online Access: | https://rrpharmacology.ru/article/90447/download/pdf/ |
| _version_ | 1797429258572791808 |
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| author | Mikhail S. Bozhokin Svetlana A. Bozhkova Julia V. Sopova Elena R. Mikhailova Daria V. Marchenko Mikhail G. Khotin |
| author_facet | Mikhail S. Bozhokin Svetlana A. Bozhkova Julia V. Sopova Elena R. Mikhailova Daria V. Marchenko Mikhail G. Khotin |
| author_sort | Mikhail S. Bozhokin |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-09T09:10:35Z |
| format | Article |
| id | doaj.art-01eb493cb72f4ad998e9decb59522010 |
| institution | Directory Open Access Journal |
| issn | 2658-381X |
| language | English |
| last_indexed | 2024-03-09T09:10:35Z |
| publishDate | 2022-12-01 |
| publisher | Belgorod National Research University |
| record_format | Article |
| series | Research Results in Pharmacology |
| spelling | doaj.art-01eb493cb72f4ad998e9decb595220102023-12-02T08:40:38ZengBelgorod National Research UniversityResearch Results in Pharmacology2658-381X2022-12-018410911410.3897/rrpharmacology.8.9044790447Effect of recombinant Sox9 protein on the expression of cartilage-specific genes in human dermal fibroblasts cell cultureMikhail S. Bozhokin0Svetlana A. Bozhkova1Julia V. Sopova2Elena R. Mikhailova3Daria V. Marchenko4Mikhail G. Khotin5Institute of Cytology of the Russian Academy of ScienceVreden National Medical Research Centre of Traumatology and OrthopedicsSaint-Petersburg branch of Vavilov Institute of General GeneticsInstitute of Cytology of the Russian Academy of ScienceInstitute of Cytology of the Russian Academy of ScienceInstitute of Cytology of the Russian Academy of ScienceIntroduction: 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.https://rrpharmacology.ru/article/90447/download/pdf/ |
| spellingShingle | Mikhail S. Bozhokin Svetlana A. Bozhkova Julia V. Sopova Elena R. Mikhailova Daria V. Marchenko Mikhail G. Khotin Effect of recombinant Sox9 protein on the expression of cartilage-specific genes in human dermal fibroblasts cell culture Research Results in Pharmacology |
| title | Effect of recombinant Sox9 protein on the expression of cartilage-specific genes in human dermal fibroblasts cell culture |
| title_full | Effect of recombinant Sox9 protein on the expression of cartilage-specific genes in human dermal fibroblasts cell culture |
| title_fullStr | Effect of recombinant Sox9 protein on the expression of cartilage-specific genes in human dermal fibroblasts cell culture |
| title_full_unstemmed | Effect of recombinant Sox9 protein on the expression of cartilage-specific genes in human dermal fibroblasts cell culture |
| title_short | Effect of recombinant Sox9 protein on the expression of cartilage-specific genes in human dermal fibroblasts cell culture |
| title_sort | effect of recombinant sox9 protein on the expression of cartilage specific genes in human dermal fibroblasts cell culture |
| url | https://rrpharmacology.ru/article/90447/download/pdf/ |
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