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Self-Assembled KLD-12/SDF-1 Polypeptide Promotes Differentiation and Migration of BMSCs via the Wnt/β-catenin
Mingyu Cao1, Yabin Hu2, Yukun Zhang3
1Department of Trauma Orthopedics, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang Uygur Autonomous Region, 830011, China.
Objective:
This study aimed to evaluate the combination of SDF-1 and KLD-12 to form self-assembling polypeptide and its effect on osteogenic differentiation.
Methods:
ELISA assay was performed to detect whether KLD-12 composite SDF-1 self-assembled polypeptide was successfully prepared. BMSCs were isolated and characterized by Flow cytometry. MTT assays, Calcein-AM/PI fluorescence staining, and Glycosaminoglycans (GAGs) measurement were carried out to detect cell viability after cells exposed to KLD-12 composite SDF-1 selfassembled polypeptide. The migration of cells induced by KLD-12 composite SDF-1 selfassembled polypeptide was also examined by transwell assay and Immunoblot. Osteogenic differentiation of cells stimulated with KLD-12 composite SDF-1 self-assembled polypeptide was analyzed by Immunoblot, Alizarin Red Staining, and Alkaline Phosphatase activity. Additionally, immunoblot and immunofluorescence assays were performed to investigate the effects of the polypeptide on the Wnt/β-catenin pathway.
Results:
KLD-12 composite SDF-1 self-assembled polypeptide was successfully prepared and identified. In addition, we isolated and characterized mouse mesenchymal stem BMSCs. Our data further revealed that KLD-12 combined with SDF-1 self-assembled polypeptide improved the survival of BMSCs and promoted cell migration. Moreover, the self-assembled polypeptide induced osteogenic differentiation of BMSCs. Mechanically, we found that the self-assembled polypeptide activated the Wnt/β-catenin pathway, therefore promoting the differentiation and migration of BMSCs.
Conclusion:
Our proposed treatment can potentially be effective for bone defects.
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