Self-Assembled KLD-12/SDF-1 Polypeptide Promotes Differentiation and Migration of BMSCs via the Wnt/β-catenin Signaling Pathways

  • Protein Pept Lett. 2022;29(10):851-858. doi: 10.2174/0929866529666220822124627.
Mingyu Cao  1 ,  Yabin Hu  2 ,  Yukun Zhang  3 ,  Jiang Xie  3 ,  Zengru Xie  1
Affiliations
  • 1. Department of Trauma Orthopedics, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang Uygur Autonomous Region, 830011, China.
  • 2. Department of Spine Surgery, The Second People's Hospital, Nanjin, Jiangsu, 210000, China.
  • 3. Department of the Spine Surgery, The Sixth Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang Uygur Autonomous Region,83-0092, China.
PMID: 35996270 DOI: 10.2174/0929866529666220822124627
Abstract

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.

Keywords
Bone defects; KLD-12; SDF-1; Wnt/β-catenin pathway; bone marrow-derived mesenchymal stem cells (BMSCs); self-assembled polypeptide.
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