A New Method to Develop Human Dental Pulp Cells and Platelet-rich Fibrin Complex

Area of Science:

• Biomaterials Science
• Regenerative Medicine
• Cell Biology

Background:

• Platelet-rich fibrin (PRF) is utilized as a scaffold in tissue regeneration.
• Previous methods for combining PRF with seed cells compromised PRF structure and increased in vitro manipulation time.

Purpose of the Study:

• To develop an optimal method for creating a PRF-human dental pulp cell (hDPC) complex.
• To maintain the structural integrity of PRF during cell integration.
• To identify the most effective component of PRF for cell incorporation.

Main Methods:

• Developed PRF-hDPC complexes by adding hDPC suspension at different stages: before centrifugation (BC), immediately after centrifugation (IAC), and 10 minutes after centrifugation (AC).
• Cultured complexes for 7 days and evaluated using histologic, immunohistochemistry, and scanning electron microscopy.
• Assessed gene expression (alkaline phosphatase, dentin sialophosphoprotein) via real-time PCR and growth factor release via ELISA from different PRF parts.

Main Results:

• Histologic and microscopic analyses confirmed hDPC presence and proliferation exclusively in the BC group.
• Real-time PCR indicated increased expression of alkaline phosphatase and dentin sialophosphoprotein in the PRF-hDPC complex.
• The lower portion of the PRF yielded the highest concentration of released growth factors.

Conclusions:

• A novel method for developing PRF-hDPC complexes successfully maintained PRF structural integrity.
• Human dental pulp cells were primarily located in the buffy coat, suggesting this is the most efficient PRF component for cell integration.