title: A New Method to Develop Human Dental Pulp Cells and Platelet-rich Fibrin Complex.
abstract: BACKGROUND
  Platelet-rich fibrin (PRF) has been used as a scaffold material in various tissue regeneration studies. In the previous methods to combine seed cells with PRF, the structure of PRF was damaged, and the manipulation time in vitro was also increased. The objective of this in vitro study was to explore an appropriate method to develop a PRF-human dental pulp cell (hDPC) complex to maintain PRF structure integrity and to find out the most efficient part of PRF.
  
  
  METHODS
  The PRF-hDPC complex was developed at 3 different time points during PRF preparation: (1) the before centrifugation (BC) group, the hDPC suspension was added to the venous blood before blood centrifugation; (2) the immediately after centrifugation (IAC) group, the hDPC suspension was added immediately after blood centrifugation; (3) the after centrifugation (AC) group, the hDPC suspension was added 10 minutes after blood centrifugation; and (4) the control group, PRF without hDPC suspension. The prepared PRF-hDPC complexes were cultured for 7 days. The samples were fixed for histologic, immunohistochemistry, and scanning electron microscopic evaluation. Real-time polymerase chain reaction was performed to evaluate messenger RNA expression of alkaline phosphatase and dentin sialophosphoprotein. Enzyme-linked immunosorbent assay quantification for growth factors was performed within the different parts of the PRF.
  
  
  RESULTS
  Histologic, immunohistochemistry, and scanning electron microscopic results revealed that hDPCs were only found in the BC group and exhibited favorable proliferation. Real-time polymerase chain reaction revealed that alkaline phosphatase and dentin sialophosphoprotein expression increased in the cultured PRF-hDPC complex. The lower part of the PRF released the maximum quantity of growth factors.
  
  
  CONCLUSIONS
  Our new method to develop a PRF-hDPCs complex maintained PRF structure integrity. The hDPCs were distributed in the buffy coat, which might be the most efficient part of PRF.
authors: Department of Operative Dentistry and Endodontology, College and Hospital of Stomatology, Guangxi Medical University, Nanning, Guangxi, China.
authors: Xuan
authors: X
authors: He
authors: Department of Operative Dentistry and Endodontology, College and Hospital of Stomatology, Guangxi Medical University, Nanning, Guangxi, China. Electronic address: angelaxiacw@163.com.
authors: Wen-Xia
authors: WX
authors: Chen
authors: Department of Operative Dentistry and Endodontology, College and Hospital of Stomatology, Guangxi Medical University, Nanning, Guangxi, China.
authors: Guifei
authors: G
authors: Ban
authors: Department of Operative Dentistry and Endodontology, College and Hospital of Stomatology, Guangxi Medical University, Nanning, Guangxi, China.
authors: Wei
authors: W
authors: Wei
authors: Department of Operative Dentistry and Endodontology, College and Hospital of Stomatology, Guangxi Medical University, Nanning, Guangxi, China.
authors: Jun
authors: J
authors: Zhou
authors: Department of Operative Dentistry and Endodontology, College and Hospital of Stomatology, Guangxi Medical University, Nanning, Guangxi, China.
authors: Wen-Jin
authors: WJ
authors: Chen
authors: Department of Operative Dentistry and Endodontology, College and Hospital of Stomatology, Guangxi Medical University, Nanning, Guangxi, China.
authors: Xian-Yu
authors: XY
authors: Li
mesh_terms: D000293:Adolescent; D000328:Adult; D001792:Blood Platelets / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002478:Cells, Cultured; D002498:Centrifugation; D003782:Dental Pulp / Q000166:cytology* / Q000000981:diagnostic imaging; D005337:Fibrin; D006801:Humans; D008964:Molar, Third; D012313:RNA / Q000096:biosynthesis; D012038:Regeneration / Q000502:physiology; D054457:Tissue Scaffolds; D055815:Young Adult
chemical_list: D012313:RNA; D005337:Fibrin
keywords: Human dental pulp cells; platelet-rich fibrin; platelet-rich fibrin–human dental pulp cell complex; scaffold