Tissue Eng Regen Med.  2021 Oct;18(5):895-904. 10.1007/s13770-021-00360-1.

Effect of Leukocyte-Platelet Rich Fibrin (L-PRF) on Tissue Regeneration and Proliferation of Human Gingival Fibroblast Cells Cultured Using a Modified Method

  • 1Department of Oral Implantology, Hospital of Stomatology, Jilin University, Changchun 130021, People’s Republic of China
  • 2Provincial Key Laboratory of Dental Development, Jaw Remodeling and Regeneration, Jilin University, Changchun 130021, People’s Republic of China
  • 3Department of Oral and Maxillofacial Surgery and Periodontology, Faculty of Dentistry, The Arab American University, Jenin 240, Palestine
  • 4Department of Dental Implantology, School and Hospital of Stomatology, Jilin University, No. 1500 Qinghua Rd, Chaoyang district, Changchun City 130021, Jilin Province, People’s Republic of China
  • 5College of Clinical Medicine, Jilin University, Changchun 130021, People’s Republic of China


An in vitro study on rapid culturing method of human gingival fibroblast cells (HGFCs) was established to investigate the potential use of the leukocyte-platelet rich fibrin (L-PRF) in tissue engineering technology, different medical fields, including periodontology and implantology.
Eight biopsies were obtained from eight different donors and a modified culturing technique was developed to obtain HGFCs. The modified 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay was used to compare the cell viability when the modified culturing method was used in comparison to the standard method. Blood samples were collected from the same patients and L-PRF was isolated using a standard protocol. The releases of platelet-derived growth factor-AA and transforming growth factor-beta1 at various time intervals were observed using enzyme-linked immunosorbent assay (ELISA) kit. The proliferative effect of L-PRF on HGFCs was assessed by the cell counting kit—8 assay.
A simple and rapid modified method for in vitro HGFC culture yielded a cellular monolayer within three to nine days after cell culture. L-PRF with three-dimensional polymer fibers released growth factors that peaked during the first three hours and continued to produce up to 10 days. The L-PRF presented a dose-dependent effect on HGFCs proliferation where HGFCs proliferation increased with an increase in L-PRF concentration.
The modified technique for the culture of HGFCs might be useful for the development of future experimental and clinical studies, besides L-PRF has great therapeutic potential in oral surgery fields.


Cell culture; Fibroblast; Growth factors; Platelet-rich fibrin; Tissue engineering
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