Journal of the Korean Association of Oral and Maxillofacial Surgeons

The Korean Association of Oral and Maxillofacial Surgeons (대한구강악안면외과학회)
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Effects of ascorbic acid augmented albumin platelet-rich fibrin on the wound healing activity of human gingival fibroblasts: an in vitro trial

  • Manjiri Kulkarni (Department of Periodontics, Bapuji Dental College and Hospital) ;
  • Sowmya NK (Department of Periodontics, Bapuji Dental College and Hospital) ;
  • Gayathri GV (Department of Periodontics, Bapuji Dental College and Hospital) ;
  • Triveni MG (Department of Periodontics, Bapuji Dental College and Hospital)
  • Received : 2024.03.14
  • Accepted : 2024.06.19
  • Published : 2024.08.31
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Abstract

Objectives: The current in vitro study aimed to assess the effects of ascorbic acid augmented albumin platelet-rich fibrin (AA Alb-PRF) on the wound healing activity of human gingival fibroblasts (HGFs) purported to be a regenerative biomaterial in surgical procedures. Materials and Methods: All assays were performed on three HGF groups, group I: complete media; group II: Alb-PRF, and group III: AA Alb-PRF. Alb-PRF was prepared following the protocol by Fujioka-Kobayashi et al. (2021). For preparation of AA Alb-PRF, 2,500 ㎍ AA was added to the blood pre-centrifugation. All groups were subjected to 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to estimate cell viability and proliferation, scratch assay for migration (0, 4, 12, and 24 hours) and transwell migration assay for chemotactic migration assessment (24 hours). Outcome variables were optical density (OD) for MTT assay, percentage of wound closure in scratch assay, and number of migrated cells in transwell migration assay. One-way ANOVA for MTT and transwell migration assays and two-way ANOVA for scratch assay with Bonferroni correction were performed with significance set at P<0.05. Results: Cell viability and proliferation (OD: 0.684±0.003 and proliferation: 28%) and wound closure (49.92%±1.62% at 4 hours and 61.39%±0.88% at 12 hours) were significantly higher in group III, while group II demonstrated the maximum number of HGFs migrating across the transwell membrane (9.25±2.49) with P<0.05. Conclusion: HGFs demonstrated a significant increase in viability and proliferation along with rapid wound closure in the presence AA Alb-PRF compared to Alb-PRF alone, indicating additional beneficial effects of AA. Thus, AA Alb-PRF potentiates the wound healing activity of HGFs and could be employed in oral, maxillofacial, and periodontal surgeries as a regenerative biomaterial.

Keywords

Acknowledgement

The authors would like to thank Dr. Kishore G Bhat and Dr. Chetana Bogar at the Central Research Laboratory of Maratha Mandal's NGH Institute of Dental Sciences, Belagavi, Karnataka, India, for their help and support with the cell culture and all the assays carried out in this study. We would also like to acknowledge Dr. Usha GV, Department of Public Health Dentistry, Bapuji Dental College and Hospital, Davangere, Karnataka, India, for the statistical analysis of the data.

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