Effects of L‐PRF and A‐PRF+ on periodontal fibroblasts in in vitro wound healing experiments

Abstract

Objective To determine whether leukocyte‐platelet‐rich fibrin (L‐PRF) and advanced platelet‐rich fibrin (A‐PRF+) differ in their in vitro capacity to induce proliferation and migration of periodontal fibroblasts. Background L‐PRF and A‐PRF + are autologous materials used in periodontal regenerative surgery. They derive from blood from patients, but have different characteristics. The literature is controversial regarding the effects of the two PRF preparations on periodontal tissue fibroblasts. Materials and methods L‐PRF and A‐PRF + membranes were prepared from eight patients and incubated in 3 mL of culture medium for 2 days. Gingival fibroblasts (G‐F) and periodontal ligament fibroblast (PDL‐F) primary cells were retrieved from 7 donors. These cells were pre‐cultured for 1 day in wound healing experiment plates leaving a gap of 500 ± 50 µm in a concentration of 3.3 x 105 cells/mL. 70 µL of the cell suspension was placed in each half of the well. Thereafter, the pre‐cultured L‐PRF and A‐PRF + supernatants were added to the experimental plates, and the fibroblasts were incubated for another 24 h. Medium alone (NEG) and fibroblast growth factor II (FGF) were used as controls. Subsequently, cell migration was registered for 24 h with live cell imaging in a time frame microscope at 5% CO2 in air at 37°C. Images were analyzed using ImageJ. Cell proliferation and cell viability were measured. Results L‐PRF and A‐PRF + induced higher cell proliferation than FGF and NEG. Both A‐PRF + and L‐PRF induced significant faster artificial wound closure than controls. Both PRF conditioned media induced faster cell migration in the initial phase (P < .01), but in the stoppage phase, the induced migration was higher for the A‐PRF+, compared with L‐PRF (P < .01). Conclusion L‐PRF and A‐PRF + have a stimulatory effect on migration and proliferation of periodontal fibroblasts, and artificial wound closure was longer sustained by A‐PRF + than L‐PRF.

Deciphering the secretome of leukocyte-platelet rich fibrin: towards a better understanding of its wound healing properties

“…In recent years, many studies have analysed in vitro the impact of PRC in order to evaluate proliferation, migration, and differentiation of different cell types [11][12][13][14] . At the same time, other groups have tried to figure out which proteins provide the regeneration properties to PRC.…”

Hyperbaric Oxygen Therapy and A-PRF Pre-Treated Implants in Severe Periodontitis: A Case Report

“…Many studies support PDGF use for periodontal and soft tissue repair, but it seems to produce good results in bone regeneration and improve the success rate in immediate implantation [ 51 ]. A-PRF seems to induce a faster proliferation and healing rate than leukocyte and platelet-rich fibrin (L-PRF), fibroblast grow factor (FGF), or a negative control group [ 52 ]. The implants used in this case were soaked in A-PRF, a platelet concentrate obtained after the patient’s blood was centrifuged, which worked as a coating to help clotting, neovascularization, and osteointegration [ 53 ].…”

Simplified protocol for horizontal and vertical post-extractive GBR with intentionally exposed PTFE membrane - Case series

“…Horizontal periosteal incisions are not provided, the soft tissues can be mobilized with the Soft Brushing Technique (J. Choukroun), disorganizing the periosteal collagen fiber as the brush tools don't cut anything avoiding conventional flap: the aim is to decrease the tension and increase the release of the tissues, which have as more as possible to cover the new graft volume. PTFE membrane should be inserted first in lingual/palatal site, since usually is simpler; a bone graft must be fit in place, obtaining it by heterologous bone mixed with A-PRF (cut into small pieces) and solidifying with PRGF 10,11,12,13 , very handy and stable to be positioned in desired volume, overcompensating by 2-3mm. Then the membrane is curved over the graft and placed to cover whole vestibular dimension; it is very important that the membrane does not touch adjacent teeth, leaving almost 1-2mm from dental surface.…”

“…Our principal aim is to validate a simplified protocol for bone regeneration in post-extractive sites, avoiding vertical release incisions and flap displacement, in order to obtain new keratinized gingiva (Figure 4,11) by secondary closure of the surgical site and correct bone volume to place prosthetically planned implants. We used Open Barrier Technique (E. Funakoshi) with intentionally exposed d-PTFE (Cytoplast) membrane to protect the bone graft (Geistlich Bio-Oss ® with A-PRF and PRGF) 1,2,3,4,5,8,9 ; membranes have nanopores less than 0,3 microns to prevent bacterial infiltration and epithelium colonization of bone graft, but allow microscopical perfusion of oxygen and nutrition.…”