IADR Abstract Archives
L-PRF Derived From Smokers/non-smokers Donors Induced Proliferation of Periodontal Cells
Objectives: Cigarette smoking has been associated with delayed wound healing. As well, in vitro evidence has reported alterations of proliferation and differentiation of periodontal cells. Clinical approaches aimed to improve wound healing have incorporated the use of platelet-derived fractions, including the use of Leucocyte platelet-rich fibrin (L-PRF) in non-smokers (NS) individuals. However, characterization of L-PRF derived from smokers (S) and its biological effects on periodontal cells have not been reported.
Objective: to evaluate the effects of L-PRF-derived conditioned media (LPRF-CM), from S or NS donors on periodontal ligament cells (PDLC) proliferation.
Methods: Methods. L-PRF clots from 4 donors (2NS and 2 S) were obtained using venous blood samples centrifuged at 400 x G for 12 minutes. L-PRF clots were incubated in DMEM at 37°C and were collected after 1 or 24 hours and 10 Days, to obtain conditioned medium (LPRF-CM). As same, periodontal ligament cells from healthy donors were cultured and stimulated with LPRF-CM. Likewise, conventional media supplemented with fetal bovine (FBS) serum 10% was included as positive control. After 24 hours of treatment, cells were fixed and immunostained for Ki67 proliferation marker and counterstained with DAPI. Images of each condition were taken and analyzed as a percentage of total cells per field. Results were analyzed statistically using ANOVA and turkey’s test for multiple comparisons.
Results: Results.
Our preliminary study showed that LPRF-CM derived from S or NS L-PRF-CM, taken at 1 or 24 hours were able to stimulate PDLC proliferation as the same level as conventional media, supplemented with FBS (80%, average). However, this proliferative effect was significantly reduced with LPRF-CM taken at 10D after the initial isolation (40%, average), from both, NS and S.
Conclusions: Conclusions
Preliminarily, our results suggested a similar proliferative effect of LPRF-CM derived from S or NS took at 1 or 24 hours after LPRF clot obtaining.
Funding Fondecyt 1181007 (CM)
Objective: to evaluate the effects of L-PRF-derived conditioned media (LPRF-CM), from S or NS donors on periodontal ligament cells (PDLC) proliferation.
Methods: Methods. L-PRF clots from 4 donors (2NS and 2 S) were obtained using venous blood samples centrifuged at 400 x G for 12 minutes. L-PRF clots were incubated in DMEM at 37°C and were collected after 1 or 24 hours and 10 Days, to obtain conditioned medium (LPRF-CM). As same, periodontal ligament cells from healthy donors were cultured and stimulated with LPRF-CM. Likewise, conventional media supplemented with fetal bovine (FBS) serum 10% was included as positive control. After 24 hours of treatment, cells were fixed and immunostained for Ki67 proliferation marker and counterstained with DAPI. Images of each condition were taken and analyzed as a percentage of total cells per field. Results were analyzed statistically using ANOVA and turkey’s test for multiple comparisons.
Results: Results.
Our preliminary study showed that LPRF-CM derived from S or NS L-PRF-CM, taken at 1 or 24 hours were able to stimulate PDLC proliferation as the same level as conventional media, supplemented with FBS (80%, average). However, this proliferative effect was significantly reduced with LPRF-CM taken at 10D after the initial isolation (40%, average), from both, NS and S.
Conclusions: Conclusions
Preliminarily, our results suggested a similar proliferative effect of LPRF-CM derived from S or NS took at 1 or 24 hours after LPRF clot obtaining.
Funding Fondecyt 1181007 (CM)