IADR Abstract Archives
LPRF conditioned media stimulates human periodontal cell proliferation.
Objectives: The main goal of our study was to evaluate the effects of conditioned media isolated from L-PRF plugs on the proliferation of periodontal ligament mesenchymal cells.
Methods: All procedures were approved by the Ethical committee of health science at Pontificia Universidad Católica de Chile. Briefly, cells isolated from human periodontal ligament (LPH) were seeded in 24-well plates and treated for 24 and 48 hours with conditioned media obtained from LPRF plugs. For LPRF plug obtention, venous blood samples from healthy donors were centrifuged at 406 Xg for 12 minutes. LPRF clots were compressed using pistons in a stainless-steel device and immersed in cell culture media. We recovered and changed this media after 3 hours, 3 days and 7 days. Likewise, exudates released after LPRF clot compression were recollected. As controls, we used conventional cell culture media, DMEM supplemented with 10% or 0.1% fetal bovine serum. Then, cells were fixed with 4% and immunostaining with the Ki67 proliferation marker was done. Total number of cells was evidenced by DAPI nuclear staining. Finally, images on epifluorescence microscopy were taken to determine the percentage of positive Ki67 cells.
Results: The conditioned media collected from LPRF plus, promote the proliferation of periodontal cells after 24 and 48 hours, even in a greater proportion than the conventional culture medium. Cells incubated with exudates similarly stimulated cell proliferation. Being greater in those treated at 40%.
Conclusions: Conditioned media from LPRF plugs were able to stimulate periodontal cell proliferation, an important biological activity during the wound healing process.
Methods: All procedures were approved by the Ethical committee of health science at Pontificia Universidad Católica de Chile. Briefly, cells isolated from human periodontal ligament (LPH) were seeded in 24-well plates and treated for 24 and 48 hours with conditioned media obtained from LPRF plugs. For LPRF plug obtention, venous blood samples from healthy donors were centrifuged at 406 Xg for 12 minutes. LPRF clots were compressed using pistons in a stainless-steel device and immersed in cell culture media. We recovered and changed this media after 3 hours, 3 days and 7 days. Likewise, exudates released after LPRF clot compression were recollected. As controls, we used conventional cell culture media, DMEM supplemented with 10% or 0.1% fetal bovine serum. Then, cells were fixed with 4% and immunostaining with the Ki67 proliferation marker was done. Total number of cells was evidenced by DAPI nuclear staining. Finally, images on epifluorescence microscopy were taken to determine the percentage of positive Ki67 cells.
Results: The conditioned media collected from LPRF plus, promote the proliferation of periodontal cells after 24 and 48 hours, even in a greater proportion than the conventional culture medium. Cells incubated with exudates similarly stimulated cell proliferation. Being greater in those treated at 40%.
Conclusions: Conditioned media from LPRF plugs were able to stimulate periodontal cell proliferation, an important biological activity during the wound healing process.