IADR Abstract Archives
Doxycycline-Doped Nanoparticles Exert Cementogenic/Osteogenic Effect on Human Periodontal Ligament Cells
Objectives: To evaluate if doxycycline-doped polymeric nanoparticles exert effects on proliferation and cementogenic/osteogenic differentiation of stem cells from human periodontal ligament.
Methods: The polymeric nanoparticles (NPs) were produced by a polymerization/precipitation process. NPs were doped with doxycycline by NPs immersion in a 40 mg/mL aqueous solution of doxycycline hyclate (Dox-NPs). NPs were suspended in phosphate buffered solution at 1 µg/mL concentration. Stem cells were isolated from human periodontal ligament (PDLSCs). Two experimental groups were established for evaluation (Undoped NPs and Dox-NPs). A negative control and a positive control with osteogenic medium (OM) were also tested. PDLSCs were cultured in the presence or absence of the different NPs and cell proliferation was assessed by MTT-assay after 48h. Cell differentiation was also assessed by osteogenic gene expression measured with real-time quantitative polymerase chain reaction (RT-qPCR) after 7 and 14 d of culturing. The measured genes were alkaline phosphatase (ALP) and Runt-related transcription factor 2 (RUNX2). Data analysis was performed by one-way ANOVA and Student Newman Keuls multiple comparisons (p<0.05).
Results: No differences in cell proliferation were found at the different studied groups (p=0.103) (Control group average was 1, while NPs and Dox-NPs groups were 0.893 and 0.806, respectively). After 1 week of treatment, the osteogenic medium significantly upregulated ALP gene expression of PDLSCs (p=0.01) but not RUNX2 (p=0.08). After 2 weeks of treatment, gene expression of ALP and RUNX2 in PDLSCs were significantly upregulated by the osteogenic medium (ALP: p<0.001; RUNX2: p=0.005). Doxycycline NPs further enhanced (2-fold) ALP gene expression of PDLSCs treated with the osteogenic medium.
Conclusions: Dox-NPs increase cementogenic/osteogenic differentiation in PDLSCs, being a potential tool to be further developed for periodontal regeneration treatments.
Grant PID2020-114694RB-I00 funded by MCIN/AEI 10.13039/501100011033. M.T.-O. is fellow FPU of Ministry of Universities [grant FPU20/00450].
Methods: The polymeric nanoparticles (NPs) were produced by a polymerization/precipitation process. NPs were doped with doxycycline by NPs immersion in a 40 mg/mL aqueous solution of doxycycline hyclate (Dox-NPs). NPs were suspended in phosphate buffered solution at 1 µg/mL concentration. Stem cells were isolated from human periodontal ligament (PDLSCs). Two experimental groups were established for evaluation (Undoped NPs and Dox-NPs). A negative control and a positive control with osteogenic medium (OM) were also tested. PDLSCs were cultured in the presence or absence of the different NPs and cell proliferation was assessed by MTT-assay after 48h. Cell differentiation was also assessed by osteogenic gene expression measured with real-time quantitative polymerase chain reaction (RT-qPCR) after 7 and 14 d of culturing. The measured genes were alkaline phosphatase (ALP) and Runt-related transcription factor 2 (RUNX2). Data analysis was performed by one-way ANOVA and Student Newman Keuls multiple comparisons (p<0.05).
Results: No differences in cell proliferation were found at the different studied groups (p=0.103) (Control group average was 1, while NPs and Dox-NPs groups were 0.893 and 0.806, respectively). After 1 week of treatment, the osteogenic medium significantly upregulated ALP gene expression of PDLSCs (p=0.01) but not RUNX2 (p=0.08). After 2 weeks of treatment, gene expression of ALP and RUNX2 in PDLSCs were significantly upregulated by the osteogenic medium (ALP: p<0.001; RUNX2: p=0.005). Doxycycline NPs further enhanced (2-fold) ALP gene expression of PDLSCs treated with the osteogenic medium.
Conclusions: Dox-NPs increase cementogenic/osteogenic differentiation in PDLSCs, being a potential tool to be further developed for periodontal regeneration treatments.
Grant PID2020-114694RB-I00 funded by MCIN/AEI 10.13039/501100011033. M.T.-O. is fellow FPU of Ministry of Universities [grant FPU20/00450].