Dental Pulp Stem Cell Conditioned Medium Enhancing Gingival Fibroblast Properties
Objectives: To investigate the biological effects of dental pulp stem cell-derived conditioned medium (DPSC-CM) on human gingival fibroblasts (HGFs). Methods: DPSC was isolated and characterized. DPSC-CM were obtained from cell culture supernatants after 2 days in serum-free medium. The total protein in DPSC-CM was determined using Bradford assay. For the cell viability assay using MTT, HGFs were treated with various concentrations of DPSC-CM (1.56, 3.12, 6.25, 12.5, 25, 50 and 100 µg/ml) in serum-free DMEM for 24 hours. For the proliferation assay using CCK-8 assay, HGFs were treated with 3.12 and 6.25 µg/ml of DPSC-CM in DMEM with serum. The proliferation of HGFs after treatment on days 1, 3, 5, and 7 was measured. For collagen production using picrosirius red staining, HGFs were treated with 3.12 and 6.25 µg/ml of DPSC-CM in DMEM with serum. The qualitative and quantitative analyses of collagen amount were conducted on days 7 and 14. The COL1A1 and COL3A1 gene expression was examined at days 7, 14 and 21 by real-time PCR. Results: HGFs treated with DPSC-CM at 3.12 and 6.25 µg/ml showed significantly higher cell viability than that of the control group without DPSC-CM. DPSC-CM at concentrations of 3.12 and 6.25 µg/ml significantly increased proliferation on day 7. Collagen amount was significantly higher in HGFs treated with 3.12 and 6.25 µg/ml of DPSC-CM at days 7 and 14, compared to the control group. Relative gene expression of COL3A1 was upregulated at days 7 and 14, compared to the control. The 6.25 µg/ml showed the highest expression of COL3A1 at both days, and of COL1A1 at day 14. At day 21, both COL3A1 and COL1A1 were down-regulated. Conclusions: DPSC-CM has the potential to promote the viability, proliferation, and collagen production in HGFs, indicating its promising properties to apply for enhancing oral wound healing.
2023 South East Asian Division Meeting (Singapore) Singapore
2023 111 Stem Cell Biology
Rueanson, Kittiphoom
( Faculty of Dentistry, Mahidol University
, Ratchathewi
, Bangkok
, Thailand
)
Tepsimanon, Kaniknun
( Faculty of Dentistry, Mahidol University
, Ratchathewi
, Bangkok
, Thailand
)
Puangprom, Nunnaphat
( Faculty of Dentistry, Mahidol University
, Ratchathewi
, Bangkok
, Thailand
)
Kaewmuangmoon, Jirattikarn
( Faculty of Dentistry, Mahidol University
, Bangkok
, --None--
, Thailand
)
Chantadul, Varunya
( Faculty of Dentistry, Mahidol University
, Bangkok
, --None--
, Thailand
)
Ruangsawasdi, Nisarat
( Faculty of Dentistry, Mahidol University
, Bangkok
, --None--
, Thailand
)
Janebodin, Kajohnkiart
( Faculty of Dentistry, Mahidol University
, Bangkok
, --None--
, Thailand
)