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- Academic Editor
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Background: Sappanwood is widely used in the prevention and treatment
in diseases due to its ability to seal blood vessels, dissipate stasis, and
relieve pain. Important monomer components of sappanwood, Protosappanin A (PA)
and Protosappanin B (PB) have anti-tumour and antimicrobial medicinal properties.
This study investigated the anti-inflammatory and osteogenic differentiation
effects of a crude extract of Sappanwood (ESP), PA and PB against periodontitis
in periodontal ligament stem cells (PDLSCs). Methods: Oil Red O staining
was used to assess the ability of adipocytes to differentiate. Alizarin Red
staining was used to assess the capacity to differentiate into osteoblasts.
Third-passage PDLSCs were grown in either basic medium alone or basic media with
varying doses of ESP (0.0625 mg/mL, 0.03125 mg/mL and 0.125 mg/mL), PA and PB
(2.5 µM, 5 µM, 10 µM). The CCK-8 assay was used to measure cell
proliferation. Real Time PCR (RT-qPCR) and Enzyme-Linked Immunosorbnent Assay
(ELISA) assay were used to measure gene expression. The capacity to differentiate
into osteoblasts was evaluated using Alizarin Red staining, and Alkaline
Phosphatase (ALP) staining and activity. Results: The development of
lipid droplets and mineralized nodules was examined using Oil Red O staining and
Alizarin Red staining. Flow cytometry revealed that PDLSCs were CD29 (98.23%)
and CD44 (98.81%) positive, but CD34 (0.16%) and CD45 (0.09%) negative. CCK-8
assay showed that ESP at three concentrations (0.03125 mg/mL, 0.0625 mg/mL and
0.125 mg/mL) and 2.5 µM, 5 µM and 10 µM PA and PB had no
cytotoxicity at 5 and 7 days (p