Abstract
Rhizomes of Reynoutria japonica Houtt. are a traditional Chinese medicinal herb (Polygoni cuspidati rhizoma, hu zhang) used for treatment of numerous diseases including wound healing support. The aim of this study was to provide evidence for the value of this herbal drug’s traditional use as a gingival healing treatment as well as to obtain the most active extract. In vitro studies were performed using primary human gingival fibroblasts (HGFs) with determination of viability (MTT assay), cell proliferation (the confocal laser scanning microscope (CLSM) was used to visualize histone 3 expression), cell migration (wound healing assay), and evaluation of the expression of collagen type III (immunocytochemical staining) after incubation with extracts from R. japonica rhizomes (25% or 40% ethanol or 60% acetone). In addition to these extracts, commercial dental rinse (containing chlorhexidine digluconate 0.2%) was tested as the gold standard of choice for gum healing in dental practice. The studied extracts were qualitatively and quantitatively characterized using the validated HPLC/DAD/ESI-HR-QTOF-MS method. Total phenols and tannins content were determined using the Folin–Ciocalteu assay. Low concentration of all extracts after 24 h incubation caused significant increase in HGF viability. This effect was most pronounced at a concentration of 50 µg/mL, which was selected for further experiments. All extracts (at 50 µg/mL) stimulated HGF to proliferate, migrate, and increase collagen III synthesis, but with different strength. The highest stimulated proliferation and migration activity was observed after incubation with 25% EtOH, which according to phytochemical analysis may be related to the highest content of resveratrol and an appropriate composition of procyanidins. The 25% EtOH extract from R. japonica rhizomes appears to be a promising gingival wound healing agent worthy of animal and clinical trials.
Highlights
We based the study on our earlier research [5], in which we showed that 70% acetone extract from rhizomes of R. japonica demonstrated antibacterial activity against dominant caries pathogen-Streptococcus mutans and increased viability of human gingival fibroblasts (MTT assay, up to 138% compared with the control) after 24 h incubation with low concentrations of extracts
These results are in accordance with our previous report [5], where incubation of normal human gingival fibroblasts with 50 or 250 μg/mL of 70% acetone extract caused a significant increase in their viability
The 25% EtOH, 40% EtOH, and 60% acetone extracts from rhizomes of R. japonica stimulated gingival fibroblasts to proliferate and migrate, as well as resulting in increased synthesis of collagen III
Summary
Fallopia japonica (Houtt.) Ronse Decr., Polygonum cuspidatum Sieb. & Zucc.; placed within the family Polygonaceae, genus Fallopia, section Reynoutria,) are a traditional Chinese medicinal herb (known in pharmacopoeias as Polygoni cuspidati rhizoma and as hu zhang in pinyin Chinese) that have long been used for treatment of numerous diseases, such as hyperlipemia, inflammation, infection, and cancer, etc. Pharmaceutics 2021, 13, 1764 ago [1]. In later monographs of Materia Medica: Dian-nan Bencao, Rihuazi Bencao, Sichuan
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