IN-VITRO ANALYSIS OF HEALING PROPERTIES OF FLAXSEED (LINUM USITATISSIMUM) EXTRACT ON HUMAN GINGIVAL FIBROBLAST (HGF) AND HUMAN OSTEOBLAST-LIKE (SAOS2) CELLS
SAMA NAZIYAH SHABAN
sama.nazih@uniq.edu.iq
Abstract:
The process of tooth extraction causes injury to the soft and hard tissues. The restoration of the damage tissues and the healing process takes few weeks in normal cases but could also extended to months in cases with complication. The bioactivities of flaxseed (linum usitatissimum) extract; also known as Biji Rami in Malay,such as antimicrobial, antioxidant and anti-inflammatory effects including the extract’s ability to enhance healing of skin wounds in-vivo have been reported. However, the effect of the extract on the socket healing process (post tooth extraction) has not been demonstrated and described. Thus, the current research might be the first in-vitro study investigating the effect of flaxseed extract on the socket healing process using two different cell lines; human gingival fibroblast (HGF) and human osteoblast-like (SaOS2) cell lines, representing the soft tissue and bone, respectively, both of which are involved in the process to mimic socket healing. Additionally, the effect of flaxseed extract on the expression of wound healing-related genes was also analysed on these cell lines. Flaxseed was extracted using soxhlet extraction method with both absolute ethanol (ABS EtOH) and 60% ethanol in water (60% EtOH) as an extraction medium. The active components of flaxseed extract from both extractions were assessed using gas chromatography mass spectrum (GC-MS) which demonstrated many active compounds such as lignan, oleic acid and linoleic acid. The activity of both cell lines against flaxseed extract in both ABS EtOH and 60% EtOH were analysed using colorimetric MTT (3- (4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay in three different time lines (24,48, and 72 hours) which revealed the HGF and SaOS2 cell proliferation activity in both extracts increasing by time. Wound healing activity was performed via scratch assay on both cell lines and the results were recorded in a time line manner (18,24,48,72, 96 and 120 hours) to closely resembled the wound healing ability of the extract. Interestingly, the results illustrated cell regeneration in the wound area occurs as early as 18 hours’ post flaxseed extract treatment which lasted for 5 days. Lastly, some wound healing related genes; EGFR, BMP2, and FGF2 were measured using real time q-PCR to confirm the wound healing activity of flaxseed extract on HGF and SaOS2 cell lines. The cells treated with flaxseed extract revealed a significant increase in the expression of the mentioned wound healing related genes. ABS EtOH flaxseed extract managed to increase the expression of these genes in both cell lines in significantly higher levels (p < 0.05) than the negative control, all the results were compared to a negative control to prove the activity of the extract. This study indicated that both absolute ethanol and ethanol/water flaxseed extract have wound healing effects on the cells that are involved in the process of socket healing, due to the major active components present in the extract such as lignan, linoleic acid and oleic acid; all of which aid in the process of wound healing, due to their high anti-inflammatory and anti-oxidant effects. This study also confirms that flaxseed extract elevates the expression of wound healing related genes hence it enhances the wound healing process. Therefore, flaxseed extract can be considered as a potential wound healing agent for oral wounds. Future in vivo analysis of oral wound in animal model will further justify the positive effects of flaxseed extract as a naturalbased wound healing agent.
