Anat Cell Biol.  2019 Sep;52(3):324-332. 10.5115/acb.19.009.

Influence of traditional medicines on the activity of keratinocytes in wound healing: an in-vitro study

• Kotian SR
• Bhat KM
• Padma D
• Pai KS

• KMID: 2459548
• DOI: http://doi.org/10.5115/acb.19.009

Abstract

Natural medicinal systems such as Ayurveda and folk medicine has remedies for wound management. However, the exact cellular and extracellular mechanisms involved in the healing process and its influence on keratinocytes is less discussed. Therefore, the present study was designed to evaluate the effect of certain natural wound healing medicines on the biology of the keratinocytes/HaCaT cells. Test materials such as honey (H), ghee (G), aqueous extracts of roots of Glycyrrhiza glabra (GG) and leaves of Nerium indicum (NI) were considered. The HaCaT cells were treated with the test materials singly and in combinations (H+G, all combined [Tot]) for a specific period (24, 48, and 72 hours). The cells were then subjected to cytotoxicity/proliferation and migration/scratch assays. All the test materials, except NI, were non-cytotoxic and showed increased cell proliferation at variable concentrations. Significant observations were made in the groups treated with honey (100 µg/ml at 48 hours, P<0.05; 1,000 µg/ml at 72 hours, P<0.05), GG (all concentrations at 48 hours, P<0.05; 750 µg/ml at 72 hours, P<0.05), H+G (250 µg/ml at 24 hours, P<0.001; 500 µg/ml at 48 and 72 hours, P<0.05), and Tot (50 µg/ml at 24, 48 and 72 hours, P<0.01). In the in-vitro wound healing assay, all the treated groups showed significant migration and narrowing of the scratch area by 24 and 48 hours (P<0.001) compared to control. The results obtained from the present study signifies the positive influence of these natural wound healing compounds on keratinocytes/HaCaT cells.

Keyword

HaCaT cells; Honey; Ghee; Glycyrrhiza glabra; Nerium indicum

MeSH Terms

Biology
Cell Proliferation
Ghee
Glycyrrhiza
Honey
Keratinocytes*
Medicine, Traditional
Nerium
Wound Healing*
Wounds and Injuries*
Ghee

Figure

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  Fig. 1 Proliferative effect of different doses of the honey (A) and ghee (B) on keratinocytes during 24, 48, and 72 hours as indicated by MTT assay. *P<0.05 in comparison with control (C, untreated group).

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  Fig. 2 Proliferative effect of different doses of Glycyrrhiza glabra (GG) (A) and Nerium indicum (NI) (B) on keratinocytes during 24, 48, and 72 hours as indicated by MTT assay. *P<0.05 and ***P<0.001 in comparison with control (C, untreated group).

• 

  Fig. 3 Proliferative effect of different doses of honey+ghee (H+G) (A) and combination of all test materials (Tot) (B) on keratinocytes during 24, 48, and 72 hours as indicated by MTT assay. *P<0.05, **P<0.01, and ***P<0.001 in comparison with control (C, untreated group).

• 

  Fig. 4 Analyzed images of Scratch assay of HaCaT cells treated with the different test materials compared to control (untreated). Yellow dotted line indicates the site of the scratch wound. GG, Glycyrrhiza glabra; NI, Nerium indicum; H+G, honey+ghee; Tot, combination of all test materials.

• 

  Fig. 5 Graphical representation of the percentage of wound closure in all the experimental groups in a scratch assay using HaCaT cells. GG, Glycyrrhiza glabra; NI, Nerium indicum; H+G, honey+ghee; Tot, combination of all test materials.***P<0.001 and ###P<0.001 in comparison with 24 and 48 hours' control (untreated) groups respectively.

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