An In Vitro Study of the Healing Potential of Black Mulberry (Morus nigra L.) Extract in a Liposomal Formulation

Abstract

Natural compounds are used in modern dermal treatments to avoid side effects commonly associated with conventional treatments. The aim of our study was to develop a liposomal formulation including black mulberry extract and to highlight its potential on the healing of normal human dermal fibroblasts (NHDF) in vitro using the scratch test. Mulberry-loaded liposomes (Mn_L) were prepared using a thin-film hydration method based on cholesterol (C) and phosphatidylcholine (PC) in a 1:3 (w/w) ratio. The liposomal formulation was characterized by analyzing its size, electric surface potential, morphology, entrapment efficiency, and in vitro healing effects. Also, the black mulberry fruits (Morus nigra L.) were characterized from point of view of polyphenolic compounds and antioxidant capacity by Ferric-Reducing Antioxidant Power (FRAP) assay. HPLC-DAD-MS (ESI+) (high performance liquid chromatography-photodiode array detection-mass spectrometry (electrospray ionization)) analysis indicated the presence of phenolic compounds namely from hydroxybenzoic and hydroxycinnamic acids and flavonols. Among flavonols, quercetin-glucoside represented 50.56%, and chlorogenic acid was the predominant compound among hydroxycinnamic acids (37.06%). In vitro fibroblast wound closure was more effective with mulberry-loaded liposomes (L_Mn) than extracts of mulberries. According to our study, mulberry-loaded liposomes have been shown to be effective in wound healing and can be used as a natural treatment.

Characterization in Terms of Phytochemical Content and Medicinal Potential of the Stellaria media Plant Extract

“…It can be seen that the SM sample after 36h shows a percentage of closure of the simulated in vitro wound area comparable to the ALA control sample and higher than that of the CT and LIPO samples (used as positive control). At the same time, the liposomal formula shows at the same time interval (T36h) a percentage of wound area closure (86.884%) more than the SM sample (81.2221%), which indicates that the created liposomal formulation is more advantageous, increasing the bioavailability of the included extract, favoring the passage through the membrane of the fibroblast type cell and due to the protection offered to the included extract [21].…”

“…The equations of the curves were used for quantitative calculation for each phenolic compound as follows: hydroxybenzoic acid was calculated as gallic acid equivalent and flavones as luteolin equivalent. The phenolic compounds were identified by comparing the retention time, UV-Vis absorption, and mass spectra with those of the standard compounds and with data from the specialized literature [21][22][23]. The quantification of phenolic compounds was done by using the calibration curve with gallic acid for hydroxybenzoic acid, respectively with luteolin for flavones [21,22,24].…”

“…The phenolic compounds were identified by comparing the retention time, UV-Vis absorption, and mass spectra with those of the standard compounds and with data from the specialized literature [21][22][23]. The quantification of phenolic compounds was done by using the calibration curve with gallic acid for hydroxybenzoic acid, respectively with luteolin for flavones [21,22,24].…”

“…Through DLS analysis, the size of the liposomes and the electrical surface charge are highlighted and the EE% is also tested using the Folin Ciocalteu method to determine the total polyphenols from the S. media plant extract before and after encapsulation by applying the formula (2) [21].…”

“…After 2 hours, the formazan dye was quantified by measuring absorbance at 492 nm using a microplate reader (Stat Fax 2100, Palm City, USA), while the reference wavelength was 630 nm. Results were expressed as a percentage of cell viability compared to the control [21].…”