Evaluation of Mitragyna speciosa (Korth.) Havil extract-based nanostructured lipid carriers incorporated into hydrogel film for wound healing

The evaluation of Mitragyna speciosa (Korth.) Havil (MS) extract as an active ingredient in nanostructured lipid carriers incorporated into hydrogel film (MSNLC hydrogel) for wound healing were reported herein. MS extract demonstrated advantageous characteristics such as low toxicity, antiinflammation, antioxidant, and antibacterial that may facilitate the mechanisms of the wound healing process but low biocompatibility and bioavailability. Overcoming the skin's robust barrier function to transport the active ingredients to the target site in an adequate and optimal concentration is the challenging element. Hence, a nanohybrid system, MS-NLC hydrogel was used because of its nanosized, stability and low toxicity that will improve the effectiveness of skin absorption and encapsulation for transdermal drug delivery. In this study, MS extract yielded (19.92%) using maceration method. The extract contained phytochemicals such as alkaloids (mitragynine, 7-hydroxymitragynine, speciogynine), flavonoids (quercetin, apigenin, kaempferol), saponins (daucosterol, quinovic acid 3-o-beta-d-quinovopyranoside, 1-o-feruloyl-beta-dglucose) and other bioactive phytochemicals (chlorogenic acid, umbelliferone, ursolic acid). The quantitative analysis showed that 52.8% of mitragynine compound was obtained from the MS extract. The flavonoid, phenolic content and DPPH assay of MS extract were 50.43 ± 0.47 mg GAE/g, 90.88 ± 0.30mg QE/g and IC50 at 0.0397±0.0035 mg/mL, respectively. Based on the screening assessment of MS-NLC, the maximum solubility in liquid lipid was 2.859 ±0.010 mg/mL whereas drug entrapped efficiency (%) in solid lipid was 53.02 ±0.39% and Tween 80 (51.53 %) was selected as the main surfactant. The optimum formulation using response surface methodology (RSM) of MS-NLC was 5.0 % (w/w) of MS extract, 3.0 % (w/w) of oleic acid: compritol 888 ATO, 60 min of reaction time and 9400 rpm of homogenizer stirring rate. The particle size, PDI and zeta potential of MS-NLC hydrogel was 130.00 ± 0.18 nm, 0.25 PDI and -33.0mV, respectively. MS-NLC hydrogel showed highest swelling ratio (349%), gel fraction (28.89%) and shear thinning behaviour which appropriate design for wound healing. The best temperature for storing with maintained nanoparticle size of MS-NLC hydrogel was at 25 ᵒC (132.55 ± 1.6 nm). In antimicrobial study, MS-NLC hydrogel showed the strongest inhibitory activity level against S. aureus (19.20 ± 0.2 mm) whereas low inhibition towards hyaluronidase activity (2.33 1.07%). After 24h scratch assay treatment using 3T3 fibroblast cells, MS-NLC hydrogel (64.21%) demonstrated improvement in the cell migration with non-toxic effect (IC50 > 500 μg/mL. The in vivo nanotoxicity test revealed that MS-NLC hydrogel were non-toxic towards zebrafish embryo with LC50 > 500 μg/mL. Besides, the in vitro permeation study of MS-NLC hydrogel (90.40 %) showed improvement of permeation and able to sustained the release of MS extract over 24 h.

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