TOPICAL DELIVERY OF QUERCETIN LOADED TRANSFERSOMES FOR WOUND TREATMENT: IN VITRO AND IN VIVO EVALUATION Original Article MARWA ABDALLAH 1* , DEMIANA I. NESSEEM 1 , OMAIMA N. ELGAZAYERLY 2 , ALY A. ABDELBARY 2,3 1 National Organization of Drug Control and Research, Pharmaceutics Department, Egypt, 2 Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Cairo University, Cairo, Egypt, 3 School of Life and Medical Sciences, University of Hertfordshire hosted by Global Academic Foundation, Cairo, Egypt Email: dr_maramiro@msn.com Received: 14 Mar 2021, Revised and Accepted: 13 Jul 2021 ABSTRACT Objective: To design topical Quercetin (Qc)-loaded transfersomes (TFs) for wound treatment. Methods: Qc-loaded TFs were prepared by thin-film hydration technique using 2 2 4 1 full factorial design and the optimum formula was selected. In vivo skin, deposition and cutaneous wound induction studies were performed for four groups of male wistar rats. At the end of the experiment, biochemical parameters were measured in the healed tissues (total proteins (TP), total antioxidant capacity (TAC), glutathione reductase (GSH), nitric oxide (NO), and malonaldehyde (MDA). Two in vivo histopathological experiments using male wistar rats were performed; the first study was done for the healed tissues of the above experiment and the second was to confirm the safety of formulations. Results: Qc optimum TFs (F6) showed EE% of 91.1%, PS of 695.35 nm, PDI of 0.592, and ZP of-11.1 mV, and spherical shape. In vivo skin deposition study showed that drug percentage retained in the skin from Qc optimum TFs was significantly higher than that from Qc suspension and Qc liposomes (p<0.05). There was no significant difference in the values of TP, TAC and MDA between the treated groups (p>0.05). GSH in TFs treated groups was significantly higher than the other groups (p<0.05) while NO in TFs treated groups was significantly lower than the other treated groups (p<0.05). Histopathological experiments showed that wounds treated by TFs healed better than those treated by both liposomes and Qc suspension. Conclusion: Qc-loaded TFs can be used as successful drug-delivery system for wound healing. Keywords: Quercetin, Transfersomes, Wound treatment, Topical formulation © 2021 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/) DOI: https://dx.doi.org/10.22159/ijap.2021v13i5.41435. Journal homepage: https://innovareacademics.in/journals/index.php/ijap INTRODUCTION Herbal drugs have been used in many diseases [1]. Plant pigments which are found in herbal drugs as flavonoids which are also found in fruits and vegetables, have many physiological activities [2]. Flavonoids are important drug candidates due to their free radical scavenging and anti-inflammatory properties [3, 4]. There is a structure-activity relationship between several flavonoids (luteolin, kaempferol, and chrysin) and their antioxidant activity and the number of OH groups in their structure so they are used in inflammatory disorders such as psoriasis and arthritis [5]. Quercetin (Qc) has high antioxidant activity which reduces the expression of matrix metalloproteinase that causes wrinkling and decreasing of the elasticity of healthy and photoaged skin [6]. Qc has antioxidant and antiradical effects due to three functional groups in its structure [7-9]. Nanotechnology is urgently needed for herbal drugs due to the poor solubility of those drugs [10]. The topical application of Qc in various formulation approaches includes permeation enhancers [11] such as lecithin and chitosan nanoparticles [12] which increase the penetration of Qc through the skin to make its topical and transdermal delivery easier, targeting the drug towards the desired tissue and concentrating the drug in it while reducing its concentration in the non-target tissues. Hence, drug action increases and side effects decrease [13]. Topical dosage forms are superior drug delivery systems because they avoid first- pass hepatic metabolism, gastric degradation and improve penetration of the drug into skin layers [14, 15]. Surface-active agent vesicles are an alternative drug delivery system to conventional liposomes. Transfersomes (TFs) are elastic nano-vesicles composed of lecithin and surface-active agent and they differ from the conventional liposomes by their ability to squeeze themselves into the intercellular region of the stratum corneum [16, 17]. This study aimed to develop topical Qc-loaded TFs for wound treatment. Optimization of production parameters to produce stable, highly concentrated Qc vesicles with a small particle size was done using full factorial statistical design. In addition, in vivo skin deposition of Qc from optimum TFs was compared with Qc liposomes and Qc suspension using male Wistar rats. In addition, wound induction was done to male Wistar rats which were then treated with Qc optimum TFs, Qc liposomes, and Qc suspension by measuring the biological parameters in the treated skin tissues of rats. Furthermore, two in vivo histopathological experiments were performed; the first to compare the wound treatment effect of Qc optimum TFs, Qc liposomes, and Qc suspension. The second in vivo histopathological experiment was performed to assess the irritation potential of Qc optimum TFs on rat skin. MATERIALS AND METHODS Materials Quercetin (Qc) dihydrate 97%, Cholesterol 95%, and Polysorbate 60 (Tween 60) were purchased from ALFA Aesar (A Johnson Matthey Company), Germany. Ethanol absolute was purchased from Sigma Aldrich, Germany. Chloroform was purchased from Sigma Aldrich, United Kingdom. Sodium Chloride, Potassium chloride, Disodium hydrogen orthophosphate, Potassium dihydrogen orthophosphate, and Tween 80 were purchased from ADWIC, ELNASR pharmaceutical chemicals company, Egypt. Span 60 and Span 40 were purchased from LOBA Chemie, India. Lecithin granular from egg yolk was purchased from Acros organics, USA. Sorbitan monooleate (Span80) was purchased from MP Biomedicals, France. Glutathione reduced (GSH), Malonaldehyde (MDA), Nitric oxide (NO), Total proteins (TP) and Total antioxidant capacity (TAC) kits colorimetric method were purchased from Biodiagnostic and research reagents, Egypt. Experimental design The design used was complete factorial 2 2 4 1, which studied the influence of different variables on Qc-loaded TFs [18]. One factor was assessed with 4 levels (X1: Type of the surface-active agent), while the others with two levels (X2: Lecithin to surface-active agent molar ratio) and (X3: Hydration volume). (Y1: EE %), (Y2: PS), (Y3: International Journal of Applied Pharmaceutics ISSN- 0975-7058 Vol 13, Issue 5, 2021