Quality by Design (QbD) Accredited Naringenin-loaded Chitosan Nanoparticles Impregnated Gel for Diabetic Wound Healing: Formulation, Optimization, In silico and In vitro Characterization

Abstract

Naringenin (NRG), a citrus-derived flavonoid with antioxidant, anti-inflammatory, and anti-diabetic properties, shows promise in treating chronic diabetic wounds distinguished by a prolonged inflammatory phase and enhanced oxidative stress. Present research investigated NRG-loaded chitosan nanoparticles (NRG/CS-NPs) incorporated into a SEPINEO™ P 600 gel for wound healing. Initially, the interaction of NRG with diabetic wound-associated biomarkers [MMP-9 (-7.655), SOD-1 (-5.164), catalase (-7.236), TNF-α (-6.459), and IL-1β (-4.549)] was evaluated through in silico molecular docking studies.

NRG/CS-NPs were fabricated via the ionic gelation method and optimized using 2⁴ factorial design, considering variables such as sodium tripolyphosphate concentration, chitosan concentration, pH, and drug content. The optimized NRG/CS-NPs exhibited a vesicular size (124.4 ± 0.93 nm), PDI (0.243 ± 0.07), zeta potential (12.3 ± 1.9 mV), and entrapment efficiency (81 ± 0.16%). The TEM imaging confirmed the morphology of the nanocarrier, aligning with DLS measurements. A sustained drug release of 76.19 ± 1.05% at 24 h followed a non-Fickian release profile. Cell viability assays demonstrated 80–90% viability at concentrations from 50 μM, indicating effective biocompatibility.

Incorporating the NRG/CS-NPs into a gel base improved retention time at the wound site. The NRG/CS-NPs-gel exhibited a smooth texture, pH (6.80 ± 0.10), and favourable spreadability (0.059 cm²/g) and viscosity (6.35 х 10⁵ ± 120331 cP) compared to a marketed product. Hemocompatibility studies confirmed the safety of the gel formulation and in vitro cell migration has demonstrated 78%, showing therapeutic potential and paving the way for further non-clinical investigations in diabetic wound healing.

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Materials

NRG and sodium tripolyphosphate anhydrous (TPP) were procured from Sisco Research Laboratories, Mumbai, India. Chitosan (5-20 mPa.s, > 70% deacetylation) was acquired from Tokyo Chemical Industry Co., Ltd., Japan. SEPINEO™ P-600 gel base was received as a gift sample from SEPPIC.

Noella Abraham, Tejaswini Kolipaka, Giriraj Pandey, Dilip Kumar Arya, Abhishek Sharma, Ganesh Vambhurkar, Rahul Kumar, P.S. Rajinikanth, Saurabh Srivastava, Quality by Design (QbD) Accredited Naringenin-loaded Chitosan Nanoparticles Impregnated Gel for Diabetic Wound Healing: Formulation, Optimization, In silico and In vitro Characterization, Journal of Drug Delivery Science and Technology, 2025, 107418, ISSN 1773-2247, https://doi.org/10.1016/j.jddst.2025.107418.

Read also our introduction article on Chitosan here:

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