Development of Cissus quadrangularis-Loaded POSS-Reinforced Chitosan-Based Bilayer Sponges for Wound Healing Applications: Drug Release and In Vitro Bioactivity

Abstract

Nowadays, antibiotic-loaded biomaterials have been widely used in wound healing applications. However, the use of natural extracts has come into prominence as an alternative to these antimicrobial agents in the recent period. Among natural sources, Cissus quadrangularis (CQ) herbal extract is used for treatment of bone and skin diseases in ayurvedic medicine due to its antibacterial and anti-inflammatory effects. In this study, chitosan-based bilayer wound dressings were fabricated with electrospinning and freeze-drying techniques. CQ extract-loaded chitosan nanofibers were coated on chitosan/POSS nanocomposite sponges using an electrospinning method. The bilayer sponge is designed to treat exudate wounds while mimicking the layered structure of skin tissue. Bilayer wound dressings were investigated with regard to the morphology and physical and mechanical properties. In addition, CQ release from bilayer wound dressings and in vitro bioactivity studies were performed to determine the effect of POSS nanoparticles and CQ extract loading on NIH/3T3 and HS2 cells. The morphology of nanofibers was investigated with SEM analysis. Physical characteristics of bilayer wound dressings were determined with FT-IR analysis, swelling study, open porosity determination, and mechanical test. The antimicrobial activity of CQ extract released from bilayer sponges was investigated with a disc diffusion method. Bilayer wound dressings' in vitro bioactivity was examined using cytotoxicity determination, wound healing assay, proliferation, and the secretion of biomarkers for skin tissue regeneration. The nanofiber layer diameter was obtained in the range of 77.9-97.4 nm. The water vapor permeability of the bilayer dressing was obtained as 4021 to 4609 g/m²day, as it is in the ideal range for wound repair. The release of the CQ extract over 4 days reached 78-80% cumulative release. The release media were found to be antibacterial against Gram-negative and Gram-positive bacteria. In vitro studies showed that both CQ extract and POSS incorporation induced cell proliferation as well as wound healing activity and collagen deposition. As a result, CQ-loaded bilayer CHI-POSS nanocomposites were found as a potential candidate for wound healing applications.