BITS Faculty Publications
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Item Chitosan-Chondroitin sulfate based polyelectrolyte complex for effective management of chronic wounds(Elsevier, 2019-07) Roy, AniruddhaAcute and chronic wound remain an unresolved clinical problem among various demographic groups. Traditional marketed products focus mainly on inhibition of bacterial growth at the wound site neglecting the tissue repair, which significantly affect the healing rate. It would be highly beneficial if a wound healing material can be developed which has both antibacterial as well as tissue regenerating potential. We have prepared a polyelectrolyte complex (PEC) using chitosan (CH) and chondroitin sulfate (CS) which can form an in-situ scaffold by spontaneous mixing. The fabrication of CH-CS PEC was optimized using Quality-By-Design (QbD) approach. The prepared PEC showed very high swelling and porosity property. It was found to be non-hemolytic with good blood compatibility and low blood clotting index. It also exhibited good antibacterial activity against both gram-positive and gram-negative bacteria. The cell proliferation study exhibited good cytocompatibility and almost four-fold increase in cell density when treated with CH-CS PEC compared to control. In summary, we demonstrated that the prepared CH-CS PEC showed good blood compatibility, high antibacterial effect, and promoted wound healing potentially by stimulating fibroblast growth, making it an ideal wound dressing material.Item Development of an in-situ forming, self-healing scaffold for dermal wound healing: in-vitro and in-vivo studies(Elsevier, 2021-09) Roy, AniruddhaThe importance of the extra-cellular matrix (ECM) for wound healing has been extensively researched. Understanding its importance, multiple ECM mimetic scaffolds have been developed. However, the majority of such scaffolds are prefabricated. Due to their stiffness, prefabricated scaffolds cannot come into direct contact with the basal skin cells at the wound bed, limiting their efficacy. We have developed a unique wound dressing, using chitosan (CH) and chondroitin sulfate (CS), that can form a porous scaffold (CH-CS PEC) in-situ, at the wound site, by simple mixing of the polymer solutions. As CH is positively and CS is negatively charged, mixing these two polymer solutions would lead to electrostatic cross-linking between the polymers, converting them to a porous, viscoelastic scaffold. Owing to the in-situ formation, the scaffold can come in direct contact with the cells at the wound bed, supporting their proliferation and biofunction. In the present study, we confirmed the cross-linked scaffold formation by solid-state NMR, XRD, and TGA analysis. We have demonstrated that the scaffold had a high viscoelastic property, with self-healing capability. Both keratinocyte and fibroblast cells exhibited significantly increased migration and functional markers expression when grown on this scaffold. In the rat skin-excisional wound model, treatment with the in-situ forming CH-CS PEC exhibited enhanced wound healing efficacy. Altogether, this study demonstrated that mixing CH and CS solutions lead to the spontaneous formation of a highly viscoelastic, porous scaffold, which can support epidermal and dermal cell proliferation and bio-function, with an enhanced in-vivo wound healing efficacy.Item Chitosan, chondroitin sulfate, and hyaluronic acid based in-situ forming scaffold for efficient cell grafting(Elsevier, 2023-01) Roy, AniruddhaCurrent cell grafting techniques are majorly dependent on seeding cells on a pre-formed scaffold. However, cells grow in a 2-dimensional (2D) space in such constructs, not mimicking the tissue's 3-dimensional (3D) architecture. The present study evaluated a unique poly-electrolyte complexation (PEC) based strategy for the 3D engraftment of cells in a porous polymeric scaffold. The scaffold was synthesized using a positively charged polysaccharide chitosan (CH) and negatively charged glycosaminoglycans chondroitin sulfate (CS) and hyaluronic acid (HA). Two different scaffolds were synthesized, one using CH and CS [CH-CS] and another using CH and CS + HA [CH-(CS-HA)]. The physicochemical characterization of both the PECs confirmed electrostatic interactions, leading to a porous and viscoelastic PEC formation. Fibroblast cells were grafted and seeded in both scaffolds to evaluate the effect of different scaffold compositions and the difference between seeded and grafted cells. Imaging studies confirmed that grafting of the fibroblast cells supports cellular proliferation. The qPCR studies demonstrated increased expression of functional markers TGF-β, α-SMA, collagen-I, and fibronectin in the CH-(CS-HA) grafted cells. In summary, it was demonstrated that an in-situ forming PEC of CH, CS, and HA had good physicochemical properties for cell grafting and supported grafted cells with improved functionItem Dual antibacterial and anti-inflammatory efficacy of a chitosan-chondroitin sulfate-based in-situ forming wound dressing(Elsevier, 2022-12) Roy, AniruddhaNone of the currently available wound dressings exhibit combined antibacterial and anti-inflammatory activity. Using polyelectrolyte complexation (PEC) between a cationic polysaccharide chitosan (CH) and an anionic glycosaminoglycan chondroitin sulfate (CS), we have developed a unique in-situ forming scaffold (CH-CS PEC), which develops at the wound site itself to influence the function of the wound bed cells. The current study demonstrated that CH-CS PEC could induce bacterial cell death through membrane pore formation and increased ROS production. Moreover, possibly due to its unique material properties including medium-soft viscoelasticity, porosity, and surface composition, CH-CS PEC could modulate macrophage function, increasing their phagocytic ability with low TNF-α and high IL-10 production. Faster wound closure and decreased CFU count was observed in an in-vivo infected wound model, with reduced NF-κB and increased VE-cadherin expression, indicating reduced inflammation and enhanced angiogenesis. In summary, this study exhibited that CH-CS PEC has substantial antibacterial and immunomodulatory properties.Item Development and evaluation of a simvastatin-loaded biopolymeric scaffold for improved angiogenesis and healing of diabetic wounds(Elsevier, 2023-09) Roy, Aniruddha; Majumder, SyamantakIn diabetic wounds, lack of angiogenesis limits the supply of oxygen and nutrients at the wound site, resulting in poor healing. A well-known lipid-lowering drug, simvastatin (SIM), exhibited pleiotropic effects in wound healing, including promotion of angiogenesis. However, its clinical application is limited due to its poor physicochemical properties, including low solubility. In this study, a Soluplus and TPGS-based mixed micelle was developed for loading SIM in an in-situ forming chitosan-chondroitin sulfate-based poly-electrolyte complex hydrogel (CH-CS PEC). The hypothesis was that CH-CS PEC would improve overall wound healing due to the favorable viscoelasticity and porosity, whereas SIM would assist neoangiogenesis. SIM-loaded CH-CS PEC exhibited good mechanical stability and viscoelastic properties and demonstrated prolonged release of SIM. The formulation promoted endothelial cell sprouting in an ex-vivo rat aortic ring assay. Applying SIM-loaded CH-CS PEC in a diabetes-induced rat wound model resulted in faster wound closure, increased collagen deposition, and enhanced neovascularization with up-regulation of vascular endothelial growth factor (VEGF) expression. In summary, we have developed a drug-loaded, in-situ forming scaffold that can be directly applied at the wound site and can improve wound healing by promoting angiogenesis and collagen deposition at the wound site. This study demonstrated the combined efficacy of a viscoelastic scaffold and a proangiogenic drug for enhanced wound healing. The easy and simple fabrication method of the drug-loaded scaffold makes it suitable for clinical translation.