Research

OUR RESEARCH

Our team works in a multidisciplinary approach using Nanotechnology to combat devastating diseases like Breast and lung cancer. One of the emphases here is overcoming lung cancer mutations induced after using various Tyrosine kinase inhibitors. Our laboratory uses various novel compounds that act as chemosensitizers and help overcome resistance in lung and breast cancers. We are also using exosomes as markers of tumor progression, enabling diagnosis and tumor progression as a liquid biopsy. Another emphasis is to use 3D printing approaches to develop tumor scaffolds and conduct cytotoxicity assays using 3D printed co-cultures of tumor cells and fibroblasts. Apart from this, our team is also printing Cornea using various bioinks.

Allergic Contact Dermatitis (ACD)

Allergic contact dermatitis (ACD) is one of the most prevalent occupational skin diseases. ACD may result from the interaction of xenobiotics with the immune system. ACD is regarded as the emergence of immunotoxicity in humans. Many chemicals have been shown to cause skin sensitization. In common with other forms of allergy, ACD develops in two phases, defined operationally as induction and elicitation. Induction of skin sensitization is initiated by topical exposure of the chemical allergen to an individual sufficient to induce a cutaneous immune response of the necessary vigor, known as sensitization. If the sensitized individual is now exposed subsequently, at the same or a different skin site, to the inducing chemical allergen, then a more vigorous secondary immune response will be provoked at the point of contact. This, in turn, initiates the cutaneous inflammatory reaction known as ACD. This model was sensitized and challenged using 2,4-dinitrofluorobenzene (DNFB).

Psoriasis-like model

Psoriasis is a chronic inflammatory skin disease characterized by erythematous plaques, excessive growth, and aberrant differentiation of keratinocytes, along with increases in angiogenesis and inflammatory cell infiltrate. The most commonly used model for investigating psoriasis is a xenograft model, in which immunodeficient mice are transplanted with human psoriasis-prone skin. However, these experiments are laborious, expensive, and require considerable expertise and technical skills. Therefore, a psoriasis plaque like the model was developed using topical application of imiquimod (IMQ) on the mice’s back skin. This model showed the most features of human psoriasis where cytokines like IL-23 and IL-17 are involved.

In vitro 3D culture model for wound healing

Presently there is no wound healing model available in vitro, and most studies are being conducted in vivo. In our laboratory, we are developing a 3D human wound healing model using the EFT-300 cultures from Mattek Corp. Currently, the model is being developed using various corrosive chemicals (e.g., Sulphuric acid, acrylic acid, potassium hydroxide) to induce a wound or heat to induce a burn wound. Various histological changes and the role of various growth factors and cytokines are also being monitored to validate such a model. EFT-300 model consists of epidermis and dermis, which are very important to understand the natural mechanism of wound healing to establish a normal equilibrium of the skin.

In vitro tumor 3D culture model

This project involves the development of an in-vitro tumor model which is more predictive of disease states and drug responses. Algimatrix obtained from Invitrogen is currently being used as the matrix to grow tumor cells as spheroids which will then be treated with various drugs/formulations and perform various mechanistic studies. The objective is to show a correlation between in vitro and in vivo studies, and currently, studies are in progress with lung tumor cells. Algimatrix is pure, non-toxic, has a better nutrient delivery without damage to cells.

CORNEAL PENETRATION STUDY

Fresh porcine eyes were obtained from Bradley Country Store in Tallahassee, FL (slaughterhouse). The anterior portion of the eye was cannulated using a 27 g needle and first flushed and inflated with keratinocyte serum-free medium (KSFM with pituitary extract and EGF) followed by the medium flow rate of 1-2 microlitre/min for the period of study at 37°C. Different formulations were prepared with Coumarin-6 as a tracer dye and applied to eyeballs. Confocal laser scanning microscopy with z-stacking (10μ intervals) was performed. Fluorescence intensities at different depths of the cornea showed significant differences in the different formulations. Nanomicelle gel showed deeper penetration and helped the drug traverse deeper in the corneal layers compared to the nanomicelle solution.

FORMULATION AND DEVELOPMENT OF DISSOLVABLE MICRONEEDLES AND TABLETS USING 3D PRINTING TECHNOLOGY

The formulation of dosage forms for the therapeutic delivery of drugs to the systemic circulation is currently seeing advancements in the pharmaceutical industry today. Variations of fabrication techniques are actively being researched, with one of the major players being the additive manufacturing of drugs using 3D printing techniques. In our lab, we are currently working on optimal designs and formulations for both the novel microneedle arrays (MNs) and the traditional tablet drug dosage forms. We have been successful in achieving well-defined 3D-printed MNs. However, we are now working on optimizing the composition of the MNs to demonstrate and advance therapeutic drug dissolution rates in addition to favorable skin permeation rates. Our goal of 3D printing a suitable tablet with favorable dissolution rates to bypass the harsh conditions of the GIT and the liver's tendency to metabolize drugs before reaching the systemic circulation is also coming along with favorable results. Our main driving force in developing this novel form of tablet manufacturing stems from a desire to enable ease of manufacturing and multi-drug tablets. We are using DLP and SLA printing technologies for this purpose and have formulated MNs and tablets by using several model drugs.

"(a): “Partially Polymerized Membrane” effect on the base on the iμ NA due to background printing from over-exposure, which peels off as the polymer matrix is not completely polymerized in the portions of the structure between the μN bases. (b) “Pancaking” effect on the edges of the iμ NA base. (c) Optimally printed iμ NA. (d-l) Optimization of the aspect ratio (AR) of the iμ NA to successfully achieve the desired ROC to penetrate human skin. (m) iμ NA dyed in methylene blue shows that the polymer matrix retains its hydrogel properties post-printing, allowing for “intelligent” release. (n) SEM image of an optimal aspect ratio iμ NA having a R_OC of ∼20 μm (o). (p) Dyeing the tips with Gentian Violet to study the effect of penetration on an artificial skin model. (q) Photomicrographs showing successful penetration of an entire 10×10 array with a close-up image in (r)." Kundu et al. (2020)

"(a): Schematic of PEGDA (blue) being mixed with 2.5 wt% Diphenyl (2, 4, 6-trimethylbenzoyl) phosphine oxide (TPO) (green) and 0.25wt% therapeutic cargo of diclofenac sodium (pink). (b) DLP 3D printing of the prepared polymer matrix with a UV light source of 385 nm onto the build platform. The iμNA printed consists of a sacrificial base plate and a raft to improve the adhesion of the 3D printed device onto the build platform. (c) Removal of the iμNA from the build platform to have the final iμNA, which may be used for ocular, acute, and chronic drug delivery via transdermal route and allergen testing on the human torso, among other applications. (d) Close-up of a singular API loaded μNA in the intelligent polymer matrix showing the stimulus-based release of the drugs upon sensing its external environment change while retaining the non-drug PEGDA matrix." Kundu et al. (2020)