Conjunctival melanoma (CοΜ) is of high malignancy that diffusely impacts the ocular surface, which is challenging to surgically remove and pose a life-threatening risk. As one of the most suitable treatment options, drug dropping drugs to the eye surface, however, is limited in its clinical application for CoM treatment due to low drug utilization. Using BRAF/MEK inhibitors (BRAF/MEKi) as model drugs, this study proposes a long-lasting eyedrop based on BRAF/MEK dual-targeted liposome-doped dynamic hydrogels with drugs (B/MLDDHs), which can overcome the tear flushing to succeed in effective ocular surface drug delivery. B/MLDDHs encapsulate drugs within lipid bilayers, enhancing drug bioavailability and biocompatibility. The aldehyde dextran and protonated chitosan-based injectable hydrogel system facilitates drug retention on the ocular surface exerting a sustained liposome release, which significantly enhances the drug bioavailability. In therapeutics, this study demonstrates significantly enhanced in vivo therapeutic effect on orthotopic mouse model of CoM. Given their ability to fulfill permeable and long-term release of hydrophobic drugs, B/MLDDHs may serve as a promising platform for non-invasive delivery of various drug molecules, thereby improving therapeutic outcomes for CoM.

Herein we develop a unique differentiated-uptake strategy capable of efficient and high-purity isolation of genuine drug-resistant (DR) cells from three types of drug-surviving cancer cells, which include paclitaxel-surviving human ovarian OVCAR-3 cancer cells and human lung carcinoma A549/Taxol cells, and doxorubicin-surviving human immortalized myelogenous leukemia K562/ADR cells. By using this strategy which relies on fluorescent glycan nanoparticle (FGNP)-based fluorescence-activated cell sorting (FACS) assays, two subpopulations with distinct fluorescences existing in drug-surviving OVCAR-3 cells were separated, and we found that the lower fluorescence (LF) subpopulation consisted of DR cells, while the higher fluorescence (HF) subpopulation was comprised of non-DR cells. Besides, the DR cells and their progenies were found distinct in their increased expression of drug-resistant genes. More intriguingly, by using the FGNP-based FACS assay to detect DR/non-DR phenotypes, we found that the DR phenotype had a potential to differentiate into the non-DR progeny, which demonstrates the differentiation feature of stem-like cancer cells. Further research disclosed that the assay can quantitatively detect the degree of drug resistance in DR cells, as well as the reversal of drug resistance that are tackled by various therapeutic methods. The strategy thus paves the way to develop theranostic approaches associated with chemotherapy-resistance and cancer stemness.