Improving Efficacy and Reducing Systemic Toxicity: An In Vitro Study on the Role of Electrospun Gelatin Nanofiber Membrane for Localized Melanoma Treatment

Abstract

Malignant melanoma is a highly metastatic skin cancer, representing about 5% of all cancer diagnoses in the United States. Conventional chemotherapy often has limited effectiveness and severe systemic side effects. This study explores a localized, topical delivery system using cisplatin-loaded nanomembranes as a safer and more targeted alternative. Cell viability assays established the safe cisplatin concentrations for tissue culture. Gelatin-based nanomembranes incorporating cisplatin were fabricated via electrospinning. Biocompatibility and therapeutic efficacy were tested by applying the membranes to cultured melanoma and normal skin cells. Controlled drug release profiles were evaluated by adjusting cross-linking times. Cisplatin concentration between 3.125 and 12.5 µg/mL were found safe. Nanomembranes with these doses effectively eliminated melanoma cells with minimal harm to healthy skin cells. Drug-free membranes showed high biocompatibility. Cross-linking duration allowed tunable and stable drug release. Cisplatin-loaded gelatin nanomembranes offer a promising topical therapy for melanoma, enhancing drug targeting while reducing systemic toxicity. This approach may serve as a cost-effective alternative to systemic treatments like immunotherapy. Future research will focus on in vivo testing and clinical application.

Keywords: chemotherapy; cisplatin; drug delivery; gelatin nanomembrane; melanoma.

Figure 1

Preparation and morphology of cisplatin-incorporated gelatin nanomembrane. (A) Schematic of the electrospinning process used to fabricate cisplatin-loaded gelatin nanofibers. (B) Schematic representation of nanofibers. SEM images showing the morphology of the nanomembranes. Smooth, interconnected fibers were observed, with average diameters of 150 ± 50 nm (scale bars = 5 µm). (a–c) Pure gelatin nanofibers; (e–f) cisplatin-loaded gelatin nanofibers.

Scheme 1

Design process flow chart.

Figure 2

SEM-EDS photomicrographs showing cisplatin crust on the nanofibers’ surface. (A) SEM-EDS mapping images of the nanofiber surface. (B) EDS spectra of 100 µg/mL cisplatin-incorporated nanomembrane. Processing option: all elements were analyzed and normalized. Note: the electron shells (e.g., K, L, M) correspond to the respective X-ray emission origins. N = 4 scans.

Figure 3

FTIR characterization of cisplatin-incorporated gelatin nanofibers, showing key spectral shifts indicative of drug–polymer interactions. Characteristic spectral changes were observed, particularly in the following regions: 3300–3200 cm⁻¹ (asymmetric and symmetric stretching of NH groups), 1600–1300 cm⁻¹ (asymmetric and symmetric bending of H-NH) region, around 800 cm⁻¹ (H-NH in-plane bending), and 499 cm⁻¹ (Pt-N stretching vibration).

Figure 4

Cell viability after treatment with varying cisplatin concentrations, measured by CCK-8 assay. Cell viability decreases with cisplatin concentration (A) and increased incubation time (B). n = 4; #: p > 0.05.

Figure 5

Cell–nanomembrane interaction at 1 h, 5 h, and 24 h post-application. (A) Low magnification: whole membrane area and junction. (B) High magnification: junction region at 3.125 µg/mL concentration. Note: pre-test control = no membrane or cisplatin; control = gelatin membrane only; white arrows (=>): shrinkage of cells. (C) Quantitative image analysis of cell coverage area. (*: p< 0.05).

Figure 5

Cell–nanomembrane interaction at 1 h, 5 h, and 24 h post-application. (A) Low magnification: whole membrane area and junction. (B) High magnification: junction region at 3.125 µg/mL concentration. Note: pre-test control = no membrane or cisplatin; control = gelatin membrane only; white arrows (=>): shrinkage of cells. (C) Quantitative image analysis of cell coverage area. (*: p< 0.05).

Figure 6

Cumulative release profile of ketoprofen and different molecules with/without cross-linking, and a schematic representation of the cross-linking. (A): (a) cross-linked and (b) non-cross-linked (scale bars = 5 µm). (c–f) Cumulative release over 7 days: (c) cumulative release over time at 12 or 48 h cross-linked for the same bioactive materials (ketoprofen); (d) cumulative release over time at 48 h cross-linked for different bioactive materials (MET: metformin, anticancer drug: 5-fluorouracil, macromolecule: exosomes); (B): (e) cumulative release over time for the same bioactive materials at different cross-linking time; (f) cumulative release over time for the same bioactive materials at different combination ratio of cross-linking time. When the same bioactive material was cross-linked at different combination ratios of cross-linking time, a controllable and stable release profile could be achieved. (Data are shown as percentage of non-cross-linked/12 h cross-linked/48 h cross-linked). All the formulations have shown more than 95% of drugs being released within 7 days; each data point represents the average of 4 samples from 4 batches (n = 4; *: p > 0.05 between 2 groups; #: p > 0.05 between all groups).

Figure 6

Cumulative release profile of ketoprofen and different molecules with/without cross-linking, and a schematic representation of the cross-linking. (A): (a) cross-linked and (b) non-cross-linked (scale bars = 5 µm). (c–f) Cumulative release over 7 days: (c) cumulative release over time at 12 or 48 h cross-linked for the same bioactive materials (ketoprofen); (d) cumulative release over time at 48 h cross-linked for different bioactive materials (MET: metformin, anticancer drug: 5-fluorouracil, macromolecule: exosomes); (B): (e) cumulative release over time for the same bioactive materials at different cross-linking time; (f) cumulative release over time for the same bioactive materials at different combination ratio of cross-linking time. When the same bioactive material was cross-linked at different combination ratios of cross-linking time, a controllable and stable release profile could be achieved. (Data are shown as percentage of non-cross-linked/12 h cross-linked/48 h cross-linked). All the formulations have shown more than 95% of drugs being released within 7 days; each data point represents the average of 4 samples from 4 batches (n = 4; *: p > 0.05 between 2 groups; #: p > 0.05 between all groups).