A new preclinical study reports that ferulic acid delivered inside nanoscale niosomes substantially reduced testicular damage in rats treated with the chemotherapy drug doxorubicin, outperforming the same compound given in its free, unencapsulated form. The work, published in Biological Research, examined how the nano-formulated antioxidant influenced oxidative stress, inflammatory signaling, cell death pathways, and hormone-producing machinery in the testes of doxorubicin-exposed animals. The researchers, led by Mahran Mohamed Abd El-Emam and Hussein I. Elbelbesy of Zagazig University in Egypt, argue that the vesicular delivery system may offer a way to protect male fertility during cancer treatment, although the findings so far rest entirely on animal data.

Doxorubicin is an anthracycline antibiotic widely used against solid tumors and blood cancers, including breast, prostate, ovarian, gastric, and childhood cancers as well as leukemias and sarcomas. Its clinical value, however, is constrained by toxicity to rapidly dividing tissues, and the testes are particularly vulnerable. According to the study’s authors, doxorubicin-driven reproductive damage is largely mediated by overproduction of reactive oxygen species, which deplete antioxidant defenses, damage membranes and mitochondria, and set off cascades of inflammation and apoptosis that impair sperm production and testosterone synthesis. Male-related factors contribute solely to roughly 20 to 30 percent of infertility cases and are implicated in up to half of affected couples, making preservation of fertility during chemotherapy a significant clinical concern.

The research team focused on ferulic acid, a plant-derived phenolic acid abundant in foods such as wheat bran, corn bran, artichokes, beans, and eggplant. Ferulic acid shows antioxidant, anti-inflammatory, and other biological activities, and earlier experimental work suggested it can improve sperm motility, viability, and counts in diabetic animal models. Its antioxidant power stems from its capacity to scavenge free radicals, inhibit lipid peroxidation, and suppress the generation of reactive oxygen and nitrogen species. Yet the compound’s usefulness is limited by poor aqueous solubility and chemical instability, which compromise its bioavailability when administered conventionally.

To overcome that limitation, the researchers encapsulated ferulic acid in niosomes, vesicular bilayer structures composed of cholesterol and non-ionic surfactants that can enhance drug solubility, stability, circulation time, and controlled release. Using a thin-film hydration technique with Span 60 and cholesterol in an equimolar ratio, they produced particles with a mean diameter of 342 plus or minus 35.17 nanometers and a polydispersity index of 0.25 plus or minus 0.03, indicating a relatively homogeneous distribution. The particles carried a strongly negative zeta potential of minus 44.47 plus or minus 2.04 millivolts, and transmission electron microscopy confirmed a spherical morphology. The formulation achieved an entrapment efficiency of 75.77 plus or minus 3.4 percent, and in vitro release testing showed sustained delivery: 41.87 percent of the ferulic acid was released after six hours and 74.41 percent after 24 hours, compared with only 27.96 percent and 28.73 percent from a free ferulic acid suspension over the same intervals.

The in vivo experiment involved forty male albino Wistar rats, aged ten weeks and weighing 250 to 260 grams, randomly assigned to four groups of ten animals each. A control group received saline orally once daily for 21 days. The remaining rats received doxorubicin intraperitoneally at 3 milligrams per kilogram of body weight on days 7, 14, and 21, and were divided into three treatment arms: doxorubicin alone, doxorubicin plus free ferulic acid at 20 milligrams per kilogram orally, and doxorubicin plus the ferulic acid-loaded niosomes at the same dose. Blinding was applied during outcome assessments that could be masked, including histopathology and immunohistochemistry, although the authors note that treatment administration was not blinded and that complete blinding was not always feasible during sample preparation. The protocol was approved by the ethics committee of the Faculty of Veterinary Medicine at Zagazig University.

The results showed that doxorubicin inflicted broad reproductive harm. Treated rats displayed a 133.87 percent increase in abnormal sperm compared with controls, a 77.77 percent drop in sperm motility, and a 70.94 percent decline in sperm count. Serum testosterone fell by 0.6-fold, with luteinizing hormone and follicle-stimulating hormone reduced by 0.8- and 0.5-fold respectively. Testicular malondialdehyde, a marker of lipid peroxidation, rose 1.8-fold while the antioxidants glutathione, superoxide dismutase, and catalase fell by 0.6-, 0.7-, and 0.5-fold. Gene expression analysis revealed an approximately six-fold upregulation of the NLRP3 inflammasome component and a 6.5-fold rise in interleukin-1 beta, alongside a nine-fold increase in the pro-apoptotic gene Bax and a downregulation of the anti-apoptotic Bcl-2. The steroidogenic genes StAR, CYP11A1, and 3-beta-HSD were suppressed. Histology showed atrophied seminiferous tubules, necrotic germinal epithelium, empty lumina, giant spermatocytes, and interstitial edema.

Both forms of ferulic acid partially reversed these changes, but the niosomal formulation consistently performed better. In the FA-NIO group, abnormal sperm percentages fell 44.13 percent relative to the doxorubicin group versus 34.48 percent with free ferulic acid, and sperm motility rose 337.5 percent versus 200 percent, with sperm count gains of 188.46 percent versus 142.30 percent. The niosomal treatment also outperformed free ferulic acid on every sperm parameter at statistical significance. Testosterone increased 1.5-fold with FA-NIO compared with 0.9-fold for free ferulic acid, and gonadotropin levels rose further. FA-NIO reduced malondialdehyde by 0.6-fold and raised glutathione, superoxide dismutase, and catalase by 1.7-, 2.3-, and 0.8-fold, exceeding the free compound’s effects. NLRP3 and interleukin-1 beta expression were suppressed more strongly, Bax fell further while Bcl-2 rose 2.3-fold, and the steroidogenic genes StAR, CYP11A1, and 3-beta-HSD were upregulated by 0.4-, 1.5-, and twofold respectively. Immunohistochemistry confirmed faint NLRP3, caspase-1, and Bax staining in the FA-NIO group, and histological examination showed near-normal seminiferous tubule architecture with only mild interstitial edema.

The authors interpret these findings as evidence that niosomal encapsulation maximizes ferulic acid’s antioxidant and endocrine-modulating actions by improving bioavailability and facilitating delivery to vulnerable tissues. They suggest that the vesicular system may promote intracellular accumulation of the compound, more effectively dampening inflammasome-associated and apoptotic signaling while restoring the transcriptional program of testosterone synthesis, in which StAR transports cholesterol into mitochondria, CYP11A1 cleaves cholesterol to pregnenolone, and 3-beta-HSD drives the subsequent steps toward androstenedione and testosterone. The parallel recovery of hormone levels, sperm quality, and tissue architecture supports, in their view, a coordinated protective mechanism spanning oxidative, inflammatory, apoptotic, and steroidogenic pathways.

Important caveats accompany the results. The study was conducted in a rat model with a specific doxorubicin regimen, and the authors acknowledge a key limitation: the design lacked control groups receiving ferulic acid alone, FA-NIO alone, or empty niosomes without the active compound. Those controls would have clarified the intrinsic effects of the compound and the carrier itself. Although no overt adverse effects were observed in the treated groups, the authors state that further studies are needed to evaluate the formulation’s safety, efficacy, and possible clinical relevance before any translation toward protecting chemotherapy patients’ reproductive health can be considered. For now, the work stands as a preclinical proof of concept that nanoscale delivery can amplify a plant antioxidant’s protective effects against doxorubicin-induced testicular injury in rats.

Subject of Research: Niosomal ferulic acid protection against doxorubicin-induced testicular toxicity in rats

Article Title: Targeting oxidative stress-NLRP3-apoptotic-steroidogenic axis in doxorubicin-induced testicular toxicity: protective efficacy of ferulic acid niosomes

Article References: El-Emam, M. M. A., Atia, A. E., Bakry, M. A., Husseini, R. A., Elrefaei, E., El-Shaer, N. O., Khater, S. I., Ali, O., Yousef, A. E.-S., & Elbelbesy, H. I. (2026). Targeting oxidative stress-NLRP3-apoptotic-steroidogenic axis in doxorubicin-induced testicular toxicity: protective efficacy of ferulic acid niosomes. Biological Research, 59(1), Article 43. https://doi.org/10.1186/s40659-026-00707-9

Image Credits: AI Generated

DOI: 10.1186/s40659-026-00707-9

Keywords: doxorubicin, ferulic acid, niosomes, testicular toxicity, oxidative stress, NLRP3 inflammasome, apoptosis, steroidogenesis, male infertility, nanomedicine, preclinical study, rats