Background/Objectives: This study explores the bioactive potential of Bovistella utriformis biomass and its polysaccharides (PsBu) through comprehensive biochemical and bioactivity analyses, focusing on their antioxidant, cytotoxic, and antihyperglycemic properties. Methods: Elemental analysis determined the biomass’s chemical composition. Antioxidant activity was assessed using ABTS and DPPH assays. Monosaccharide composition was analyzed via gas chromatography-mass spectrometry (GC-MS). In vitro cytotoxicity assays were conducted on cancer and normal cell lines to determine IC₅₀ values and selectivity indices (SI). Zebrafish embryo toxicity was evaluated for teratogenic effects, and an in vivo antihyperglycemic study was performed in diabetic rat models. Results: The biomass exhibited high carbon content (around 41%) and nitrogen levels, with a balanced C/N ratio nearing 5. Protein content exceeded 50%, alongside significant carbohydrate, fiber, and ash levels. Antioxidant assays revealed inhibition rates of approximately 89% (ABTS) and 64% (DPPH). GC-MS analysis identified glucose as the predominant sugar (>80%), followed by galactose and mannose. Additionally, HPLC detected a photoprotective compound, potentially a mycosporin-like amino acid. Cytotoxicity assays demonstrated PsBu’s selective activity against colon, lung, and melanoma cancer cell lines (IC₅₀: 100–500 µg·mL⁻¹), while effects on normal cell lines were lower (IC₅₀ > 1300 µg·mL⁻¹ for HaCaT, >2500 µg·mL⁻¹ for HGF-1), with SI values approaching 27, supporting PsBu’s potential as a targeted anticancer agent. Zebrafish embryo assays yielded LC₅₀ values ranging from 1.4 to 1.8 mg·mL⁻¹. In vivo, PsBu reduced fasting blood glucose levels in hyperglycemic rats (approximately 210 mg·dL⁻¹ vs. 230 mg·dL⁻¹ in controls) and preserved pancreatic β-cell integrity (around 80% vs. 65% in controls). Conclusions: These findings suggest that B. utriformis biomass and PsBu exhibit strong antioxidant activity, selective cytotoxicity against cancer cells, and antihyperglycemic potential, making them promising candidates for further biomedical applications.

Bovistella utriformis (Bull.) Demoulin & Rebriev is a cosmopolitan puffball mushroom traditionally recognized for its anti-inflammatory and wound-healing properties. This study aimed to evaluate the cytotoxic profile, regenerative potential, and molecular effects of polysaccharides extracted from B. utriformis (PsBU) using complementary in vitro and in vivo approaches. Thermogravimetric analysis revealed good thermal stability up to 300 °C with 26.91% residual mass at 800 °C. Human HaCaT keratinocytes were used to assess cell viability, proliferation, and cell-cycle distribution through MTT assays and flow cytometry, while wound-healing activity was examined using in vitro scratch assays. PsBU exhibited no cytotoxic effects across concentrations from 19.53 to 10,000 µg mL⁻¹, with cell viability remaining above 68%. At 5000 and 10,000 µg mL⁻¹, viability increased to 130% and 126%, respectively. The optimal in vitro wound-healing effect was observed at 500 µg mL⁻¹, achieving 40% wound closure at 32 h. Label-free quantitative proteomic analysis by UHPLC-HRMS identified 83 differentially expressed proteins, including upregulation of tissue repair-related factors such as plasminogen and FHL2, alongside modulation of cell-cycle regulation and mRNA transport pathways. In vivo zebrafish caudal fin regeneration assays demonstrated maximal regenerative activity at 200 µg mL⁻¹ (p < 0.001 vs. control). Overall, these findings demonstrate that B. utriformis polysaccharides are safe bioactive compounds that promote key biological processes involved in tissue regeneration, supporting their potential application as natural wound-healing agents.

Natural fungal polysaccharides are increasingly explored as bioactive compounds capable of orchestrating complex regenerative responses during tissue repair. This study aimed to evaluate the in vivo wound-healing efficacy and molecular mechanisms of a topical polysaccharide formulation derived from Bovistella utriformis (Calvatin 2%) using complementary murine, zebrafish, and proteomic approaches. Phylogenetic analysis based on ITS sequences confirmed the taxonomic identity of the Chilean specimen. In a murine full-thickness excisional wound model, Calvatin 2% significantly accelerated wound contraction and re-epithelialization compared to both saline and base-cream controls, achieving near-complete closure by day 10. Label-free quantitative proteomic analysis of wound tissue by UHPLC-HRMS identified 2432 high-confidence proteins, with 171 upregulated and 153 downregulated proteins in the Calvatin versus control comparison (p < 0.01). Functional enrichment revealed strong activation of innate immune response, complement activation, coagulation cascades, and acute-phase response pathways, while lipid metabolism, mitochondrial energy production, and muscle-related processes were significantly downregulated. KEGG pathway analysis further highlighted complement and coagulation cascades and neutrophil extracellular trap formation as the most prominently affected pathways. In a zebrafish laser-induced wound model, Calvatin induced early and sustained regenerative responses, reaching over 93% wound closure by 18 days post-lesion, significantly outperforming both PBS and vehicle-treated groups. Chronic oral administration of polysaccharides did not induce major hepatic inflammatory responses, supporting systemic safety. Overall, these findings indicate that B. utriformis polysaccharides are associated with modulation of immune- and repair-related pathways together with tissue reprogramming processes that may contribute to accelerated cutaneous regeneration, positioning Calvatin as a promising bioactive formulation for wound-healing applications.

Plantago lanceolata L. is a herb with diverse pharmacological properties attributed to its rich phytochemical composition, including iridoids, flavonoids, caffeic acid derivatives, and polyphenols with significant antioxidant potential. This study investigated how defoliation frequency (extended leaf length, ELL: 15, 25, and 35 cm) and intensity (residual height, RH: 5 and 8 cm) affect bioactive compound content and antioxidant activity in P. lanceolata extracts. Six defoliation combinations were evaluated in a 3 × 2 factorial arrangement (three pots per combination; n = 18 pots). Total phenolic content (TPC), antioxidant activity (DPPH radical scavenging), lipid content, the iridoids aucubin (AU) and catalpol (CAT), and the phenylpropanoid glycoside acteoside (AC) were determined in the aerial biomass, and high- and low-molecular-weight water-soluble carbohydrate (WSC) content in the aerial and root biomass. Functional group profiling was performed using Fourier transform infrared (FT-IR) spectroscopy; lipophilic compounds were putatively identified by gas chromatography–mass spectrometry (GC-MS) in pooled samples; and AU, CAT, and AC were quantified by high-performance liquid chromatography (HPLC). Total phenolic content depended on the ELL × RH interaction, while antioxidant activity was higher at 5 cm than at 8 cm RH (43.0 vs. 30.3%). Lipid content was not significantly affected by defoliation. GC-MS profiles were dominated by n-alkanes; putative neophytadiene exceeded the reporting threshold only at 15 cm ELL/5 cm RH, and tentatively identified α-tocospiro compounds (α-tocopherol oxidation products) showed their highest relative abundance at 35 cm ELL. Among the targeted bioactive compounds, acteoside was higher under the more frequent defoliations (15 and 25 cm ELL) and at 5 cm RH, catalpol remained unchanged across treatments, and aucubin was below the limit of quantification. High- and low-molecular-weight WSC contents in the aerial and root biomass accumulated most under the less frequent (25 and 35 cm ELL) defoliation regimes, driven mainly by greater herbage and root mass rather than by changes in concentration. Root low-molecular-weight WSC content was also higher at 8 than at 5 cm RH. Overall, more intense defoliation (5 cm RH) increased antioxidant activity and acteoside concentration, whereas less frequent defoliation increased WSC content, suggesting that defoliation management could be adjusted to balance the phytochemical quality and regrowth potential of P. lanceolata.

This study presents the first comprehensive lipophilic chemical profile of Bovistella utriformis, integrated with a multimodal physicochemical characterization of its whole biomass collected from Chilean Patagonia, complementing recent studies that demonstrate polysaccharide bioactivity, wound-healing efficacy, and molecular mechanisms of action. Scanning electron microscopy (SEM) revealed a heterogeneous microstructure comprising spheroidal particles (2–5 µm) embedded in a porous, fibrous matrix. Fourier transform infrared spectroscopy with attenuated total reflectance (FTIR-ATR) revealed characteristic bands of polysaccharides, proteins, and chitin, with C–O/C–O–C bands typical of fungal cell-wall polysaccharides. Thermogravimetric analysis (TGA) revealed three degradation events at 75.4 °C, 212.0 °C, and 306.0 °C, with a residual mass of 25.9% at 800 °C under nitrogen. Gas chromatography–mass spectrometry (GC-MS) analysis showed that the underivatized hexane, ethyl acetate and chloroform extracts are dominated by an unresolved envelope of free fatty acids (78–81% of total ion current) accompanied by 29 resolved minor compounds, mainly fatty acid methyl and ethyl esters, C8 alcohols, niacinamide and an ergostane sterol; acid-catalyzed transesterification collapsed this profile into six fatty acid methyl esters and three sterols. Remarkably, linoleic acid accounted for 90–92% of total fatty acids in all derivatized extracts, with palmitic acid (5–7%) as the only other major component and oleic acid at just 1.2–1.4%; ergosterol and ergosta-7,22-dien-3β-ol (0.9–3.5%) were the accompanying sterols. This represents the highest reported linoleic acid content in puffball mushrooms and distinguishes B. utriformis from most other edible fungi studied, supporting its valorization as an exceptional source of bioactive fatty acids and fungal sterols for nutraceutical, pharmaceutical, and biotechnological applications.