Chemical Profile, Bioactive Constituents and In Vitro Growth Stimulation Properties of Cold-Pressed Hemp Seed Oils from Romanian Varieties: In Vitro and In Silico Evaluation

Industrial hemp (<i>Cannabis sativa</i> L.; <i>Cannabaceae</i>), traditionally cultivated for fiber, also represents a valuable source of nutrient-rich seed oil. In this study, cold-pressed hemp seed oils from three Romanian varieties (Teodora, Silvana, and Armanca) were evaluated for their fatty acid composition, minor bioactive constituents, antioxidant activity, growth-promoting property toward probiotic strains in vitro, and molecular docking interactions with probiotic targets. Gas chromatography revealed a fatty acid profile dominated by linoleic (49.4-51.9%), oleic (16.3-22.8%), and α-linolenic acids (9.8-14.4%), resulting in favorable PUFA/SFA ratios (5.17-6.39) and ω-6/ω-3 ratios (3.93-5.53). The oils also contained phenolics (118-160 mg GAE/kg), chlorophylls (6.18-8.31 mg/kg), and carotenoids (2.58-3.37 mg/kg), which contributed to their antioxidant activity (DPPH inhibition 35.92 µM TE/100 g-43.37 µM TE/100 g). Broth microdilution assays against <i>Lacticaseibacillus rhamnosus</i> GG, <i>L. paracasei</i> ATCC BAA-52, and <i>L. acidophilus</i> ATCC 4356 demonstrated strain- and dose-dependent potential to promote probiotic growth under in vitro conditions. While <i>L. rhamnosus</i> and <i>L. paracasei</i> were inhibited at low concentrations and only mildly stimulated at higher levels, <i>L. acidophilus</i> showed robust growth promotion, reaching +54.7% effect and CP = 1.55 with Teodora oil at 16 mg/mL. Molecular docking highlighted strong binding affinities of γ-linolenic and linoleic acids with key metabolic enzymes involved in probiotic metabolism (hydratase, enolase, glyceraldehyde-3-phosphate dehydrogenase, ribonucleoside hydrolase), forming stable hydrophilic and hydrophobic interactions which are explored in defining the stability of the ligand-protein complexes. These results indicate that both major fatty acids and minor bioactive constituents contribute to the nutritional and antioxidant value of Romanian hemp seed oils and reveal a potential to promote probiotic growth under in vitro conditions, as supported by complementary in silico evidence.