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<p>This study evaluates the bioaccessibility and anti-osteoporotic efficacy of a green leafy vegetable-based food formulation rich in vitamins D<sub>2</sub> (ergocalciferol) and K<sub>1</sub> (phylloquinone). Freeze-drying preserved nutrient content most effectively (D<sub>2</sub>: 97.1%; K<sub>1</sub>: 95.9% w/w in <i>Murraya koenigii</i>), and an optimized static gastrointestinal digestion model demonstrated high, concentration-independent bioaccessibility (D<sub>2</sub>: 76.6%; K<sub>1</sub>: 72.7%). Pharmacokinetic analysis in rabbits revealed a strong correlation between in vitro and in vivo results (relative bioavailability: D<sub>2</sub> 68.7%; K<sub>1</sub> 64.4%; Pearson <i>r</i> = 1.0000, <i>p</i> &lt; 0.0001). In ovariectomized rats, the high-dose formulation (2&#xa0;g/kg, n = 6) significantly restored mineral homeostasis (serum calcium: + 8.2&#xa0;mg/dL vs. OVX control), reduced bone turnover markers (alkaline phosphatase − 42%, osteocalcin − 38%, <i>p</i> &lt; 0.05), and restored femoral bone architecture comparable to alendronate (5&#xa0;mg/kg) and standard vitamin D<sub>2</sub>–K<sub>1</sub> supplementation. The low-dose formulation (1&#xa0;g/kg) demonstrated limited efficacy, with only an 18% reduction in alkaline phosphatase, indicating a dose-dependent therapeutic response. Histological analysis confirmed nearly complete trabecular bone repair in the high-dose group (<i>p</i> &lt; 0.05) compared to OVX disease control. These findings support the use of this food-based formulation as an effective adjunct for the management of postmenopausal osteoporosis.</p>

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Evaluating the bioaccessibility and osteoprotective effects of a vitamin-rich green leafy vegetable formulation: insights from in vitro and in vivo methodologies

  • 期刊论文

  • 发表日期：

    2026年4月15日

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  • Anoop Karthika,
  • Kowmudi Gullapalli,
  • Rashmi Venkatesh,
  • Sai Varshini Magham,
  • Krishnaveni Nagappan,
  • Praveen Thaggikuppe Krishnamurthy,
  • Sreenath Konanki,
  • Anilakumar Kandangath Raghavan

摘要

This study evaluates the bioaccessibility and anti-osteoporotic efficacy of a green leafy vegetable-based food formulation rich in vitamins D2 (ergocalciferol) and K1 (phylloquinone). Freeze-drying preserved nutrient content most effectively (D2: 97.1%; K1: 95.9% w/w in Murraya koenigii), and an optimized static gastrointestinal digestion model demonstrated high, concentration-independent bioaccessibility (D2: 76.6%; K1: 72.7%). Pharmacokinetic analysis in rabbits revealed a strong correlation between in vitro and in vivo results (relative bioavailability: D2 68.7%; K1 64.4%; Pearson r = 1.0000, p < 0.0001). In ovariectomized rats, the high-dose formulation (2 g/kg, n = 6) significantly restored mineral homeostasis (serum calcium: + 8.2 mg/dL vs. OVX control), reduced bone turnover markers (alkaline phosphatase − 42%, osteocalcin − 38%, p < 0.05), and restored femoral bone architecture comparable to alendronate (5 mg/kg) and standard vitamin D2–K1 supplementation. The low-dose formulation (1 g/kg) demonstrated limited efficacy, with only an 18% reduction in alkaline phosphatase, indicating a dose-dependent therapeutic response. Histological analysis confirmed nearly complete trabecular bone repair in the high-dose group (p < 0.05) compared to OVX disease control. These findings support the use of this food-based formulation as an effective adjunct for the management of postmenopausal osteoporosis.