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Purpose <p>This study investigates the incorporation of curcumin, resveratrol, and vitamin D3 into polymer-integrated calcium phosphate (CaP) scaffolds to assess their effects on osteogenic gene expression and osteosarcoma cell viability. Polymeric micelles and a polycaprolactone-polyethylene glycol (PCL-PEG) system were utilized to enhance drug loading, release, and bioactivity.</p> Methods <p>The scaffolds were functionalized with curcumin, resveratrol, and vitamin D3 using different polymeric carriers for controlled release. The release profile of these drugs was evaluated at physiological and acidic pH conditions. Osteoblast proliferation and differentiation were assessed. An osteoblast and osteoclast co-culture and RT-qPCR were performed to investigate osteogenic and osteoclastic gene expression. Resorption pit formation and Tartrate-Resistant Acid Phosphatase (TRAP) assays were conducted to analyze osteoclast activity. Lastly, osteosarcoma cell viability and morphology were examined at multiple time points to determine the anti-osteosarcoma efficacy of the drugs in vitro.</p> Results <p>The drug release study shows sustained release over a period of three days. At pH 5.0, the cumulative release of curcumin, resveratrol, and vitamin D3 reached 64%, 100%, and 80%, respectively. At pH 7.4, the corresponding release values were 25% for curcumin, 69% for vitamin D3, and 92% for resveratrol. In vitro, treated scaffolds enhance osteoblast proliferation by 1.1- to 1.3-fold and upregulate key osteogenic markers. Osteoclast activity was significantly reduced, with smaller resorption pits and decreased TRAP activity. Osteosarcoma viability decreases by 2.5- to 2.8-fold by day 11, indicating anti-cancer efficacy of the drugs.</p> Conclusion <p>The scaffolds successfully delivered bioactive compounds that promote osteoblast growth while reducing osteoclast activity and osteosarcoma cell proliferation. This multifunctional approach demonstrates the potential for enhancing bone regeneration and minimizing tumor recurrence post-surgery.</p> Lay Summary <p>Bone injuries often require implants that support healing and prevent complications like tumor recurrence. This study integrates curcumin, resveratrol, and vitamin D3 into CaP scaffolds to enhance bone regeneration and inhibit cancer cell growth. Using polymeric micelles and PCL-PEG, the compounds exhibit controlled release, improving osteoblast proliferation while reducing osteoclast activity. The scaffolds also suppress osteosarcoma cell viability, demonstrating potential for post-surgical cancer prevention. These findings suggest that drug-loaded CaP scaffolds could serve as multifunctional bone implants for bone tissue engineering applications.</p> Graphical Abstract <p></p>

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Multiple Drug Delivery Ability of Calcium Phosphate Scaffolds promotes Osteogenic Gene Expression in In Vitro Co-culture Model

  • 期刊论文

  • 发表日期：

    2025年9月2日

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  • Naboneeta Sarkar,
  • Yongdeok Jo,
  • Priya Kushram,
  • Susmita Bose

摘要

Purpose

This study investigates the incorporation of curcumin, resveratrol, and vitamin D3 into polymer-integrated calcium phosphate (CaP) scaffolds to assess their effects on osteogenic gene expression and osteosarcoma cell viability. Polymeric micelles and a polycaprolactone-polyethylene glycol (PCL-PEG) system were utilized to enhance drug loading, release, and bioactivity.

Methods

The scaffolds were functionalized with curcumin, resveratrol, and vitamin D3 using different polymeric carriers for controlled release. The release profile of these drugs was evaluated at physiological and acidic pH conditions. Osteoblast proliferation and differentiation were assessed. An osteoblast and osteoclast co-culture and RT-qPCR were performed to investigate osteogenic and osteoclastic gene expression. Resorption pit formation and Tartrate-Resistant Acid Phosphatase (TRAP) assays were conducted to analyze osteoclast activity. Lastly, osteosarcoma cell viability and morphology were examined at multiple time points to determine the anti-osteosarcoma efficacy of the drugs in vitro.

Results

The drug release study shows sustained release over a period of three days. At pH 5.0, the cumulative release of curcumin, resveratrol, and vitamin D3 reached 64%, 100%, and 80%, respectively. At pH 7.4, the corresponding release values were 25% for curcumin, 69% for vitamin D3, and 92% for resveratrol. In vitro, treated scaffolds enhance osteoblast proliferation by 1.1- to 1.3-fold and upregulate key osteogenic markers. Osteoclast activity was significantly reduced, with smaller resorption pits and decreased TRAP activity. Osteosarcoma viability decreases by 2.5- to 2.8-fold by day 11, indicating anti-cancer efficacy of the drugs.

Conclusion

The scaffolds successfully delivered bioactive compounds that promote osteoblast growth while reducing osteoclast activity and osteosarcoma cell proliferation. This multifunctional approach demonstrates the potential for enhancing bone regeneration and minimizing tumor recurrence post-surgery.

Lay Summary

Bone injuries often require implants that support healing and prevent complications like tumor recurrence. This study integrates curcumin, resveratrol, and vitamin D3 into CaP scaffolds to enhance bone regeneration and inhibit cancer cell growth. Using polymeric micelles and PCL-PEG, the compounds exhibit controlled release, improving osteoblast proliferation while reducing osteoclast activity. The scaffolds also suppress osteosarcoma cell viability, demonstrating potential for post-surgical cancer prevention. These findings suggest that drug-loaded CaP scaffolds could serve as multifunctional bone implants for bone tissue engineering applications.

Graphical Abstract