SFRP1-Silencing GapmeR-Loaded Lipid-Polymer Hybrid Nanoparticles for Bone Regeneration in Osteoporosis: Effect of Dosing and Targeting Strategy

Int J Nanomedicine. 2024 Nov 20:19:12171-12188. doi: 10.2147/IJN.S476546. eCollection 2024.

Abstract

Introduction: Osteoporosis is a metabolic disorder characterized by the loss of bone mass and density. Nucleic acid-based therapies are among the most innovative approaches for osteoporosis management, although their effective delivery to bone tissue remains a challenge. In this work, SFRP1-silencing GampeR loaded-nanoparticles were prepared and functionalized with specific moieties to improve bone targeting and, consequently, therapeutic efficacy. SFRP1-silencing would promote osteoblastic differentiation by enhancing the WNT/β-catenin pathway and thus diminishing the progression of osteoporosis.

Methods: A nucleic acid-based delivery system consisting of lipid-polymer hybrid nanoparticles (LPNPs) loading a GapmeR for SFRP1 silencing was developed and further functionalized with two bone-targeting moieties: a specific aptamer (Apt) for murine mesenchymal stem cells and an antiresorptive drug, namely alendronate (ALD). These systems were tested in vivo in osteoporotic mice at different dosage regimens to analyze dose dependence in bone-forming activity and potential toxicity. The quality of trabecular and cortical bone was assessed by both micro computed tomography (micro-CT) and histological and histomorphometric analyses. Early and late osteogenesis were quantified by immunohistochemistry.

Results: Results showed that functionalizing LPNPs loaded with an SFRP1-silencing GapmeR using both Apt and ALD improved bone quality and enhanced osteogenesis following a dose-effect relationship, as revealed by micro-CT, histological and immunohistochemical analyses. In contrast, non-functionalized LPNPs did not produce these effects.

Conclusion: These findings highlight the relevance of proper targeting and dosage in nucleic acid-based therapeutics, proving to be crucial for exerting their therapeutic effect: a deficient targeting strategy and/or dosage may result in the therapeutic failure of an adequate gene therapy agent.

Keywords: alendronate; aptamer; bone targeting; gene therapy; micro-CT; nanoparticle dose-effect; osteoporosis.

MeSH terms

  • Alendronate* / administration & dosage
  • Alendronate* / chemistry
  • Alendronate* / pharmacokinetics
  • Alendronate* / pharmacology
  • Animals
  • Aptamers, Nucleotide / administration & dosage
  • Aptamers, Nucleotide / chemistry
  • Aptamers, Nucleotide / pharmacology
  • Bone Density Conservation Agents / administration & dosage
  • Bone Density Conservation Agents / pharmacology
  • Bone Regeneration* / drug effects
  • Female
  • Gene Silencing / drug effects
  • Intercellular Signaling Peptides and Proteins / administration & dosage
  • Intercellular Signaling Peptides and Proteins / pharmacology
  • Lipids / chemistry
  • Membrane Proteins* / genetics
  • Mesenchymal Stem Cells / drug effects
  • Mice
  • Mice, Inbred C57BL
  • Nanoparticles* / chemistry
  • Osteogenesis / drug effects
  • Osteoporosis* / drug therapy
  • Polymers / chemistry
  • Wnt Signaling Pathway / drug effects
  • X-Ray Microtomography

Substances

  • Membrane Proteins
  • Alendronate
  • Lipids
  • Sfrp1 protein, mouse
  • Intercellular Signaling Peptides and Proteins
  • Polymers
  • Aptamers, Nucleotide
  • Bone Density Conservation Agents

Grants and funding

This work was supported by the Spanish Ministry of Science and Innovation (MCIN/AEI/10.13039/501100011033/FEDER, UE, PID2021-127493OB-C21 and PID2021-127493OA-C22). E.B. thanks the Canary Islands Agency for Research, Innovation and Information Society (ACIISI) of the Counseling of Economy, Knowledge and Employment and the European Social Fund (ESF) Integrated Operational Program of the Canary Islands 2014–2020 for his predoctoral grant (TESIS2022010094).