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Local Application of Pyrophosphorylated Simvastatin Prevents Experimental Periodontitis

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Abstract

Purpose

Simvastatin (SIM), a HMG-CoA reductase inhibitor widely prescribed for hypercholesterolemia, has been reported to ameliorate inflammation and promote osteogenesis. Its clinical applications on these potential secondary indications, however, have been hampered by its lack of osteotropicity and poor water solubility. To address this challenge, we propose to design and evaluate the therapeutic efficacy of a novel simvastatin prodrug with better water solubility and bone affinity.

Method

The prodrug (SIM-PPi) was synthesized by directly conjugating a SIM trimer to a pyrophosphate (PPi). It was characterized and evaluated in vitro for its water solubility, osteotropicity, toxicity, anti-inflammatory and osteoinductive properties. It was then tested for anti-inflammatory and osteoinductive properties in vivo by three weekly injections into gingiva of a ligature-induced experimental periodontitis rat model.

Results

In vitro studies showed that SIM-PPi has greatly improved water-solubility of SIM and shows strong binding to hydroxyapatite (HA). In macrophage culture, SIM-PPi inhibited LPS-induced pro-inflammatory cytokines (IL-1β, IL-6). In osteoblast culture, it was found to significantly increase alkaline phosphatase (ALP) activity with accelerated mineral deposition, confirming the osteogenic potential of SIM-PPi. When tested in vivo on an experimental periodontal bone-loss model, SIM-PPi exhibited a superior prophylactic effect compared to dose equivalent SIM in reducing inflammatory cells and in preserving alveolar bone structure, as shown in the histological and micro-CT data.

Conclusion

SIM-PPi may have the potential to be further developed for better clinical management of bone loss associated with periodontitis.

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Abbreviations

ABC:

Alveolar bone crest

ALP:

Alkaline phosphatase

BMP-2 :

Bone morphogenetic protein-2

CEJ:

Cementoenamel junction

EDTA:

Ethylenediaminetetraacetic acid

GCF:

Gingival crevicular fluid

HA:

Hydroxyapatite

HMG-CoA:

3-Hydroxy-3-methyl-glutaryl-coenzyme A

LPS:

Lipopolysaccharide

M1:

First molar

M2:

Second molar

MCP-1 :

Monocyte chemoattractant protein-1

MMPs:

Matrix metalloproteinases

MTT:

3-(4,5-Dimethyl- thiazol-2yl)-2,5- diphenyltetrazoliumbromide

OPG:

Osteoprotegerin

PPi:

Pyrophosphate

RANK:

Receptor activator of nuclear factor-kappa B

RANKL:

Receptor activator of nuclear factor-kappa B ligand

ROI:

Region of interest

SIM:

Simvastatin

SIM-PPi:

Simvastatin-pyrophosphate

Tb.N:

Trabecular number

Tb.Sp:

Trabecular spacing or separation

Tb.Th:

Trabecular thickness

μ-CT:

Micro-computed tomography

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ACKNOWLEDGMENTS AND DISCLOSURES

This work was financially supported in part by the University of Nebraska Medical Center College of Pharmacy. Yosif Almoshari was supported by the Ministry of Education Scholarship, Jazan University (Jazan, Saudi Arabia). DW, RAR, XBW and ZSJ are co-inventors of a patent application of the SIM-PPi prodrug technology. The other authors report no conflicts of interest related to this study.

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Author notes

  1. Xiaobei Wang, Zhenshan Jia and Yosif Almoshari contributed equally to this work.

Authors and Affiliations

  1. The Department of Pharmaceutical Sciences, College of Pharmacy, University of Nebraska Medical Center, Omaha, Nebraska, 68198-6125, USA

    Xiaobei Wang, Zhenshan Jia, Yosif Almoshari & Dong Wang

  2. The Department of Pharmaceutics, College of Pharmacy, Jazan University, Jazan, 45142, Saudi Arabia

    Yosif Almoshari

  3. The Department of Pathology & Microbiology, College of Medicine, University of Nebraska Medical Center, Omaha, Nebraska, 6819-3135, USA

    Subodh M. Lele

  4. The Department of Surgical Specialties, College of Dentistry, University of Nebraska Medical Center, Lincoln, Nebraska, 68583-0740, USA

    Richard A. Reinhardt

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  1. Xiaobei Wang
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  2. Zhenshan Jia
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Correspondence to Dong Wang.

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Wang, X., Jia, Z., Almoshari, Y. et al. Local Application of Pyrophosphorylated Simvastatin Prevents Experimental Periodontitis. Pharm Res 35, 164 (2018). https://doi.org/10.1007/s11095-018-2444-z

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