Reduction of inflammation in a chronic periodontitis model in rats by TNF-α gene silencing with a topically applied siRNA-loaded calcium phosphate paste

被引：23

作者：

Tenkumo, Taichi ^{[1
]}

Rojas-Sanchez, Leonardo ^{[2
,3
]}

Saenz, Juan Ramon Vanegas ^{[4
]}

Ogawa, Toru ^{[1
]}

Miyashita, Makiko ^{[1
]}

Yoda, Nobuhiro ^{[1
]}

Prymak, Oleg ^{[2
,3
]}

Sokolova, Viktoriya ^{[2
,3
]}

Sasaki, Keiichi ^{[1
]}

Epple, Matthias ^{[2
,3
]}

机构：

[1] Tohoku Univ, Grad Sch Dent, Div Adv Prosthet Dent, Aoba Ku, 4-1 Seiryo Machi, Sendai, Miyagi 9808575, Japan

[2] Univ Duisburg Essen, Inorgan Chem, Univ Str 5-7, D-45117 Essen, Germany

[3] Univ Duisburg Essen, Ctr Nanointegrat Duisburg Essen CeNIDE, Univ Str 5-7, D-45117 Essen, Germany

[4] Univ Amer UAM, Costado Noroeste Camino Oriente, Fac Odontol, Mat Dentales, Managua, Nicaragua

来源：

ACTA BIOMATERIALIA | 2020年 / 105卷

基金：

日本学术振兴会;

关键词：

Calcium phosphate; Nanoparticles; Gene silencing; TNF-alpha; Periodontitis; Anti-inflammation; IN-VIVO; NUCLEIC-ACIDS; NANOPARTICLES; TRANSFECTION; DELIVERY; DNA; MACROPHAGES; EXPRESSION; CARRIERS; RELEASE;

D O I：

10.1016/j.actbio.2020.01.031

中图分类号：

R318 [生物医学工程];

学科分类号：

0831 ;

摘要：

We developed a calcium phosphate-based paste containing siRNA against TNF-alpha and investigated its anti-inflammatory and bone-healing effects in vitro and in vivo in a rat periodontitis model. The bioactive spherical CaPIPElisiRNA/SiO2 nanoparticles had a core diameter of 40-90 nm and a positive charge (+23 mV) that facilitated cellular uptake. The TNF-alpha gene silencing efficiency of the nanoparticles in J774.2 monocytes, gingival-derived cells, and bone marrow-derived cells was 12 +/- 2%, 36 +/- 8%, and 35 +/- 22%, respectively. CaP/PElisiRNA/SiO2 nanoparticles cancelled the suppression of alkaline phosphatase (ALP) activity in LPS-stimulated bone marrow-derived cells. In vivo, ALP mRNA was up-regulated, TNF-alpha mRNA was down-regulated, and the amount of released TNF-alpha was significantly reduced after topical application of the calcium phosphate-based paste containing siRNA-loaded nanoparticles. The number of TNF-alpha-positive cells in response to CaP/PElisiRNA/SiO2 nanoparticle application was lower than that observed in the absence of siRNA. Elevated ALP activity and numerous TRAP-positive cells (osteoclasts) were observed in response to the application of all calcium phosphate pastes. These results demonstrate that local application of a paste consisting of siRNA-loaded calcium phosphate nanoparticles successfully induces TNF-alpha silencing in vitro and in vivo and removes the suppression of ALP activity stimulated by inflammation. Statement of significance We developed a calcium phosphate-based paste containing nanoparticles loaded with siRNA against TNF-alpha. The nanoparticles had a core diameter of 40-90 nm and positive charge (+23 my). The antiinflammatory and osteoinductive effects of the paste were investigated in vitro and in vivo in a rat periodontitis model. In vitro, the TNF-alpha gene silencing efficiency of the nanoparticles in J774.2 monocytes, gingival-derived cells, and bone marrow-derived cells was 12 +/- 2%, 36 +/- 8%. and 35 +/- 22%, respectively. The ALP activity of bone marrow-derived cells was recovered. In vivo, TNF-alpha mRNA was down-regulated and the amount of released TNF-alpha was significantly reduced, whereas the ALP mRNA was up-regulated. Elevated ALP activity and TRAP-positive cells were observed by immunohistochemistry. (C) 2020 Published by Elsevier Ltd on behalf of Acta Materialia Inc.

引用

页码：263 / 279

页数：17

共 59 条

[1]

[Anonymous], HOST PARASITE INTERA

[2]

[Anonymous], PULPAL REACTIONS CAR

[3] Systematic review: genetic biomarkers associated with anti-TNF treatment response in inflammatory bowel diseases [J].