GNAS1 and PHD2 short-interfering RNA support bone regeneration in vitro and in an in vivo sheep model.
Clin Orthop Relat Res
; 470(9): 2541-53, 2012 Sep.
Article
em En
| MEDLINE
| ID: mdl-22833384
ABSTRACT
BACKGROUND:
Our ability to guide cells in biomaterials for in vivo bone repair is limited and requires novel strategies. Short-interfering RNA (siRNA) allows the regulation of multiple cellular pathways. Core binding factor alpha 1 (Cbfa1) and hypoxia-inducible factor 1 (HIF-1) pathways can be modulated to direct bone formation via siRNA against guanine nucleotide-binding protein alpha-stimulating activity polypeptide 1 (siGNAS1) and prolyl hydroxylase domain-containing protein 2 (siPHD2), respectively. QUESTIONS/PURPOSES:
We determined whether the administration of siGNAS1 and siPHD2 in mesenchymal stem cells (MSCs) promotes osteogenic phenotype, the dose-dependent effects of siGNAS1 on MSC differentiation to osteogenic phenotype, and whether the two siRNAs promote bone formation in vivo.METHODS:
siRNAs were administered to MSCs at Day 0, and protein expression of bone-specific markers was assessed at Days 1, 2, and 4 (n = 3/group/time point). In an in vivo model using seven sheep, chambers containing silk fibroin-chitosan (SFCS) scaffolds with siRNA were implanted over the periosteum and harvested at Days 7, 21, 36, and 70 (n = 4/group/time point, except at Day 70 [n = 2]) to assess bone formation.RESULTS:
siGNAS1 promoted collagen I and osteopontin expression, whereas siPHD2 had no effect in vitro. Dose-dependent effects of siGNAS1 on ALP expression were maximal at Day 1 for 10 µg/mL and Day 4 for 100 µg/mL. In vivo, by Day 70, mean bone volume increased compared to Day 7 for siGNAS1-SFCS (47.8 versus 1.8 mg/mL) and siPHD2-SFCS (61.3 versus 1.5 mg/mL).CONCLUSIONS:
Both siPHD2 and siGNAS1 support bone regeneration in vivo, whereas only siGNAS1 regulates bone phenotype in MSCs in vitro.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Periósteo
/
Regeneração Óssea
/
Terapia Genética
/
Pró-Colágeno-Prolina Dioxigenase
/
Subunidades alfa Gs de Proteínas de Ligação ao GTP
/
RNA Interferente Pequeno
/
Interferência de RNA
/
Células-Tronco Mesenquimais
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article