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An ectopic study of tissue-engineered bone with Nell-1 gene modified rat bone marrow stromal cells in nude mice / 中华医学杂志(英文版)
Chinese Medical Journal ; (24): 972-979, 2009.
Artigo em Inglês | WPRIM | ID: wpr-279799
ABSTRACT
<p><b>BACKGROUND</b>Tissue engineering techniques combined with gene therapy have been recently used to improve osteogenesis. NEL-like molecule-1 (Nell-1), a novel growth factor, has been reported to have specificity for osteochondral lineage. The study assessed the osteogenic differentiation of rat bone marrow stromal cells (bMSCs) after Nell-1 gene modification and examined its ectopic bone formation ability in a nude mice model with tissue engineering technique.</p><p><b>METHODS</b>bMSCs obtained from Fischer 344 rats were transduced with either AdNell-1 (Nell-1 group) or Ad-beta-galactosidase (AdLacZ, LacZ group) or left untransduced (untransduced group). The expression of Nell-1 protein was determined by Western blotting and transfer efficiency was assessed. mRNA expressions of osteopontin (OP), bone sialoprotein (BSP) and osteocalcin (OC) were assessed by real-time PCR 0, 3, 7, 14, and 21 days after gene transfer. Alkaline phosphatase (ALP) activity was measured and von Kossa test was also conducted. Finally, with a tissue engineering technique, gene transduced bMSCs, combining with beta-tricalcium phosphate (beta-TCP) at a concentration of 2 x 10(7) cells/ml, were implanted at subcutaneous sites on the back of nude mice. Four weeks after surgery, the implants were evaluated with histological staining and computerized analysis of new bone formation.</p><p><b>RESULTS</b>Under current transduction conditions, gene transfer efficiency reached (57.9 +/- 6.8)%. Nell-1 protein was detected in Nell-1 group but not in untransduced group and LacZ group. Induced by Nell-1, BSP and OP expression were increased at intermediate stage and OC expression was increased at later stage. ALP activity and the number of calcium nodules were highest in Nell-1 group. Four weeks after implanted into nude mice subcutaneously, the percentage of new bone area in Nell-1 group was (18.1 +/- 5.0)%, significantly higher than those of untransduced group (11.3 +/- 3.2)% and LacZ group (12.3 +/- 3.1)% (P < 0.05).</p><p><b>CONCLUSIONS</b>This study has demonstrated the ability of Nell-1 to induce osteogenic differentiation of rat bMSCs in vitro and to enhance bone formation with a tissue engineering technique. The results suggest that Nell-1 may be a potential osteogenic gene to be used in bone tissue engineering.</p>
Assuntos
Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Osteogênese / Ratos Endogâmicos F344 / Sialoglicoproteínas / Células da Medula Óssea / Microscopia Eletrônica de Varredura / Osteocalcina / Western Blotting / Células Estromais / Reação em Cadeia da Polimerase Via Transcriptase Reversa / Biologia Celular Tipo de estudo: Estudo prognóstico Limite: Animais Idioma: Inglês Revista: Chinese Medical Journal Ano de publicação: 2009 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Osteogênese / Ratos Endogâmicos F344 / Sialoglicoproteínas / Células da Medula Óssea / Microscopia Eletrônica de Varredura / Osteocalcina / Western Blotting / Células Estromais / Reação em Cadeia da Polimerase Via Transcriptase Reversa / Biologia Celular Tipo de estudo: Estudo prognóstico Limite: Animais Idioma: Inglês Revista: Chinese Medical Journal Ano de publicação: 2009 Tipo de documento: Artigo