AI Article Synopsis
- The study aimed to explore how lyophilization (a freeze-drying process) affects the biological activity of recombinant lentiviral vectors carrying bone morphogenetic protein 2 (BMP-2).
- Prior to and after lyophilization, tests showed no significant change in how the lyophilized lenti-BMP-2 affected the proliferation of rat bone marrow stromal cells (BMSCs) or the expression of key proteins like BMP-2.
- The results indicate that using trehalose in the lyophilization process effectively preserves the activity of lenti-BMP-2, making it a reliable method for long-term storage.
Article Abstract
Purpose: To investigate the influence of lyophilization on the biological activity of recombinant lentiviral vectors of bone morphogenetic protein 2(BMP-2).
Methods: Recombinant lenti-BMP-2 was constructed. lenti-BMP-2 was transfected with rat bone marrow stromal cells (BMSCs) by multiplicity infection (MOI) of 10, 25, 50, 100, 200. The infection efficiency was observed by X-gal staining. Under suitable conditions, the lenti-BMP-2 and 10% trehalose ratio of lyophilized protective agent was mixed into the lyophilization form. Before and after lyophilization, the effect of lenti-BMP-2 on the proliferation of BMSCs was evaluated by MTS assay. The expression of BMP-2 protein in the cells of lyophilized lenti-BMP-2 was detected by ELISA method. The expression of Runx-2, OCN, Col1 and OPN in BMSCs was detected by real-time PCR after transfection of lyophilized lenti-BMP-2. SPSS13.0 software package was used for statistical analysis.
Results: X-gal staining showed an MOI of 100 pfu/cell, and stable transfection efficiency. Before and after lyophilization, no significant change was observed in regard to the effect of lyophilized lenti-BMP-2 on BMSCs proliferation (P>0.05). ELISA method showed that BMSCs transfected by lyophilized lenti-BMP-2 could express BMP-2 protein continuously and stably at a high level. Before and after lyophilization, the result of real-time PCR showed that no significant difference in the expression of OPN,Col1,OCN and Runx-2 in BMSCs (P>0.05).
Conclusions: Lyophilized lenti-BMP-2 with trehalose can maintain high activity for a long time as an effective and reliable storage method.
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[Effects of lyophilized on the biological activities of lentiviral vector of bone morphogenetic protein 2].
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Article Synopsis
- The study aimed to explore how lyophilization (a freeze-drying process) affects the biological activity of recombinant lentiviral vectors carrying bone morphogenetic protein 2 (BMP-2).
- Prior to and after lyophilization, tests showed no significant change in how the lyophilized lenti-BMP-2 affected the proliferation of rat bone marrow stromal cells (BMSCs) or the expression of key proteins like BMP-2.
- The results indicate that using trehalose in the lyophilization process effectively preserves the activity of lenti-BMP-2, making it a reliable method for long-term storage.
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Department of Orthodontics, College of Stomatology, Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
To enhance biocompatibility and osseointegration between titanium implants and surrounding bone tissue, numerous efforts have been made to modify the surface topography and composition of Ti implants. In this paper, Lenti-BMP-2-loaded TiO2 nanotube coatings were fabricated by lyophilization in the presence of trehalose to functionalize the surface. We characterized TiO2 nanotube layers in terms of the following: surface morphology; Lenti-BMP-2 and trehalose release; their ability to induce osteogenesis, proliferation, and anti-inflammation in vitro; and osseointegration in vivo.
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