Simvastatin With PRP Promotes Chondrogenesis of Bone Marrow Stem Cells In Vitro and Wounded Rat Achilles Tendon-Bone Interface Healing In Vivo.

BACKGROUND: Tendons and ligaments are joined to bone in a specialized interface that transmits force from muscle to bone and permits body movement. Tendon/ligament injuries always occur in the interface areas, and injured tendons/ligaments have a limited healing response because the insertion site is composed of a fibrocartilaginous zone. PURPOSE: To study the effect of simvastatin with platelet-rich plasma (PRP) on chondrogenesis of rat bone marrow stem cells (BMSCs) in vitro and wounded rat Achilles tendon-bone interface healing in vivo. STUDY DESIGN: Controlled laboratory study. METHODS: The in vitro model was performed by the culture of rat BMSCs with various concentrations of simvastatin (0, 10, 50, 100 nM) for 2 weeks. The effect of simvastatin on the chondrogenic differentiation of the BMSCs was examined by histochemical analysis and real-time quantitative reverse transcription polymerase chain reaction. The in vivo model was carried out by testing the healing effect of simvastatin with PRP on 12 wounded rat Achilles tendon-bone interfaces. RESULTS: Simvastatin induced chondrogenic differentiation of rat BMSCs in a concentration-dependent manner as evidenced by histological staining and real-time quantitative reverse transcription polymerase chain reaction. The wounds treated with simvastatin alone or with simvastatin-containing PRP gel healed much faster than the wounds treated with saline alone or PRP alone. Histological analysis showed that higher percentages of healed tissues were positively stained with safranin O and fast green in wounds treated with simvastatin-containing PRP gel than in the other 3 groups. Immunohistochemical analysis further demonstrated these findings, as evidenced by more positively stained healed tissues with collagen I and II antibodies in the wound areas treated with simvastatin-containing PRP gel than the other 3 groups. CONCLUSION: The combination of simvastatin with PRP induced chondrogenesis of BMSCs in vitro and enhanced fibrocartilage formation in vivo. The simvastatin-PRP gel treatment promotes wounded tendon-bone interface healing in clinical treatment. CLINICAL RELEVANCE: The combination of simvastatin with PRP may be a good clinical treatment for wounded tendon/ligament junction healing, especially for acute sports-related tendon/ligament injuries.

Effects of allicin on the proliferation and cell cycle of chondrocytes.

The present study demonstrates the effect of allicin on the proliferation and the cell cycle distribution of the chondrocytes. MTT assay and flow cytometry were used for the evaluation of the effect of allicin on cell proliferative and the cell cycle distribution, respectively of the chondrocytes. The reverse transcription polymerase chain reaction (RT-PCR) and western blot analysis were respectively used for the analysis of mRNA and protein expression levels of cyclin D1, CDK4 and CDK6. The results revealed that exposure of the chondrocytes to allicin at a concentration of 40 µM significantly promoted the cell viability. Treatment of the cells with 10, 20, 30, 40, and 50 µg/mL of allicin enhanced the cell viability by 2.5.47 ± 0.86, 5.43 ± 0.66, 10.74 ± 1.48, 35.89 ± 3.78, and 32.21 ± 2.92%, respectively after 36 h compared to control cells. Allicin exposure caused a marked decrease in the percentage of cells in G0/G1 phase with a subsequent increase in the S phase population. Furthermore, allicin treatment enhanced the expression of cyclin D1, CDK4 and CDK6. Therefore, allicin treatment enhances the proliferation of chondrocytes by promoting the transition from G1 to S phase of the cell cycle through increase in the expression of cyclin D1, CDK4 and CDK6 levels.

Preparation and characterization of nano-liposome-mediated FL gene in the Lovo cells.

AIMS: The purpose of the present work was to formulate and evaluate cationic nano-liposomes as novel nonviral gene delivery for colon cancer treatment. METHODS: Recombinant pEGFP-c1-Fms-like tyrosine kinase receptor 3 ligand (FL) plasmids containing human FL gene and green fluorescent protein (GFP) reporter genes were constructed. FL and GFP Gene-carrying cationic nano-liposomes were prepared based on the electrostatic adherence principle and then transfected into Lovo cells. The morphology, particle size, and zeta potential of gene-carrying cationic nano-liposomes were observed using an electron microscope. GFP expression was observed by fluorescence microscopy to assay the transfection efficiency. The cytotoxicity of FL/nano-liposomes was evaluated by the MTT method. RESULTS: Recombinant plasmids pEGFP-c1-FL are successfully constructed using gene cloning methods and confirmed by restriction enzyme digestion and sequencing. The cationic nano-liposomes carrying pEGFP-cl-FL were observed by an electron micrograph and showed uniform spherical or elliptical shapes and many pores. The fluorescence microscopy images of gene-carrying cationic nano-liposomes showed good expression of GFP in pEGFP and pEGFP-cl-FL groups. The MTT assay of cell death indicated a significantly higher level of cell death between the FL group and the control group at 24, 48, and 96 hours after transplantation. CONCLUSION: Cationic nano-liposomes show safe and high-performance transfection as gene carriers. Gene therapy has significant implications for colon cancer treatment in future.