Biomechanical testing to assess shear strength of repair On Day 14, shear strength of repair between the outer ring and sellckchem inner core of meniscal repair explants was measured with a push out test using an Elec troforce 3200 materials testing system. Briefly, explants were centered in a custom made appa ratus, such that the 3 mm inner core was centered over a 4 mm concentric hole in the bottom of the dish. A 2 mm diameter rod attached to a load cell displaced the inner core at a rate of 0. 0833 mm s until the inner core was dislodged from the outer ring. The force required for displacement was recorded over time. Following the push out test, the inner core was imaged using a digital video camera with a 94 mm video lens to measure the inner core thickness using LabVIEW Vision Builder AI.

Shear strength of repair was calculated by dividing the peak force measured during the push out test by the surface area of the interface. Histological staining of meniscal explants On Day 12 of the meniscal repair model explant cul ture, 0. 05% nitroblue Inhibitors,Modulators,Libraries tetrazolium chloride was added to the explant culture media for histological analyses. NBT is a cell permeable com pound that Inhibitors,Modulators,Libraries is reduced by live cells to form a blue for mazan product that remains stable to histological processing and paraffin embedding and has been docu mented as a live cell marker for chondrocytes. At Day 14, explants were fixed overnight in 4% parafor maldehyde, containing 100 mM sodium cacodylate trihydrate, pH 7. 4 at 4 C. Samples were dehydrated Inhibitors,Modulators,Libraries in EtOH, infiltrated with xylene, and paraffin embedded.

Sections were stained with 0. 02% aqueous fast green to label collagens and Accustain Safranin O solution to identify proteoglycans. Statistical analyses Statistical analyses were performed using Statistica 7. 0. A factorial analysis of variance and the Newman Keuls post hoc test were performed Inhibitors,Modulators,Libraries to determine significant differences and the interactive effect of time and treatment in the micro wounding experiments. In the meniscal repair model explant studies, the interactive effect of treatment and tissue zone in the surface images and push out test and of treatment, tissue zone and cross section layer in the cross section images were also determined using a factorial ANOVA Inhibitors,Modulators,Libraries and Newman Keuls post hoc test.

Results The effects of serum on inner and outer zone micro wound repair Serum treatment of meniscal cells from both the inner and outer zones resulted in increased accumulation of proliferated cells in the micro wound. For inner zone meniscal cells, 10% serum increased the total number of cells in the wound as compared to the control, increased selleck Navitoclax the percentage of proliferated cells in the wound com pared to all other treatments, and enhanced cellular proliferation away from the wound over the control and 1% serum treatments. Addition ally, 5% serum promoted cellular proliferation in the wound over the control treatment.