Human bone marrow stromal cells (HBMSCs) are multipotent and can form bone, cartilage or other tissues under different inductive conditions. The aim of this study was to investigate the effects of enamel matrix derivative (EMD) on the growth and osteogenic differentiation of HBMSCs. HBMSCs were cultured in monolayer with EMD (1, 10, 50,100, 250μg/ml) in aMEM supplemented with 2% FBS for 3 days. Cells cultured in aMEM supplemented with 2% FBS (basal medium) served as the control group. Double-stranded DNA was quantified by PicoGreen assay. Quantitative RT-PCR was performed to determine the expression levels of RUNX2, osteopontin (OPN) and osteocalcin (OCN), dentin matrix protein1 (DMP1) and dentin sialophosphoprotein (DSPP) at different time points (day 0, 5 and 10) when exposed to 10μg/ml EMD or basal medium. Alkaline phosphatase specific activity (ALPSA) was determined after 5 and 10 days culture. Mineral deposition (as calcium) was visualised using Alizarin Red staining.Objective
Methods
It was aimed to investigate the isolated effect of hydrostatic pressure on chondrocyte metabolism. Chondrocytes obtained from bovine metatarso-phalangeal joints were cultured in cylindrical 2% agarose gels. A special apparatus which was designed and constructed, allowed the application of hydrostatic pressure of either 2 MPa or 5 MPa on the chondrocytes for 4 hours either in a pulsatile (1Hz) or a static manner. Changes in the syntheses of glycosaminoglycan (GAG) and DNA during and after the application of the hydrostatic pressure were analysed with 35S-sulphate and 3H-thymidine incorporation, respectively. Radiolabelling was carried out for the following conditions: (a) 4 hours during the application of hydrostatic pressure; (b) 4 hours and (c) 20 hours immediately after the application of load. In addition, the experiments were carried out at 2 days, 7 days and 14 days after embedding the chondrocytes in agarose gels. Static hydrostatic pressure of 5 MPa caused a significant increase by 13% on average in the GAG synthesis during the load application on Day 2 7 and 14 (p <
0.05). On the contrary, pulsatile pressure of 2 MPa caused a significant decrease by 17% in the GAG synthesis measured at 20 hours after the loading on Day 14 (p <
0.01). In addition, there was a significant decrease by 29% in the DNA synthesis measured at 4 hours after the pulsatile loading of 5 MPa on Day 7 (p <
0.01). The results suggest that hydrostatic pressure alone, which causes no cell deformation, can affect the GAG synthesis and proliferation of chondrocytes. In addition, the effect of hydrostatic pressure on the chondrocyte metabolism varies according to the regimes of loading and with the period of cell culture.
There are various methods of measuring proprioception at the knee. Beard et al (1993) have described a delay in reflex hamstring contraction in anterior cruciate deficient knees. We have repeated their experiment and were unable to detect any significant difference in reflex hamstring contraction between the injured and uninjured legs. We discuss possible neurophysiological and biomechanical causes for the conflicting results and conclude that this method may not be a valid measure of proprioception.
We describe a new method for the reconstruction of ruptures of the patellar ligament or the quadriceps tendon, using the flexible open-weave polyester Leeds-Keio prosthetic ligament. Of 25 operations performed since 1985, we were able to review 18 patients (19 knees) with a mean follow-up of 3.5 years (3 to 6.5). No patient had required immobilisation of the knee after operation and the average period to return to normal activities without a walking aid was 10.5 weeks. The average range of motion was 146 degrees, and four patients could squat fully in the Japanese style. There was an extension lag (<
10 degrees) in only four patients; eight patients had some patellar crepitus which was mildly painful in three. There were no cases of infection, persistent joint effusion or rerupture of the extensor apparatus.
The healing of anterior cruciate ligaments reconstructed with the Leeds-Keio artificial ligament was observed by arthroscopy in 42 knees and biopsy in 19 knees at intervals from 3 to 24 months after implantation. By three months the implant was covered with immature new tissue, and a dense vascular network crossed its surface. At 12 months a new ligament had developed and matured, looking like the natural one in most cases. Histology at this stage showed abundant collagenous fibres running parallel and longitudinally, while the synovial membrane showed no more than very slight inflammatory changes. By 18 to 24 months, the new ligament often had the arthroscopic appearance of a normal anterior cruciate ligament. These results suggest that this scaffold type of artificial ligament is effective for cruciate reconstruction, giving satisfactory healing without significant complications.
The effects of advancement of the tibial tuberosity by inserting bony wedges was studied on cadaveric specimens of the knee. The geometry, the contact areas and the forces acting on the patellofemoral joint were investigated, and the forces acting on the tibiofemoral compartment were calculated. A 1 cm advancement was found to be optimal in reducing the high patellofemoral joint forces occurring at 90 degrees and 110 degrees of flexion, whilst causing least reduction of the contact areas; the stresses on the joint were reduced significantly. Advancement by 2 cm and by 3 cm drastically reduced the congruity and the size of the contact area. It was also shown that 2 cm and 3 cm advancement caused an increase in forces at the patellofemoral joint and also in the tibiofemoral joint in a direction tangential to the articular surfaces.