Alterations in subchondral bone plate, trabecular bone and articular cartilage properties of rabbit femoral condyles at 4 weeks after anterior cruciate ligament transection.

OBJECTIVE: To quantify early osteoarthritic-like changes in the structure and volume of subchondral bone plate and trabecular bone and properties of articular cartilage in a rabbit model of osteoarthritis (OA) induced by anterior cruciate ligament transection (ACLT). METHODS: Left knee joints from eight skeletally mature New Zealand white rabbits underwent ACLT surgery, while the contralateral (CTRL) right knee joints were left unoperated. Femoral condyles were harvested 4 weeks after ACLT. Micro-computed tomography imaging was applied to evaluate the structural properties of subchondral bone plate and trabecular bone. Additionally, biomechanical properties, structure and composition of articular cartilage were assessed. RESULTS: As a result of ACLT, significant thinning of the subchondral bone plate (P < 0.05) was accompanied by significantly reduced trabecular bone volume fraction and trabecular thickness in the medial femoral condyle compartment (P < 0.05), while no changes were observed in the lateral compartment. In both lateral and medial femoral condyles, the equilibrium modulus and superficial zone proteoglycan (PG) content were significantly lower in ACLT than CTRL joint cartilage (P < 0.05). Significant alterations in the collagen orientation angle extended substantially deeper into cartilage from the ACLT joints in the lateral femoral condyle relative to the medial condyle compartment (P < 0.05). CONCLUSIONS: In this model of early OA, significant changes in volume and microstructure of subchondral bone plate and trabecular bone were detected only in the femoral medial condyle, while alterations in articular cartilage properties were more severe in the lateral compartment. The former finding may be associated with reduced joint loading in the medial compartment due to ACLT, while the latter finding reflects early osteoarthritic changes in the lateral compartment.

Failure mode and effects analysis in designing a virtual reality-based training simulator for bilateral sagittal split osteotomy.

BACKGROUND: Virtual reality-based simulators offer a cost-effective and efficient alternative to traditional medical training and planning. Developing a simulator that enables the training of medical skills and also supports recognition of errors made by the trainee is a challenge. The first step in developing such a system consists of error identification in the real procedure, in order to ensure that the training environment covers the most significant errors that can occur. This paper focuses on identifying the main system requirements for an interactive simulator for training bilateral sagittal split osteotomy (BSSO). METHODS: An approach is proposed based on failure mode and effects analysis (FMEA), a risk analysis method that is well structured and already an approved technique in other domains. RESULTS: Based on the FMEA results, a BSSO training simulator is currently being developed, which centres upon the main critical steps of the procedure (sawing and splitting) and their main errors. CONCLUSIONS: FMEA seems to be a suitable tool in the design phase of developing medical simulators. Herein, it serves as a communication medium for knowledge transfer between the medical experts and the system developers. The method encourages a reflective process and allows identification of the most important elements and scenarios that need to be trained.

Estimation of the forces generated by the thigh muscles for transtibial amputee gait.

The forces generated by the muscles with origin on the human femur play a major role in transtibial amputee gait, as they are the most effective of the means that the body can use for propulsion. By estimating the forces generated by the thigh muscles of transtibial amputees, and comparing them to the forces generated by the thigh muscles of normal subjects, it is possible to better estimate the energy output needed from prosthetic devices. The purpose of this paper is to obtain the forces generated by the thigh muscles of transtibial amputees and compare these with forces obtained from the same muscles in the case of normal subjects. Two transtibial amputees and four normal subjects similar in size to the amputees were investigated. Level ground walking was chosen as the movement to be studied, since it is a common activity that most amputees engage in. Inverse dynamics and a muscle recruitment algorithm (developed by AnyBody Technology(®)) were used for generating the muscle activation patterns and for computing the muscle forces. The muscle forces were estimated as two sums: one for all posterior muscles and one for the anterior muscles, based on the position of the muscles of the thigh relative to the frontal plane of the human body. The results showed that a significantly higher force is generated by the posterior muscles of the amputees during walking, leading to a general increase of the metabolic cost necessary for one step.

C-peptide stimulates ERK1/2 and JNK MAP kinases via activation of protein kinase C in human renal tubular cells.

AIMS/HYPOTHESIS: Accumulating evidence indicates that replacement of C-peptide in type 1 diabetes ameliorates nerve and kidney dysfunction, but the molecular mechanisms involved are incompletely understood. C-peptide shows specific binding to a G-protein-coupled membrane binding site, resulting in Ca(2+) influx, activation of mitogen-activated protein kinase signalling pathways, and stimulation of Na(+), K(+)-ATPase and endothelial nitric oxide synthase. This study examines the intracellular signalling pathways activated by C-peptide in human renal tubular cells. METHODS: Human renal tubular cells were cultured from the outer cortex of renal tissue obtained from patients undergoing elective nephrectomy. Extracellular-signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK) and Akt/protein kinase B (PKB) activation was determined using phospho-specific antibodies. Protein kinase C (PKC) and RhoA activation was determined by measuring their translocation to the cell membrane fraction using isoform-specific antibodies. RESULTS: Human C-peptide increases phosphorylation of ERK1/2 and Akt/PKB in a concentration- and time-dependent manner in renal tubular cells. The C-terminal pentapeptide of C-peptide is equipotent with the full-length C-peptide, whereas scrambled C-peptide has no effect. C-peptide stimulation also results in phosphorylation of JNK, but not of p38 mitogen-activated protein kinase. MEK1/2 inhibitor PD98059 blocks the C-peptide effect on ERK1/2 phosphorylation. C-peptide causes specific translocation of PKC isoforms delta and epsilon to the membrane fraction in tubular cells. All stimulatory effects of C-peptide were abolished by pertussis toxin. The isoform-specific PKC-delta inhibitor rottlerin and the broad-spectrum PKC inhibitor GF109203X both abolish the C-peptide effect on ERK1/2 phosphorylation. C-peptide stimulation also causes translocation of the small GTPase RhoA from the cytosol to the cell membrane. Inhibition of phospholipase C abolished the stimulatory effect of C-peptide on phosphorylation of ERK1/2, JNK and PKC-delta. CONCLUSIONS/INTERPRETATION: C-peptide signal transduction in human renal tubular cells involves the activation of phospholipase C and PKC-delta and PKC-epsilon, as well as RhoA, followed by phosphorylation of ERK1/2 and JNK, and a parallel activation of Akt.

C-peptide stimulates Na+, K+-ATPase via activation of ERK1/2 MAP kinases in human renal tubular cells.

Proinsulin-connecting peptide (C-peptide) exerts physiological effects partially via stimulation of Na(+), K(+)-ATPase. We determined the molecular mechanism by which C-peptide stimulates Na(+), K(+)-ATPase in primary human renal tubular cells (HRTCs). Incubation of the cells with 5 nM human C-peptide at 37 degrees C for 10 min stimulated (86)Rb(+) uptake by 40% (p<0.01). The carboxy-terminal pentapeptide was found to elicit 57% of the activity of the intact molecule. In parallel with ouabain-sensitive (86)Rb(+) uptake, C-peptide increased alpha subunit phosphorylation and basolateral membrane (BLM) abundance of the Na(+), K(+)-ATPase alpha(1) and beta(1) subunits. The increase in BLM abundance of the Na(+), K(+)-ATPase alpha(1) and beta(1) subunits was accompanied by depletion of alpha(1) and beta(1) subunits from the endosomal compartments. C-peptide action on Na(+), K(+)-ATPase was ERK1/2-dependent in HRTCs. C-peptide-stimulated Na(+), K(+)-ATPase activation, phosphorylation of alpha(1)-subunit and translocation of alpha(1) and beta(1) subunits to the BLM were abolished by a MEK1/2 inhibitor (20 muM PD98059). C-peptide stimulation of (86)Rb(+) uptake was also abolished by preincubation of HRTCs with an inhibitor of PKC (1 muM GF109203X). C-peptide stimulated phosphorylation of human Na(+), K(+)-ATPase alpha subunit on Thr-Pro amino acid motifs, which form specific ERK substrates. In conclusion, C-peptide stimulates sodium pump activity via ERK1/2-induced phosphorylation of Thr residues on the alpha subunit of Na(+), K(+)-ATPase.