Anterior knee pain after total knee arthroplasty: A multifactorial analysis.

PURPOSE: Dissatisfaction and an impaired quality of life after total knee arthroplasty (TKA) is often associated with postsurgical anterior knee pain (AKP). The underlying pathological mechanisms are not yet fully understood. Therefore, a multifactorial approach encompassing clinical and radiological parameters seemed reasonable and promising to investigate postsurgical AKP. METHODS: In this cross-sectional study, 25 patients without and 25 patients with postsurgical AKP after unilateral TKA were randomly recruited from a larger cohort of patients. Multiple clinical and radiological parameters-including real-time shear wave elastography (SWE) to measure the patellar and quadriceps tendon elasticity-were acquired and subsequently associated with AKP. For statistical analysis, SPSS (IBM, version 25) was used. RESULTS: In total 50 participants (58.0% men, mean age 63.42 years, mean body mass index 29.75 kg/m2), having different prosthetic designs implanted, were included. Independently of key covariates, the strength of the quadriceps muscle (p = 0.021), a thinner inlay (p = 0.041), and a lower position of the patella (p = 0.041) were associated with AKP. Although no correlation with AKP was found (p = 0.346, resp. p = 0.154), we observed significantly decreased Young's modulus of the patellar and quadriceps tendons for the involved knee compared to the uninvolved knee (p < 0.001). CONCLUSION: In conclusion, quadriceps muscle strength, inlay thickness, and the patella position might be of particular relevance in avoiding postsurgical AKP. Future studies with larger sample sizes are needed to clarify the impact of quadriceps muscle strength and the postoperative patella position as well as the role of SWE as a personalized modifiable prediction marker.

Evaluation of Postoperative Changes in Patellar and Quadriceps Tendons after Total Knee Arthroplasty-A Comprehensive Analysis by Shear Wave Elastography, Power Doppler and B-mode Ultrasound.

RATIONALE AND OBJECTIVES: Up to now, the diagnosis of tendinopathies is based on conventional B-mode-ultrasound (B-US), Power Doppler-ultrasound (PD-US), and magnetic resonance imaging. In the past decade, Shear Wave Elastography (SWE) has been introduced in tendon imaging, for example in athletes or patients suffering from tendinopathy. SWE allows real-time quantification of tissue stiffness, and, by this, the assessment of the mechanical properties of a tendon and its changes during acute disease and tendon healing. So far there are no ultrasound-based studies that have evaluated postoperative tendon changes, anatomical and mechanical properties and tendon healing of the patellar, and quadriceps tendon following Total Knee Arthroplasty (TKA). The purpose of this prospective study was two-fold: first to analyze morphologic, vascular, and mechanical properties of patellar and quadriceps tendons in patients following TKA; and, second to evaluate possible changes thereof and their visibility in the course of time. MATERIALS AND METHODS: Observational cross-sectional, IRB-approved study in 63 postoperative patients with a total of 76 total knee arthroplasties (50 unilateral, 13 bilateral) and 50 nonoperated knees for comparison, resulting in 152 postoperative patellar- and quadriceps and 100 nonoperated patellar- and quadriceps-tendons for comparative analysis. For further examination, we divided the 63 patients into two groups according to the duration since surgery (group A < 24 months; group B > 24 months). All patients completed a standardized questionnaire, furthermore the Knee Society score and the Knee Society function score. The amount of experienced pain was assessed using the ordinal numeric rating scale and the presence of anterior knee pain was examined. Subsequently every participant underwent a standardized multimodal ultrasound protocol consisting of B-US, PD-US, and SWE of the left and right patellar and quadriceps tendons. RESULTS: Using the different US-modalities, operated patellar, and quadriceps tendons (n = 152) were significantly more frequent classified as pathological (B-US) (p < 0.001), the mean Ohberg score was significantly higher (PD-US) (p < 0.001), and the tendons were significantly softer (SWE) than their nonoperated counterparts (n = 100). Mean SWE-value of postoperative patellar tendons was 45.66 ± 14.84 kPa versus 60.08 ± 19.13 kPa in nonoperated knees (p < 0.001). Mean SWE-value of postoperative quadriceps tendons was 35.73 ± 15.66 kPa versus 52.69 ± 16.20 kPa in nonoperated knees (p < 0.001). Comparing the two postoperative groups (group A and B), we recognized a significant decrease of pathologically classified patellar and quadriceps tendons (B-US and PD-US) in group B. The early postoperatively reduced SWE values slightly increased during the course of time. CONCLUSION: After TKA, patellar, and quadriceps tendons show significant measurable alterations in B-US, PD-US, and SWE. Especially a significant decrease of tendon stiffness in operated knees, as assessed by SWE, might be a surrogate marker for changed mechanical properties. These alterations improve, the longer ago the surgery was. The quantitative information obtained by SWE could be of particular interest in follow-up and therapy monitoring after TKA. Knowledge about tendon stiffness and it's varieties in different population groups (e.g. athletes, elderly, postoperative patients) is crucial to sonographically rate a tendon as "healthy" or "diseased."

Repetitive magnetic stimulation of human-derived neuron-like cells activates cAMP-CREB pathway.

Repetitive transcranial magnetic stimulation (rTMS) is a non-invasive neurostimulatory technique widely used in research, diagnostics, and neuro-psychiatric therapy. Despite its growing popularity, basic molecular mechanisms underlying the clinical effects of rTMS have remained largely under-researched. Here, we present a human-derived neuronal cell culture system responsive to rTMS effects. SH-SY5Y neuroblastoma cells were differentiated by retinoic acid treatment for 10 days, resulting in a neuronal phenotype characterized by upregulation of neuronal marker proteins and generation of an action potential in response to depolarizing current step injection. Repetitive magnetic stimulation of these cells resulted in increased intracellular cAMP levels and increased phosphorylation of transcription factor CREB. Pretreatment with ketamine (1 µM) potentiated, while pretreatment with lithium (2 mM) attenuated this cellular response to repetitive magnetic stimulation. In conclusion, we introduce here a novel in vitro system responding to rTMS at the level of second messenger signaling. The use of human-derived cells with neuron-like properties will prove useful for further studies on the cellular effects of rTMS.

Chronic ethanol exposure changes dopamine D2 receptor splicing during retinoic acid-induced differentiation of human SH-SY5Y cells.

There is evidence for ethanol-induced impairment of the dopaminergic system in the brain during development. The dopamine D2 receptor (DRD2) and the dopamine transporter (DAT) are decisively involved in dopaminergic signaling. Two splice variants of DRD2 are known, with the short one (DRD2s) representing the autoreceptor and the long one (DRD2l) the postsynaptic receptor. We searched for a model to investigate the impact of chronic ethanol exposure and withdrawal on the expression of these proteins during neuronal differentiation. RA-induced differentiation of human neuroblastoma SH-SY5Y cells seems to represent such a model. Our real-time RT-PCR, Western blot, and immunocytochemistry analyses of undifferentiated and RA-differentiated cells have demonstrated the enhanced expression of both splice variants of DRD2, with the short one being stronger enhanced than the long one under RA-treatment, and the DRD2 distribution on cell bodies and neurites under both conditions. In contrast, DAT was down-regulated by RA. The DAT is functional both in undifferentiated and RA-differentiated cells as demonstrated by [(3)H]dopamine uptake. Chronic ethanol exposure during differentiation for up to 4 weeks resulted in a delayed up-regulation of DRD2s. Ethanol withdrawal caused an increased expression of DRD2l and a normalization of DRD2s. Thus the DRD2s/DRD2l ratio was still disturbed. The dopamine level was increased by RA-differentiation compared to controls and was diminished under RA/ethanol treatment and ethanol withdrawal compared to RA-only treated cells. In conclusion, chronic ethanol exposure impairs differentiation-dependent adaptation of dopaminergic proteins, specifically of DRD2s. RA-differentiating SH-SY5Y cells are suited to study the impact of chronic ethanol exposure and withdrawal on expression of dopaminergic proteins during neuronal differentiation.

9-Methyl-beta-carboline has restorative effects in an animal model of Parkinson's disease.

In a previous study, a primary culture of midbrain cells was exposed to 9-methyl-beta-carboline for 48 h, which caused an increase in the number of tyrosine hydroxylase-positive cells. Quantitative RT-PCR revealed increased transcription of genes participating in the maturation of dopaminergic neurons. These in vitro findings prompted us to investigate the restorative actions of 9-methyl-beta-carboline in vivo. The compound was delivered for 14 days into the left cerebral ventricle of rats pretreated with the neurotoxin 1-methyl-4-phenyl-pyridinium ion (MPP+) for 28 days applying a dose which lowered dopamine by approximately 50%. Interestingly, 9-methyl-beta-carboline reversed the dopamine-lowering effect of the neurotoxin in the left striatum. Stereological counts of tyrosine hydroxylase-immunoreactive cells in the substantia nigra revealed that the neurotoxin caused a decrease in the number of those cells. However, when treated subsequently with 9-methyl-beta-carboline, the number reached normal values. In search of an explanation for the restorative activity, we analyzed the complexes that compose the respiratory chain in striatal mitochondria by 2-dimension gel electrophoresis followed by MALDI-TOF peptide mass fingerprinting.We found no changes in the overall composition of the complexes. However, the activity of complex I was increased by approximately 80% in mitochondria from rats treated with MPP+ and 9-methyl-beta-carboline compared to MPP+ and saline and to sham-operated rats, as determined by measurements of nicotinamide adenine dinucleotide dehydrogenase activity. Microarray technology and single RT-PCR revealed the induction of neurotrophins: brain-derived neurotrophic factor, conserved dopamine neurotrophic factor, cerebellin 1 precursor protein, and ciliary neurotrophic factor. Selected western blots yielded consistent results. The findings demonstrate restorative effects of 9-methyl-beta-carboline in an animal model of Parkinson's disease that improve the effectiveness of the respiratory chain and promote the transcription and expression of neurotrophin-related genes.

Raf kinase inhibitor protein enhances neuronal differentiation in human SH-SY5Y cells.

Neuronal differentiation has evolved as an essential process even in the adult brain, once disturbed being associated with the pathogenesis of several psychiatric disorders. To study the effects of Raf kinase inhibitor protein (RKIP) on neuronal differentiation, we generated neuroblastoma cell lines overexpressing RKIP (RKIP(+)) and expressing RKIP-directed short hairpin RNA for downregulation of RKIP (RKIP(-)). During a 4-week time course of continuous differentiation by retinoic acid (RA), expression of neuronal and glial markers, intracellular cyclic adenosine monophosphate (cAMP) levels, protein kinase C (PKC) signal transduction to extracellular signal-regulated kinase 1/2 (ERK-1/2) and cellular morphology were investigated in relation to RKIP levels. RKIP(+) cells showed accelerated neurite outgrowth, formation of elaborated neuronal networks and increased neuronal marker expression both in RA-induced differentiation and to some extent even in non-RA-treated cells. RKIP(-) cells showed glial-like cell bodies and increased glial fibrillary acidic protein, suggesting a shift from neuronal to glial phenotype. With respect to differentiation-inducing signal pathways, PKC-mediated ERK-1/2 activation significantly correlated with RKIP levels. Furthermore, basal and forskolin-stimulated intracellular cAMP was potently increased in RKIP(+) cells versus controls. In conclusion, the conserved signal transduction modulator RKIP was shown to enhance several aspects of neuronal differentiation via enhanced crosstalk from PKC to ERK-1/2 and enhancement of G-protein-coupled receptor signaling.

Long-term ethanol exposure impairs neuronal differentiation of human neuroblastoma cells involving neurotrophin-mediated intracellular signaling and in particular protein kinase C.

BACKGROUND: Revealing the molecular changes in chronic ethanol-impaired neuronal differentiation may be of great importance for understanding ethanol-related pathology in embryonic development but also in the adult brain. In this study, both acute and long-term effects of ethanol on neuronal differentiation of human neuroblastoma cells were investigated. We focused on several aspects of brain-derived neurotrophic factor (BDNF) signaling because BDNF activates the extracellular signal-regulated kinase (ERK) cascade, promoting neuronal differentiation including neurite outgrowth. METHODS: The effects of ethanol exposure on morphological differentiation, cellular density, neuronal marker proteins, basal ERK activity, and ERK responsiveness to BDNF were measured over 2 to 4 weeks. qRT-PCR and Western blotting were performed to investigate the expression of neurotrophin receptor tyrosin kinase B (TrkB), members of the ERK-cascade, protein kinase C (PKC) isoforms and Raf-Kinase-Inhibitor-Protein (RKIP). RESULTS: Chronic ethanol interfered with the development of a neuronal network consisting of cell clusters and neuritic bundles. Furthermore, neuronal and synaptic markers were reduced, indicating impaired neuronal differentiation. BDNF-mediated activation of the ERK cascade was found to be continuously impaired by ethanol. This could not be explained by expressional changes monitored for TrkB, Raf-1, MEK, and ERK. However, BDNF also activates PKC signaling which involves RKIP, which finally leads to ERK activation as well. Therefore, we hypothesized that ethanol impairs this branch of BDNF signaling. Indeed, both PKC and RKIP were significantly down-regulated. CONCLUSIONS: Chronic ethanol exposure impaired neuronal differentiation of neuroblastoma cells and BDNF signaling, particularly the PKC-dependent branch. RKIP, acting as a signaling switch at the merge of the PKC cascade and the Raf/MEK/ERK cascade, was associated with neuronal differentiation and significantly reduced in ethanol treatment. Moreover, PKC expression itself was even more strongly reduced. In contrast, members of the Raf-1/MEK/ERK cascade were less affected and the observed changes were not associated with impaired differentiation. Thus, reduced RKIP and PKC levels and subsequently reduced positive feedback on ERK activation provide an explanation for the striking effects of long-term ethanol exposure on BDNF signal transduction and neuronal differentiation, respectively.