Neurodegeneration in the centrally-projecting Edinger-Westphal nucleus contributes to the non-motor symptoms of Parkinson's disease in the rat.

BACKGROUND: The neuropathological background of major depression and anxiety as non-motor symptoms of Parkinson's disease is much less understood than classical motor symptoms. Although, neurodegeneration of the Edinger-Westphal nucleus in human Parkinson's disease is a known phenomenon, its possible significance in mood status has never been elucidated. In this work we aimed at investigating whether neuron loss and alpha-synuclein accumulation in the urocortin 1 containing (UCN1) cells of the centrally-projecting Edinger-Westphal (EWcp) nucleus is associated with anxiety and depression-like state in the rat. METHODS: Systemic chronic rotenone administration as well as targeted leptin-saporin-induced lesions of EWcp/UCN1 neurons were conducted. Rotarod, open field and sucrose preference tests were performed to assess motor performance and mood status. Multiple immunofluorescence combined with RNAscope were used to reveal the functional-morphological changes. Two-sample Student's t test, Spearman's rank correlation analysis and Mann-Whitney U tests were used for statistics. RESULTS: In the rotenone model, besides motor deficit, an anxious and depression-like phenotype was detected. Well-comparable neuron loss, cytoplasmic alpha-synuclein accumulation as well as astro- and microglial activation were observed both in the substantia nigra pars compacta and EWcp. Occasionally, UCN1-immunoreactive neuronal debris was observed in phagocytotic microglia. UCN1 peptide content of viable EWcp cells correlated with dopaminergic substantia nigra cell count. Importantly, other mood status-related dopaminergic (ventral tegmental area), serotonergic (dorsal and median raphe) and noradrenergic (locus ceruleus and A5 area) brainstem centers did not show remarkable morphological changes. Targeted partial selective EWcp/UCN1 neuron ablation induced similar mood status without motor symptoms. CONCLUSIONS: Our findings collectively suggest that neurodegeneration of urocortinergic EWcp contributes to the mood-related non-motor symptoms in toxic models of Parkinson's disease in the rat.

Pilot Study of Cartilage Repair in the Knee Joint with Multiply Incised Chondral Allograft.

BACKGROUND: Focal cartilage lesions in the knee joint have limited capacity to heal. Current animal experiments show that incisions of the deep zone of a cartilage allograft allow acceptable integration for the graft. QUESTIONS/PURPOSES: We performed this clinical study to determine (1) if the multiply incised cartilage graft is surgically applicable for focal cartilage lesions, (2) whether this allograft has a potential to integrate to the repair site, and (3) if patients show clinical improvement. PATIENTS AND METHODS: Seven patients with 8 chondral lesions were enrolled into the study. Symptomatic lesions between 2 and 8 cm(2) were accepted. Additional injuries were allowed but were addressed simultaneously. Grafts were tailored to match and the deep zone of the cartilage was multiply incised to augment the basal integration before securing in place. Rigorous postoperative physiotherapy followed. At 12 and 24 months the patients' satisfaction were measured and serial magnetic resonance imaging (MRI) was performed in 6 patients. RESULTS: Following the implantations no adverse reaction occurred. MRI evaluation postoperatively showed the graft in place in 5 out of 6 patients. In 1 patient, MRI suggested partial delamination at 1 year and graft degeneration at 2 years. Short Form-36 health survey and the Lysholm knee score demonstrated a significant improvement in the first year; however, by 2 years there was a noticeable drop in the scores. Conclusions. Multiply incised pure chondral allograft used for cartilage repair appears to be a relatively safe method. Further studies are necessary to assess its potential in cartilage repair before its clinical use.

Increased chondrocyte death after steroid and local anesthetic combination.

BACKGROUND: Hyaline articular cartilage has limited repair and regeneration capacity. Intraarticular administration of glucocorticoid and local anesthetic injections play an important role in the therapy of osteoarthritis. Glucocorticoids and anesthetics reportedly enhance apoptosis in chondrocytes, but effects of the combined use of glucocorticoids and local anesthetics are unknown. QUESTIONS/PURPOSES: We asked whether glucocorticoid and local anesthetic agents combined had any synergistic effects on chondrocyte apoptosis. METHODS: Cell viability and apoptosis/necrosis assessment of human articular chondrocytes were performed in vitro (chondrocyte cell cultures) and ex vivo (osteochondral specimens) using flow cytometry and TUNEL analysis, respectively. RESULTS: Glucocorticoids and local anesthetics induce apoptosis in chondrocytes at various rates. When used in combination, the percentage of dead chondrocytes was increased in in vitro chondrocyte cell cultures and osteochondral ex vivo specimens. CONCLUSIONS: We observed a time-dependent decrease in chondrocyte viability after concurrent steroid and local anesthetic exposure. CLINICAL RELEVANCE: The combination of glucocorticoids and local anesthetics has an adverse effect on articular chondrocytes, and it raises a question regarding whether concomitant administration should be used in treating osteoarthritis.

Osteochondral integration of multiply incised pure cartilage allograft: repair method of focal chondral defects in a porcine model.

BACKGROUND: A focal cartilage lesion has limited capacity to heal, and the repair modalities used at present are still unable to provide a universal solution. Pure cartilage graft implantation appears to be a simple option, but it has not been applied widely as cartilage will not reattach easily to the subchondral bone. HYPOTHESIS: We used a multiple-incision technique (processed chondrograft) to increase cartilage graft surface. We hypothesized that pure cartilage graft with augmented osteochondral fusion capacity may be used for cartilage repair and we compared this method with other repair techniques. STUDY DESIGN: Controlled laboratory study. METHODS: Full-thickness focal cartilage defects were created on the medial femoral condyle of 9-month-old pigs; defects were repaired using various methods including bone marrow stimulation, autologous chondrocyte implantation, and processed chondrograft. After the repair, at weeks 6 and 24, macroscopic and histologic evaluation was carried out. RESULTS: Compared with other methods, processed chondrograft was found to be similarly effective in cartilage repair. Defects without repair and defects treated with bone marrow stimulation appeared slightly irregular with fibrocartilage filling. Autologous chondrocyte implantation produced hyalinelike cartilage, although its cellular organization was distinguishable from the surrounding articular cartilage. Processed chondrograft demonstrated good osteochondral integration, and the resulting tissue appeared to be hyaline cartilage. CONCLUSION: The applied cartilage surface processing method allows acceptable osteochondral integration, and the repair tissue appears to have good macroscopic and histologic characteristics. CLINICAL RELEVANCE: If further studies confirm its efficacy, this technique could be considered for human application in the future.

[Legal regulation in the field of tissue engineering]. / A sejt- és szövettenyésztés jogi vonatkozásai. Hazai és külföldi tapasztalatok.

In the past few years, biotechnology and tissue engineering have developed a great deal. Legislation, however, has not been able to keep pace with the rapid changes in the research and treatment using tissue-engineered products. The authors tried to explore and compare all the regulatory factors concerning cell and tissue engineering in the Hungarian and foreign law. The thorough study elucidated that the Hungarian regulation in this area is fairly deficient when compared with the regulations of other European countries, where quite extensive directives are applied. A currently accepted regulation fills this void efficiently in the European Union providing strong base for tissue engineering-related research and therapy for the future.

[The comparison of in vivo and in vitro elution of gentamycin from bone cement]. / Gentamicin csontcementbol történo in vivo és in vitro emissziójának összehasonlítása.

INTRODUCTION: The authors report their in vivo and in vitro results of the elution characteristics of gentamycin sulfate from bone cement, which is the most commonly used way of local antibiotic prophylaxis in Europe in the field of orthopedic surgery. AIM: The aim of this study was to investigate the elution of the gentamycin sulfate from bone cement, describe the dynamics of the emission in time and evaluate the relationship between the minimal inhibitory concentration (MIC) and the eluted concentration of the antibiotic. METHODS: The in vivo investigation samples were taken from 9 patient from drain fluids to evaluate the eluted antibiotic concentration by fluorescent polarisation immunoassay method (FPI). The in vitro emission-dynamics of two different bone cement-Gentamycin sulphate complex were analysed by plate diffusion method during one-year period after mixing. RESULTS: Their results showed that 24 hours after the operation the gentamycin concentrations in the drain fluid taken from around the endoprosthesis implanted with Palacos-R bone cement diminished, yet remained above the MIC level. High but rapidly decreasing antibiotic level was detected by the in vitro method within the first week, reading an almost steadily low concentration by the end of the first month. Surprisingly, after one year it was still possible to demonstrate the inhibitory effect of the drug from both tested types of cements. CONCLUSIONS: It is concluded that the gentamycin is able to elute from the bone cement in useful concentration after the implantation of endoprosthesis. However, this in vitro method is a useful and reproducible technique for the measurement of the efficacy of antibiotic emission from bone cement, the conversation of the results to the in vivo remains to be obscured. Nevertheless, the usage of local antibiotic prophylaxis seems to be useful during orthopaedic major intervention.