1. Osteolýza (OL) je považována za největší problém endoprotetiky kyčle/kolena. Pro indukci OL jsou nejdůležitější DAMP receptory makrofágů a fibroblastů, které reagují na protetické částice. 2. O rozsahu OL rozhodují amplifikační prozánětlivé/protizánětlivé dráhy a zejména stupeň osteoklastogeneze. Pokud se rozvine adaptace na částice, nerozvine se ani výrazný zánět, ani osteoklastogeneze, tj. nevzniknou rozsáhlé kostní defekty. 3. Projekt je zaměřen na analýzu expresních profilů (mRNA, proteiny) vybraných kandidátních molekul (KM) v periprotetických tkáních v různých časových obdobích po implantaci endoprotézy. Analýza expresních profilů bude doplněna imunohistochemickou detekcí, resp. vyšetřením vybraných genových polymorfismů regulujících expresi KM. 4. Srovnání expresního profilu u pacientů v různých obdobích po operaci/s různým rozsahem OL umožní identifikovat molekuly/dráhy, které jsou sdruženy s nevýznamnou/významnou OL/předčasným selháním. Projekt umožní určit KM pro diagnostiku/léčbu OL.; 1.Osteolysis(OL) is considered a major cause of hip/knee arthroplasty failure. DAMP receptors on macrophages, fibroblasts stimulated by particles induce increased inflammatory chemokine/cytokine synthesis. 2.The extent of OL depends on regulatory molecules (RM) of pro-inflammatory pathways and increased osteoclastogenesis. If the tissues do not adapt to particles then OL develops and vice versa. 3.Project is focused on analysis of expression profiles (EP) - mRNA, proteins - of selected RM in periprosthetic tissues obtained from patients with different extent of OL/time to surgery. EP will be supplemented with immunohistochemical/joint fluid analysis/ investigation of candidate gene polymorphism. 4. Comparing EPs and further data between cases with different extent of OL/time to reoperation we will be able to identify molecules/pathways connected with insignificant/ significant OL/early prosthetic failure. Project can identify candidate molecules for diagnostics/treatment of OL.

• MeSH
• Gene Amplification MeSH
• Surgical Procedures, Operative MeSH
• Gene Expression MeSH
• Genetic Testing MeSH
• Immunohistochemistry MeSH
• Joint Capsule immunology   MeSH
• RNA, Messenger MeSH
• Arthroplasty, Replacement, Hip MeSH
• Osteolysis diagnosis   etiology   immunology   therapy   MeSH
• Polymorphism, Genetic MeSH
• Arthroplasty, Replacement, Knee MeSH
• Inflammation genetics   immunology   MeSH

• Conspectus
• Ortopedie. Chirurgie. Oftalmologie
• NML Fields
• ortopedie
• biochemie
• NML Publication type
• závěrečné zprávy o řešení grantu IGA MZ ČR

• Publication type
• Meeting Abstract MeSH

• Publication type
• Meeting Abstract MeSH

PURPOSE OF THE STUDY: Aseptic loosening (AL) and periprosthetic osteolysis (PPOL) in total hip (THA) and knee (TKA) arthroplasty are linked to an inflammatory process initiated by wear debris released from artificial joints. There is still limited information about the contribution of Toll-like receptors (TLRs) and distinct regulatory cytokines to AL/PPOL in both joints. METHODS: In this study, we investigated mRNA expression of TLR-1,-2,-4 and cytokines/receptors (IL-2,-2R,-10,-10R, TGFb1) in pseudosynovial tissue obtained from 55 patients with aseptically failed THAs/TKAs and 37 control patients with hip/knee primary osteoarthritis (OA) using quantitative RT-PCR. Immunohistochemical staining was used to detect the corresponding proteins. Non-parametric Kruskal-Wallis and Mann-Whitney tests were used to determine differences between the patient groups. RESULTS: When comparing expression profiles between patients with aseptically failed THA and TKA, higher amounts of TLR-1,- 2,-4 and IL-2R mRNA transcripts were detected in THA patients. The mRNA expression of studied molecules (TLR-1,-2,-4, IL-2, IL-10, IL-2R, IL-10R, TGFb1) did not differ between THA and OA hip tissues. Lower mRNA expression of TLR-1,-2,- 4, IL-10, and IL-10R was detected in TKA when compared to control knee OA. Similar mRNA profiles of IL-2, IL-2R, and TGFb1 were observed in TKA and knee OA. Using immunohistochemistry, we detected low expression of TLR-1 protein in failed THA/TKA, whereas TLR-2 protein levels were higher in TKA/THA patients than in OA controls. High individual variability in TLR-4 protein levels was detected among patients with aseptically loosened THA and TKA. IL-10 protein levels were similar in THA and TKA patient subgroups and control subjects, whereas IL-10R protein level was higher in failed TKAs and OA controls than in THAs. No difference in IL-2 protein levels was detected between patients with THA/TKA and those with OA. DISCUSSION: Our data indicate close similarity between the expression patterns in aseptically failed THA and TKA. However, certain differences were observed which also suggest unique pathways associated with the end-stage of aseptic loosening in THA and TKA. For instance, differences in the size, shape and load of polyethylene particles between THA and TKA could play some role. The composition of THA and TKA and differences in terms of mechanical forces might also be involved. CONCLUSIONS: This is the fist study comparing the gene expression profile of a particular set of innate immunity regulatory molecules between tissues from aseptically failed THA and TKA. Low expression of TLR-1,-2,-4 and cytokines/receptors (IL-2, IL-2R, IL-10, IL-10R, and TGFb1) was observed in pseudosynovial tissues obtained from aseptically failed THAs and TKAs. Higher amount of TLR transcripts was detected in THA as compared to TKA. These findings indicate certain differences in the mechanism of aseptic loosening occurring at the site of THA and TKA. Further research is warranted.

• MeSH
• Cytokines biosynthesis   genetics   MeSH
• Gene Expression immunology   MeSH
• Hip Prosthesis MeSH
• Humans MeSH
• RNA, Messenger genetics   MeSH
• Arthroplasty, Replacement, Hip MeSH
• Immunity, Innate MeSH
• Knee Prosthesis * MeSH
• Receptors, Cytokine biosynthesis   genetics   MeSH
• Reoperation MeSH
• Prosthesis Failure * MeSH
• Case-Control Studies MeSH
• Synovial Membrane immunology   MeSH
• Toll-Like Receptors biosynthesis   genetics   MeSH
• Arthroplasty, Replacement, Knee * MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• English Abstract MeSH
• Journal Article MeSH

• Publication type
• Meeting Abstract MeSH

Myelodysplastický syndrom je heterogenní skupina onemocnění, jejichž typickým projevem je neefektivní krvetvorba a závislost na krevních transfuzích. Chelátory železa v současné době představují důležitou léčebnou modalitu u těch pacientů s myelodysplastickým syndromem, u kterých v důsledku přítomnosti anémie a opakovaných krevních transfuzí dochází k hromadění toxického železa v orgánech. Bylo prokázáno, že chelatační terapie u pacientů s mye-lodysplastickým syndromem významně prodlužuje celkové přežití a oddaluje leukemickou transformaci. Kromě vlastní chelatace iontů železa bylo u těchto chelatačních látek prokázáno i výrazné antiproliferační a proapoptotické působení na nádorové buňky. Předpokládanými mechanismy protinádorových účinků chelátorů železa jsou inhibice progrese buněčného cyklu, vyvolání stresu endoplazmatického retikula, akumulace poškozené DNA specificky u nádorových buněk i modulace protinádorové imunitní odpovědi. Přesný mechanismus působení chelatačních látek na nádorové buňky však prozatím není zcela objasněn a i naše výsledky naznačují, že bude velmi komplexní.

Myelodysplastic syndrome represents a heterogeneous group of diseases, typically characterised by ineffective haematopoiesis and transfusion dependency. Iron chelators currently represent an important treatment modality in patients with myelodysplastic syndrome (MDS), who given the presence of anaemia, are dependent on repeated blood transfusions leading to the accumulation of toxic iron in organs. It has been shown that chelation therapy significantly improves overall survival and leukaemia-free survival in patients with MDS. Besides iron chelation, iron chelators also exhibit significant antiproliferative and pro-apoptotic effects on cancer cells. The suggested mechanisms of antitumor effects of iron chelators include inhibition of cell cycle progression, induction of endoplasmic reticulum stress, accumulation of DNA damage specifically in cancer cells and modulation of antitumor immune response. The exact mechanism of action of chelating agents on cancer cells is not yet fully understood and even our data suggest that it is likely to be very complex.

• Keywords
• deferoxamin mesylát, ionty železa,
• MeSH
• Apoptosis drug effects   MeSH
• Immunity, Cellular drug effects   MeSH
• Cell Cycle drug effects   MeSH
• Iron Chelating Agents * pharmacology   metabolism   therapeutic use   MeSH
• Deferasirox MeSH
• Deferiprone MeSH
• Deferoxamine therapeutic use   MeSH
• Ions MeSH
• Humans MeSH
• Myelodysplastic Syndromes * drug therapy   metabolism   MeSH
• Iron Overload drug therapy   prevention & control   MeSH
• Antineoplastic Agents * pharmacology   therapeutic use   MeSH
• Signal Transduction drug effects   MeSH
• Endoplasmic Reticulum Stress drug effects   MeSH
• Iron metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH

Esophageal adenocarcinoma (EAC) is highly aggressive malignancy that frequently develops from Barrett's esophagus (BE), a premalignant pathologic change occurring in the lower end of the esophagus. MicroRNAs (miRNAs) are small, non-coding RNAs that function as posttranscriptional regulators of gene expression and were repeatedly proved to play key roles in pathogenesis of BE as well as EAC. In our study, we used Affymetrix GeneChip miRNA arrays to obtain miRNA expression profiles in total of 119 tissue samples [24 normal esophageal mucosa (EM), 60 BE and 35 EAC]. We identified a number of miRNAs, that showed altered expression progressively in sequence EM, BE and EAC, including for instance miR-21, miR-25, miR-194 and miR-196a with increasing levels (P < 0.0015) and miR-203, miR-205, miR-210 and miR-378 with decreasing levels (P < 0.0001). The subsequent analysis revealed four diagnostic miRNA signatures enabling to distinguish EM and BE [12 miRNAs, area under curve (AUC) = 0.971], EM and EAC (13 miRNAs, AUC = 1.0), BE without and BE with dysplasia (21 miRNAs, AUC = 0.856) and BE without dysplastic changes and BE with dysplasia together with EAC (2 miRNAs, AUC = 0.886). We suggest that miRNA expression profiling expands current knowledge in molecular pathology of Barrett's-based carcinogenesis and enables identification of molecular biomarkers for early detection of BE dysplasia and progression to EAC.

• MeSH
• Adenocarcinoma genetics   pathology   MeSH
• Barrett Esophagus genetics   pathology   MeSH
• Esophagus metabolism   pathology   MeSH
• Middle Aged MeSH
• Humans MeSH
• Lymphatic Metastasis MeSH
• MicroRNAs genetics   MeSH
• Biomarkers, Tumor genetics   MeSH
• Esophageal Neoplasms genetics   pathology   MeSH
• Follow-Up Studies MeSH
• Prognosis MeSH
• Disease Progression MeSH
• Gene Expression Regulation, Neoplastic MeSH
• Retrospective Studies MeSH
• ROC Curve MeSH
• Oligonucleotide Array Sequence Analysis MeSH
• Aged MeSH
• Neoplasm Staging MeSH
• Gene Expression Profiling * MeSH
• Case-Control Studies MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Clinical Trial MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH

PURPOSE OF THE STUDY A consensual classification of the periprosthetic interface membrane obtained at revision total joint arthroplasty was published by Morawietz et al. in 2006. Based on histomorphological criteria, four types of periprosthetic membrane were proposed: type I, aseptic failure; type II, septic failure; type III, combined type (carrying signs of both type I and II); and type IV, indeterminate type. The aim of this study was to find out whether and to what extent the Morawietz system would be suitable for use at an independent institution involved in the evaluation of periprosthetic membranes for a long time. Should it appear that the institution achieved an equally good or even better agreement between the clinical diagnosis and the histopathological finding, this consensus classification could be recommended for routine use. MATERIAL AND METHODS The samples of periprosthetic tissue evaluated in this study were obtained during surgery from the following groups of patients: 66 patients with aseptic loosening of total hip (THA) or knee arthroplasty, 15 patients with infection of THA, 16 patients with THA without any signs of aseptic loosening, osteolysis or infection; 8 patients with hip osteoarthritis and 8 patients with knee osteoarthritis. Sample collection and processing (for purposes of histomorphological evaluation and immunohistochemical staining) was performed according to the established protocol. The tissue samples evaluation was made by an experienced pathologist hand in hand with the method described in the original paper by Morawietz et al. For a more detailed tissue analysis, selected antibodies (CD4, CD8, CD20, IFN-γ and Hsp-60) were visualized by immunohistochemistry. RESULTS The majority of samples from aseptic reoperations were classified as membranes of the type I (79%) and III (16%). Specimens retrieved from septic cases were mostly classified as membranes of type II and III (60% together). The septic membranes showed a significantly higher expression of CD20 protein when compared with both the aseptic (p < 0.0001) and control THA samples (p = 0.003). The membranes retrieved from the surroundings of a stable THA without osteolysis and infection had lower expression levels of Hsp60 and IFN-γ, when compared with those from both aseptic and septic loosening. Finally, Hsp-60 expression was significantly higher in osteoarthritic tissue than in samples from stable THA (p = 0.041). DISCUSSION Morawietz et al. proposed a standardized classification system for evaluation of periprosthetic tissue. As any attempt at generalization of a complex issue, this proposal has certain shortcomings. One of these is poor detection of chronic and low-grade infections. A method that would improve the conventional counting of polymorphonuclear leukocytes is still being sought. In this connection, immunostaining for CD20 combined with an assessment of antimicrobial peptides may be a promising option. The supplementary specimen staining showed that pseudosynovial tissue is much more active in patients carrying infection and the least active in samples from stable THA in which certain tolerance and thus tissue homeostasis might be expected. CONCLUSIONS 1. In this study the distribution of findings classified according to the Morawietz system was similar to the results published in the original study from 2006. 2. The definition of an aseptic membrane (type I) in the Morawietz system meets the requirements of clinical practice (agreement, about 80%). 3. An increased sensitivity for infectious membrane detection can be achieved by using supplementary immunohistochemical staining effective particularly in chronic and low-grade infections. 4. Painless and stable THAs typically have very low expression levels of CD4, CD20 and Hsp-60 proteins, and interferon- -gamma (IFN-γ) as well. Key words: total hip arthroplasty, total knee arthroplasty, aseptic loosening, prosthetic joint infection, tissue analysis, membranes, CD receptors, Hsp-60 protein, IFN-γ.

• MeSH
• Antigens, CD20 metabolism   MeSH
• CD4 Antigens metabolism   MeSH
• Chaperonin 60 metabolism   MeSH
• Immunohistochemistry MeSH
• Prosthesis-Related Infections pathology   MeSH
• Interferon-gamma metabolism   MeSH
• Knee Joint pathology   MeSH
• Hip Joint pathology   MeSH
• Humans MeSH
• Membranes metabolism   pathology   MeSH
• Mitochondrial Proteins metabolism   MeSH
• Arthroplasty, Replacement, Hip adverse effects   MeSH
• Foreign-Body Reaction pathology   MeSH
• Bone-Implant Interface pathology   MeSH
• Prosthesis Failure MeSH
• Arthroplasty, Replacement, Knee adverse effects   MeSH
• Check Tag
• Humans MeSH
• Publication type
• English Abstract MeSH
• Journal Article MeSH
• Validation Study MeSH

• Publication type
• Meeting Abstract MeSH

OBJECTIVE: To identify expression profiles (EP) associated with aseptic loosening of total knee arthroplasty (TKA) and to compare them with EP observed in total hip arthroplasty (THA), and primary knee and hip osteoarthritis (OA). DESIGN: Gene EP of TNF, IL-6, IL-8, CHIT1, BMP4, CCL3, CCL18, MMP9, RANKL, OPG, DC-STAMP and SOCS3 were assessed using quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) on tissues retrieved from patients with aseptically failed TKA (n = 21), THA (n = 41) and primary knee (n = 20) and hip (n = 17) OA. Immunohistochemistry was applied to localize the proteins. RESULTS: When compared to knee OA, the pseudosynovial tissue in TKA exhibit (1) elevation of alternative macrophage activation marker (CHIT1), chemokine (IL-8), and a proteolytic enzyme (MMP9); (2) downregulation of pro-inflammatory cytokine (TNF), osteoclastic regulator (OPG) and a stimulator of bone formation (BMP4); (3) no difference in IL-6, CCL3, CCL18, RANKL, DC-STAMP and SOCS3. The EP in TKA differed from EP in aseptically failed THA by lower CCL3 and DC-STAMP mRNA and protein expression. EP of all studied inflammatory and osteoclastogenic molecules were similar in knee and hip OA. CONCLUSIONS: Comparing to OA, aseptic loosening of TKA is associated with upregulated expression of CHIT1, IL-8 and MMP9, dysregulated RANKL:OPG ratio and low levels of inflammatory cytokines. Similar cytokine profiles were associated with primary knee and hip OA. Further research is required to explain the differences in CCL3 and DC-STAMP expression between failed TKA and THA.

• MeSH
• Adaptor Proteins, Signal Transducing genetics   metabolism   MeSH
• Arthroplasty, Replacement methods   MeSH
• Osteoarthritis, Knee genetics   metabolism   surgery   MeSH
• Osteoarthritis, Hip genetics   metabolism   surgery   MeSH
• Chemokine CCL3 biosynthesis   genetics   MeSH
• Cytokines biosynthesis   genetics   MeSH
• Immunohistochemistry MeSH
• Knee Joint metabolism   surgery   MeSH
• Hip Joint metabolism   surgery   MeSH
• Middle Aged MeSH
• Humans MeSH
• Membrane Proteins genetics   metabolism   MeSH
• RNA, Messenger genetics   MeSH
• Arthroplasty, Replacement, Hip MeSH
• Reverse Transcriptase Polymerase Chain Reaction MeSH
• Gene Expression Regulation * MeSH
• Retrospective Studies MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Arthroplasty, Replacement, Knee MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH