1 630 76 BIOLOGICAL AND MECHANICAL FACTORS AND EPIGENETIC REGULATION INVOLVED IN TENDON HEALING. TENDONS ARE AN IMPORTANT PART OF THE MUSCULOSKELETAL SYSTEM. CONNECTING MUSCLES TO BONES, TENDONS CONVERT FORCE INTO MOVEMENT. TENDON INJURY CAN BE ACUTE OR CHRONIC. NOTICEABLY, TENDON HEALING REQUIRES A LONG TIME SPAN AND INCLUDES INFLAMMATION, PROLIFERATION, AND REMODELING PROCESSES. THE MISMATCH BETWEEN ENDOGENOUS AND EXOGENOUS HEALING MAY LEAD TO ADHESION CAUSING FURTHER NEGATIVE EFFECTS. MANAGEMENT OF TENDON INJURIES AND COMPLICATIONS SUCH AS SUBSEQUENT ADHESION FORMATION ARE STILL CHALLENGES FOR CLINICIANS. DUE TO NUMEROUS FACTORS, TENDON HEALING IS A COMPLEX PROCESS. THIS REVIEW INTRODUCES THE ROLE OF VARIOUS BIOLOGICAL AND MECHANICAL FACTORS AND EPIGENETIC REGULATION PROCESSES INVOLVED IN TENDON HEALING. 2023 2 1724 15 DYSREGULATION OF EPIGENETIC RELATED GENES IN DIABETIC TRIGGER FINGER PATIENTS; PRELIMINARY ANALYSIS OF PATIENT-DERIVED SAMPLES. BACKGROUND: TRIGGER FINGER (TF), A PAINFUL CONDITION INVOLVING A FINGER FLEXOR TENDON, IS A COMMON PROBLEM WITH A PREVALENCE OF ~2-3% IN THE GENERAL POPULATION. HOWEVER, THE TF PREVALENCE IS HIGHER AMONG DIABETIC PATIENTS-RANGES FROM 6.7% TO 10%. WE HAVE ANALYZED THE EXPRESSION OF THE EXTRACELLULAR MATRIX, INFLAMMATION, AND EPIGENETIC RELATED GENES IN DIABETIC AND NON-DIABETES TF. WE HYPOTHESIZED THAT DIABETES CONDITION INDUCES ALTER THE EXPRESSION OF EPIGENETIC MODIFICATION GENES IN DIABETIC PATIENTS AND ONE OF THE UNDERLYING DETERMINANTS FOR MORE PREVALENCE OF TF IN DIABETIC PATIENTS. METHOD: TISSUES FROM THE FINGERS OF PATIENTS WITH SYMPTOMATIC TRIGGER FINGERS WERE COLLECTED. WE HAD THREE GROUPS: CARPAL TUNNEL SYNDROME (AS A CONTROL), TRIGGER FINGER, AND DIABETIC TRIGGER FINGER. A QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION WAS PERFORMED. THE GENE EXPRESSION OF EXTRACELLULAR MATRIX (ECM) COMPONENTS [COL-I, COL-II, COL-X, AGGRECAN], DNA METHYLTRANSFERASES ENZYMES (DNMT1, DNMT3), GROWTH FACTORS (TGF-B, IGF), AND HISTONE DEACETYLASE ENZYMES (HDAC1, HDAC2) WERE EVALUATED IN ALL GROUPS. RESULTS: THE MRNA EXPRESSION OF COL-I, COL-II, AGGRECAN WAS SIGNIFICANTLY HIGHER IN THE PULLY A1 OF DIABETIC PATIENTS (P= 0.0164, P=0.0351, P=0.0399, RESPECTIVELY) AS COMPARED TO NON-DIABETIC TF PATIENTS. DIABETES WAS ASSOCIATED WITH A SIGNIFICANT INCREASE IN THE DNMT3 EXPRESSION COMPARED TO NON-DIABETIC TF PATIENTS (P=0.0485). HDAC1 AND HDAC2 GENE EXPRESSION WERE UP-REGULATED IN DIABETIC TF THAN NON-DIABETIC TF. CONCLUSION: THE CHRONIC STATE OF HYPERGLYCEMIA INDUCES EPIGENETIC MODIFICATION OF GENE EXPRESSIONS IN TRIGGER FINGERS. THIS SEEMS TO HAVE A SIGNIFICANT IMPACT ON THE DEVELOPMENT, RECURRENCE, AND PROGRESSION OF TRIGGER FINGER IN DIABETIC PATIENTS. 2020 3 1210 21 COX2 ENHANCES NEOVASCULARIZATION OF INFLAMMATORY TENOCYTES THROUGH THE HIF-1ALPHA/VEGFA/PDGFB PATHWAY. TENDON INJURIES ARE AMONG THE MOST CHALLENGING IN ORTHOPEDICS. DURING THE EARLY TENDON REPAIR, NEW BLOOD VESSEL FORMATION IS NECESSARY. HOWEVER, EXCESSIVE ANGIOGENESIS ALSO EXACERBATES SCAR FORMATION, LEADING TO PAIN AND DYSFUNCTION. A SIGNIFICANTLY WORSE OUTCOME WAS ASSOCIATED WITH HIGHER EXPRESSION LEVELS OF HYPOXIA-INDUCIBLE FACTOR-1 ALPHA (HIF-1ALPHA), AND ITS TRANSCRIPTIONAL TARGETS VASCULAR ENDOTHELIAL GROWTH FACTOR A (VEGFA) AND PLATELET-DERIVED GROWTH FACTOR B (PDGFB), BUT THE UNDERLYING MOLECULAR MECHANISMS REMAIN UNCLEAR. IN THIS STUDY, LIPOPOLYSACCHARIDE (LPS) WAS USED TO INDUCE AN INFLAMMATORY RESPONSE IN TENOCYTES. LPS INCREASED THE TENOCYTES' INFLAMMATORY FACTOR COX2 EXPRESSION AND ACTIVATED THE HIF-1ALPHA/VEGFA/PDGFB PATHWAY. MOREOVER, THE CONDITIONED MEDIUM FROM THE TENOCYTES BOOSTED RAT AORTIC VASCULAR ENDOTHELIAL CELL (RAOEC) ANGIOGENESIS. FURTHERMORE, TRICHOSTATIN A (TSA), AN INHIBITOR OF HISTONE DEACETYLASE, WAS USED TO TREAT INFLAMMATORY TENOCYTES. THE EXPRESSION LEVELS OF HIF-1ALPHA AND ITS TRANSCRIPTIONAL TARGETS VEGFA AND PDGFB DECREASED, RESULTING IN RAOEC ANGIOGENESIS INHIBITION. FINALLY, THE DUAL-LUCIFERASE REPORTER GENE ASSAY AND CHROMATIN IMMUNOPRECIPITATION (CHIP) ASSAY PROVED THAT THE HIF-1ALPHA/PDGFB PATHWAY PLAYED A MORE CRITICAL ROLE IN TENOCYTE ANGIOGENESIS THAN THE HIF-1ALPHA/VEGFA PATHWAY. TSA COULD ALLEVIATE ANGIOGENESIS MAINLY THROUGH EPIGENETIC REGULATION OF THE HIF-1ALPHA/PDGFB PATHWAY. TAKEN TOGETHER, TSA MIGHT BE A PROMISING ANTI-ANGIOGENESIS DRUG FOR ABNORMAL ANGIOGENESIS, WHICH IS INDUCED BY TENDON INJURIES. 2021 4 6351 23 THE ROLE OF EPIGENOMICS IN MAPPING POTENTIAL PRECURSORS FOR FOOT AND ANKLE TENDINOPATHY: A SYSTEMATIC REVIEW. TENDINOPATHY OF THE FOOT AND ANKLE IS A COMMON CLINICAL PROBLEM FOR WHICH THE EXACT ETIOLOGY IS POORLY UNDERSTOOD. THE FIELD OF EPIGENETICS HAS BEEN A RECENT FOCUS OF THIS INVESTIGATION. THE PURPOSE OF THIS ARTICLE WAS TO REVIEW THE GENOMIC ADVANCES IN FOOT AND ANKLE TENDINOPATHY THAT COULD POTENTIALLY BE USED TO STRATIFY DISEASE RISK AND CREATE PREVENTATIVE OR THERAPEUTIC AGENTS. A MULTI-DATABASE SEARCH OF PUBMED, COCHRANE, GOOGLE SCHOLAR, AND CLINICALTRIALS.GOV FROM JANUARY 1, 2000 TO JULY 1, 2022 WAS PERFORMED. A TOTAL OF 18 ARTICLES MET INCLUSION AND EXCLUSION CRITERIA FOR THIS REVIEW. THE MAJORITY OF SUCH RESEARCH UTILIZED CASE-CONTROL CANDIDATE GENE ASSOCIATION TO IDENTIFY DIFFERENT GENETIC RISK FACTORS ASSOCIATED WITH CHRONIC TENDINOPATHY. POLYMORPHISMS IN COLLAGEN GENES COL5A1, COL27A1, AND COL1A1 WERE NOTED AT A SIGNIFICANTLY HIGHER FREQUENCY IN ACHILLES TENDINOPATHY VERSUS CONTROL GROUPS. OTHER ALLELIC VARIATIONS THAT WERE OBSERVED AT AN INCREASED INCIDENCE IN ACHILLES TENDINOPATHY WERE TNC AND CASP8. THE EXTRACELLULAR MATRIX (ECM) DEMONSTRATED MACROSCOPIC CHANGES IN ACHILLES TENDINOPATHY, INCLUDING AN INCREASE IN AGGRECAN AND BIGLYCAN MRNA EXPRESSION, AND INCREASED EXPRESSION OF MULTIPLE MATRIX METALLOPROTEINASES. CYTOKINE EXPRESSION WAS ALSO INFLUENCED IN PATHOLOGY AND ABERRANTLY DEMONSTRATED DYNAMIC RESPONSE TO MECHANICAL LOAD. THE PATHOLOGIC ACCUMULATION OF ECM PROTEINS AND CYTOKINE EXPRESSION ALTERS THE ADAPTIVE RESPONSE NORMAL TENDON HAS TO PHYSIOLOGIC STRESS, FURTHER PROPAGATING THE RISK FOR TENDINOPATHY. BY IDENTIFYING AND UNDERSTANDING THE EPIGENETIC MEDIATORS THAT LEAD TO TENDINOPATHY, THERAPEUTIC AGENTS CAN BE DEVELOPED TO TARGET THE EXACT UNDERLYING ETIOLOGY AND MINIMIZE SIDE EFFECTS.LEVEL OF EVIDENCE: LEVEL IV: SYSTEMATIC REVIEW OF LEVEL II-IV STUDIES. 2023 5 2349 21 EPIGENETIC REGULATION OF MYOFIBROBLAST DIFFERENTIATION AND EXTRACELLULAR MATRIX PRODUCTION IN NASAL POLYP-DERIVED FIBROBLASTS. BACKGROUND: NASAL POLYPOSIS IS A MULTI-FACTORIAL DISEASE ASSOCIATED WITH CHRONIC INFLAMMATORY CONDITION OF THE PARANASAL SINUSES. MYOFIBROBLAST DIFFERENTIATION AND EXTRACELLULAR MATRIX (ECM) ACCUMULATION ARE INVOLVED IN THE PATHOGENESIS OF NASAL POLYPOSIS. OBJECTIVE: THE AIM OF THIS STUDY WAS TO STUDY THE EFFECT OF TRICHOSTATIN A (TSA), A HISTONE DEACETYLASE (HDAC) INHIBITOR, ON TRANSFORMING GROWTH FACTOR (TGF)-BETA1-INDUCED MYOFIBROBLAST DIFFERENTIATION AND ECM ACCUMULATION IN NASAL POLYP-DERIVED FIBROBLASTS (NPDFS). METHODS: NASAL POLYP-DERIVED FIBROBLASTS WERE ISOLATED FROM NASAL POLYPS OF PATIENTS WHO HAVE CHRONIC RHINOSINUSITIS WITH NASAL POLYP. TSA WAS TREATED IN TGF-BETA1-INDUCED NPDFS. EXPRESSION LEVELS OF HDAC2, ALPHA-SMOOTH MUSCLE ACTIN (SMA), TGF-BETA1, COLLAGEN TYPE I, ACETYLATED HISTONE H3, ACETYLATED HISTONE H4, PHOSPHORYLATED SMAD2/3 AND SMAD7 WERE DETERMINED BY RT-PCR, WESTERN BLOT AND/OR IMMUNOFLUORESCENT STAINING. THE TOTAL COLLAGEN AMOUNT PRODUCTION WAS ANALYSED BY SIRCOL SOLUBLE COLLAGEN ASSAY AND CONTRACTILE ACTIVITY WAS MEASURED BY COLLAGEN GEL CONTRACTION ASSAY. HDAC2 INHIBITION BY TSA OR HDAC2 SILENCING WAS ESTABLISHED BY RT-PCR AND WESTERN BLOT. THE EPIGENETIC EFFECT ON ALPHA-SMA GENE INACTIVATION WAS EXAMINED BY CHROMATIN IMMUNOPRECIPITATION ASSAY. PROLIFERATION WAS DETERMINED BY KI67-POSITIVE CELL STAINING AND CYTOTOXICITY WAS ASSESSED BY 3-(4,5- DIMETHYLTHIAZOL-2YL)-2,5-DIPHENYL-2H-TETRAZOLIUM BROMIDE (MTT) ASSAY. RESULTS: THE EXPRESSION LEVELS OF HDAC2, ALPHA-SMA AND TGF-BETA1 WERE INCREASED IN NASAL POLYP TISSUES COMPARED TO NORMAL INFERIOR TURBINATE TISSUES. TSA AND HDAC2 SILENCING INHIBITED EXPRESSION LEVELS ALPHA-SMA, COLLAGEN AND HDAC2. TSA INDUCED HYPERACETYLATION OF HISTONE AND SUPPRESSED OPENING OF ALPHA-SMA GENE PROMOTER IN TGF-BETA1-INDUCED NPDFS. TSA INHIBITED TGF-BETA1-INDUCED SMAD 2/3 AND RESCUED TGF-BETA1-SUPPRESSED SMAD7 SIGNALLING PATHWAY. FINALLY, TSA BLOCKED PROLIFERATION IN TGF-BETA1-INDUCED NPDFS AND HAS NO CYTOTOXIC EFFECT IN NPDFS. CONCLUSIONS AND CLINICAL RELEVANCE: THESE RESULTS SUGGEST THAT HDAC INHIBITION IS ASSOCIATED WITH MYOFIBROBLAST DIFFERENTIATION AND EXTRACELLUAR MATRIX ACCUMULATION IN NASAL POLYPOSIS. TSA MAY BE USEFUL AS AN INHIBITOR OF NASAL POLYP GROWTH, AND THUS HAS POTENTIAL TO BE USED AS A NOVEL TREATMENT OPTION FOR NASAL POLYPOSIS. 2012 6 2784 22 EZH2 PROMOTES EXTRACELLULAR MATRIX DEGRADATION VIA NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) AND P38 SIGNALING PATHWAYS IN PULPITIS. PULPITIS IS A COMPLICATED CHRONIC INFLAMMATORY PROCESS WHICH CAN BE IN A DYNAMIC BALANCE BETWEEN DAMAGE AND REPAIR. THE EXTRACELLULAR MATRIX PLAYS AN IMPORTANT REGULATORY ROLE IN WOUND HEALING AND TISSUE REPAIR. THE AIM OF THIS STUDY WAS TO EXPLORE THE ROLE OF THE EPIGENETIC MARK, ENHANCER OF ZESTE HOMOLOG 2 (EZH2) ON THE DEGRADATION OF EXTRACELLULAR MATRIX DURING PULPITIS. QUANTITATIVE POLYMERASE CHAIN REACTION WAS USED TO ASSESS THE EXPRESSION OF MATRIX METALLOPROTEINASES (MMPS) AND TYPE I COLLAGEN IN HUMAN DENTAL PULP CELLS (HDPCS) UPON EZH2 AND EI1 (EZH2 INHIBITOR) STIMULATION. THE MECHANISM OF EZH2 AFFECTING EXTRACELLULAR MATRIX WAS EXPLORED THROUGH QUANTITATIVE POLYMERASE CHAIN REACTION AND WESTERN BLOT. A RAT MODEL OF DENTAL PULP INFLAMMATION WAS ESTABLISHED, AND THE EXPRESSION OF TYPE I COLLAGEN IN DENTAL PULP UNDER EZH2 STIMULATION WAS DETECTED BY IMMUNOHISTOCHEMICAL STAINING. EZH2 UPREGULATED THE EXPRESSION OF MMP-1, MMP-3, MMP-8, AND MMP-10 AND DECREASED THE PRODUCTION OF TYPE I COLLAGEN IN HDPCS, WHILE EI1 HAD THE OPPOSITE EFFECT. EZH2 ACTIVATED THE NUCLEAR FACTOR-KAPPA B (NF-KAPPAB) AND P38 SIGNALING PATHWAYS IN HDPCS, THE INHIBITION OF WHICH REVERSED THE INDUCTION OF MMPS AND THE SUPPRESSION OF TYPE I COLLAGEN. EZH2 CAN DOWNREGULATE THE TYPE I COLLAGEN LEVELS IN AN EXPERIMENTAL MODEL OF DENTAL PULPITIS IN RATS. EZH2 PROMOTES EXTRACELLULAR MATRIX DEGRADATION VIA NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) AND P38 SIGNALING PATHWAYS IN PULPITIS. EZH2 CAN DECREASE THE TYPE I COLLAGEN LEVELS IN VIVO AND IN VITRO. 2021 7 854 21 CHONDROPROTECTIVE EFFECTS OF A HISTONE DEACETYLASE INHIBITOR, PANOBINOSTAT, ON PAIN BEHAVIOR AND CARTILAGE DEGRADATION IN ANTERIOR CRUCIATE LIGAMENT TRANSECTION-INDUCED EXPERIMENTAL OSTEOARTHRITIC RATS. OSTEOARTHRITIS (OA) IS THE MOST COMMON ARTICULAR DEGENERATIVE DISEASE CHARACTERIZED BY CHRONIC PAIN, JOINT INFLAMMATION, AND MOVEMENT LIMITATIONS, WHICH ARE SIGNIFICANTLY INFLUENCED BY ABERRANT EPIGENETIC MODIFICATIONS OF NUMEROUS OA-SUSCEPTIBLE GENES. RECENT STUDIES REVEALED THAT BOTH THE ABNORMAL ACTIVATION AND DIFFERENTIAL EXPRESSION OF HISTONE DEACETYLASES (HDACS) MIGHT CONTRIBUTE TO OA PATHOGENESIS. IN THIS STUDY, WE INVESTIGATED THE CHONDROPROTECTIVE EFFECTS OF A MARINE-DERIVED HDAC INHIBITOR, PANOBINOSTAT, ON ANTERIOR CRUCIATE LIGAMENT TRANSECTION (ACLT)-INDUCED EXPERIMENTAL OA RATS. THE INTRA-ARTICULAR ADMINISTRATION OF 2 OR 10 MICROG OF PANOBINOSTAT (EACH GROUP, N = 7) PER WEEK FROM THE 6TH TO 17TH WEEK ATTENUATES ACLT-INDUCED NOCICEPTIVE BEHAVIORS, INCLUDING SECONDARY MECHANICAL ALLODYNIA AND WEIGHT-BEARING DISTRIBUTION. HISTOPATHOLOGICAL AND MICROCOMPUTED TOMOGRAPHY ANALYSIS SHOWED THAT PANOBINOSTAT SIGNIFICANTLY PREVENTS CARTILAGE DEGENERATION AFTER ACLT. MOREOVER, INTRA-ARTICULAR PANOBINOSTAT EXERTS HYPERTROPHIC EFFECTS IN THE CHONDROCYTES OF ARTICULAR CARTILAGE BY REGULATING THE PROTEIN EXPRESSIONS OF HDAC4, HDAC6, HDAC7, RUNT-DOMAIN TRANSCRIPTION FACTOR-2, AND MATRIX METALLOPROTEINASE-13. THE STUDY INDICATED THAT HDACS MIGHT HAVE DIFFERENT MODULATIONS ON THE CHONDROCYTE PHENOTYPE IN THE EARLY STAGES OF OA DEVELOPMENT. THESE RESULTS PROVIDE NEW EVIDENCE THAT PANOBINOSTAT MAY BE A POTENTIAL THERAPEUTIC DRUG FOR OA. 2021 8 4300 19 MICRORNA-19A CONTRIBUTES TO THE EPIGENETIC REGULATION OF TISSUE FACTOR IN DIABETES. BACKGROUND: DIABETES MELLITUS IS CHARACTERIZED BY CHRONIC VASCULAR DISORDER AND PRESENTS A MAIN RISK FACTOR FOR CARDIOVASCULAR MORTALITY. IN PARTICULAR, HYPERGLYCAEMIA AND INFLAMMATORY CYTOKINES INDUCE VASCULAR CIRCULATING TISSUE FACTOR (TF) THAT PROMOTES PRO-THROMBOTIC CONDITIONS IN DIABETES. IT HAS RECENTLY BECOME EVIDENT THAT ALTERATIONS OF THE POST-TRANSCRIPTIONAL REGULATION OF TF VIA SPECIFIC MICRORNA(MIR)S, SUCH AS MIR-126, CONTRIBUTE TO THE PATHOGENESIS OF DIABETES AND ITS COMPLICATIONS. THE ENDOTHELIAL MIR-19A IS INVOLVED IN VASCULAR HOMEOSTASIS AND ATHEROPROTECTION. HOWEVER, ITS ROLE IN DIABETES-RELATED THROMBOGENICITY IS UNKNOWN. UNDERSTANDING MIR-NETWORKS REGULATING PROCOAGULABILITY IN DIABETES MAY HELP TO DEVELOP NEW TREATMENT OPTIONS PREVENTING VASCULAR COMPLICATIONS. METHODS AND RESULTS: PLASMA OF 44 PATIENTS WITH KNOWN DIABETES WAS ASSESSED FOR THE EXPRESSION OF MIR-19A, TF PROTEIN, TF ACTIVITY, AND MARKERS FOR VASCULAR INFLAMMATION. HIGH MIR-19A EXPRESSION WAS ASSOCIATED WITH REDUCED TF PROTEIN, TF-MEDIATED PROCOAGULABILITY, AND VASCULAR INFLAMMATION BASED ON EXPRESSION OF VASCULAR ADHESION MOLECULE-1 AND LEUKOCYTE COUNT. WE FOUND PLASMA EXPRESSION OF MIR-19A TO STRONGLY CORRELATE WITH MIR-126. MIR-19A REDUCED THE TF EXPRESSION ON MRNA AND PROTEIN LEVEL IN HUMAN MICROVASCULAR ENDOTHELIAL CELLS (HMEC) AS WELL AS TF ACTIVITY IN HUMAN MONOCYTES (THP-1), WHILE ANTI-MIR-19A INCREASED THE TF EXPRESSION. INTERESTINGLY, MIR-19A INDUCED VCAM EXPRESSION IN HMEC. HOWEVER, MIR-19A AND MIR-126 CO-TRANSFECTION REDUCED TOTAL ENDOTHELIAL VCAM EXPRESSION AND EXHIBITED ADDITIVE INHIBITION OF A LUCIFERASE REPORTER CONSTRUCT CONTAINING THE F3 3'UTR. CONCLUSIONS: WHILE BOTH MIRS HAVE DIFFERENTIAL FUNCTIONS ON ENDOTHELIAL VCAM EXPRESSION, MIR-19A AND MIR-126 COOPERATE TO EXHIBIT ANTI-THROMBOTIC PROPERTIES VIA REGULATING VASCULAR TF EXPRESSION. MODULATING THE POST-TRANSCRIPTIONAL CONTROL OF TF IN DIABETES MAY PROVIDE A FUTURE ANTI-THROMBOTIC AND ANTI-INFLAMMATORY THERAPY. 2018 9 908 20 CHRONIC EXPOSURE TO ENVIRONMENTALLY RELEVANT CONCENTRATION OF FLUORIDE IMPAIRS OSTEOBLAST'S COLLAGEN SYNTHESIS AND MATRIX MINERALIZATION: INVOLVEMENT OF EPIGENETIC REGULATION IN SKELETAL FLUOROSIS. GLOBALLY, 200 MILLION PEOPLE ARE SUFFERING FROM TOXIC MANIFESTATIONS OF FLUORIDE(F), DENTAL AND SKELETAL FLUOROSIS; UNFORTUNATELY, THERE IS NO TREATMENT. TO UNRAVEL THE PATHOGENESIS OF SKELETAL FLUOROSIS, WE ESTABLISHED FLUOROSIS MICE BY TREATING ENVIRONMENTALLY RELEVANT CONCENTRATION OF F (15 PPM NAF) THROUGH DRINKING WATER FOR 4 MONTHS. AS IN SKELETAL FLUOROSIS, LOCOMOTOR DISABILITY, CRIPPLING DEFORMITIES OCCUR AND THUS, OUR HYPOTHESIS WAS F MIGHT ADVERSELY AFFECTS COLLAGEN WHICH GIVES THE BONE TENSILE STRENGTH. THIS WORK INEVITABLY HAD TO BE CARRIED OUT ON OSTEOBLAST CELLS, RESPONSIBLE FOR SYNTHESIS, DEPOSITION, AND MINERALIZATION OF BONE MATRIX. ISOLATED OSTEOBLAST CELLS WERE CONFIRMED BY ALP ACTIVITY AND MINERALIZED NODULES FORMATION. EXPRESSION OF COLLAGEN COL1A1, COL1A2, COL1A1 WAS SIGNIFICANTLY REDUCED IN TREATED MICE. FURTHER, A STUDY REVEALED THE INVOLVEMENT OF EPIGENETIC REGULATION BY PROMOTER HYPERMETHYLATION OF COL1A1; EXPRESSIONAL ALTERATIONS OF TRANSCRIPTION FACTORS, CALCIUM CHANNELS AND OTHER GENES E.G., CBFA-1, TGF-BETA1, BMP1, SP1, SP7, NF-(K)B P65, BMP-2, BGLAP, GPRC6A AND CAV(1.2) ARE ASSOCIATED WITH IMPAIRMENT OF COLLAGEN SYNTHESIS, DEPOSITION AND DECREASED MINERALIZATION THUS, ENFEEBLING BONE HEALTH. THIS STUDY INDICATES THE POSSIBLE ASSOCIATION OF EPIGENETIC REGULATION IN SKELETAL FLUOROSIS. HOWEVER, NO ASSOCIATION WAS FOUND BETWEEN POLYMORPHISMS IN THE COL1A1 (RSAI, HINDIII) AND COL1A2 (RSAI, HINDIII) GENES WITH FLUOROSIS IN MICE. 2023 10 1826 18 EFFECTS OF HISTONE DEACETYLASE INHIBITOR ON EXTRACELLULAR MATRIX PRODUCTION IN HUMAN NASAL POLYP ORGAN CULTURES. BACKGROUND: NASAL POLYPOSIS IS ASSOCIATED WITH A CHRONIC INFLAMMATORY CONDITION OF THE SINONASAL MUCOSA AND INVOLVES MYOFIBROBLAST DIFFERENTIATION AND EXTRACELLULAR MATRIX (ECM) ACCUMULATION. EPIGENETIC MODULATION BY HISTONE DEACETYLASE (HDAC) INHIBITORS INCLUDING TRICHOSTATIN A (TSA) HAS BEEN REPORTED TO HAVE INHIBITORY EFFECTS ON MYOFIBROBLAST DIFFERENTIATION IN LUNG AND RENAL FIBROBLASTS. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE THE INHIBITORY EFFECT OF TSA ON MYOFIBROBLAST DIFFERENTIATION AND ECM PRODUCTION IN NASAL POLYP ORGAN CULTURES. METHODS: NASAL POLYP TISSUES FROM 18 PATIENTS WERE ACQUIRED DURING ENDOSCOPIC SINUS SURGERY. AFTER ORGAN CULTURE, NASAL POLYPS WERE STIMULATED WITH TGF-BETA1 AND THEN TREATED WITH TSA. ALPHA-SMOOTH MUSCLE ACTIN (ALPHA-SMA), FIBRONECTIN, AND COLLAGEN TYPE I EXPRESSION LEVELS WERE EXAMINED BY REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION (PCR), REAL-TIME PCR, WESTERN BLOT, AND IMMUNOFLUORESCENT STAINING. HDAC2, HDAC4, AND ACETYLATED H4 EXPRESSION LEVELS WERE ASSAYED BY WESTERN BLOT. CYTOTOXICITY WAS ANALYZED BY THE TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE BIOTIN-DUTP NICK END LABELING ASSAY. RESULTS: THE EXPRESSION LEVELS OF ALPHA-SMA, FIBRONECTIN, AND COLLAGEN TYPE 1 WERE INCREASED IN NASAL POLYP AFTER TRANSFORMING GROWTH FACTOR (TGF) BETA1 TREATMENT. TSA-INHIBITED TGF-BETA1 INDUCED THESE GENE AND PROTEIN EXPRESSION LEVELS. FURTHERMORE, TSA SUPPRESSED PROTEIN EXPRESSION LEVELS OF HDAC2 AND HDAC4. HOWEVER, TSA INDUCED HYPERACETYLATION OF HISTONES H4. TREATMENT WITH TGF-BETA1 WITH OR WITHOUT TSA DID NOT HAVE CYTOTOXIC EFFECT. CONCLUSION: THESE FINDINGS PROVIDE NOVEL INSIGHTS INTO THE EPIGENETIC REGULATION IN MYOFIBROBLAST DIFFERENTIATION AND ECM PRODUCTION OF NASAL POLYP. TSA COULD BE A CANDIDATE OF A THERAPEUTIC AGENT FOR REVERSING THE TGF-BETA1-INDUCED ECM SYNTHESIS THAT LEADS TO NASAL POLYP DEVELOPMENT. 2013 11 3355 22 HISTONE EXTRACTION FROM HUMAN ARTICULAR CARTILAGE FOR THE STUDY OF EPIGENETIC REGULATION IN OSTEOARTHRITIS. OSTEOARTHRITIS (OA) IS A CHRONIC DISEASE THAT AFFECTS ARTICULAR CARTILAGE, CAUSING ITS DEGENERATION. ALTHOUGH OA IS ONE OF THE MOST PREVALENT PATHOLOGIES GLOBALLY, THERE ARE NO DEFINITIVE TREATMENTS AVAILABLE. RECENTLY, RESEARCH HAS FOCUSED ON ELUCIDATING THE COMPLEX INTERPLAY THAT TAKES PLACE BETWEEN INFLAMMATORY PROCESSES AND EPIGENETIC REGULATION, SHOWING THAT HISTONE POST-TRANSLATIONAL MODIFICATIONS (PTMS) CAN EXERT A PRONOUNCED EFFECT ON THE EXPRESSION OF OA-RELATED GENES. OA CHONDROCYTES ENHANCE THE PRODUCTION OF INTERLEUKIN 1BETA (IL-1BETA) AND INTERLEUKIN 8 (IL-8), WHICH ARE EPIGENETICALLY REGULATED. THESE CYTOKINES UPREGULATE THE SYNTHESIS OF MATRIX METALLOPROTEINASES (MMPS) AND AGGRECANASES, WHICH PROMOTE THE EXTRACELLULAR MATRIX (ECM) DESTRUCTION. THIS MOTIVATES THE STUDY OF HISTONE PTMS TO INVESTIGATE THE EPIGENETIC REGULATION OF PROINFLAMMATORY MOLECULES, BUT THE ABSENCE OF SPECIFIC PROTOCOLS TO EXTRACT HISTONES FROM HUMAN ARTICULAR CARTILAGE HAS COMPLICATED THIS TASK. THE LACK OF EFFECTIVE METHODS CAN BE EXPLAINED BY THE STRUCTURAL COMPLEXITY AND LOW CELLULARITY OF THIS TISSUE, WHICH ARE RESPONSIBLE FOR THE BIOMECHANICAL PROPERTIES THAT ALLOW THE MOVEMENT OF THE JOINT BUT ALSO COMPLICATE HISTONE ISOLATION. HERE, WE PROVIDE A HISTONE EXTRACTION PROCEDURE SPECIFICALLY ADAPTED FOR CRYOPRESERVED HUMAN ARTICULAR CARTILAGE THAT CAN BE USEFUL TO UNDERSTAND EPIGENETIC REGULATION IN OA AND ACCELERATE THE SEARCH FOR NOVEL STRATEGIES. 2022 12 814 15 CHANGES IN SMALL NONCODING RNA EXPRESSION DURING CHONDROCYTE SENESCENCE. OBJECTIVE: OSTEOARTHRITIS (OA) IS CHARACTERIZED BY THE CHRONIC AND PROGRESSIVE DETERIORATION OF ARTICULAR CARTILAGE. CHONDROCYTE SENESCENCE COULD LEAD TO A SHIFT IN THE BALANCE BETWEEN EXTRACELLULAR MATRIX (ECM) COMPONENT SYNTHESIS AND DEGRADATION. SMALL NONCODING RNAS (SNCRNAS), INCLUDING MICRORNAS (MIRNAS), P-ELEMENT-INDUCED WIMPY TESTIS-(PIWI-) INTERACTING RNAS (PIRNAS), SMALL NUCLEOLAR RNAS (SNORNAS), SMALL NUCLEAR RNAS (SNRNAS), AND REPEAT-ASSOCIATED SIRNAS (RASIRNAS), ARE A CLASS OF IMPORTANT EPIGENETIC MOLECULES. WE AIMED TO GAIN INSIGHTS INTO THE CHANGES AND ROLES OF SNCRNA IN CHONDROCYTE SENESCENCE. DESIGN: HEALTHY MOUSE POSTNATAL CHONDROCYTES WERE ISOLATED, AND A REPLICATIVE AGING MODEL WAS CONSTRUCTED. WE USED SMALL RNA SEQUENCING (SMALL RNA-SEQ) TO GENERATE EXTENSIVE SMALL RNA DATA. WE IDENTIFIED DIFFERENTIALLY EXPRESSED SNCRNAS AND PERFORMED TISSUE-SPECIFIC ANALYSIS USING REAL-TIME QUANTITATIVE POLYMERASE CHAIN REACTION (QRT-PCR). BETA-GALACTOSIDASE STAINING WAS USED TO DETECT CHONDROCYTE SENESCENCE. THE RESULTS SHOWED THAT THE EXPRESSION PROFILES OF SNCRNA IN PASSAGE 5 CHONDROCYTES WERE SIGNIFICANTLY DIFFERENT FROM THOSE IN PASSAGE 0 CHONDROCYTES. THE EXPRESSION OF SNCRNA WAS TISSUE SPECIFIC. WE FOUND THAT 40 MIRNAS WERE UPREGULATED AND 70 MIRNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE, AND THAT MIR-132-5P EXPRESSION INHIBITION PREVENTED CHONDROCYTE SENESCENCE. WE FOUND THAT 8 PIRNAS WERE UPREGULATED AND 17 PIRNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE, AND THAT PIRNA PIR_025576 OVEREXPRESSION DELAYED CHONDROCYTE SENESCENCE. WE FOUND THAT 24 SNORNAS WERE UPREGULATED AND 28 SNORNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE, AND THAT SNORNA ENSMUSG00000087935 OVEREXPRESSION DELAYED CHONDROCYTE SENESCENCE. WE FOUND THAT 5 SNRNAS WERE UPREGULATED AND 6 SNRNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE, AND THAT SNRNA ENSMUSG00000064682 OVEREXPRESSION DELAYED CHONDROCYTE SENESCENCE. WE FOUND THAT 1 RASIRNA WAS UPREGULATED AND 4 RASIRNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE. CONCLUSIONS: THESE FINDINGS MIGHT PROVIDE NOVEL INSIGHTS INTO OA PATHOGENESIS AND CONTRIBUTE TO THE DEVELOPMENT OF CANDIDATES FOR TARGETED THERAPEUTICS IN OA. 2022 13 6218 22 THE JOINT SYNOVIUM: A CRITICAL DETERMINANT OF ARTICULAR CARTILAGE FATE IN INFLAMMATORY JOINT DISEASES. THE SYNOVIUM CONSTITUTES THE ENVELOPE OF ARTICULAR JOINTS AND IS A CRITICAL PROVIDER OF SYNOVIAL FLUID COMPONENTS AND ARTICULAR CARTILAGE NUTRIENTS. ITS INFLAMMATION IS A PREDOMINANT FEATURE AND CAUSE OF JOINT DEGENERATION IN DISEASES AS DIVERSE AS RHEUMATOID, PSORIATIC, JUVENILE AND IDIOPATHIC ARTHRITIS, AND LUPUS, GOUT AND LYME DISEASE. THESE INFLAMMATORY JOINT DISEASES (IJDS) ARE DUE TO A WIDE VARIETY OF GENETIC, EPIGENETIC AND ENVIRONMENTAL FACTORS THAT TRIGGER, PROMOTE, AND PERPETUATE JOINT DESTABILIZATION. IN SPITE OF THIS VARIETY OF CAUSES, IJDS SHARE MAIN PATHOLOGICAL FEATURES, NAMELY INFLAMMATION OF THE JOINT SYNOVIUM (SYNOVITIS) AND PROGRESSIVE DEGENERATION OF ARTICULAR CARTILAGE. IN ADDITION TO BEING A DRIVING FORCE BEHIND THE DESTRUCTION OF ARTICULAR CARTILAGE IN IJD, SYNOVITIS IS ALSO INCREASINGLY BEING RECOGNIZED AS A SIGNIFICANT CONTRIBUTOR OF ARTICULAR CARTILAGE DEGENERATION IN OSTEOARTHRITIS, A DISEASE PRIMARILY DUE TO AGING- OR TRAUMA-RELATED WEAR AND TEAR OF CARTILAGE SURFACES. IN VIEW OF THIS IMPORTANT ROLE OF THE SYNOVIUM IN DETERMINING THE FATE OF ARTICULAR CARTILAGE, THIS REVIEW FOCUSES ON ITS UNDERLYING MECHANISMS IN THE PATHOLOGY OF IJD. WE ADDRESS THE ROLES OF SYNOVIAL FIBROBLASTS, MACROPHAGES AND ENDOTHELIAL CELLS IN THE MAINTENANCE OF JOINT HEALTH AND IN THE DESTRUCTION OF ARTICULAR CARTILAGE INTEGRITY DURING IJD. MOLECULAR MECHANISMS THAT HAVE BEEN RECENTLY SHOWN TO GOVERN THE PATHOLOGICAL ACTIVITIES OF THE RESIDENT SYNOVIAL CELLS ARE HIGHLIGHTED. FINALLY, ADVANTAGES AND DISADVANTAGES OF TARGETING THESE NEW MOLECULAR MECHANISMS FOR PREVENTING CARTILAGE DEGENERATION DUE TO CHRONIC INFLAMMATION ARE ALSO DISCUSSED. 2017 14 2694 28 EVOLVING SPECTRUM OF DIABETIC WOUND: MECHANISTIC INSIGHTS AND THERAPEUTIC TARGETS. DIABETES MELLITUS IS A CHRONIC METABOLIC DISORDER RESULTING IN AN INCREASED BLOOD GLUCOSE LEVEL AND PROLONGED HYPERGLYCEMIA, CAUSES LONG TERM HEALTH CONSE-QUENCES. CHRONIC WOUND IS FREQUENTLY OCCURRING IN DIABETES PATIENTS DUE TO COMPROMISED WOUND HEALING CAPABILITY. MANAGEMENT OF WOUNDS IN DIABETIC PATIENTS REMAINS A CLINICAL CHALLENGE DESPITE MANY ADVANCEMENTS IN THE FIELD OF SCIENCE AND TECHNOLOGY. INCREASING EVIDENCE INDICATES THAT ALTERATION OF THE BIOCHEMICAL MILIEU RESULTING FROM ALTERATION IN INFLAMMATORY CYTOKINES AND MATRIX METALLOPROTEINASE, DECREASE IN FIBROBLAST AND KERATINOCYTE FUNCTIONING, NEUROPATHY, ALTERED LEUKOCYTE FUNCTIONING, INFECTION, ETC., PLAYS A SIGNIFICANT ROLE IN IMPAIRED WOUND HEALING IN DIABETIC PEOPLE. APART FROM THE CURRENT PHARMACOTHERAPY, DIFFERENT OTHER APPROACHES LIKE THE USE OF CONVENTIONAL DRUGS, ANTIDIABETIC MEDICATION, ANTIBIOTICS, DEBRIDEMENT, OFFLOADING, PLATELET-RICH PLASMA, GROWTH FACTOR, OXYGEN THERAPY, NEGATIVE PRESSURE WOUND THERAPY, LOW-LEVEL LASER, EXTRACORPOREAL SHOCK WAVE BIOENGINEERED SUBSTITUTE CAN BE CONSIDERED IN THE MANAGEMENT OF DIABETIC WOUNDS. DRUGS/THERAPEUTIC STRATEGY THAT INDUCE ANGIOGENESIS AND COLLAGEN SYNTHESIS, INHIBITION OF MMPS, REDUCTION OF OXIDATIVE STRESS, CONTROLLING HYPERGLYCEMIA, INCREASE GROWTH FACTORS, REGULATE INFLAMMATORY CYTOKINES, CAUSE NO INDUCTION, INDUCE FIBROBLAST AND KERATINOCYTE PROLIFERATION, CONTROL MICROBIAL INFECTIONS ARE CONSIDERED IMPORTANT IN CONTROLLING DIABETIC WOUND. FURTHER, MEDICINAL PLANTS AND/OR PHYTOCONSTITUENTS ALSO OFFER A VIABLE ALTERNATIVE IN THE TREATMENT OF DIABETIC WOUND. THE FOCUS OF THE PRESENT REVIEW IS TO HIGHLIGHT THE MOLECULAR AND CELLULAR MECHANISMS, AND DISCUSS THE DRUG TARGETS AND TREATMENT STRATEGIES INVOLVED IN THE DIABETIC WOUND. 2022 15 5922 22 TARGETING DNA METHYLATION AND DEMETHYLATION IN DIABETIC FOOT ULCERS. BACKGROUND: POOR WOUND HEALING IS A SIGNIFICANT COMPLICATION OF DIABETES, WHICH IS COMMONLY CAUSED BY NEUROPATHY, TRAUMA, DEFORMITIES, PLANTAR HYPERTENSION AND PERIPHERAL ARTERIAL DISEASE. DIABETIC FOOT ULCERS (DFU) ARE DIFFICULT TO HEAL, WHICH MAKES PATIENTS SUSCEPTIBLE TO INFECTIONS AND CAN ULTIMATELY CONDUCE TO LIMB AMPUTATION OR EVEN DEATH IN SEVERE CASES. AN INCREASING NUMBER OF STUDIES HAVE FOUND THAT EPIGENETIC ALTERATIONS ARE STRONGLY ASSOCIATED WITH POOR WOUND HEALING IN DIABETES. AIM OF REVIEW: THIS WORK PROVIDES SIGNIFICANT INSIGHTS INTO THE DEVELOPMENT OF THERAPEUTICS FOR IMPROVING CHRONIC DIABETIC WOUND HEALING, PARTICULARLY BY TARGETING AND REGULATING DNA METHYLATION AND DEMETHYLATION IN DFU. KEY SCIENTIFIC CONCEPTS OF REVIEW: DNA METHYLATION AND DEMETHYLATION PLAY AN IMPORTANT PART IN DIABETIC WOUND HEALING, VIA REGULATING CORRESPONDING SIGNALING PATHWAYS IN DIFFERENT BREEDS OF CELLS, INCLUDING MACROPHAGES, VASCULAR ENDOTHELIAL CELLS AND KERATINOCYTES. IN THIS REVIEW, WE DESCRIBE THE FOUR MAIN PHASES OF WOUND HEALING AND THEIR ABNORMALITY IN DIABETIC PATIENTS. FURTHERMORE, WE PROVIDED AN IN-DEPTH SUMMARY AND DISCUSSION ON HOW DNA METHYLATION AND DEMETHYLATION REGULATE DIABETIC WOUND HEALING IN DIFFERENT TYPES OF CELLS; AND GAVE A BRIEF SUMMARY ON RECENT ADVANCES IN APPLYING CELLULAR REPROGRAMMING TECHNIQUES FOR IMPROVING DIABETIC WOUND HEALING. 2023 16 4880 19 OVERVIEW OF MMP-13 AS A PROMISING TARGET FOR THE TREATMENT OF OSTEOARTHRITIS. OSTEOARTHRITIS (OA) IS A COMMON DEGENERATIVE DISEASE CHARACTERIZED BY THE DESTRUCTION OF ARTICULAR CARTILAGE AND CHRONIC INFLAMMATION OF SURROUNDING TISSUES. MATRIX METALLOPROTEINASE-13 (MMP-13) IS THE PRIMARY MMP INVOLVED IN CARTILAGE DEGRADATION THROUGH ITS PARTICULAR ABILITY TO CLEAVE TYPE II COLLAGEN. HENCE, IT IS AN ATTRACTIVE TARGET FOR THE TREATMENT OF OA. HOWEVER, THE DETAILED MOLECULAR MECHANISMS OF OA INITIATION AND PROGRESSION REMAIN ELUSIVE, AND, CURRENTLY, THERE ARE NO INTERVENTIONS AVAILABLE TO RESTORE DEGRADED CARTILAGE. THIS REVIEW FULLY ILLUSTRATES THE INVOLVEMENT OF MMP-13 IN THE INITIATION AND PROGRESSION OF OA THROUGH THE REGULATION OF MMP-13 ACTIVITY AT THE MOLECULAR AND EPIGENETIC LEVELS, AS WELL AS THE STRATEGIES THAT HAVE BEEN EMPLOYED AGAINST MMP-13. THE AIM OF THIS REVIEW IS TO IDENTIFY MMP-13 AS AN ATTRACTIVE TARGET FOR INHIBITOR DEVELOPMENT IN THE TREATMENT OF OA. 2021 17 1257 24 CURRENT TRENDS IN EPIGENETIC, CELLULAR AND MOLECULAR PATHWAYS IN MANAGEMENT OF RHEUMATOID ARTHRITIS. RHEUMATOID ARTHRITIS IS A SYSTEMIC CHRONIC POLYARTICULAR AUTOIMMUNE DISORDER OF JOINTS AND JOINT MEMBRANE MAINLY AFFECTING FEET AND HANDS. THE PATHOLOGICAL MANIFESTATION OF THE DISEASE INCLUDES INFILTRATION OF IMMUNE CELLS, HYPERPLASIA OF THE LINING OF SYNOVIUM, FORMATION OF PANNUS AND BONE AND CARTILAGE DESTRUCTION. IF LEFT UNTREATED, THE APPEARANCE OF SMALL FOCAL NECROSIS, ADHESION OF GRANULATION, AND FORMATION OF FIBROUS TISSUE ON THE SURFACE OF ARTICULAR CARTILAGE IS NOTED. THE DISEASE PRIMARILY AFFECTS NEARLY 1% OF THE POPULATION GLOBALLY, WOMEN BEING MORE AFFECTED THAN MEN WITH A RATIO 2:1 AND CAN INITIATE REGARDLESS OF ANY AGE. THE SYNOVIAL FIBROBLAST IN RHEUMATOID ARTHRITIS INDIVIDUALS EXHIBITS AN AGGRESSIVE PHENOTYPE WHICH UPREGULATES THE MANIFESTATION OF PROTOONCOGENES, ADHESIVE COMPOUNDS, INFLAMMATORY CYTOKINES AND MATRIX-DETERIORATING ENZYMES. APART FROM THE INFLAMMATORY EFFECTS OF CYTOKINES, CHEMOKINES ARE ALSO NOTED TO INDUCE SWELLING AND PAIN IN ARTHRITIC INDIVIDUALS BY RESIDING IN SYNOVIAL MEMBRANE AND FORMING PANNUS. THE CURRENT TREATMENT OF RHEUMATOID ARTHRITIS INCLUDES TREATMENT WITH NON-STEROIDAL ANTI-INFLAMMATORY DRUGS, DISEASE-MODIFYING ANTIRHEUMATIC DRUGS, TREATMENT WITH BIOLOGICS SUCH AS INHIBITORS OF TNF-ALPHA, INTERLEUKINS, PLATELET ACTIVATING FACTOR, ETC. WHICH PROVIDES SIGNIFICANT RELIEF FROM SYMPTOMS AND AIDS IN MANAGEMENT OF THE DISEASE. THE CURRENT REVIEW HIGHLIGHTS THE PATHOGENESIS INVOLVED IN THE ONSET OF RHEUMATOID ARTHRITIS AND ALSO COVERS EPIGENETIC, CELLULAR AND MOLECULAR PARAMETERS ASSOCIATED WITH IT TO AID BETTER AND ADVANCED THERAPEUTIC APPROACHES FOR MANAGEMENT OF THE DEBILITATING DISEASE. 2023 18 5850 18 SUBEROYLANILIDE HYDROXAMIC ACID (SAHA) REDUCES FIBROSIS MARKERS AND DEACTIVATES HUMAN STELLATE CELLS VIA THE EPITHELIAL-MESENCHYMAL TRANSITION (EMT). HEPATIC FIBROSIS IS KNOWN AS THE ACCUMULATION OF CONNECTIVE TISSUE SECONDARY TO CHRONIC DAMAGE TO THE LIVER. EPITHELIAL-MESENCHYMAL TRANSITION (EMT) CORRESPONDING INCREASE IN LIVER FIBROGENESIS WAS SHOWN WITH IMMUNOHISTOCHEMISTRY AND PCR-BASED STUDIES. SUBEROYLANILIDE HYDROXAMIC ACID (SAHA), A SYNTHETIC COMPOUND APPROVED AS A HISTONE DEACETYLASE INHIBITOR (HDAC) BY THE FDA TO TREAT CUTANEOUS T-CELL LYMPHOMA IS UNDER INVESTIGATION FOR THE TREATMENT OF LUNG AND RENAL FIBROSIS. EXPERIMENTAL MODELING FOR HEPATIC FIBROSIS CAN BE CONSTRUCTED WITH AN LX2 CELL LINE ISOLATED FROM HUMAN HEPATIC STELLATE CELLS (HSCS). IN THIS STUDY, WE AIMED TO INVESTIGATE THE MODULATION OF SAHA IN THE PATHOGENESIS OF LIVER FIBROSIS BY DETECTING THE LEVELS OF PROTEINS; (E-CADHERIN (E-CAD), N-CADHERIN (N-CAD), VIMENTIN (VIM), AND GENES; E-CAD, N-CAD, VIM, TRANSFORMING GROWTH FACTOR-BETA (TGF-BETA), ALPHA-SMOOTH MUSCLE ACTIN (ALPHA-SMA), TYPE 1 COLLAGEN (COL1A1), TYPE 3 COLLAGEN (COL3A1)) THAT PLAY A SIGNIFICANT ROLE IN EMT WITH THE LX2 CELL LINE. WE ALSO EVALUATED THE ACTION OF SAHA WITH CELL PROLIFERATION, CLONOGENIC, AND MIGRATION ASSAY. CELL PROLIFERATION WAS PERFORMED BY FLOW CYTOMETRY. ALL THE PROTEIN LEVELS WERE DETERMINED BY WESTERN BLOT ANALYSIS, AND GENE EXPRESSION LEVELS WERE MEASURED BY REAL-TIME PCR. OUR STUDY OBSERVED THAT SAHA TREATMENT DECREASED CELL VIABILITY, COLONY FORMATION AND MIGRATION IN LX2 CELLS. WE FOUND THAT SAHA INCREASED E-CAD EXPRESSION LEVEL, WHILE IT DECREASED N-CAD, VIM, COL1A1, COL3A1, ALPHA-SMA TGF-BETA GENES EXPRESSION LEVELS. SAHA DECREASED THE LEVEL OF E-CAD, N-CAD, AND VIM PROTEIN LEVELS. WE THOUGHT THAT SAHA POSSESSES POTENT ANTIFIBROTIC AND ANTI-EMT PROPERTIES IN LX2. 2021 19 4873 22 OVER-EXPRESSION OF PROTHYMOSIN-ALPHA ANTAGONIZES TGFBETA SIGNALLING TO PROMOTE THE DEVELOPMENT OF EMPHYSEMA. EMPHYSEMA, A MAJOR CONSEQUENCE OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD), IS CHARACTERIZED BY THE PERMANENT AIRFLOW RESTRICTION RESULTING FROM ENLARGEMENT OF ALVEOLAR AIRSPACE AND LOSS OF LUNG ELASTICITY. TRANSFORMING GROWTH FACTOR-BETA (TGFBETA) SIGNALLING REGULATES THE BALANCE OF MATRIX METALLOPROTEINASE (MMP)/TISSUE INHIBITOR OF MATRIX METALLOPROTEINASE (TIMP) TO CONTROL MATRIX HOMEOSTASIS. PATIENTS WITH COPD HAVE DYSREGULATED TGFBETA SIGNALLING AND REDUCED HISTONE DEACETYLASE (HDAC) ACTIVITY THROUGH EPIGENETIC UP-REGULATION OF HISTONE ACETYLATION IN THE PROMOTERS OF PRO-INFLAMMATORY GENES. HOWEVER, THE POTENTIAL LINK BETWEEN DECREASED HDAC ACTIVITY AND DYSREGULATED TGFBETA SIGNALLING IN EMPHYSEMA PATHOGENESIS REMAINS TO BE DETERMINED. PROTHYMOSIN ALPHA (PROT), A HIGHLY CONSERVED ACIDIC NUCLEAR PROTEIN, PLAYS A ROLE IN THE ACETYLATION OF HISTONE AND NON-HISTONE PROTEINS. THE AIM OF THIS STUDY WAS TO TEST THE HYPOTHESIS THAT PROT INHIBITS TGFBETA-SMAD SIGNALLING THROUGH SMAD7, THEREBY CONTRIBUTING TO EMPHYSEMA PATHOGENESIS. WE SHOW THAT PROT ENHANCES SMAD7 ACETYLATION BY DECREASING ITS ASSOCIATION WITH HDAC AND THEREBY DOWN-REGULATES TGFBETA-SMAD SIGNALLING. PROT CAUSED AN IMBALANCE BETWEEN MMP AND TIMP THROUGH ACETYLATED SMAD7 IN FAVOUR OF MMP EXPRESSION. IN ADDITION TO INTERFERING WITH R-SMAD ACTIVATION AND TARGETING RECEPTORS FOR DEGRADATION IN THE CYTOPLASM, ACETYLATED SMAD7 POTENTIATED BY PROT COMPETITIVELY ANTAGONIZED BINDING OF THE PSMAD2/3-SMAD4 COMPLEX TO THE TIMP-3 PROMOTER, RESULTING IN REDUCED TIMP-3 EXPRESSION. THESE EFFECTS WERE DETECTED IN PROT-OVER-EXPRESSING CELLS, LUNGS OF PROT TRANSGENIC MICE DISPLAYING AN EMPHYSEMA PHENOTYPE AND IN EMPHYSEMA PATIENTS. IMPORTANTLY, INCREASED SMAD7 AND REDUCED TIMP-3 WERE FOUND IN THE LUNGS OF EMPHYSEMA PATIENTS AND MICE WITH CIGARETTE SMOKE EXTRACT (CSE)-INDUCED EMPHYSEMA. SUCH EFFECTS COULD BE ABROGATED BY SILENCING ENDOGENOUS PROT EXPRESSION. COLLECTIVELY, OUR RESULTS UNCOVER ACETYLATED SMAD7 REGULATED BY PROT AS AN IMPORTANT DETERMINANT IN DYSREGULATED TGFBETA SIGNALLING THAT CONTRIBUTES TO EMPHYSEMA PATHOGENESIS. 2016 20 5585 30 ROLE OF OXIDATIVE STRESS AND GENETIC POLYMORPHISM OF MATRIX METALLOPROTEINASE-2 AND TISSUE INHIBITOR OF METALLOPROTEINASE-2 IN COPD. CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD), A COMPLAINT DESCRIBED BY PROGRESSIVE AND INADEQUATELY REVERSIBLE LIMITATION IN LUNGS WITH SYSTEMIC INFLAMMATION, IS LARGELY CURRENT IN INDIA. THERE'S NO REMEDY AVAILABLE SO FAR IT IS, THUS, IMPERATIVE TO UNDERSTAND THE UNDERPINNING PATHOGENESIS OF THE COMPLAINANT. A SET OF PROTEASES KNOWN AS MATRIX METALLOPROTEINASE (MMPS) ARE ESPECIALLY INVOLVED IN THE PROCESS OF ALVEOLAR DESTRUCTION AND MUCUS HYPERSECRETION. THERE ARE RESPONSIBLE FACTORS IN AN INHERITABLE POSITION TO CONTROL COPD LIKE MMPS AND TIMPS (TISSUE INHIBITOR OF METALLOPROTEINASES). MMPS DEGRADE EXTRACELLULAR MATRIX AND LEAD TO THE PATHOGENESIS OF COPD [1]. TIMPS PROTEINS THAT HELP TO INHIBIT THE MATRIX METALLOPROTEINASES. [2]. THIS REVIEW SUMMARIZES THE IMPLICIT PART OF CRUCIAL MMP-2 AND TIMP-2 IN COPD DISEASE. THOUGH THE CONCEPT SEEMS PROMISING, LIMITED KNOWLEDGE ABOUT THE EXACT FUNCTIONS OF A PARTICULAR MMP IN COPD AND THE COMPLICATIONS OF MMP IN SUBSTRATE AFFINITY MAKES THIS A GRUELING TASK. MMP2 AND TIMP2 BOTH ARE DIRECTLY OR INDIRECTLY REGULATED BY OXIDATIVE STRESS AND EPIGENETIC MECHANISM WHICH REGULATES THEIR EXPRESSIONS. COPD IS A SEDITIOUS RESPONSE TO FACTORS LIKE DUST, SMOKE, ETC., AND TRIGGERS EXTRA-PULMONARY GOODS WHICH CAUSE INFLAMMATION. [3]. THIS REVIEW EXPLAINS THE RELATIONSHIP BETWEEN MMP2 AND TIMP2 IN COPD PATIENTS WITH OXIDATIVE STRESS, ITS IMPACT ON COPD PATHOGENESIS, AND GENE EXPRESSION OF TIMP2 AND MMP2 WITH THEIR DOWNSTREAM EFFECTS. THIS ALSO GIVES SOME INSIGHTS INTO THERAPEUTIC INTERVENTIONS FOR TARGETING THESE ENZYMES. MMP2 AND TIMP2 BOTH PLAY A ROLE IN THE DEVELOPMENT OF COPD AND THEY NEED TO BE STUDIED WITH THE UTMOST FOCUS. 2023