Tissue Inhibitor Of Metalloproteinase Research Articles

Exosomal therapy mitigates silver nanoparticles-induced neurotoxicity in rats

Introduction Our investigation aims to appraise the neuroprotective impact of Bone Marrow-Mesenchymal Stem Cells (BM-MSCs) derived exosomes against Ag NPs-inducing neurotoxicity in rats. Materials and Methods Twenty-four albino rats were divided into 3 groups. Group I (control negative), Group II (intraperitoneally injected with Ag NPs for 28 days, whereas Group III (intraperitoneally injected with Ag NP and BM-MSCs derived exosomes. Results There was a marked elevation of Malondialdehyde (MDA) along with a reduction of brain antioxidants, Gamma-aminobutyric acid (GABA) and Monoamine Oxidase (MAO) in the Ag NPs receiving group. Ag NPs upregulated c-Jun N-terminal Kinases (JNK) genes and c-Myc and downregulated the tissue inhibitors of metalloproteinases (TIMP-1) and Histone deacetylase 1 (HDAC1) genes. Otherwise, the co-treatment of BM-MSCs derived exosomes with Ag NPs could markedly increase the rat’s body weight, activity and learning while, decreasing anxiety, restoring all the toxicological parameters and improving the microscopic appearance of different brain areas. Conclusion BM-MSCs-derived exosomes downregulated both apoptotic and inflammatory mediators and upregulated the antiapoptotic genes. BM-MSCs-derived exosomes exhibit a great therapeutic effect against the neurotoxic effects of Ag NPs.

Bone and periosteum protein analysis via tandem mass tag quantitative proteomics in pediatric patients with osteomyelitis.

Bone healing is crucial in managing osteomyelitis after fracture fixation. Understanding the mechanism of extensive callus formation in pediatric osteomyelitis is highly important. This study aims to analyze bone and periosteum samples from pediatric patients to elucidate the essential processes involved in callus formation during osteomyelitis. The study included eight patients from our hospital: four with positive microbial culture who underwent osteomyelitis debridement and four who had osteotomy surgery as contral. We used tandem mass tag quantitative proteomics to investigate proteomic changes in bone and periosteum tissues obtained from these patients. Differential expression proteins were analyzed for their pathways through Gene Ontology (GO) annotation, GO enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, and protein-protein interaction networks. A total of 4737 proteins were successfully identified. About 2224 differentially expressed proteins were detected in the bone tissues group and periosteum tissues group. Among the differentially expressed proteins, 10 protein genes in the bone group were associated with inflammation and osteogenesis, while in the periosteum group were nine. Cytochrome b-245, beta polypeptide (CYBB), nicotinamide phosphoribosyltransferase (NAMPT), tissue inhibitor of metalloproteinases 1 (TIMP-1), Raf-1 proto-oncogene, serine/threonine kinase (RAF-1), RELA proto-oncogene, NF-KB subunit (RELA), and sphingomyelin synthase 2 (SGMS2) may play an important role in callus formation in patients with osteomyelitis. This study provides novel clues for understanding callus formation in pediatric patients with osteomyelitis.

Modification of transvaginal polypropylene mesh with co-axis electrospun nanofibrous membrane to alleviate complications following surgical implantation

BackgroundSurgeries for treating pelvic organ prolapse involving the utilization of synthetic mesh have been associated with complications such as mesh erosion, postoperative pain, and dyspareunia. This work aimed to reduce the surgical implantation-associated complications by nanofibrous membranes on the surface of the polypropylene mesh. The nanofiber of the nanofibrous membrane, which was fabricated by co-axial electrospinning, was composed of polyurethane as fiber core and gelatin as the fiber out layer. The biocompatibility of the modified mesh was evaluated in vitro by cell proliferation assay, immunofluorescence stain, hematoxylin-eosin (HE) staining, and mRNA sequencing. Polypropylene mesh and modified mesh were implanted in a rat pelvic organ prolapse model. Mesh-associated complications were documented. HE and Picro-Sirius red staining, immunohistochemistry, and western blotting were conducted to assess the interactions between the modified mesh and vaginal tissues.ResultsThe modified mesh significantly enhanced the proliferation of fibroblasts and exerted a positive regulatory effect on the extracellular matrix anabolism in vitro. When evaluated in vivo, no instances of mesh exposure were observed in the modified mesh group. The modified mesh maintained a relatively stable histological position without penetrating the muscle layer or breaching the epidermis. The collagen content in the vaginal wall of rats with modified mesh was significantly higher, and the collagen I/III ratio was lower, indicating better tissue elasticity. The expression of metalloproteinase was decreased while the expression levels of tissue inhibitor of metalloproteinase were increased in the modified mesh group, suggesting an inhibition of collagen catabolism. The expression of TGF-β1 and the phosphorylation levels of Smad3, p38 and ERK1/2 were significantly increased in the modified mesh group. NM significantly improved the biocompatibility of PP mesh, as evidenced by a reduction in macrophage count, decreased expression levels of TNF-α, and an increase in microvascular density.ConclusionsThe nanofibrous membrane-coated PP mesh effectively reduced the surgical implantation complications by inhibiting the catabolism of collagen in tissues and improving the biocampibility of PP mesh. The incorporation of co-axial fibers composed of polyurethane and gelatin with polypropylene mesh holds promise for the development of enhanced surgical materials for pelvic organ prolapse in clinical applications.

Effect of Laparoscopic Inguinal Hernia Repairs on Inflammatory Factors, Oxidative Stress Levels and Postoperative Recovery.

To unravel the effects of different laparoscopic inguinal hernia repairs (IHRs) on inflammatory factors, oxidative stress, and postoperative recovery of inguinal hernia (IH) patients. A total of 88 patients for laparoscopic treatment of IH were studied in a retrospective cohort, and were randomized into the transabdominal preperitoneal (TAPP) repair group (n = 44) and the total extraperitoneal (TEP) repair group (n = 44) according to the method of treatment. Patients in the TAPP group underwent laparoscopic TAPP IHR, while patients in the TEP group underwent laparoscopic TEP IHR. Visual analogue scores (VAS) were measured at 1, 3, and 7 d after repair. Gastrin (GAS), motilin (MTL), matrix metalloproteinase (MMP)-2, MMP-9, and tissue inhibitor of metalloproteinases-1 (TIMP-1) were assessed by enzyme-linked immunosorbent assay. Superoxide dismutase (SOD) was tested by spectrophotometry and malondialdehyde (MDA) and β-endorphin (β-EP) were examined by radioimmunoprecipitation. Total antioxidant capacity (T-AOC) was evaluated by chemical colorimetry, quality of life was evaluated by SF-36, and fibrinogen (FIB), C-reactive protein (CRP), antitrypsin (al-AT), and interleukin-6 (IL-6) levels were determined by immunoturbidimetry. Complications and postoperative recurrence were recorded. VAS scores in the TEP group were lower than those in the TAPP group. Operative time, hospital stay, time for postoperative activity out of bed, and time taken to exhaust were shorter in the TEP group than in the TAPP group. Intraoperative blood loss was less in the TEP group than in the TAPP group. GAS, MTL, SOD, and T-AOC were higher, and MDA, β-EP, CRP, FIB, alAT, IL-6, MMP-2, MMP-9, and TIMP-1 were lower in the TEP group than the TAPP group. Quality of life was improved in the TEP group compared with the TAPP group. There was no significant difference in the rate of postoperative complications and recurrence rates between both groups of patients. For IHs, laparoscopic TEP repair can effectively relieve pain, reduce inflammatory factors and oxidative stress indices, and facilitate postoperative recovery.

A comprehensive study on mechanisms of action of fibroin, aloe vera, and ginger extracts through histochemical, inflammation biomarkers, and matrix metalloproteinases analysis against diabetic wounds

IntroductionDiabetes causes complications like delayed wound healing for a long time. Fibroin, aloe vera, and ginger extracts along with their combinations are used for diabetic wound healing. MethodsAfter induction of diabetes, The wound healing effects of fibroin (50 mg/ml), aloe vera gel (50 mg/ml), and ginger extract (30 mg/ml), individually and in combination, were assessed. The pro-inflammatory cytokines including tumor narcosis factor-α (TNF-α) interleukin (IL-6, IL-8, IL-1β), matrix metalloproteinases (MMP 2, MMP7, MMP 9), vascular endothelial growth factor (VEGF) and tissue inhibitors of metalloproteinases (TIMPs) levels were analyzed in the serum. ResultsA combination of fibroin + aloe vera gel + ginger extract (Fi + Al + Gi) healed the wounds in 11 days via wound contraction of 98.5 ± 0.9 % as compared to diabetic control (58.2 ± 0.7 %) and positive control (73.3 ± 0.6 %) groups. However, the wounds of the Polyfax and the diabetic control groups were healed in 17 and 19 days, corresponding to a contraction of: 96.7 ± 1.4 % and 96.3 ± 1.1 %. The histological assay showed that the Fi + Al + Gi group indicated an increased growth of collagen fibers, fibroblasts, keratinocytes and blood vessels with lessened inflammation. The Fi + Al + Gi group alleviated the serum level of TNF-α (12.7 ± 0.9 pg/ml), IL-6 (9.6 ± 0.9 pg/ml), IL-8 (19.6 ± 1.0 pg/ml), MMP2 (217.0 ± 9.2 pg/ml), MMP7 (279.0 ± 9.8 pg/ml), and MMP9 (156.0 ± 11.6 pg/ml) significantly as compared to the diabetic control (P ≤ 0.05). TIMP serum level (202.0 ± 6.9 pg/ml) was significantly elevated as compared to the diabetes control group. ConclusionThe present study concludes that the biomaterials in their combinations possess high regenerative and healing abilities.

Use of multiparametric MRI to noninvasively assess iodinated contrast-induced acute kidney injury

PurposeTo gauge the utility of multiparametric MRI in characterizing pathologic changes after iodinated contrast-induced acute kidney injury (CI-AKI) in rats. MethodsWe randomly grouped 24 rats injected with 8 g iodine/kg of body weight (n = 6 each) and 6 rats injected with saline as controls. All rats underwent T1, T2 mapping and diffusion kurtosis imaging (DKI) after contrast injection at 0 (control), 1, 3, 7, 13 days. T1, T2, and mean kurtosis (MK) values were performed in renal outer/inner stripes of outer medulla (OSOM and ISOM) and cortex (CO), and their diagnosis performance for CI-AKI also been evaluated. Serum creatinine (SCr), insulin-like growth factor-binding protein 7 (IGFBP7), tissue inhibitor metalloproteinase 2 (TIMP-2), aquaporin-1 (AQP1), α-smooth muscle actin (α-SMA), and histologic indices were examined. ResultsCompared with controls, urinary concentrations of both TIMP-2 and IGFBP7 were obviously elevated from Day 1 to Day 13 (all p < 0.05). T2 values were significantly higher than control group for Days 1 and 3, and T1 and MK increased were more remarkable at all time points (Days 1–13) in CI-AKI (all p < 0.05) than control group. Changes in T1 and MK strongly correlated with renal injury scores of all anatomical compartments and with expression levels of AQP1 and moderately correlated with α-SMA. Changes in T2 values correlating moderately with renal scores of CO, ISOM and OSOM and AQP1. The MK obtained the highest area under the receiver operating characteristic (ROC) curve of 0.846 with a sensitivity of 70.8 % and specificity of 88.9 %. ConclusionsCombined use of multiparametric MRI could be a valid noninvasive method for comprehensive monitoring of CI-AKI. Among these parameters, MK may achieve the best diagnostic performance for CI-AKI.

Alpha(α)-mangostin (Xanthone of Garcinia mangostana L.): Augmenting Macrophages Activity for an Effective Diabetic Wound Healing

Diabetic wounds are particularly difficult to treat medically because they heal at a slower pace than regular wounds. Macrophages are essential in all stages of normal wound healing, including inflammation, proliferation and rem odelling. When wound healing is affected, macrophages can reduce the level of growth factor and increase the level of interleukin-6 (IL-6), disrupt the balance between tissue inhibitors of metalloproteinase (TIMPs) and matrix metalloproteinase (MMPs), which can slow down healing. Alpha(α)-mangostin, a natural xanthone derived from the pericarp of the mangosteen, has gained considerable attention due to its anti-inflammatory properties, suggesting its potential to promote wound healing. However, its exact role in healing diabetic foot ulcers, common in diabetes, remains unclear. Hence, this study aims to explore how α-mangostin might affect diabetic wound healing by evaluating its impact on PDGF, CTGF, BFGF, VEGF, TGF-β, MMP-9, TIMP-2 and IL-6 secretion in macrophage cells. Human monocytic macrophages (THP-1) were incubated with a 35 mM glucose solution for 72 h to create a glucose-enriched medium. The cells were then incubated with α-mangostin (0.15, 2.5 and 5 µg/mL) together with 35 mM glucose. Carboxymethyl cellulose (CMC) served as positive controls; glucose-enriched media and media-alone served as negative controls. Protein expression was measured using ELISA. α-mangostin (2.5 µg/mL) increased the levels of PDGF and VEGF and decreased the level of MMP-9 compared to glucose controls. There was no significant difference in other growth factors, TIMP-2 and IL-6 protein levels across any of the treatment groups compared to glucose controls. In conclusion, α-mangostin particularly at 2.5 µg/mL demonstrated a significant increase in PDGF and VEGF levels while simultaneously reducing MMP-9 in macrophage cells under glucose-induced conditions. These findings suggest that α-mangostin holds the potential for enhancing the healing of chronic wounds in diabetic conditions. HIGHLIGHTS Treatment of macrophages (THP-1 cells) with 2.5 µg/mL Alpha (α)-mangostin in a high glucose medium led to a significant increase in PDGF and VEGF secretion compared to glucose control. Treatment of the cells with 2.5 µg/mL Alpha (α)-mangostin in a high glucose medium led to a significant reduction of MMP-9 secretion compared to glucose control. There is no significant effects of Alpha (α)-mangostin on IL-6 and TIMP secretion compared to controls This study suggests that Alpha (α)-mangostin may be beneficial in promoting diabetic wound healing via stimulation of PDGF and VEGF secretion and reducing MMP-9 levels. GRAPHICAL ABSTRACT

Study of biomarkers to determine severity and lung damages in COVID-19 patients.

Identifying inflammation and lung damage markers is crucial in reducing morbidity and mortality of coronavirus disease 2019 (COVID-19). This study aimed to examine the validity and reliability of severity and post-infection lung damage and analyse their relationship. This was a prospective analysis study at the Airlangga University Hospital, Surabaya, Indonesia, from March to August 2021. The infection`s severity was measured by examining angiotensin-converting enzyme 2 (ACE2) levels and complete blood count. Lung damage was estimated by reviewing Krebs von de Lungen (KL)-6, matrix metalloproteinase (MMP)-9, tissue inhibitor metalloproteinase (TIMP)-1, and MMP-9/TIMP-1. Two-factor confirmatory factor analysis (CFA) and canonical correlation were calculated using Lisrel and SPSS (version 25). The research sample included 76 patients. The t count loading factor values were calculated: ACE2 (6.00), neutrophils (-0.80), lymphocytes (-0.63), neutrophil-lymphocyte ratio (NLR, 1.27), eosinophils (-1.52), basophils (1.72), monocytes (0.05), platelets (0.53), leukocytes (-0.51), platelet-lymphocyte ratio (PLR, -1.15), KL-6 (10.47), MMP-9 (11.91), TIMP-1 (11.79), and MMP-9/TIMP-1 (-0.24). The t values were: neutrophil covariance error (6.11), lymphocytes (6.12), NLR (6.10), eosinophils (6.08), basophils (6.07), monocytes (6.12), platelets (6.12), leukocytes (6.12), PLR (6.10), ACE2 (0.97), KL-6 (5.63), MMP-9 (2.08), TIMP-1 (2.77), and MMP-9/TIMP-1 (6.12). t value canonical correlation of 7.04 (t count > 1.96) indicated a correlation between the severity of the patient and post-infection lung damage. The severity was adequately measured through ACE2, IL-6, IL-10, neutrophils, lymphocytes, leukocytes, and NLR. Lung damage was measured with KL-6, MMP-9, and TIMP-1. There was a correlation between disease severity and lung damage.

Proteolytic imbalance in plasma of patients with multiple sclerosis following COVID-19

Background. The present research was conducted with the following objectives: 1) to determine the plasma levels of five matrix metalloproteinases (MMPs), namely MMP-1, -2, -3, -8, -10, and tissue inhibitor of metalloproteinase-1 (TIMP-1); 2) to analyze protease activity profiles in plasma using a zymographic method; and 3) to perform preliminary analysis on plasma peptide pool composition in patients with multiple sclerosis (MS) with and without COVID-19 history. Materials and methods. We examined 97 patients with MS: 41 had been diagnosed with COVID-19 in the past 4–6 months (MS + COVID group), and 56 did not suffer from SARS-CoV-2 infection previously (MS group). The plasma of healthy volunteers (n = 30) with no evidence of disease was used as control. The enzyme-linked immunosorbent assay was used to measure MMP and TIMP-1 concentrations. Plasma MMP activity was verified by gelatin-substrate zymography. Peptide pools were extracted from the plasma of MS patients and healthy subjects. Then size exclusion chromatography was used to identify separate fractions present in peptide pools. Results. We found that plasma concentration of MMP-2 was remarkably increased in the MS group compared with healthy controls, while in the MS + COVID patients, the levels of two other MMPs, MMP-1 and -10, were elevated. Zymography showed four dominant gelatinolytic bands of 92, 84, 72, and 62 kDa in MS plasma samples, whereas only traces of MMP were detected in healthy subjects. Most of MS plasma samples showed MMP-2 lytic activity, but only a few contained MMP-9. Finally, we determined the concentration of circulating peptides. The levels of plasma peptides were higher in patients from both the MS and MS + COVID group compared to control subjects. According to our results, the development of MS was accompanied by changes in both quantity and quality of peptide pool composition compared to healthy controls. Conclusions. Thus, an advanced understanding of the role of MMPs in MS pathogenesis following infection is important in developing optimized interventions to improve health and clinical outcomes during COVID-19.

Serum matrix metallopeptidase-9 levels in infantile epileptic spasms syndrome of unknown etiology

PurposeEpileptic spasms are the primary symptom of infantile epileptic spasms syndrome (IESS); however, their direct impact on blood–brain barrier (BBB) function is unknown. Matrix metallopeptidase-9 (MMP-9), degrades type IV collagen, a key component of the blood-brain barrier, while tissue inhibitor of metalloproteinase-1 (TIMP-1) suppresses its activity, protecting BBB integrity. This study aimed to assess serum MMP-9 and TIMP-1 levels in patients with IESS of unknown etiology. MethodsWe prospectively assessed serum MMP-9 and TIMP-1 levels prior to administering vigabatrin or adrenocorticotropic hormone therapy in patients with IESS of unknown etiology at Saitama Children’s Medical Center between February 2012 and December 2023. We compared these biomarkers between patients with epileptic spasms and age-matched controls and performed a curve regression analysis between the biomarkers and the frequency of epileptic spasms. Additionally, we assessed whether MMP-9 and TIMP-1 levels were diagnostic predictors of IESS. ResultsThis study included 22 patients with IESS (11 males) and 12 controls. Serum MMP-9 and MMP-9/TIMP-1 ratios were higher in patients with IESS than in controls (p < 0.001 and p = 0.002, respectively). A high frequency of epileptic spasms also led to higher serum MMP-9 levels (y = 0.0871x2 + 0.195x + 195.15, R² = 0.77, p < 0.001). Using MMP >188 ng/mL as the cutoff level, the sensitivity for diagnosing IESS was 95.5 %, the specificity was 75.0 %, the positive likelihood ratio was 3.82 (95 % confidence interval (CI) 1.43–10.22), and the relative risk was 8.75 (95 % CI 1.36–56.5). ConclusionPatients with IESS had elevated serum MMP-9 levels, suggesting an association between epileptic spasms and blood–brain barrier dysfunction. MMP-9 level measurement may be useful for diagnosing suspected patients.

Regulation of alternative polyadenylation isoforms of Timp2 is an effector event of RAS signaling in cell transformation.

Alternative polyadenylation (APA) generates mRNA isoforms with different lengths of the 3' untranslated region (3' UTR). The tissue inhibitor of metalloproteinase 2 (TIMP2) plays a key role in extracellular matrix remodeling under various developmental and disease conditions. Both human and mouse genes encoding TIMP2 contain two highly conserved 3'UTR APA sites, leading to mRNA isoforms that differ substantially in 3'UTR size. APA of Timp2 is one of the most significantly regulated events in multiple cell differentiation lineages. Here we show that Timp2 APA is highly regulated in transformation of NIH3T3 cells by the oncogene HRAS G12V . Perturbations of isoform expression with long 3'UTR isoform-specific knockdown or genomic removal of the alternative UTR (aUTR) region indicate that the long 3'UTR isoform contributes to the secreted Timp2 protein much more than the short 3'UTR isoform. The short and long 3'UTR isoforms differ in subcellular localization to endoplasmic reticulum (ER). Strikingly, Timp2 aUTR enhances secreted protein expression but no effect on intracellular proteins in reporter assays. Furthermore, downregulation of Timp2 long isoform mitigates gene expression changes elicited by HRAS G12V . Together, our data indicate that regulation of Timp2 protein expression through APA isoform changes is an integral part of RAS-mediated cell transformation and 3'UTR isoforms of Timp2 can have distinct impacts on expression of secreted vs. intracellular proteins.

Amelioration Potency of a Nano-Therapeutic Drug in Rats with Uninephrectomy and Cisplatin-induced Toxicity

While physicians describe drugs to treat diseases, these medications may have cytotoxic effects on certain organs, necessitating the use of some drugs to ameliorate such adverse effects. The study was conducted to investigate the protective behavior of nanoemulsified sodium salicylate on uninephrectomized rats injected with cisplatin to induce nephrotoxicity. Fifty adult male albino rats, aged five weeks and weighing approximately 100-120 g, were divided into five groups. The first group received 200 mg/kg/day i.p normal saline for 30 days. The second group was administrated 200 mg/kg/day of nanoemulsified salt of salicylic acid for 30 days. The third group, comprising uninephrectomized rats, was injected with two doses of cisplatin (20 mg/kg body weight) on alternate days from the start of the experiment to induce nephrotoxicity. The fourth group, also uninephrectomized, received 200 mg/kg/day i.p of nanoemulsified sodium salicylate for 30 days. The fifth group, uninephrectomized and treated with 200 mg/kg/day sodium salicylate nanoemlusion for 21 days, was subsequently injected with two doses of cisplatin, followed by continued nanoemulsified sodium salicylate treatment until day 30 from the start of the study. The results showed a significant increase in tissue inhibitor metalloproteinase 1 (TIMP-1), Hyaluronic acid (HA), malondialdehyde, kidney injury molecule -1(KIM-1), and nitric oxide in the nephrotoxic group injected with cisplatin compared to the control group. Additionally, there was an elevation in the mRNA expression of nephrotoxic group with uninephrectomy. However, nephrotoxic rats treated with nanoemulsified sodium salicylate exhibited only a modest increase in TIMP-1, HA, and KIM-1 levels, along with elevated expressions of podocin and nephrin compared to the healthy control group. These findings suggest that nanoemulsified sodium salicylate exerts a protective effect against cisplatin-induced nephrotoxicity in uninephrectomized.

Tissue inhibitor of metalloproteinase (TIMP)-1 predicts failure of recovery of ejection fraction in acute heart failure with reduced ejection fraction

BackgroundHeart failure (HF) with improved ejection fraction (HFimpEF) is a recently identified phenotype of HF, which had better cardiovascular outcomes compared with persistent HF with reduced ejection fraction (HFrEF). The...

Improving Circulation Half-Life of Therapeutic Candidate N-TIMP2 by Unfolded Peptide Extension.

Matrix metalloproteinases (MMPs) are significant drivers of many diseases, including cancer, and are established targets for drug development. Tissue inhibitors of metalloproteinases (TIMPs) are endogenous MMP inhibitors and are being pursued for the development of anti-MMP therapeutics. TIMPs possess many attractive properties for drug candidates, such as complete MMP inhibition, low toxicity, low immunogenicity, and high tissue permeability. However, a major challenge with TIMPs is their rapid clearance from the bloodstream due to their small size. This study explores a method for extending the plasma half-life of the N-terminal domain of TIMP2 (N-TIMP2) by appending it with a long, intrinsically unfolded tail containing Pro, Ala, and Thr (PATylation). We designed and produced two PATylated N-TIMP2 constructs with tail lengths of 100 and 200 amino acids (N-TIMP2-PAT100 and N-TIMP2-PAT200). Both constructs demonstrated higher apparent molecular weights and retained high inhibitory activity against MMP-9. N-TIMP2-PAT200 significantly increased plasma half-life in mice compared to the non-PATylated variant, enhancing its therapeutic potential. PATylation offers distinct advantages for half-life extension, such as fully genetic encoding, monodispersion, and biodegradability. It can be easily applied to N-TIMP2 variants engineered for high affinity and selectivity toward individual MMPs, creating promising candidates for drug development against MMP-related diseases.

Fibulin-3 Regulates Tissue Inhibitor of Metalloproteinases 3 to Inhibit Senescence in Intervertebral Disc Nucleus Pulposus Cells

To investigate the effect of fibulin-3 on the senescence of intervertebral disc nucleus pulposus cells (NPCs) through the regulation of tissue inhibitor of metalloproteinases 3 (TIMP-3) expression and to elucidate the molecular mechanisms involved. 1). The nucleus pulposus tissues and imaging data of 37 patients who had undergone intervertebral disc surgery were collected. The degree of degeneration of the intervertebral discs were classified according to the Pfirrmann grading system. The senescence degree of NPCs was determined using senescence-associated β-galactosidase (SA-β-gal) staining. Fibulin-3 expression levels were determined using Western blot and ELISA. The relationship between fibulin-3 and disc degeneration and NPCs senescence was investigated. 2). Human intervertebral disc NPCs were cultured in vitro. The proliferation and senescence of NPC across continuous passage were observed via CCK-8 assay and SA-β-gal staining, respectively. Fibulin-3 expression levels and the expression of inflammatory cytokines and matrix metalloproteinases were assessed. Exogenous fibulin-3 was added to verify its effect on the proliferation and senescence of NPCs. 3). The effect of fibulin-3 on the apoptosis and proliferation of NPCs was verified through gene overexpression, which was used in combination with an apoptosis inhibitor for bidirectional verification. 4). Bioinformatics analysis was performed to explore the relationship between fibulin-3 and the TIMP family. Experiments overexpressing fibulin-3 and silencing the TIMP-3 gene were performed to verify their role in NPCs senescence. 1). The intervertebral disc degeneration samples from 37 patients were classified according to the Pfirrmann grading system. The higher the degeneration grade, the lower fibulin-3 expression. Spearman correlation analysis showed that the disc grade was negatively correlated with the NPC senescence grade (r=－0.87, P<0.001) and fibulin-3 expression (r=－0.79, P<0.001). 2). As the passage number of NPCs increased, fibulin-3 expression gradually decreased, cell proliferation ability weakened, and the expression of inflammatory cytokines and matrix metalloproteinases increased. After exogenous fibulin-3 was added, cell morphology and growth status were maintained, cell senescence was significantly inhibited, and the expression of inflammatory cytokines and matrix metalloproteinases was markedly reduced. 3). Gene overexpression experiments showed that fibulin-3 reduced NPC apoptosis and promoted cell proliferation, thereby inhibiting NPC senescence. 4). Bioinformatics analysis revealed a significant association between fibulin-3 and TIMP-3 of the TIMP family. Further experiments confirmed that overexpressing fibulin-3 enhanced TIMP-3 expression, while silencing the TIMP-3 gene significantly weakened the inhibitory effect of fibulin-3 on NPCs senescence. This indicates that, through regulating TIMP-3, fibulin-3 inhibits the activity of matrix metalloproteinases, affects the synthesis and degradation of the extracellular matrix, and ultimately inhibits NPCs senescence. This study demonstrates that fibulin-3 plays a crucial role in inhibiting the senescence of intervertebral disc NPCs by regulating TIMP-3. The specific mechanisms involved are as follows, fibulin-3 upregulates TIMP-3 expression, inhibits matrix metalloproteinase activity, and reduces extracellular matrix degradation, thereby promoting extracellular matrix synthesis. Additionally, fibulin-3 inhibits NPCs senescence by reducing apoptosis and promoting cell proliferation. Therefore, fibulin-3 and TIMP-3 have potential therapeutic significance in maintaining intervertebral disc health and delaying degeneration.

Exploring Genetic Drug Targets in Acne Vulgaris: A Comprehensive Proteome-Wide Mendelian Randomization Study.

Acne vulgaris presents a substantial clinical challenge due to its complex pathophysiology and significant impact on quality of life. Identification of novel therapeutic targets for acne using genetic tools can guide the development of more effective treatments. Utilizing a dataset comprising 35 559 Icelandic individuals, we performed proteomic analyses to quantify 4709 circulating proteins. We integrated these data with acne-specific genome-wide association studies (GWAS) encompassing 34 422 acne patients and 364 991 controls. Mendelian randomization (MR) analyses employed the TwoSampleMR tool and Summary-data-based Mendelian Randomization (SMR) to estimate the causal effects of identified proteins on acne risk. Colocalization analyses assessed the likelihood of shared genetic etiology between protein levels and acne using the "coloc" R package. Our proteome-wide MR analysis initially identified 128 proteins potentially associated with acne risk. Following multiple testing corrections using the Benjamini-Hochberg method, fatty acid synthase (FASN) and tissue inhibitor of metalloproteinases 4 (TIMP4) remained significantly associated with acne risk. FASN exhibited a protective effect against acne (OR = 0.768, 95% CI: 0.676-0.872, p = 4.685E-05), while TIMP4 was associated with an increased risk (OR = 1.169, 95% CI: 1.103-1.241, p = 1.956E-07). Colocalization analysis supported a shared genetic basis for these protein-acne associations, with posterior probabilities indicating strong evidence of shared causal variants. Our findings highlight the utility of integrative genomic approaches in identifying potential therapeutic targets for acne. FASN and TIMP4, in particular, demonstrate strong potential as targets for therapeutic intervention, pending further validation through clinical research. These results offer a foundation for targeted acne treatment development, aligning with personalized medicine principles.

Differential effects of macrophage subtype-specific cytokines on fibroblast proliferation and endothelial cell function in co-culture system.

Macrophages are involved in several critical activities associated with tissue repair and regeneration. Current approaches in regenerative medicine are focusing on leveraging the innate immune response to accelerate tissue regeneration and improve long-term healing outcomes. Of particular interest in this regard are the currently known, four main M2 macrophage subtypes: M2interleukin (IL)-4,IL-13, M2IC, M2IL-10, M2non-selective adenosine receptor agonists (NECA) (M2IL-4,IL-13 → M2NECA). In this study, rat bone marrow-derived macrophages (M0) were polarized to each of the four subtypes M2IL-4,IL-13 → M2NECA and cultured for 72 h in vitro. Luminex assay results highlighted increased production of tissue inhibitor of metalloproteinases-1 (TIMP-1) for M2IL-4,IL-13, higher amounts of transforming growth factor-beta 1 (TGF-β1) for M2IL-10, and elevated vascular endothelial growth factor A (VEGF-A) from M2NECA. Co-culture experiments performed with M2IL-10 macrophages and L929 fibroblasts highlighted the increased production of soluble collagen within the media as well as higher amounts of collagen in the extracellular matrix. Human umbilical vein endothelial cells (HUVECs) were co-cultured with M2NECA macrophages, which demonstrated an increase in intercellular adhesion molecule (ICAM) and platelet endothelial cell adhesion molecule (PECAM), as well as increased formation of endothelial tubes. The findings of this study emphasize a critical demand for further characterization and analyses of distinct M2 subtypes and careful selection of specific macrophage populations for regeneration of specific tissue types. The current, broad classification of "M2" may be sufficient in many general tissue engineering applications, but, as conditions are constantly in flux within the microenvironment in vivo, a higher degree of specificity and control over the initial M2 subtype could result in more consistent long-term outcomes where macrophages are utilized as part of an overall regenerative strategy.

Comprehensive biomarker assessment for predicting severe acute kidney injury and need of kidney replacement therapy in liver transplantation patients

Background Renal dysfunction is a common complication following liver transplantation (LT). This study aimed to determine whether a comprehensive assessment of kidney function using nineteen serum and urinary biomarkers (BMs) within the first 48 h post-LT could enhance the prediction of severe acute kidney injury (AKI) and the need of kidney replacement therapy (KRT) during the first postoperative week. Methods Blood and urine (U) samples were collected during the pre- and postoperative periods. Nineteen BMs were evaluated to assess kidney health in the first 48 h after LT. Classification and regression tree (CART) cross-validation identified key predictors to determine the best BM combination for predicting outcomes. Results Among 100 LT patients, 36 developed severe AKI, and 34 required KRT within the first postoperative week. Preoperative assessment of U neutrophil gelatinase-associated lipocalin (NGAL) and liver-type fatty acid-binding protein (L-FABP) predicted the need for KRT with 75% accuracy. The combined assessment of U osmolality (OSM), U kidney injury molecule 1 (KIM-1), and tissue inhibitor of metalloproteinase (TIMP-1) within 48 h post-LT predicted severe AKI with 80% accuracy. U-OSM alone, measured within 48 h post-LT, had an accuracy of 83% for predicting KRT need, outperforming any BM combination. Conclusions Combined BM analysis can accurately predict severe AKI and KRT needs in the perioperative period of LT. U-OSM alone proved to be an effective tool for monitoring the risk of severe AKI, available in most centers. Further studies are needed to assess its impact on AKI progression postoperatively. Registered at Clinical Trials (clinicaltrials.gov) in March 24th, 2014 by title ‘Acute Kidney Injury Biomarkers: Diagnosis and Application in Pre-operative Period of Liver Transplantation (AKIB)’ and identifier NCT02095431.

Doxycycline versus Curcumin for Inhibition of Matrix Metalloproteinase Expression and Activity Following Chemically Induced Inflammation in Corneal Cells.

Sulfur mustard (SM) is a potent blistering agent. This alkylating chemical agent has extremely toxic effects on the eye. MMP-2 and MMP-9 are the two most important matrix metalloproteinase enzymes involved in the pathology of chemical eye injuries. Curcumin is regarded as a natural anti-inflammatory agent. This study aims to compare the anti-inflammatory effects of curcumin versus doxycycline on chemically induced corneal injuries. The HCE-2 cell line was used as a model for corneal cells. The effective concentrations of 2-chloroethyl ethyl sulfide (CEES) - as an analog of SM - doxycycline, and curcumin were determined using the MTT assay. The gene expression of MMP-2, MMP-9, and tissue inhibitors of metalloproteinase (TIMP-1) was evaluated by the real-time PCR method. Also, the activity of MMP-2 and MMP-9 enzymes was determined by zymography. The expression of the MMP-2 and MMP-9 genes increased 5- and 3.3-fold after exposure to CEES, respectively. Following the treatment with curcumin and doxycycline, MMP-2 expression decreased significantly. Also, after treatment with curcumin and doxycycline, the MMP-9 expression decreased 2.5- and 1.6-fold, respectively. The reduction in activity was 32% for MMP-2 and 56% for MMP-9 after treatment with curcumin. The corresponding values were 12% and 40% following doxycycline treatment. There was no significant difference between the effects of curcumin and doxycycline on reducing MMP-2 expression, but the difference was statistically significant in the case of MMP-9. Doxycycline and curcumin can inhibit MMP expression and activity in chemically exposed corneal cells. Curcumin has a greater ability than doxycycline to inhibit MMP-2 and MMP-9 enzymes; however, the difference is statistically significant only in the case of MMP-9. After further validation, these substances can be introduced as anti- inflammatory agents to treat corneal chemical burns.

Efficacy of Picosecond Laser in Clinical, Histopathology, and Immunohistochemistry Examination in Indonesian Wrinkled Skin.

Introduction: The elderly population wants to improve skin function and maintain a youthful appearance without wrinkles. Laser is one of the highly developed and widely used therapeutic modalities of skin aging by stimulating dermal collagen formation. Picosecond laser therapy is performed on skin with wrinkles to determine the mechanism of wrinkle improvement with a picosecond laser through an increasing in tissue inhibitor of metalloproteinases 1 (TIMP-1), and a decrease in matrix metalloproteinase-1 (MMP-1), which was confirmed by collagen density. Methods: The study was started from March 2021 to July 2021 at URJ Dr. Soetomo Hospital Surabaya and Surabaya Skin Centre. Twenty patients meeting the inclusion criteria of being 36-55 years of age and having a wrinkle in the forearm were enrolled by random sampling. Each sample data was recorded, the average TIMP-1, MMP-1, collagen density, and clinical wrinkles were calculated, and the laser was performed by using a picosecond 755 nm laser. Results: All patients showed a moderate pre-test wrinkle scale. The figure for TIMP-1 and MMP-1 after therapy was higher. The MMP-1/TIMP-1 ratio in wrinkles treated by using a picosecond laser was lower but insignificant than before. The picture of collagen density indicates that collagen density after laser therapy was higher than before. Conclusion: The result of clinical examination, histopathology, and immunohistochemistry in this research explained that a picosecond 755 nm laser with DLA is an effective therapy for wrinkles in Indonesian patients without any adverse effects.