Tissue Inhibitors Of Matrix Metalloproteinases Research Articles

Associations of serum and tissue TIMP1 with host response and survival in colorectal cancer

Tissue inhibitor of matrix metalloproteinase 1 (TIMP1) is a multifaceted, cytokine-like bioactive molecule whose levels are elevated in a wide range of inflammatory diseases and are associated with prognosis. Additionally, TIMP1 may play a role in driving systemic inflammation. TIMP1 immunohistochemistry and TIMP1 serum concentrations were analyzed in a cohort of 776 colorectal cancer patients. TIMP1 histoscore by cell type (tumor cell, other) was quantified using digital image analysis. Serum TIMP1 levels were evaluated for correlations with tumor TIMP1 expression, and their associations with tumor characteristics, inflammation, and prognosis were investigated. High serum TIMP1 concentrations associated with shorter overall survival (multivariable HR 1.85, 95% CI 1.30–2.65). Serum TIMP1 levels positively correlated with markers of systemic inflammation and tumor necrosis percentage but not with TIMP1 expression in tumor tissue. High TIMP1 intensity in tumor stroma associated with longer cancer-specific and overall survival in univariable analysis but not in multivariable models. T cell densities in tumor tissue positively correlated with tumor stromal TIMP1 expression and negatively with tumor epithelial TIMP1 expression. Serum TIMP1 levels show promise as a prognostic marker for colorectal cancer and correlate with systemic inflammatory markers, but do not correlate with TIMP1 expression in tumor tissue.

Evaluation the tissue inhibitor of matrix metalloproteinases first type concentration in various platelet-related products

Purpose: to evaluate the content of tissue inhibitor of matrix metalloproteinase type 1 (TIMP-1) in platelet preparations obtained in different methods.Material and methods. Platelet-rich plasma (PRP), platelet-poor plasma, and a suspension of platelets washed from plasma (WP) were isolated from the blood of 10 volunteer donors, and a morphofunctional analysis of platelets was performed. After that, a platelet lysate was prepared from PRP, WP, and platelet-poor plasma. The concentration of TIMP-1 in the lysates was determined using enzyme immunoassay.Results. The level of TIMP-1 in the lysates of PRP and WP did not differ significantly; in platelet-poor plasma, the level of TIMP-1 was significantly lower than in the lysates of PRP and WP (p = 0.003). In all types of preparations, the concentration of TIMP-1 was 2-4 times higher than similar values in the blood serum of healthy people, as reported in other studies. A weak correlation was found between the TIMP-1 concentration and the platelet number in PRP, and there was no correlation between the content of platelets with granules and the TIMP-1 level in the lysates. The presence of leukocytes in the initial PRP also did not affect the TIMP level in the final lysates. A strong correlation was found between the TIMP-1 concentration in the WP lysate and in platelet-poor plasma, as well as a direct correlation between the total platelet content and the TIMP-1 level in the PRP lysate.Conclusions. Preparations with high concentrations of TIMP-1 can be obtained both from concentrated platelet suspensions and from platelet-poor plasma. To optimize the method, it is necessary to study the effect of platelet preparations with TIMP on various biological models.

Greater TIMP-1 protein levels and neointimal formation represent sex-dependent cellular events limiting aortic vessel expansion in female rats.

Fragmentation/loss of the structural protein elastin represents the precipitating event translating to aortic expansion and subsequent aneurysm formation. The present study tested the hypothesis that greater protein expression of tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) and neointimal growth secondary to a reduction of medial elastin content represent sex-dependent events limiting aortic vessel expansion in females. TIMP-1 protein levels were higher in the ascending aorta of female versus male patients diagnosed with a bicuspid aortic valve (BAV). The latter paradigm was recapitulated in the aorta of adult male and female rats complemented by greater TIMP-2 expression in females. CaCl2 (0.5 M) treatment of the infrarenal aorta of adult male and female rats increased the in situ vessel diameter and expansion was significantly smaller in females despite a comparable reduction of medial elastin content. The preferential appearance of a neointimal region of the CaCl2-treated infrarenal aorta of female rats may explain in part the smaller in situ expansion and neointimal growth correlated positively with the % change of the in situ diameter. Neointimal formation was secondary to a significant increase in the density of medial/neointimal vascular smooth muscle cells (VSMCs) that re-entered the G2-M phase whereas VSMC cell cycle re-entry was attenuated in the CaCl2-treated infrarenal aorta of male rats. Thus, greater TIMP-1 expression in the aorta of female BAV patients may prevent excessive elastin fragmentation and preferential neointimal growth following CaCl2-treatment of the infrarenal aorta of female rats represents a sex-dependent biological event limiting vessel expansion secondary to a significant loss of the structural protein.

Fucoxanthin Enhances the Antifibrotic Potential of Placenta-derived Mesenchymal Stem Cells in a CCl4-induced Mouse Model of Liver.

The effectiveness of fucoxanthin (Fx) in liver diseases has been reported due to its anti-inflammatory and antifibrotic effects. Mesenchymal stem cells (MSCs)-based therapy has also been proposed as a promising strategy for liver fibrosis treatment. Recent studies have shown that the co-administration of MSCs and drugs demonstrates a pronounced effect on liver fibrosis. This study aimed to determine the therapeutic potential of placenta-derived MSCs (PD-MSCs) in combination with Fx to treat liver fibrosis and evaluate their impact on the main links of liver fibrosis pathogenesis. After PD-MSCs isolation and identification, outbred ICR/CD1 mice were divided into five groups: Control group, CCl4 group (CCl4), Fx group (CCl4+Fx), PD-MSCs group (CCl4+MSCs) and cotreatment group (CCl4+MSCs+Fx). Biochemical histopathological investigations were performed. Semiquantitative analysis of the alpha-smooth muscle actin (α-SMA+), matrix metalloproteinases (MMP-9+, MMP-13+), tissue inhibitor of matrix metalloproteinases-1 (TIMP-1+) areas, and the number of positive cells in them were studied by immunohistochemical staining. Transforming growth factor-beta (TGF-β), hepatic growth factor (HGF), procollagen-1 (COL1α1) in liver homogenate and proinflammatory cytokines in blood serum were determined using an enzyme immunoassay. Compared to the single treatment with PD-MSCs or Fx, their combined administration significantly reduced liver enzyme activity, the severity of liver fibrosis, the proinflammatory cytokine levels, TGF-β level, α-SMA+, TIMP-1+ areas and the number of positive cells in them, and increased HGF level, MMP-13+, and MMP-9+ areas. Fx enhanced the therapeutic potential of PD-MSCs in CCl4-induced liver fibrosis, but more investigations are necessary to understand the mutual impact of PD-MSCs and Fx.

The Preventive Effect of Ulinastatin on Blood-Brain Barrier Dysfunction in Rats with Postoperative Cognitive Dysfunction After General Anaesthesia with Isoflurane.

This study evaluated the effect of ulinastatin on blood-brain barrier (BBB) dysfunction in rats with postoperative cognitive dysfunction (POCD) following general anaesthesia with isoflurane. Specifically, we examined BBB permeability and the expression of tissue inhibitor of matrix metalloproteinase-1 (TIMP-1). Rats in the ulinastatin group received the drug intraperitoneally (50,000 U/mL), while controls received normal saline (1 mL) administered before general anaesthesia. Isoflurane (1.5% volume) anaesthesia was induced for 2 h. Cognitive function was assessed using the Y-maze test. Two days after anaesthesia, BBB permeability was measured using Evans blue, and TIMP-1 expression was evaluated. Both groups experienced cognitive decline following anaesthesia. However, the ulinastatin group showed a more limited decrease (control group, 64.2 ± 19.3 → 30.2 ± 16.2, p = 0.008; ulinastatin group, 70.0 ± 15.7 → 66.5 ± 12.0, p = 0.67). The ulinastatin group showed a significantly lower permeability of the BBB (0.034 ± 0.003 µg/g in control group vs. 0.005 ± 0.002 µg/g in ulinastatin group, p = 0.0001), and also showed a significantly higher value of TIMP-1 expression (5.81 ± 1.94% in control group vs. 13.97 ± 2.59% in ulinastatin group, p = 0.0001). Administration of ulinastatin before general anaesthesia mitigated cognitive decline in rats with POCD, likely through the prevention of BBB dysfunction, as evidenced by the lower BBB permeability and increased TIMP-1 expression.

Unveiling the depths of pelvic organ prolapse: From risk factors to therapeutic methods (Review).

Pelvic organ prolapse (POP) is a condition where one or more pelvic organs (such as the uterus, bladder and rectum) descend from their normal anatomical positions into the vagina, primarily due to the weakening of the pelvic floor support structures. While not life-threatening, POP can substantially diminish the patient's quality of life and lead to serious social and psychological complications. Researchers have explored novel directions regarding the etiology, mechanism and treatment of POP. However, existing literature on the subject often lacks comprehensive and systematic overviews. To address this gap and enhance researchers' understanding of POP, the present study reviewed the risk factors and molecular mechanisms of POP [including matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs, transforming growth factor β, advanced glycation end products (AGEs)/receptor for AGE, phosphoinositide 3-kinase/protein kinase B, fibulin, lysyl oxidase-like 1, homeobox A11, collagen α-1 (XVIII) chain, Wnt signaling pathways and estrogen receptor α], as well as therapeutic approaches, such as lifestyle interventions, physical methods, pharmacotherapy, stem cell transplantation and surgical techniques. The present review aims to provide new insights for future research and contribute to the advancement of diagnosis and treatment strategies for POP.

Muscle Tissue Response to Vipera berus berus and Vipera berus nikolskii Venoms: A Study on Matrix Metalloproteinases and Cytokine Modulation

Vipera berus, also known as the common adder, is a snake species widely distributed in Europe and parts of Asia. Its venom is a mix of enzymatic and non-enzymatic components causing both local and systemic effects including edema, hemorrhage, myonecrosis, gastrointestinal disorders, and rarely – rhabdomyolysis, coagulopathy and hypovolemic shock. Matrix metalloproteinases (MMPs) and cytokines, being inflammatory mediators, participate both in the development of these reactions and in tissue regeneration after acute damage. The current study was aimed at analyzing the concentrations of important mediators of tissue damage, inflammation and regeneration, namely, MMP-1, -2, -3, -8, -10, TIMP-1 (a tissue inhibitor of MMPs), and pro- and anti-inflammatory cytokines, in rat muscle after intraperitoneal injection of venoms from two subspecies of adders – V. berus berus and V. berus nikolskii — to clarify their role in the local response of skeletal muscle to the studied venoms. Changes in muscle tissue were revealed, namely, significant increase in almost all MMPs including MMP-2, as well as TIMP-1, IFN-γ and anti-inflammatory IL-4 and IL-10 content. Moreover, a tendency for some pro-inflammatory cytokines levels to decline 24 h after venom administration was observed, which may be a sign of inflammation suppression and the beginning of reparative processes associated with extracellular matrix remodeling and formation of an optimal environment for muscle regeneration. However, to establish the exact mechanisms of the changes we found, further studies are needed. The ability of V. berus nikolskii venom to induce more pronounced changes was also shown. Our results may be useful for the development of differentiated and more effective treatment strategies for the consequences of bites by these snakes, which include myonecrosis

Особливості показників гуморального імунітету в новонароджених дітей із маркерами недиференційованої дисплазії сполучної тканини

Connective tissue dysplasia can affect the immune response in children. The study of humoral immunity indicators in newborns with locomotor and biochemical markers of undifferentiated connective tissue dysplasia (UCTD) is very relevant. Aim - study the correlations between anthropometric, laboratory markers of UCTD and indicators of the B-cell link of immunity in newborns. Materials and methods. We examined 122 newborns: 79 full-term and 43 premature infants. In them, the perinatal history data were analyzed, anthropometric and biochemical (matrix metalloproteinase-1 (MMP-1), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1)) markers of UCTD were studied, and the ratio of MMP-1/TIMP-1 was calculated. The absolute number of B-lymphocytes and the levels of immunoglobulins of classes A (IgA), M (IgM), G (IgG) in the blood were also investigated. The main group (n=82) consisted of newborns with elevated values of ≥2 anthropometric markers of UCTD, the control group (n=40) included children without deviation from the norm of any anthropometric index. Results. Children with signs of UCTD are more likely to be born from ≥2 births, with a body weight of 2499-2000 g and ≤1999 g, and have lower levels of serum IgA, IgM than children of the control group. Such perinatal factors as operative delivery and parity of more than two births can be considered prognostically unfavorable for a lower absolute number of B-lymphocytes, lower levels of IgA and IgM in the blood of newborns. Serum content of MMP-1≥4.51 ng/ml, values of MMP-1/TIMP-1≥0.31 are associated with a higher absolute number of B-lymphocytes and lower level of IgA, also the concentration of TIMP-1≥16.08 ng/ml is associated with higher levels of IgA, IgM, IgG. The content of serum immunoglobulins correlates with the gestational age. Conclusions. Therefore, newborns with anthropometric markers of UCTD and an imbalance of biochemical markers (MMP-1, TIMP-1, index MMP-1/TIMP-1) have lower levels of serum IgA and IgM. The concentration of MMP-1≥4.51 ng/ml, MMP-1/TIMP-1≥0.31, TIMP-1≤16.07 ng/ml are prognostically unfavorable factors regarding changes in the B-cell immunity in newborns. The lowest concentrations of immunoglobulins of all classes were recorded in children with a gestational age of 31-28 weeks. The study was carried out according to the principles of the Declaration of Helsinki. The study protocol was accepted by the Local Ethical Committee of these institutions. The informed consent of the children's parents was obtained for the research. The authors declare no conflict of interest.

Effect of Silymarin on Expression of micro-RNA-21 and Matrix Metalloproteinase (MMP) 2 and 9 and Tissue Inhibitors of Matrix Metalloproteinase (TIMP) 1 and 2 in Hepatocellular Carcinoma Cell Line (HepG2).

Silymarin is a flavonolignan that has various medicinal properties such as liver protection, antioxidant, anti-inflammatory, anti-cancer and heart protection activities. The aim of this study was to investigate the effect of silymarin on the expression level of mir-21, matrix metalloproteinase(MMP), and their tissue inhibitors (TIMPs) in liver cancer HepG2 cell line. An in-vitro experimental study was conducted on the human HepG2 cells prepared from Pasteur Institute, Tehran, Iran. Four concentrations of 0 (control), 50, 100, and 150 µM of silymarin were considered as the study groups according to the MTT assay. Gene expression study was performed using real-time PCR. The studied genes were mir-21, MMP-2, MMP-9, TIMP-1 and TIMP-2. In addition, some apoptosis-related genes including BAX, BCL2 and Caspase3 (CAS3) were investigated. GAPDH was used as an internal control. Relative expression was calculated by REST program using t-test on the logarithm of expression considering a significance level of 0.05. The significant up-regulations consisted of TIMP genes for doses 100 µM and 150 µM, and the apoptosis activating genes CAS3 and BAX (P < 0.05). The significant down-regulations consisted of MMP-9 in all concentrations, MMP-2 in concentration 100 µM, and the apoptosis inhibitory gene BCL2 in concentrations 50 µM and 100 µM (P < 0.05). In addition, mir-21 as an oncogenic micro-RNA showed significant down-regulation for all doses (P < 0.05). All the comparisons were with the control group. The present study showed that silymarin could affect the HepG2 cell line at the gene expression level via increasing apoptosis and changing the expression of MMP-2, MMP-9, TIMP-1, TIMP-2 and mir-21. These findings were in line with each other and in favor of suppression of tumoral activity in this cell line.

Genetic removal of Nlrp3 protects against sporadic and R345W Efemp1-induced basal laminar deposit formation.

Chronic, unresolved inflammation has long been speculated to serve as an initiating and propagating factor in numerous neurodegenerative diseases, including a leading cause of irreversible blindness in the elderly, age-related macular degeneration (AMD). Intracellular multiprotein complexes called inflammasomes in combination with activated caspases facilitate production of pro-inflammatory cytokines such as interleukin 1 beta. Specifically, the nucleotide-binding oligomerization (NOD)-like receptor protein 3 (NLRP3) has received heightened attention due to the wide range of stimuli to which it can respond and its potential involvement in AMD. In this study, we directly tested the role of Nlrp3 and its downstream effector, caspase 1 (Casp1) in mediating early AMD-like pathology (i.e., basal laminar deposits [BLamDs]) in wild-type (WT) mice and the Malattia Leventinese/Doyne honeycomb retinal dystrophy (ML/DHRD) mouse model (p.R345W mutation in Efemp1). Compared to aged-matched controls, R345W+/+ knockin mice demonstrated increased Muller cell gliosis, subretinal Iba-1+ microglial cells, higher Nlrp3 immunoreactivity in the retina, as well as significant transcriptional upregulation of complement component 3, Nlrp3, pro-Il1b, pro-caspase-1, and tissue inhibitor of matrix metalloproteinase 3 in the retinal pigmented epithelium (RPE)/choroid. These findings were accompanied by an age-related increase in BLamD formation in the R345W+/+ mice. Genetic elimination of either Nlrp3 or Casp1 significantly reduced both the size and coverage of BLamDs in the R345W+/+ background, highlighting an important and underappreciated pathway that could affect ML/DHRD onset and progression. Moreover, Nlrp3 knockout reduced spontaneous, idiopathic BLamDs in WT mice, suggesting translatability of our findings not only to rare inherited retinal dystrophies, but also potentially to AMD itself.

Clinical and morphological features of the myometrium in patients with placental adherent and invasive pathology

BACKGROUND: The fast increase in the frequency of placental adherent and invasive pathology worldwide accounts for the growing interest in studying the pathogenesis of placenta accreta. According to the literature, the clinical and morphological features of the myometrium from the placental attachment area in placental adherent and invasive pathology are described in single articles. Study of morphological features using proteolytic markers in the myometrium from the placenta accreta area could help in understanding the pathogenesis of placental adherent and invasive pathology. For the first time in this study, the clinical and morphological features of the myometrium from the placental attachment area in patients with uterine scar and placental adherent and invasive pathology are compared with the myometrium of women with uterine scar without placenta accreta and intact myometrium. AIM: The aim of this study was to evaluate the expression of matrix metalloproteinase 2 and tissue inhibitors of matrix metalloproteinases 1 and 2 in myometrial biopsies in placental adherent and invasive pathology. MATERIALS AND METHODS: This study included 15 myometrial biopsies from the placental site, which were divided into three groups according to the clinical diagnosis of the patients: group 1 (main group), with a uterine scar after cesarean section and placental adherent and invasive pathology (n = 5); group 2 (comparison group), with a uterine scar after cesarean section (n = 5); group 3 (control group), with a normal pregnancy without a uterine scar (n = 5). Histological examination was carried out using the standard procedure. Immunohistochemical study was performed using antibodies to matrix metalloproteinase 2 and tissue inhibitors of matrix metalloproteinases 1 and 2 (Abcam, USA). Morphometry was carried out using the VideoTesT-Morphology 5.2 program (Videotest Ltd., Russia). The statistical analysis was performed using the IBM SPSS Statistics 26.0 software. RESULTS: In the biopsies of the main group, we verified terminal chorionic villi with hypervascularization and uneven plethora of the vascular bed among hypertrophied muscle fibers, a basal plate with lumen ectasia of unevenly full-blooded vessels, and the absence of a decidual membrane, in contrast to groups 2 and 3. The matrix metalloproteinase 2 expression area in the main group was higher than in the comparison and control groups (p = 0.008; p1–2 = 0.049*, p1–3 = 0.011), the matrix metalloproteinase 2 optical density not differing between the groups (p = 0.122). The tissue inhibitors of matrix metalloproteinases 1 expression area was higher in the comparison group compared to the main group (p = 0.035; p2–3 = 0.032), and the tissue inhibitors of matrix metalloproteinases 1 and 2 optical density was higher in the comparison group compared to control (p = 0.008; p2–3 = 0.005). CONCLUSIONS: In myometrial biopsies of patients with a uterine scar and placental adherent and invasive pathology, we verified pathology of the basal plate of the placenta, increased matrix metalloproteinase 2 expression and decreased tissue inhibitors of matrix metalloproteinases 1 expression, which may indicate the peculiarities of the inflammatory response in the myometrium in placental adherent and invasive pathology.

Elevated MMP-9, Survivin, TGB1 and Downregulated Tissue Inhibitor of TIMP-1, Caspase-3 Activities are Independent of the Low Levels miR-183 in Endometriosis.

This study aimed to measure the correlation between miR-183 and gene expression that regulates apoptosis and adhesion mechanism that may be linked to the pathogenesis of endometriosis. Forty-four subjects, including 22 control subjects, participated in this study. We collected ectopic endometriosis and endometrial samples. For the control, the sample was taken from endometrial tissue through pipelle biopsy. RNA was extracted from all tissues using RNA mini kit, and the expression was assessed using quantitative-real time PCR. Relative mRNA and miRNA expression were presented using the formula of the Livak method. The data were statistically analyzed using GraphPad Prism 8. The expression of Caspase-3, Survivin, Integrin β1 (ITGB1), matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) (adhesion- and apoptosis-related gene) were calculated using the relative expression method. We found significant differences in Caspase-3, Survivin, ITGB1, MMP-9, and TIMP-1 expression between ectopic endometriosis tissues of women with endometriosis compared to healthy endometrium. MMP-9, Survivin, and ITGB1 was significantly increased in the endometriosis group, while Caspase-3, TIMP-1, and miR-183 were significantly reduced in the endometriosis group. No correlation was found between the expression level of miR-183 and Caspase3, Survivin, ITGB1, and Cadherin in both tissue types. Despite the difference in expression levels of miR-183 and associated adhesion- and apoptosis-related genes, there was no significant association between miR-183 with specific adhesion and apoptosis genes in endometriosis tissue.

Protein Kinase C and Matrix Metalloproteinases Expression Using Phorbol Myristate Acetate in Degenerative Intervertebral Disc Cells.

Degeneration of nucleus pulposus (NP) cells involves multiple factors. The relationship between the canonical Wnt/β-catenin signaling pathway and matrix metalloproteinases (MMPs) is important in cellular senescence. Protein kinase C (PKC), an intermediate of the non-canonical Wnt pathway stimulated by phorbol myristate acetate (PMA), possibly prevents NP cell senescence, although not yet demonstrated in human-based studies. This study aimed to investigate the effect of PMA stimulation on the non-canonical and canonical Wnt pathways and MMP expression in human NP cells to ascertain its inhibitory effects on the senescence of NP cells. Human disc tissues of Pfirrmann grades 1 and 2 were collected from patients during spinal surgery and subsequently cultured. Protein and ribonucleic acid (RNA) were isolated from NP cells treated with PMA (400 nM) for 24 hours. Expression of MMP1, MMP13, tissue inhibitor of matrix metalloproteinase 1 (TIMP1), a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5), transient receptor potential vanilloid 4 (TRPV4), interleukin-6 (IL-6), and β-catenin were detected using western blot analysis. Messenger RNA (mRNA) expression of type II collagen and glycosaminoglycan (GAG) were analyzed using reverse transcription polymerase chain reaction. IL-6 and prostaglandin E2 (PGE2) levels were measured using enzyme-linked immunosorbent assay. Expression of PKC-δ (intermediate of the non-canonical Wnt pathway) and β-catenin (intermediate of the canonical Wnt pathway) was increased by PMA treatment. The mRNA levels of type II collagen and GAG increased; however, their protein levels were not altered. PMA treatment increased the expression of MMP1, TIMP1, ADAMTS5, IL-6, PGE2, and TRPV4; however, the expression of MMP13 and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) was unaltered. PMA activated PKC-δ, affecting the non-canonical Wnt pathway; however, its effect on β-catenin in the canonical Wnt pathway was limited. β-catenin activation through the TRPV4 channel led to increased expression of MMP1 and ADAMTS5 and that of IL-6 and PGE2 owing to NF-κB expression. Consequently, the degeneration of NP cells was not prevented.

Coenzyme Q10 and Vitamin E Regulate the Bioactivity of Human Corneal Fibroblast Cells.

Purpose: Corneal fibroblasts are involved in the wound healing of the cornea with proliferation, migration, and differentiation processes. Coenzyme Q10 (CoQ10) and vitamin E can enhance corneal wound healing when applied after a corneal lesion as an eye drop. Thus, this study was performed to determine the potential efficiency of a CoQ10 ophthalmical solution containing a CoQ10 and vitamin E D-α-tocopherol polyethylene glycol 1000 succinate (TPGS)-derived formulation in human corneal fibroblasts (HCFs) in vitro. Methods: Primary HCFs were obtained from cadaveric corneal tissue, and cell viability was determined using MTT assay at 24 and 72 h. Cell migration was evaluated using an in vitro wound healing assay, and mRNA expressions of collagen type I (COL-I), collagen type III (COL-III), lumican, hyaluronan, matrix metalloproteinase (MMP)-1, MMP-2, MMP-9, tissue inhibitors of MMP (TIMP)-1, TIMP-2, interleukin (IL)-1β, IL-6, IL-8, and IL-10 were assessed using reverse transcription polymerase chain reaction at 24 and 72 h. Results: At various concentrations of CoQ10 ophthalmical solution (CoQ10-os), cell viability and wound healing rates of HCFs increased compared with the control group. The expressions of COL-I, COL-III, lumican, and hyaluronan were increased by CoQ10-os, whereas those of MMP-1, MMP-2, MMP-9, TIMP-1, TIMP-2, and TIMP-3 were not affected by CoQ10-os at 24 and 72 h. In treating HCFs with a CoQ10-os medium, IL-1β, IL-6, and IL-8 decreased, whereas IL-10 was significantly increased in a time- and dose-dependent manner. Conclusions: The findings indicate that CoQ10 and vitamin E-TPGS are potent regulators of the bioactivity of HCFs, thus supporting their potential application as ophthalmical solutions in therapies aimed at the fast regeneration of damaged cornea tissues.

Regulation of Cell Viability, p53 Promoter Activity, and Expression of Interleukin-8, Matrixmetalloproteinase-1 and Tissue Inhibitor of Matrixmetalloproteinase-1 in Non-Irradiated or UV-Irradiated Fibroblasts and Melanoma Cells

Regulation of Cell Viability, p53 Promoter Activity, and Expression of Interleukin-8, Matrixmetalloproteinase-1 and Tissue Inhibitor of Matrixmetalloproteinase-1 in Non-Irradiated or UV-Irradiated Fibroblasts and Melanoma Cells

The Effect of Tissue Inhibitor of Metalloproteinases on Scar Formation after Spinal Cord Injury.

Spinal cord injury (SCI) often results in permanent loss of motor and sensory function. After SCI, the blood-spinal cord barrier (BSCB) is disrupted, causing the infiltration of neutrophils and macrophages, which secrete several kinds of cytokines, as well as matrix metalloproteinases (MMPs). MMPs are proteases capable of degrading various extracellular matrix (ECM) proteins, as well as many non-matrix substrates. The tissue inhibitor of MMPs (TIMP)-1 is significantly upregulated post-SCI and operates via MMP-dependent and MMP-independent pathways. Through the MMP-dependent pathway, TIMP-1 directly reduces inflammation and destruction of the ECM by binding and blocking the catalytic domains of MMPs. Thus, TIMP-1 helps preserve the BSCB and reduces immune cell infiltration. The MMP-independent pathway involves TIMP-1's cytokine-like functions, in which it binds specific TIMP surface receptors. Through receptor binding, TIMP-1 can stimulate the proliferation of several types of cells, including keratinocytes, aortic smooth muscle cells, skin epithelial cells, corneal epithelial cells, and astrocytes. TIMP-1 induces astrocyte proliferation, modulates microglia activation, and increases myelination and neurite extension in the central nervous system (CNS). In addition, TIMP-1 also regulates apoptosis and promotes cell survival through direct signaling. This review provides a comprehensive assessment of TIMP-1, specifically regarding its contribution to inflammation, ECM remodeling, and scar formation after SCI.

Liposome-encapsulated dextrazide modifies spleen extracellular matrix composition in mice with chronic BCG-induced inflammation

Fibrosis of parenchymal organs is a common complication of tuberculosis. In a model of BCG-induced inflammation in mice, changes in the metabolism of the extracellular matrix (ECM) of the spleen were demonstrated with the introduction of a liposome-encapsulated dextrazide (LEDZ) containing isoniazid and oxidized dextran.The mice were divided into 4 groups: 1 – intact animals; 2 – infected mice after a single intravenous injection of BCG vaccine. 6 mo after infection, a solution of LEDZ was administered intraperitoneally to mice of group 3 for 3 mo, and inhaled to mice of group 4. Group 2 mice showed the signs of pronounced spleen fibrosis (increased content of hyaluronan, hydroxyproline fractions) with activation of hyaluronidases, matrix metalloproteinases (MMP), α2-macroglobulin and an increased content of tissue inhibitors of MMP (TIMP-1 and TIMP-2) with respect to group 1 data. In group 3, changesin the structure of proteoglycans were noted (an increase in the content of uronic acids and galactose), a decrease in the content of hyaluronan and free hydroxyproline, an increase in the activity of hyaluronidases. The MMP activity and the TIMP content corresponded to the data of group 2. In group 4, the content of uronic acids and galactose in proteoglycans also increased, but peptide-bound hydroxyproline decreased and the hyaluronan content more noticeably decreased. The activity of all enzymes regulating the ECM metabolism reduced with respect to the data of group 2.Thus, intraperitoneal administration of LEDZ to infected mice led to activating hyaluronidases, changing the structure of proteoglycans, and decreasing the free hydroxyproline content. Inhalation administration of LEDZ, along with changes in the structure of proteoglycans, reduced the activity of MMP, hyaluronidases, α2-macroglobulin, the content of TIMP-1 and TIMP-2, peptide-bound hydroxyproline. The antifibrotic effect of LEDZ with inhalation administration was manifested in a decrease in peptide-bound hydroxyproline and in a more significant decrease in hyaluronan compared with intraperitoneal administration.Thus, intraperitoneal administration of LEDZ to infected mice led to activating hyaluronidases, changing the structure of proteoglycans, and decreasing the free hydroxyproline content. Inhalation administration of LEDZ, along with changes in the structure of proteoglycans, reduced the activity of MMP, hyaluronidases, α2-macroglobulin, the content of TIMP-1 and TIMP-2, peptide-bound hydroxyproline. The antifibrotic effect of LEDZ with inhalation administration was manifested in a decrease in peptide-bound hydroxyproline and in a more significant decrease in hyaluronan compared with intraperitoneal administration.

AMMP-8 POCT vs. Other Potential Biomarkers in Chair-Side Diagnostics and Treatment Monitoring of Severe Periodontitis.

This study aimed to compare several potential mouthrinse biomarkers for periodontitis including active matrix-metalloproteinase-8 (aMMP-8), total MMP-8, and other inflammatory biomarkers in diagnosing and monitoring the effects of nonsurgical periodontal therapy. Thirteen patients with stage III/IV periodontitis were recruited, along with thirteen periodontally and systemically healthy controls. These 13 patients were representative of the number of outpatients visiting any dentist in a single day. Full-mouth clinical periodontal parameters and biomarkers (the aMMP-8 point-of-care-test [POCT], total MMP-8, tissue inhibitor of MMPs (TIMP)-1, the aMMP-8 RFU activity assay, Myeloperoxidase, PMN elastase, calprotectin, and interleukin-6) were recorded at baseline and after nonsurgical therapy at 6 weeks. The aMMP-8 POCT was the most efficient and precise discriminator, with a cut-off of 20 ng/mL found to be optimal. Myeloperoxidase, MMP-8's oxidative activator, was also efficient. Following closely in precision was the aMMP-8 RFU activity assay and PMN elastase. In contrast, the total MMP-8 assay and the other biomarkers were less efficient and precise in distinguishing patients with periodontitis from healthy controls. aMMP-8, MPO, and PMN elastase may form a proteolytic and pro-oxidative tissue destruction cascade in periodontitis, potentially representing a therapeutic target. The aMMP-8 chair-side test with a cut-off of 20 ng/mL was the most efficient and precise discriminator between periodontal health and disease. The aMMP-8 POC test can be effectively used by dental professionals in their dental practices in online and real-time diagnoses as well as in monitoring periodontal disease and educating and encouraging good oral practices among patients.

UV-spectrophotometric and spectroscopic observed Vachellia nilotica and Nigella sativa formulations regularized the histopathological and biochemical parameters during wound contraction.

Diabetes mellitus causes impaired diabetic wounds which is linked to a number of pathological alterations that impede the healing of wounds. In the current research, Swiss albino mice were given alloxan monohydrate to induce diabetes and excision wounds of approximately 6 mm using biopsy punch. The diabetic wounds were treated with various biomaterials including Vachellia nilotica extract (VN), Nigella sativa extract (NS), V. nilotica nanoparticles (VNNPs) and N. sativa nanoparticles (NSNPs). Their effects were determined by evaluating the percent wound contraction, healing time, and histopathological analysis. The serum level of various biochemical parameters that is, pro-inflammatory cytokines, Matrix metalloproteinases (MMPs) and tissue inhibitor matrix metalloproteinases (TIMPs) were also determined. VNNPs group provided the best outcomes, with wound contraction 100% on 12th day. According to histopathological examination, VNNPs group reduced inflammation and encouraged the formation of blood vessels, fibroblasts, and keratinocytes. VNNPs group significantly alleviated the serum level of pro-inflammatory cytokines that are, TNF-α (19.4 ± 1.5 pg/mL), IL-6 (13.8 ± 0.6 pg/mL), and IL-8 (24.8 ± 1.2 pg/mL) as compared with the diabetic mice. The serum level of MMP2 (248.2 ± 7.9 pg/mL), MMP7 (316 ± 5.2 pg/mL), and MMP9 (167.8 ± 12.1 pg/mL) in the same group VNNPs were also observed much less than the diabetic mice. The serum level of TIMPs (176.8 ± 2.9 pg/mL) in the VNNPs group was increased maximally with respect to diabetic mice. It is concluded that nanoparticles and biomaterials possess healing properties and have the ability to repair the chronic/diabetic wound. RESEARCH HIGHLIGHTS: UV-spectrophotometric and Fourier transform infrared spectroscopy observation for functional group analysis and possible linkage between conjugates Optimization of the histopathological and biochemical markers after application of the formulations Microscopic analysis of epithelial tissues for evaluation of healing mechanisms Speedy contraction of wounds as the alleviation of the inflammatory and necrotic factors.

The effects of SGLT-2 inhibitors on echocardiographic indices and antioxidative properties in patients with heart failure with reduced ejection fraction and diabetes mellitus.

Sodium-glucose co-transporter-2 inhibitors (SGLT-2i) are a new class of drugs that lower blood glucose and reduce mortality in heart failure patients with reduced ejection fraction (HFrEF). They also have antioxidant effects. The exact mechanism of SGLT-2i is unknown. This study investigated the effects of SGLT-2i on asprosin, matrix metalloproteinase (MMP), and tissue inhibitor of MMP (TIMP-1) concentrations and echocardiographic measurements of strain in the left heart chamber. This prospective follow-up study included 56 patients with HFrEF and diabetes mellitus (DM) who did not initially receive SGLT-2 inhibitors. The control group consisted of 30 healthy individuals. Patients with HFrEF were administered either empagliflozin (n=28) or dapagliflozin (n=28) in addition to their treatment. The patient group was evaluated for left ventricular global longitudinal strain (LVGLS), left atrial (LA) strain, and LA volumes at the beginning and third month of the study. The control group had blood collected once, while the patient group had it twice: at the start of the trial, on the same day as the echocardiographic evaluation, and at the end of the third month after starting an SGLT-2i. Serum levels of asprosin, MMP-1 and TIMP-1 were assessed. LVGLS increased significantly in HFrEF patients at the third-month assessment compared to baseline (-8.6±2.3% vs. -9±2.5%, respectively; p<0.001), but there was no significant difference in LVEF (p=0.593). A substantial increase was observed in the left atrial ejection fraction (LAEF) compared to baseline values (36.3±9.4% vs. 42.1±8.7%, respectively; p<0.001), driven by a reduction in minimal LA volume [32.5 (19-96) ml vs. 32 (20-86) ml, respectively; p=0.018]. Compared to baseline evaluation, LA reservoir [13 (6-25) vs. 16.5 (2-26), respectively; p<0.001] and contraction strain (7.7±4.3 vs. 9.4±5.6, respectively; p=0.014) values were also enhanced at the third month. Between the baseline and the 3rd month, the patient group's LA conduit strain (p=0.122) and LA maximum volume (p=0.716) remained unchanged. Serum asprosin significantly increased (11.7±5.1 ng/mL vs. 14±9.4 ng/mL, respectively; p=0.032); however, no statistically significant alteration was detected in MMP (p=0.278) and TIMP-1 levels (p=0.401). SGLT-2i are associated with elevated levels of LVGLS, LAEF, LA contraction strain, and LA reservoir strain. SGLT-2i medications may improve plasma asprosin levels to boost energy metabolism, reduce oxidative stress and reactive oxygen radicals.