Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.24.3 (collagenase)
 18,340 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The haemolytic power of isolated nematocysts from the scyphozoan Pelagia noctiluca was studied with attention to the effect of osmotic protectants as carbohydrates at different MW, cations as Mg2+, Ca2+, Ba2+,Cu2+, K+; proteases as collagenase, trypsin, alpha-chymotrypsin, papain; and antioxidants. Crude venom was at first obtained by sonication of holotrichous-isorhiza nematocysts previously isolated from oral arms of P. noctiluca and then haemolytically tested upon human erythrocytes. Osmotic protectants were effective in inhibiting the haemolytic power depending on their molecular weight so that total inhibition of crude venom-induced haemolysis was observed after PEG treatment (polyethyleneglycol 6000Da). Amongst divalent cations only Ba2+ and Cu2+ significantly inhibited the haemolytic power of crude venom. Proteases seem not to alter the haemolytic activity while antioxidant compounds only slightly reduced the haemolytic power. Such findings may suggest a pore-forming mechanism for P. noctiluca crude venom rather than an oxidative damage to the cell membrane.
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PMID:Effect of various factors on Pelagia noctiluca (Cnidaria, Scyphozoa) crude venom-induced haemolysis. 1861 52

Chondrocytes were encapsulated in non-degrading and partially degrading poly(ethylene glycol) (PEG) gels in apposition to native cartilage layers in order to examine the effects of gel degradation on the integration of regenerated cartilaginous matrix with native tissue. In addition, the effect of collagenase predigestion of the native cartilage surfaces on this integration was examined in studies with partially degrading co-polymer gels. Integration was quantitatively assessed by mechanical measurements of adhesive strength, and visualized by histological staining and non-destructive ultrasound analysis. Constructs with encapsulated chondrocytes and a non-degrading gel layer had significantly higher adhesive strength than partially degrading gel constructs and non-degrading gel constructs without cells. In addition, better maintenance of proper cell morphology was observed near the gel-cartilage interface in non-degrading gel constructs than in partially degrading gel constructs after 8 weeks of in vitro culture. Facile collagen distribution in the degrading gels appeared to have a significant effect on mechanical adhesion measurements only when the native cartilage surface was predigested with collagenase. Ultrasound analysis provided qualitative evidence of cartilaginous matrix evolution and non-destructive imaging of developing constructs and the interface between newly formed matrix and existing cartilage tissue.
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PMID:Effects of directed gel degradation and collagenase digestion on the integration of neocartilage produced by chondrocytes encapsulated in hydrogel carriers. 1872 35

The use of exogenous superoxide dismutase (SOD) and catalase (CAT) has been previously evaluated against various reactive oxygen species-mediated brain injuries, especially those associated with ischemia/ reperfusion. In this study, we investigated effects of these enzymatic antioxidants on intracerebral hemorrhage (ICH)-induced brain injury. A total of 65 male Sprague-Dawley rats (300-380 g) were divided into a sham group, an untreated ICH group, 3 groups of ICH rats treated with lecithinized SOD (PC-SOD) at doses of 0.1, 0.3, and 1 mg/kg, and a group treated with polyethylene glycol conjugated CAT (PEG-CAT) at a dose of 10,000 U/kg. An additional group of ICH rats received a combination of PC-SOD (1 mg/kg) and PEG-CAT (10,000 U/kg). ICH was induced by collagenase injection. All drugs were administered intravenously immediately after ICH induction. Brain injury was evaluated by scoring neurological function and measuring brain edema at 24 h after ICH induction. Our results demonstrated that ICH caused significant neurological deficit associated with remarkable brain edema. Treatment with PC-SOD, PEG-CAT, or PC-SOD in combination with PEG-CAT did not reduce brain edema or neurological deficit after ICH. We conclude that intravenously administered PC-SOD and/or PEG-CAT do not reduce brain injury in the collagenase-induced ICH rat model.
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PMID:Effects of superoxide dismutase and catalase derivates on intracerebral hemorrhage-induced brain injury in rats. 1906 78

A biointeractive collagen-phospholipid corneal substitute was fabricated from interpenetrating polymeric networks comprising 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide and N-hydroxysuccinimide crosslinked porcine atelocollagen, and poly(ethylene glycol) diacrylate crosslinked 2-methacryloyloxyethyl phosphorylcholine (MPC). The resulting hydrogels showed an overall increase in mechanical strength beyond that of either original component and enhanced stability against enzymatic digestion (by collagenase) or UV degradation. More strikingly, these hydrogels retained the full biointeractive, cell friendly properties of collagen in promoting corneal cell and nerve in-growth and regeneration (despite MPC's known anti-adhesive properties). Measurements of refractive indices, white light transmission and backscatter showed the optical properties of collagen-MPC are comparable or superior to those of the human cornea. In addition, the glucose and albumin permeability were comparable to those of human corneas. Twelve-month post-implantation results of collagen-MPC hydrogels into mini-pigs showed regeneration of corneal tissue (epithelium, stroma) as well as the tear film and sensory nerves. We also show that porcine collagen can be substituted with recombinant human collagen, resulting in a fully-synthetic implant that is free from the potential risks of disease transmission (e.g. prions) present in animal source materials.
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PMID:Collagen-phosphorylcholine interpenetrating network hydrogels as corneal substitutes. 1909 43

The collagenase, produced extracellular by Bacillus pumilus Col-J, was purified by ammonium sulfate precipitation followed by two gel filtrations, involving Sephadex G-100 column and Sepharose Fast Flow column. Purified collagenase has a 31.53-fold increase in specific activity of 87.33 U/mg and 7.00% recovery. The collagenase has a relative molecular weight of 58.64 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimal temperature for the enzyme reaction was 45 degrees C. More than 50% of the original activity still remained after 5 min of incubation at 70 degrees C or 10 min at 60 degrees C. The maximal enzyme activity of collagenase was obtained at pH 7.5, and it was stable over a pH range of 6.5-8.0. The collagenase activity was strongly inhibited by Mn(2+), Pb(2+), ethylenediamine tetraacetic acid, ethylene glycol tetraacetic acid, and beta-mercaptoethanol. However, Ca(2+) and Mg(2+) greatly increased its activity. The collagenase from B. pumilus Col-J showed highly specific activity towards the native collagen from calf skin. The K(m) and V(max) of the enzyme for collagen were 0.79 mg/mL and 129.5 U, respectively.
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PMID:Purification and characterization of a novel collagenase from Bacillus pumilus Col-J. 1947 15

6-Methacryloyl-alpha-D-galactopyranose (MG) was synthesized, and characterized by Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR) spectrometry, and single-crystal X-ray diffraction. A series of interpenetrating polymer network (IPN) hydrogels was fabricated by simultaneously photocuring MG crosslinked by poly(ethylene glycol) diacrylate and chemically crosslinking type I collagen with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide. The successful incorporation of the glycopolymer, polymer MG, into collagen hydrogel was confirmed by FTIR and solid-state (13)C NMR. The optical characteristics of the IPN hydrogels are comparable to those of human corneas. The tensile strength and modulus of the hydrogels are enhanced by incorporation of polymer MG in comparison to that of the control collagen hydrogel. Biodegradation results indicated that polymer MG enhanced the stability of the composite hydrogels against collagenase. In vitro results demonstrated that the IPN hydrogel supported the adhesion and proliferation of human corneal epithelial cells and outperformed human cornea in blocking bacteria adhesion. Taken together, the IPN hydrogel might be a promising material for use in corneal lamellar keratoplasty.
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PMID:Collagen and glycopolymer based hydrogel for potential corneal application. 1963 59

The development and use of functional tissue-engineered products is currently limited by the challenge of incorporating microvasculature. To this end, we have investigated strategies to facilitate vascularization in scaffold materials, in this case poly(ethylene glycol) (PEG) hydrogels. These hydrogels are hydrophilic and resist protein adsorption and subsequent non-specific cell adhesion, but can be modified to contain cell-adhesive ligands and growth factors to support cell and tissue function. Additionally, the hydrogel matrix can include proteolytically degradable peptide sequences in the backbone of the structure to allow cells to control scaffold biodegradation, allowing three-dimensional migration. Vascular endothelial growth factor (VEGF), a potent angiogenic signal, and the cell-adhesive peptide RGDS were each covalently attached to PEG monoacrylate linkers. PEGylated RGDS and VEGF were then covalently immobilized in PEG-diacrylate (PEGDA) hydrogels in 2D and 3D. Immobilized VEGF increased endothelial cell tubulogenesis on the surface of non-degradable PEGDA hydrogels 4-fold compared to controls without the growth factor. Endothelial cell behavior in 3D collagenase-degradable hydrogels modified with RGDS and VEGF was observed using time-lapse confocal microscopy. Bulk immobilization of VEGF in 3D collagenase-degradable RGDS-modified hydrogels increased endothelial cell motility 14-fold and cell-cell connections 3-fold. Covalent incorporation of PEGylated VEGF in PEG hydrogels can be a useful tool to promote endothelial cell migration, cell-cell contact formation and tubulogenesis in an effort to produce vascularized tissue-engineered constructs.
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PMID:Covalently-immobilized vascular endothelial growth factor promotes endothelial cell tubulogenesis in poly(ethylene glycol) diacrylate hydrogels. 1972 40

Synthetic hydrogels based on poly(ethylene glycol) (PEG) have been used as biomaterials for cell biology and tissue engineering investigations. Bioactive PEG-based gels have largely relied on heterobifunctional or multi-arm PEG precursors that can be difficult to synthesize and characterize or expensive to obtain. Here, we report an alternative strategy, which instead uses inexpensive and readily available PEG precursors to simplify reactant sourcing. This new approach provides a robust system in which to probe cellular interactions with the microenvironment. We used the step-growth polymerization of PEG diacrylate (PEGDA, 3400Da) with bis-cysteine matrix metalloproteinase (MMP)-sensitive peptides via Michael-type addition to form biodegradable photoactive macromers of the form acrylate-PEG-(peptide-PEG)(m)-acrylate. The molecular weight (MW) of these macromers is controlled by the stoichiometry of the reaction, with a high proportion of resultant macromer species greater than 500kDa. In addition, the polydispersity of these materials was nearly identical for three different MMP-sensitive peptide sequences subjected to the same reaction conditions. When photopolymerized into hydrogels, these high MW materials exhibit increased swelling and sensitivity to collagenase-mediated degradation as compared to previously published PEG hydrogel systems. Cell-adhesive acrylate-PEG-CGRGDS was synthesized similarly and its immobilization and stability in solid hydrogels was characterized with a modified Lowry assay. To illustrate the functional utility of this approach in a biological setting, we applied this system to develop materials that promote angiogenesis in an ex vivo aortic arch explant assay. We demonstrate the formation and invasion of new sprouts mediated by endothelial cells into the hydrogels from embedded embryonic chick aortic arches. Furthermore, we show that this capillary sprouting and three-dimensional migration of endothelial cells can be tuned by engineering the MMP-susceptibility of the hydrogels and the presence of functional immobilized adhesive ligands (CGRGDS vs. CGRGES peptide). The facile chemistry described and significant cellular responses observed suggest the usefulness of these materials in a variety of in vitro and ex vivo biologic investigations, and may aid in the design or refinement of material systems for a range of tissue engineering approaches.
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PMID:Bioactive hydrogels made from step-growth derived PEG-peptide macromers. 2013 64

Adipose tissue engineering requires biomaterials that promote the differentiation of seeded adipocytes. Here, we report on the development and characterization of in situ forming, poly(ethylene glycol) (PEG) based hydrogels for soft tissue augmentation. Branched PEG-amines were modified with collagenase-sensitive peptides and cross-linked with branched PEG-succinimidyl propionates without the use of free-radical initiators (enzymatically degradable hydrogels). Alanine-modified PEG-amines were used for the preparation of non-degradable gels. Depending on the used polymer concentration, the strength of degradable gels after swelling ranged from 1708 to 7412 Pa; the strength of non-degradable hydrogels varied between 1496 and 7686 Pa. Enzyme mediated gel degradation occurred within 10, 16, and 19 days (5%, 10%, and 15% initial polymer content). To evaluate their suitability as scaffold materials for adipose tissue engineering, the hydrogels were functionalized with the laminin-derived adhesion peptide YIGSR, and seeded with 3T3-L1 preadipocytes. Compared to a standard two-dimensional cell culture model, the developed hydrogels significantly enhanced the intracellular triglyceride accumulation of encapsulated adipocytes. Functionalization with YIGSR further enhanced lipid synthesis within differentiating adipocytes. Long-term studies suggested that enzymatically degradable hydrogels furthermore promote the formation of coherent adipose tissue-like structures featuring many mature unilocular fat cells.
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PMID:Enzymatically degradable poly(ethylene glycol) based hydrogels for adipose tissue engineering. 2017 Sep 51

A method to functionalize collagen-based biomaterials with free amine groups was established in an attempt to improve their potential for tethering of bioactive molecules. Collagen sponges were incorporated with amine-terminated multifunctional polyethylene glycol (PEG) derivatives after N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide and N-hydroxysuccinimide (EDC/NHS) cross-linking. The extent of the incorporation of different amounts and different numbers of active moieties of amine-terminated PEG systems into the collagen scaffolds was evaluated using ninhydrin assay, Fourier transform infrared spectrophotometry (FTIR), collagenase degradation assay, denaturation temperature measurements, and in vitro cell studies. A 3% 8-arm amine-terminated PEG was found to be the minimum required effective concentration to functionalize EDC/NHS stabilized collagen scaffolds. EDC/NHS stabilized scaffolds treated with 3% 8-arm amine-terminated PEG exhibited significantly improved denaturation temperature and resistance to collagenase degradation over non-cross-linked scaffolds (p < 0.002). Biological evaluation using 3T3 cells demonstrated that the produced scaffolds facilitated maintenance of the cells' morphology, metabolic activity, and ability to proliferate in vitro. Overall, our results indicate that amine-terminated PEG systems can be used as means to enhance the functionality of collagenous structures.
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PMID:Amine functionalization of collagen matrices with multifunctional polyethylene glycol systems. 2094 84


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