scholarly journals Chondroitin Sulfate-Degrading Enzymes as Tools for the Development of New Pharmaceuticals

Catalysts ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 322 ◽  
Author(s):  
Raúl Benito-Arenas ◽  
Sandra Zárate ◽  
Julia Revuelta ◽  
Agatha Bastida
Keyword(s):  
Extracellular Matrix ◽  
Pharmaceutical Industry ◽  
Chondroitin Sulfate ◽  
Biological Tissues ◽  
Sulfated Polysaccharides ◽  
Specific Time ◽  
Time Points ◽  
Therapeutic Applications ◽  
Degrading Enzymes ◽  
Chondroitin Sulfates

Chondroitin sulfates are linear anionic sulfated polysaccharides found in biological tissues, mainly within the extracellular matrix, which are degraded and altered by specific lyases depending on specific time points. These polysaccharides have recently acquired relevance in the pharmaceutical industry due to their interesting therapeutic applications. As a consequence, chondroitin sulfate (CS) lyases have been widely investigated as tools for the development of new pharmaceuticals based on these polysaccharides. This review focuses on the major breakthrough represented by chondroitin sulfate-degrading enzymes and their structures and mechanisms of function in addition to their major applications.

Detection of chondroitin sulfates and decorin in developing fetal and neonatal rat lung

2002 ◽  
Vol 282 (3) ◽  
pp. L484-L490 ◽  
Author(s):  
Yiqiong Wang ◽  
Kaori Sakamoto ◽  
Jody Khosla ◽  
Philip L. Sannes
Keyword(s):  
Extracellular Matrix ◽  
Chondroitin Sulfate ◽  
Wide Distribution ◽  
Neonatal Rat ◽  
Chondroitin Sulfate Proteoglycan ◽  
Sulfate Proteoglycan ◽  
Rat Lung ◽  
Adult Rats ◽  
Chondroitin Sulfates ◽  
Monospecific Antibodies

Chondroitin sulfates and their related proteoglycans are components of extracellular matrix that act as key determinants of growth and differentiation characteristics of developing lungs. Changes in their immunohistochemical distribution during progressive organ maturation were examined with monospecific antibodies to chondroitin sulfate, a nonbasement membrane chondroitin sulfate proteoglycan, and the specific chondroitin sulfate-containing proteoglycan decorin in whole fetuses and lungs from newborn and adult rats. Alveolar and airway extracellular matrix immunostained heavily in the prenatal rat for both chondroitin sulfate and chondroitin sulfate proteoglycan, whereas decorin was confined to developing airways and vessels. These sites retained their respective levels of reactivity with all antibodies through 1–10 days postnatal but thereafter became progressively more diminished and focal in alveolar regions. The heavy staining seen early in development was interpreted to reflect a significant and wide distribution of chondroitin sulfates, chondroitin sulfate proteoglycans, and decorin in rapidly growing tissues, whereas the reduced and more focal reactivity observed at later time points coincided with known focal patterns of localization of fibrillar elements of the extracellular matrix and a more differentiated state.

scholarly journals THU0435 CALCIUM PYROPHOSPHATE DIHYDRATE (CPPD) CRYSTALS BUT NOT BASIC CALCIUM PHOSPHATE (BCP) CRYSTALS INDUCE SYNDECAN-4 EXPRESSION IN CARTILAGE

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 454.2-455
Author(s):  
S. Nasi ◽  
J. Bertrand ◽  
M. Bollmann ◽  
R. Stange ◽  
T. Pap
Keyword(s):  
Extracellular Matrix ◽  
Calcium Phosphate ◽  
Calcium Pyrophosphate ◽  
Calcium Pyrophosphate Dihydrate ◽  
Basic Calcium Phosphate ◽  
Time Points ◽  
Qrt Pcr ◽  
Degrading Enzymes ◽  
Pyrophosphate Dihydrate ◽  
Matrix Degrading Enzymes

Background:Chondrocalcinosis is a painful rheumatic condition caused by the deposition of calcium pyrophosphate dihydrate crystals (CPPD) in joint tissues, and especially in cartilage. It is known that CPPD crystals cause inflammation and degenerative changes in joint, but the underlying mechanisms remain poorly understood. In particular, nothing is known about how these crystals regulates transmembrane heparan sulphate proteoglycans (HSPGs). Our attention focused on one family of HSPGs called syndecans as they have important roles both as adhesion molecules, by mediating chondrocyte-extracellular matrix interactions, and as modulators of intracellular signaling triggered by cytokines and growth factors.Objectives:The aim of this study was to evaluate how CPPD crystals modulates syndecan expression in chondrocytes and in cartilage, and how this modulation can be ultimately linked to cartilage damage during chondrocylcinosis.Methods:Murine chondrocitic ATDC5 cells were stimulated with 0,1 ng/ml CPPD crystals or with 0,1 ng/ml basic-calcium phosphate crystals (BCP), a family of calcium-containing crystals found in other rheumatic conditions such as osteoarthritis (OA). Cytotoxicity was evaluated by lactate dehydrogenase (LDH) release in the supernatant at 30 minutes, and 3, 6, 24 hours after stimulation. At the same time-points, mRNA expression levels of syndecans (Synd-1, -2, -3, -4) and of matrix-degrading enzymes (Mmp-3, -9, -13; Adamts-4, -5) was analysed by qRT-PCR. Finally, Syndecan-4 protein expression was studied by immunohistochemistry (IHC) in cartilage samples of patients with chondrocalcinosis and in samples of patients with severe OA without chondrocalcinosis as control.Results:LDH assay revealed no increased cytotoxicity by CPPD or BCP at any time-point. qRT-PCR indicated that CPPD crystals but not BCP crystals induced Synd-2 and -3 upregulation at 30 minutes after stimulation and Synd-4 upregulation at 3 hours, while no modulation of syndecans was seen at later time-points. CPPD also induced Adamts-4 expression at 3 hours after stimulation, and Mmp-9 expression at 3 and 6 hours. The expression of the other matrix-degrading enzymes was not affected. Human chondrocalcinosis cartilage exhibited enhanced Synd-4 expression compared to severe OA cartilage containing BCP calcification. Interestingly, Synd-4 expression was observed in the extracellular matrix but not on cell membrane, suggesting that maybe Synd-4 undergoes shedding (Figure 1).Figure 1.Representative Synd-4 IHC in control patients (Ctrl, severe OA) and chondrocalcinosis patients (CPPD). Note increased Synd-4 expression in extracellular matrix of CPPD patients compared to Ctrl ones.Conclusion:BCP and CPPD crystals seem to trigger differential effects in terms of modulation of syndecans in chondrocitic cells. CPPD crystals induce Synd-4 and Adamts-4 and Mmp-9 which are not induced by BCP crystals. It remains to be clarified whether the two events are interlinked. In particular, further studies are required to investigate if Adamts-4 and Mmp-9 are involved in Synd-4 shedding or if vice versa Synd-4 regulates Adamts-4 and Mmp-9 activation and downstream cartilage breakdown in chondrocalcinosis.Disclosure of Interests: :Sonia Nasi: None declared, Jessica Bertrand Grant/research support from: Pfizer, Speakers bureau: Pfizer, Miriam Bollmann: None declared, Richard Stange: None declared, Thomas Pap: None declared

Effects of oversulfated and fucosylated chondroitin sulfates on coagulation

2010 ◽  
Vol 103 (05) ◽  
pp. 994-1004 ◽  
Author(s):  
Roberto Fonseca ◽  
Stephan-Nicollas Oliveira ◽  
Vitor Pomin ◽  
André Mecawi ◽  
Iracema Araujo ◽  
...  
Keyword(s):  
Chondroitin Sulfate ◽  
Anticoagulant Activity ◽  
Coagulation System ◽  
In Vitro Assays ◽  
Sulfated Polysaccharides ◽  
Activate Factor ◽  
Factor Xii ◽  
Chondroitin Sulfates ◽  
Oversulfated Chondroitin Sulfate

SummaryWe report the effects of a chemically oversulfated chondroitin sulfate and a naturally fucosylated chondroitin sulfate on the coagulation system. The former has been recently identified as a contaminant of heparin preparations and the latter has been proposed as an alternative anticoagulant. The mechanism of action of these polymers on coagulation is complex and target different components of the coagulation system. They have serpin-independent anticoagulant activity, which preponderates in plasma. They also have serpin-dependent anticoagulant activity but differ significantly in the target coagulation protease and preferential serpin. Their anticoagulant effects differ even more markedly when tested as inhibitors of coagulation proteases using plasma as a source of serpins. It is possible that the difference is due to the high availability of fucosylated chondroitin sulfate whereas over-sulfated chondroitin sulfate has strong unspecific binding to plasma protein and low availability for the binding to serpins. When tested using a venous thrombosis experimental model, oversulfated chondroitin sulfate is less potent as an antithrombotic agent than fucosylated chondroitin sulfate. These highly sulfated chondroitin sulfates activate factor XII in in vitro assays, based on kallikrein release. However, only fucosylated chondroitin sulfate induces hypotension when intravenously injected into rats. In conclusion, the complexity of the regulatory mechanisms involved in the action of highly sulfated polysaccharides in coagulation requires their analysis by a combination of in vitro and in vivo assays. Our results are relevant due to the urgent need for new anticoagulant drugs or alternative sources of heparin.

scholarly journals Optical Microscopy and the Extracellular Matrix Structure: A Review

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1760
Author(s):  
Joshua J. A. Poole ◽  
Leila B. Mostaço-Guidolin
Keyword(s):  
Extracellular Matrix ◽  
Confocal Microscopy ◽  
Laser Scanning ◽  
Biological Tissues ◽  
The Body ◽  
Stimulated Emission ◽  
Substantial Part ◽  
Fluorescence Lifetime Imaging ◽  
Structured Illumination ◽  
Structured Illumination Microscopy

Biological tissues are not uniquely composed of cells. A substantial part of their volume is extracellular space, which is primarily filled by an intricate network of macromolecules constituting the extracellular matrix (ECM). The ECM serves as the scaffolding for tissues and organs throughout the body, playing an essential role in their structural and functional integrity. Understanding the intimate interaction between the cells and their structural microenvironment is central to our understanding of the factors driving the formation of normal versus remodelled tissue, including the processes involved in chronic fibrotic diseases. The visualization of the ECM is a key factor to track such changes successfully. This review is focused on presenting several optical imaging microscopy modalities used to characterize different ECM components. In this review, we describe and provide examples of applications of a vast gamut of microscopy techniques, such as widefield fluorescence, total internal reflection fluorescence, laser scanning confocal microscopy, multipoint/slit confocal microscopy, two-photon excited fluorescence (TPEF), second and third harmonic generation (SHG, THG), coherent anti-Stokes Raman scattering (CARS), fluorescence lifetime imaging microscopy (FLIM), structured illumination microscopy (SIM), stimulated emission depletion microscopy (STED), ground-state depletion microscopy (GSD), and photoactivated localization microscopy (PALM/fPALM), as well as their main advantages, limitations.

Glycosaminoglycans from growing antlers of wapiti (Cervus elaphus)

1997 ◽  
Vol 77 (4) ◽  
pp. 715-721 ◽  
Author(s):  
H. H. Sunwoo ◽  
L. Y. M. Sim ◽  
T. Nakano ◽  
R. J. Hudson ◽  
J. S. Sim
Keyword(s):  
Hyaluronic Acid ◽  
Chondroitin Sulfate ◽  
Cervus Elaphus ◽  
Molecular Size ◽  
Enzymatic Digestion ◽  
Gel Chromatography ◽  
Cellulose Acetate Electrophoresis ◽  
Cartilaginous Tissues ◽  
Chondroitin Sulfates ◽  
Velvet Antler

The emerging wapiti industry in North America is based largely on markets for velvet antlers which are used in oriental medicine. Despite the economic opportunity, enthusiasm has been dampened by incomplete understanding of the chemical and pharmacological properties of velvet antler. This study characterizes polysaccharide constituents of glycosaminoglycans in growing antler of wapiti (Cervus elaphus). Glycosaminoglycans were isolated from four sections (tip, upper, middle and base) of growing antlers, and were studied using cellulose acetate electrophoresis, gel electrophoresis, enzymatic digestion and gel chromatography. The tip and upper sections of the antler which are rich in cartilaginous tissues contained chondroitin sulfate as a major glycosaminoglycan with small amounts of hyaluronic acid. In the middle and base sections containing bone and bone marrow, chondroitin sulfate was also a major glycosaminoglycan with small amounts of hyaluronic acid and chondroitinase-ACI resistant materials. More than half of chondroitin sulfate from the middle and base sections had larger molecular size than did the chondroitin sulfates from the tip and upper sections. Key words: Glycosaminoglycans, chondroitin sulfate, antler, wapiti

Reconstituted Extracellular Matrix Improve Osteoblastic Differentiation onto Titanium Surfaces

2012 ◽  
Vol 706-709 ◽  
pp. 584-588
Author(s):  
Lia Rimondini ◽  
Federica Demarosi ◽  
Ismaela Foltran ◽  
Nadia Quirici
Keyword(s):  
Extracellular Matrix ◽  
Electrostatic Force ◽  
Biological Tissues ◽  
Osteoblastic Differentiation ◽  
Cell Phenotype ◽  
Tissue Engineering Scaffolds ◽  
Electrospinning Technique ◽  
Differentiation Potential ◽  
Oral Implants

Electrospinning technique is an efficient processing method to manufacture micro-and nanosized fibrous structures by electrostatic force for different applications. In biomaterial field, electrospinning technique has been successfully utilized to prepare new drug delivery materials and tissue engineering scaffolds. Fiber mats of biodegradable polymers having a diameter in the nanoto submicro-scale can be considered to mimic the nanofibrous structure of native extracellular matrix (ECM). Native extracellular matrix, constituted of proteins and polysaccharides improving cells growth in its nanofibrous porous structure, controls not only the cell phenotype, but the whole structure of the biological tissues. In the present study we investigated the effect of electrospun reconstituted collagen fibers onto metals for oral implants devices manufacturing as far as the osteoblastic differentiation potential of stem cells and cytofunctionality of osteoblasts in-vitro. The cells cultured onto titanium samples coated with ECM constituents showed faster osteoblastic differentiation and more efficient deposition of mineralized matrix in comparison with those onto uncoated substrates.

Proteoglycan-substrate gel zymography for the detection of chondroitin sulfate-degrading enzymes

2019 ◽  
Vol 568 ◽  
pp. 51-52
Author(s):  
Takashi Kobayashi ◽  
Ikuko Kakizaki ◽  
Toshiya Nakamura
Keyword(s):  
Chondroitin Sulfate ◽  
Degrading Enzymes

scholarly journals Strain variation in Bacillus cereus biofilms and their susceptibility to extracellular matrix-degrading enzymes

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0245708
Author(s):  
Eun Seob Lim ◽  
Seung-Youb Baek ◽  
Taeyoung Oh ◽  
Minseon Koo ◽  
Joo Young Lee ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Bacillus Cereus ◽  
Biofilm Formation ◽  
Dnase I ◽  
Extracellular Dna ◽  
Proteinase K ◽  
Strain Variation ◽  
Protein Ratio ◽  
Degrading Enzymes ◽  
Matrix Degrading Enzymes

Bacillus cereus is a foodborne pathogen and can form biofilms on food contact surfaces, which causes food hygiene problems. While it is necessary to understand strain-dependent variation to effectively control these biofilms, strain-to-strain variation in the structure of B. cereus biofilms is poorly understood. In this study, B. cereus strains from tatsoi (BC4, BC10, and BC72) and the ATCC 10987 reference strain were incubated at 30°C to form biofilms in the presence of the extracellular matrix-degrading enzymes DNase I, proteinase K, dispase II, cellulase, amyloglucosidase, and α-amylase to assess the susceptibility to these enzymes. The four strains exhibited four different patterns in terms of biofilm susceptibility to the enzymes as well as morphology of surface-attached biofilms or suspended cell aggregates. DNase I inhibited the biofilm formation of strains ATCC 10987 and BC4 but not of strains BC10 and BC72. This result suggests that some strains may not have extracellular DNA, or their extracellular DNA may be protected in their biofilms. In addition, the strains exhibited different patterns of susceptibility to protein- and carbohydrate-degrading enzymes. While other strains were resistant, strains ATCC 10987 and BC4 were susceptible to cellulase, suggesting that cellulose or its similar polysaccharides may exist and play an essential role in their biofilm formation. Our compositional and imaging analyses of strains ATCC 10987 and BC4 suggested that the physicochemical properties of their biofilms are distinct, as calculated by the carbohydrate to protein ratio. Taken together, our study suggests that the extracellular matrix of B. cereus biofilms may be highly diverse and provides insight into the diverse mechanisms of biofilm formation among B. cereus strains.

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