Biotechnological Chondroitin a Novel Glycosamminoglycan With Remarkable Biological Function on Human Primary Chondrocytes

Abstract

Cartilage tissue engineering, with in vitro expansion of autologus chondrocytes, is a promising technique for tissue regeneration and is a new potential strategy to prevent and/or treat cartilage damage (e.g., osteoarthritis). The aim of this study was (i) to investigate and compare the effects of new biotechnological chondroitin (BC) and a commercial extractive chondroitin sulfate (CS) on human chondrocytes in vitro culture; (ii) to evaluate the anti‐inflammatory effects of the innovative BC compared to extractive CS. A chondrogenic cell population was isolated from human nasoseptal cartilage and in vitro cultures were studied through time‐lapse video microscopy (TLVM), immunohistochemical staining and cytometry. In order to investigate the effect of BC and CS on phenotype maintainance, chondrogenic gene expression of aggrecan (AGN), of the transcriptor factor SOX9, of the types I and II collagen (COL1A1 and COL1A2), were quantified through transcriptional and protein evaluation at increasing cultivation time and passages. In addition to resemble the osteoarthritis‐like in vitro model, chondrocytes were treated with IL‐1β and the anti‐inflammatory activity of BC and CS was assessed using cytokines quantification by multiplex array. BC significantly enhances cell proliferation also preserving chondrocyte phenotype increasing type II collagen expression up to 10 days of treatment and reduces inflammatory response in IL‐1β treated chondrocytes respect to CS treated cells. Our results, taken together, suggest that this new BC is of foremost importance in translational medicine because it can be applied in novel scaffolds and pharmaceutical preparations aiming at cartilage pathology treatments such as the osteoarthritis. J. Cell. Biochem. 117: 2158–2169, 2016. © 2016 The Authors. Journal of Cellular Biochemistry Published by Wiley Periodicals, Inc.

Hybrid Complexes of High and Low Molecular Weight Hyaluronans Highly Enhance HASCs Differentiation: Implication for Facial Bioremodelling

“…The fold-change of mRNA expression of the genes under evaluation was calculated by using the 2 −∆∆ Ct comparative threshold method (∆Ct = difference of ∆Ct between treated cells and non-treated cells used as controls). The results were expressed as normalized fold expression, calculated by the ratio of crossing points of amplification curves of several genes and internal standard, by using the Bio-Rad iQTM5 software (Bio-Rad Laboratories Srl) as previously reported [24,26].…”

A Modular Approach to a Library of Semi‐Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns

“…[19] After further purification steps using ethanol and repeated alcoholicp recipitations, the unsulfated, defructosylated polysaccharide was obtainedi n9 0% purity grade, ase valuated by NMR analysis andc apillary electrophoresis. [21] The transformation of CS-0i nto eight different fC andf CS polysaccharides was planned through am odular approach (Scheme1). [21] The transformation of CS-0i nto eight different fC andf CS polysaccharides was planned through am odular approach (Scheme1).…”

“…[25];( b) see ref. The benzylidene ring of derivative 7 was then cleaved under oxidative conditions (NaBrO 3 and Na 2 S 2 O 3 in aH 2 O/ethyl acetate mixture) [21] to afford polysaccharide acceptor 8 in 63 %y ield (over two steps from 1) [28] respectively;s ee also the Supporting Information), assuming that the signals to be compared displayed similar 1 J CH coupling constants and that ad ifferenceo fa round 5-8 Hz from the experimental set value did not cause as ubstantial variation in the integratedp eak volumes. tected by acetylation with Ac 2 Oa nd Et 3 Ni nC H 3 CN in the presence of ac atalytic amount of 4-dimethylaminopyridine (DMAP) to afford derivative 7 with aquantitative degree of substitution (DS = 2.00), as calculatedb yi ntegration of the H-2 and H-3 carbinolics ignals (d = 4.65 and 5.04 ppm, respectively) with respect to benzylidene methine one (d = 5.44 ppm) in the 1 HNMR spectrum (see also the SupportingI nformation).…”

Chondroprotective and anti‐inflammatory effects of amurensin H by regulating TLR4/Syk/NF‐κB signals

“…22 Chondrocytes grown on 6-well plates were stained with LIVE/DEAD Cell Imaging Kit (Invitrogen) according to the manufacturer's protocol and analysed by fluorescence Cell viability was determined using MTT (Sigma) assay with OD value measured at 570 nm by a microplate reader.…”