scholarly journals Addition of High Molecular Weight Hyaluronic Acid to Fibroblast-Like Stromal Cells Modulates Endogenous Hyaluronic Acid Metabolism and Enhances Proteolytic Processing and Secretion of Versican

Cells ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 1681
Author(s):  
Jiapeng Xue ◽  
Jinnan Chen ◽  
Quan Shen ◽  
Deva Chan ◽  
Jun Li ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Hyaluronic Acid ◽  
High Molecular Weight ◽  
Stromal Cells ◽  
De Novo ◽  
Linear Chain ◽  
Primary Cultures ◽  
Culture Conditions ◽  
Proteolytic Processing

We have examined the effect of exogenous linear chain high molecular weight hyaluronic acid (HMW HA) on endogenously synthesized hyaluronic acid (HA) and associated binding proteins in primary cultures of fibroblast-like stromal cells that were obtained by collagenase digestion of the murine peripatellar fat pad. The cultures were expanded in DMEM that was supplemented with fetal bovine serum and basic fibroblast growth factor (bFGF) then exposed to macrophage-colony-stimulating factor (MCSF) to induce macrophage properties, before activation of inflammatory pathways using E. coli lipopolysaccharide (LPS). Under all culture conditions, a significant amount of endogenously synthesized HA localized in LAMP1-positive lysosomal vesicles. However, this intracellular pool was depleted after the addition of exogenous HMW HA and was accompanied by enhanced proteolytic processing and secretion of de novo synthesized versican, much of which was associated with endosomal compartments. No changes were detected in synthesis, secretion, or proteolytic processing of aggrecan or lubricin (PRG4). The addition of HMW HA also modulated a range of LPS-affected genes in the TLR signaling and phagocytosis pathways, as well as endogenous HA metabolism genes, such as Has1, Hyal1, Hyal2, and Tmem2. However, there was no evidence for association of endogenous or exogenous HMW HA with cell surface CD44, TLR2 or TLR4 protein, suggesting that its physiochemical effects on pericelluar pH and/or ionic strength might be the primary modulators of signal transduction and vesicular trafficking by this cell type. We discuss the implications of these findings in terms of a potential in vivo effect of therapeutically applied HMW HA on the modification of osteoarthritis-related joint pathologies, such as pro-inflammatory and degradative responses of multipotent mesenchymal cells residing in the synovial membrane, the underlying adipose tissue, and the articular cartilage surface.

Selection of a Mutant Strain Streptococcus equi SH-109 and Optimization of Culture Conditions for the Production of High Molecular Weight Hyaluronic Acid

2014 ◽  
Vol 915-916 ◽  
pp. 909-912 ◽  
Author(s):  
Jian Cheng Feng ◽  
Rong Hu Zhang
Keyword(s):  
Molecular Weight ◽  
Hyaluronic Acid ◽  
High Molecular Weight ◽  
Culture Conditions ◽  
Culture Broth ◽  
Agitation Speed ◽  
Wild Type ◽  
Streptococcus Equi ◽  
Optimal Ph ◽  
Selection Of

A high molecular weight hyaluronic acid-producing mutant, designated asStreptococcus equiSH-109 that did not produce streptolysin or hyaluronidase was selected from wild-typeStreptococcus equiSH-0 by NTG treatment. The selected mutant could produce high molecular weight HA and had perfect genetic stability after reproduction. Several culture conditions were optimized in a 5-l fermentor for the production of hyaluronic acid. It was found that the optimal pH and agitation speed were 7.6 and 550rpm, respectively. Under the optimized culture condition, hyaluronic acid with a molecular weight of 4.0 x 106was accumulated in the culture broth.

Compound Mutation Breeding of Streptococcus equi JC-63 for Producing High Molecular Weight Hyaluronic Acid and Optimization of Culture Conditions

2016 ◽  
Vol 703 ◽  
pp. 337-342
Author(s):  
Rong Hu Zhang ◽  
Jian Cheng Feng
Keyword(s):  
Molecular Weight ◽  
Hyaluronic Acid ◽  
High Molecular Weight ◽  
Corn Steep Liquor ◽  
Fermentation Medium ◽  
Mutation Breeding ◽  
Culture Conditions ◽  
Compound Mutation ◽  
Steep Liquor ◽  
Orthogonal Tests

A strain of mutant Streptococcus equi JC-63, which can produce high molecular weight hyaluronic acid, was obtained from compound mutation breeding of Streptococcus equi JC-0 using 5-Bromouracil, UV rays and nitrosoguanidine as mutagenic agents. After inoculation for 25 generations, Streptococcus equi JC-63 showed inheritance stability, but not hemolysis. The culture conditions for producing HMW-HA were optimized in a 1-L shake flask through multi-factor orthogonal tests. Results showed the optimum ratio of fermentation medium comprised 50 g/L sucrose, 80 g/L corn steep liquor, 10 g/L glucose, 2 g/L MgSO4, 2 g/L NaHCO3, 2 g/L Na2HPO4, and 0.1 g/L UTP. Under these fermentation conditions, the molecular weight of the produced HA was 2.76 × 106 Da, and HA yield reached 4.12 g/L.

Preparation and Characterization of the High Molecular Weight [3H]Hyaluronic Acid

1992 ◽  
Vol 57 (10) ◽  
pp. 2151-2156 ◽  
Author(s):  
Peter Chabreček ◽  
Ladislav Šoltés ◽  
Hynek Hradec ◽  
Jiří Filip ◽  
Eduard Orviský
Keyword(s):  
Molecular Weight ◽  
Hyaluronic Acid ◽  
High Molecular Weight ◽  
Methyl Bromide ◽  
High Performance ◽  
Hydrogen Atoms ◽  
Isolation And Characterization ◽  
Labelling Procedure ◽  
Enzymatic Depolymerization

Two methods for the preparation of high molecular weight [3H]hyaluronic acid were investigated. In the first one, hydrogen atoms in the molecule were replaced by tritium. This isotopic substitution was performed in aqueous solution using Pd/CaCO3 as the catalyst. In the second method, the high molecular weight hyaluronic acid was alkylated with [3H]methyl bromide in liquid ammonia at a temperature of -33.5 °C. High-performance gel permeation chromatographic separation method was used for the isolation and characterization of the high molecular weight [3H]hyaluronic acid. Molecular weight parameters for the labelled biopolymers were Mw = 128 kDa, Mw/Mn = 1.88 (first method) and Mw = 268 kDa, Mw/Mn = 1.55 (second method). The high molecular weight [3H]hyaluronic acid having Mw = 268 kDa was degraded further by specific hyaluronidase. Products of the enzymatic depolymerization were observed to be identical for both, labelled and cold biopolymer. This finding indicates that the described labelling procedure using [3H]methyl bromide does not induce any major structural rearrangements in the molecule.

Linear and high-molecular-weight poly-porphyrins for efficient photodynamic therapy

2021 ◽  
Author(s):  
Nan Zheng ◽  
Xiahui Li ◽  
Shangwei Huangfu ◽  
Kangkai Xia ◽  
Ruofei Yue ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Photodynamic Therapy ◽  
Quantum Yield ◽  
Singlet Oxygen ◽  
High Molecular Weight ◽  
Linear Poly ◽  
High Molecular Weight Poly

A linear poly-porphyrin with high Mw and conjugated by PEG and acetazolamide was developed with enhanced singlet oxygen quantum yield, improved photo-toxicity and excellent in vivo photodynamic therapy.

scholarly journals Identification of two intermediates during processing of profilaggrin to filaggrin in neonatal mouse epidermis.

1984 ◽  
Vol 99 (4) ◽  
pp. 1372-1378 ◽  
Author(s):  
K A Resing ◽  
K A Walsh ◽  
B A Dale
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Tandem Repeats ◽  
Peptide Mapping ◽  
Two Dimensional ◽  
Major Event ◽  
Mouse Epidermis ◽  
Cell Stage ◽  
Processing Steps

A major event in the keratinization of epidermis is the production of the histidine-rich protein filaggrin (26,000 mol wt) from its high molecular weight (greater than 350,000) phosphorylated precursor (profilaggrin). We have identified two nonphosphorylated intermediates (60,000 and 90,000 mol wt) in NaSCN extracts of epidermis from C57/Bl6 mice by in vivo pulse-chase studies. Results of peptide mapping using a two-dimensional technique suggest that these intermediates consist of either two or three copies of filaggrin domains. Each of the intermediates has been purified. The ratios of amino acids in the purified components are unusual and essentially identical. The data are discussed in terms of a precursor containing tandem repeats of similar domains. In vivo pulse-chase experiments demonstrate that the processing of the high molecular weight phosphorylated precursor involves dephosphorylation and proteolytic steps through three-domain and two-domain intermediates to filaggrin. These processing steps appear to occur as the cell goes through the transition cell stage to form a cornified cell.

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