Chondroitin Sulfate Content Research Articles

Postnatal lethality and chondrodysplasia in mice lacking both chondroitin sulfate N-acetylgalactosaminyltransferase-1 and -2.

Chondroitin sulfate (CS) is a sulfated glycosaminoglycan (GAG) chain. In cartilage, CS plays important roles as the main component of the extracellular matrix (ECM), existing as side chains of the major cartilage proteoglycan, aggrecan. Six glycosyltransferases are known to coordinately synthesize the backbone structure of CS; however, their in vivo synthetic mechanism remains unknown. Previous studies have suggested that two glycosyltransferases, Csgalnact1 (t1) and Csgalnact2 (t2), are critical for initiation of CS synthesis in vitro. Indeed, t1 single knockout mice (t1 KO) exhibit slight dwarfism and a reduction in CS content in cartilage compared with wild-type (WT) mice. To reveal the synergetic roles of t1 and t2 in CS synthesis in vivo, we generated systemic single and double knockout (DKO) mice and cartilage-specific t1 and t2 double knockout (Col2-DKO) mice. DKO mice exhibited postnatal lethality, whereas t2 KO mice showed normal size and skeletal development. Col2-DKO mice survived to adulthood and showed severe dwarfism compared with t1 KO mice. Histological analysis of epiphyseal cartilage from Col2-DKO mice revealed disrupted endochondral ossification, characterized by drastic GAG reduction in the ECM. Moreover, DKO cartilage had reduced chondrocyte proliferation and an increased number of apoptotic chondrocytes compared with WT cartilage. Conversely, primary chondrocyte cultures from Col2-DKO knee cartilage had the same proliferation rate as WT chondrocytes and low GAG expression levels, indicating that the chondrocytes themselves had an intact proliferative ability. Quantitative RT-PCR analysis of E18.5 cartilage showed that the expression levels of Col2a1 and Ptch1 transcripts tended to decrease in DKO compared with those in WT mice. The CS content in DKO cartilage was decreased compared with that in t1 KO cartilage but was not completely absent. These results suggest that aberrant ECM caused by CS reduction disrupted endochondral ossification. Overall, we propose that both t1 and t2 are necessary for CS synthesis and normal chondrocyte differentiation but are not sufficient for all CS synthesis in cartilage.

A comparative study of chondroitin sulfate and heparan sulfate for directing three-dimensional chondrogenesis of mesenchymal stem cells

BackgroundMesenchymal stem cells (MSCs) hold great promise for cartilage repair given their relative abundance, ease of isolation, and chondrogenic potential. To enhance MSC chondrogenesis, extracellular matrix components can be incorporated into three-dimensional (3D) scaffolds as an artificial cell niche. Chondroitin sulfate (CS)-containing hydrogels have been shown to support 3D chondrogenesis, but the effects of varying CS concentration and hydrogel stiffness on 3D MSC chondrogenesis remains elusive. Heparan sulfate (HS) is commonly used as a growth factor reservoir due to its ability to sequester growth factors; however, how it compares to CS in supporting 3D MSC chondrogenesis remains unknown.MethodsWe fabricated photocrosslinkable hydrogels containing physiologically relevant concentrations (0–10%) of CS or HS with two stiffnesses (~7.5 kPa and ~ 36 kPa) as a 3D niche for MSC chondrogenesis.ResultsCS is a more potent factor in enhancing MSC chondrogenesis, especially in soft hydrogels (~ 7.5 kPa). A moderate dosage of CS (5%) led to the highest amount of neocartilage deposition. Stiff hydrogels (~ 36 kPa) generally inhibited neocartilage formation regardless of the biochemical cues.ConclusionsTaken together, the results from this study demonstrated that CS-containing hydrogels at low mechanical stiffness can provide a promising scaffold for enhancing MSC-based cartilage tissue regeneration.

Ageing affects chondroitin sulfates and their synthetic enzymes in the intervertebral disc

The depletion of chondroitin sulfates (CSs) within the intervertebral disc (IVD) during degenerative disc disease (DDD) results in a decrease in tissue hydration, a loss of fluid movement, cell apoptosis, a loss of nerve growth inhibition and ultimately, the loss of disc function. To date, little is known with regards to the structure and content of chondroitin sulfates (CSs) during IVD ageing. The behavior of glycosaminoglycans (GAGs), specifically CSs, as well as xylosyltransferase I (XT-I) and glucuronyltransferase I (GT-I), two key enzymes involved in CS synthesis as a primer of glycosaminoglycan (GAG) chain elongation and GAG synthesis in the nucleus pulposus (NP), respectively, were evaluated in a bovine ageing IVD model. Here, we showed significant changes in the composition of GAGs during the disc ageing process (6-month-old, 2-year-old and 8-year-old IVDs representing the immature to mature skeleton). The CS quantity and composition of annulus fibrosus (AF) and NP were determined. The expression of both XT-I and GT-I was detected using immunohistochemistry. A significant decrease in GAGs was observed during the ageing process. CSs are affected at both the structural and quantitative levels with important changes in sulfation observed upon maturity, which correlated with a decrease in the expression of both XT-I and GT-I. A progressive switch of the sulfation profile was noted in both NP and AF tissues from 6 months to 8 years. These changes give an appreciation of the potential impact of CSs on the disc biology and the development of therapeutic approaches for disc regeneration and repair.

Chondroitin Sulfate-Based Biomineralizing Surface Hydrogels for Bone Tissue Engineering.

Chondroitin sulfate (CS) is the major component of glycosaminoglycan in connective tissue. In this study, we fabricated methacrylated PEGDA/CS-based hydrogels with varying CS concentration (0, 1, 5, and 10%) and investigated them as biomineralizing three-dimensional scaffolds for charged ion binding and depositions. Due to its negative charge from the sulfate group, CS exhibited an osteogenically favorable microenvironment by binding charged ions such as calcium and phosphate. Particularly, ion binding and distribution within negatively charged hydrogel was dependent on CS concentration. Furthermore, CS dependent biomineralizing microenvironment induced osteogenic differentiation of human tonsil-derived mesenchymal stem cells in vitro. Finally, when we transplanted PEGDA/CS-based hydrogel into a critical sized cranial defect model for 8 weeks, 10% CS hydrogel induced effective bone formation with highest bone mineral density. This PEGDA/CS-based biomineralizing hydrogel platform can be utilized for in situ bone formation in addition to being an investigational tool for in vivo bone mineralization and resorption mechanisms.

Molecular characterization of CHST11 and its potential role in nacre formation in pearl oyster Pinctada fucata martensii

Background : C4ST-1 catalyses the transfer of sulfate groups in the sulfonation of chondroitin during chondroitin sulfate synthesis. Chondroitin sulfate consists of numerous copies of negatively charged sulfonic acid groups that participate in the nucleation process of biomineralization. In the present study, we obtained two CHST11 genes ( PmCHST11a and PmCHST11b ) which encoded the C4ST-1 and explored the functions of these genes in the synthesis of chondroitin sulfate and in the formation of the nacreous layer of shells. Results: Both PmCHST11a and PmCHST11b had a sulfotransferase-2 domain, a signal peptide and a transmembrane domain. These properties indicated that these genes localise in the Golgi apparatus. Real-time PCR revealed that both PmCHST11a and PmCHST11b were highly expressed in the central zone of the mantle tissue. Inhibiting PmCHST11a and PmCHST11b via RNA interference significantly decreased the expression levels of these genes in the central zone of the mantle tissue and the concentration of chondroitin sulfate in extrapallial fluid. Moreover, shell nacre crystallised irregularly with a rough surface after RNA interference. Conclusions: This study indicated that PmCHST11a and PmCHST11b are involved in the nacre formation of P. fucata martensii through participating in the synthesis of chondroitin sulfate. Normal 0 false false false EN-US ZH-CN X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:Tabla normal; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.5pt; mso-bidi-font-size:11.0pt; font-family:Calibri,sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Times New Roman; mso-bidi-theme-font:minor-bidi; mso-font-kerning:1.0pt; mso-fareast-language:ZH-CN;}

Features of metabolic disorders in patients with hernias of lumbar spine associated with arterial hypertension

The combination of arterial hypertension (AH) with the spine pathology is a natural manifestation of connected structural-functional disorders.Aim is to determine the features of metabolic disorders in the organism of patients with hernias of lumbar spine, associated with arterial hypertension (AH), on the base of clinical-biochemical and immunological markers.Materials and methods. The study was carried out in 2016 on the base of SI “Institute of spine and joints pathology, named after prof. M.I. Sitenko, NAMS of Ukraine”. There were examined 37 patients with lumbar spine hernias: 23 – without concomitant pathologies, 14 – in association with AH of 1–2 degrees of І–ІІ stage. The control group – 30 persons, 13 men and 17 women. General protein, glycoproteins, chondroitin sulfates, C-reactive protein, glucose, urea, creatinine, β-lipoproteins, calcium, ALAT, ASAT, CPK, LDG, IL-1, IL-4 and IL-6 were determined in blood serum.Results of research. The content of interleukins in the group of patients with AH was higher than in the group of patients without it. In the group of patients with AH IL-1 was increased in 10,5 times, IL-6 – in 5,3 times that testifies to heavier immunologic disorders in patients with AH. The content of glycoproteins in patients without AH was increased by 55,8 %, in patients with AH – in 2,1 times comparing with control. The content of chondroitin sulfates in patients with hernias without AH was increased in 2,8 times, in ones with AH – in 4 times. LDG was increased, CPK – decreased in both groups. β-lipoproteins in blood of patients without concomitant AH was increased by 44,6 %, in patients with AH – by 67 %.Conclusions. In patients with lumbar spine hernias, associated with AH were observed heavier inflammatory-destructive disorders in the cartilage tissue. The increased content of β-lipoproteins in blood of patients with lumbar spine hernias, associated with AH comparing with patients without it may indicate age disorders of lipids metabolism. IL-1 and IL-6 уin patients with lumbar spine hernias, associated with AH were higher comparing with patients without it

A practical method for determining chondroitin sulfate content in shark cartilage materials and products

A practical method for determining chondroitin sulfate content in shark cartilage materials and products

ВЫСОКИЙ УРОВЕНЬ ГЛИКОЗАМИНОГЛИКАНОВ СЫВОРОТКИ КРОВИ КАК НЕЗАВИСИМЫЙ ПРЕДИКТОР РАЗВИТИЯ СПАЕЧНОЙ БОЛЕЗНИ БРЮШИНЫ

Peritoneal commissures is the phenomenon of adhesions formation with apparent symptom group of violations in functioning of the gastrointestinal tract and small pelvis. Unlike other postoperative complications adhesive disease is characterized by high risk for life manifestations in the form of intestinal obstruction, infertility and chronic pain. It is known that in the process of restoring the integrity of the peritoneum the decisive role is given to makrophages, peritoneal immune system, processes of angiogenesis, increasing fibroblasts and collagen production. However, the biochemical processes taking place in the intercellular matrix of connective tissue, particularly, the exchange of glycosaminoglycans in the formation of peritoneal adhesions have not been studied. In this context, the main objective of this work was to study levels of glycosaminoglycans of the blood in the development of adhesive disease and justification of the use of indicators to exchange connective tissue as additional prognostic criteria for the development of postoperative adhesions. An open prospective randomized trial analyzed the data of 67 patients that were admitted for the planned reconstructive surgery on the anterior abdominal wall to eliminate postoperative ventral hernia, at the surgery department of State Clinical Hospital No. 8, city ofUFAduring the period of 2005-2008. The severity of adhesions was assessed and contrasted to the results of determination of glycosaminoglycans in blood serum. The total content of glycosaminoglycans, hyaluronic acid, chondroitin sulfates and heparan sulfates was determined. Postoperative monitoring stages of the studied indicators -3, 5 and 7 day after herniotomy. The group of patients with adhesive peritoneum desease showed total content of glycosaminoglycans by 19.2% (p < 0.0001) higher than reference values. It should be noted that there was continuous growth of all factions of glycosaminoglycans during all seven days of the postoperative monitoring. The growth of the overall content of the glycosaminoglycans made 261% (p < 0.0001) compared with the control group. Method of ROC-analysis established that the area under the ROC curve is a common glycosaminoglycans is equal 0.824 ± 0.055, 95% confidence interval-0.906 0.711. Sensitivity-70.4% (54.8 -86.0), specificity is 97.5% (82.4 -98.6). The optimum point for the division amounted to 30 μmol/l. Positive predicting level amounted to 96.8%, negative 75.1%. Thus, the assessment of the level of glycosaminoglycans can serve as an additional prognostic test to diagnose disturbance in intercellular matrix that can be used in the diagnosis of commissural process and in monitoring the effectiveness of preventive measures.

Chondroitin Sulfate Inhibits Monocyte Chemoattractant Protein-1 Release From 3T3-L1 Adipocytes: A New Treatment Opportunity for Obesity-Related Inflammation?

Monocyte chemoattractant protein-1 (MCP-1) overproduction from inflamed adipose tissue is a major contributor to obesity-related metabolic syndromes. 3T3-L1 embryonic fibroblasts were cultured and differentiated into adipocytes using an established protocol. Adipocytes were treated with lipopolysaccharide (LPS) to induce inflammation and thus MCP-1 release. At the same time, varying concentrations of chondroitin sulfate (CS) were added in a physiologically relevant range (10-200 µg/mL) to determine its impact on MCP-1 release. Chondroitin sulfate, a natural glycosaminoglycan of connective tissue including the cartilage extracellular matrix, was chosen on the basis of our previous studies demonstrating its anti-inflammatory effect on macrophages. Because the main action of MCP-1 is to induce monocyte migration, cultured THP-1 monocytes were used to test whether CS at the highest physiologically relevant concentration could inhibit cell migration induced by human recombinant MCP-1. Chondroitin sulfate (100-200 µg/mL) inhibited MCP-1 release from inflamed adipocytes in a dose-dependent manner (P < .01, 95% confidence interval [CI]: −5.89 to −3.858 at 100 µg/mL and P < .001, 95% CI: −6.028 to −3.996 at 200 µg/mL) but had no effect on MCP-1–driven chemotaxis of THP-1 monocytes. In summary, CS could be expected to reduce macrophage infiltration into adipose tissue by reduction in adipocyte expression and release of MCP-1 and as such might reduce adipose tissue inflammation in response to pro-inflammatory stimuli such as LPS, now increasingly recognized to be relevant in vivo.

Serum levels of endothelial glycocalyx constituents in women at 20 weeks' gestation who later develop gestational diabetes mellitus compared to matched controls: a pilot study

ObjectivesThe aim of this pilot study was to determine the serum concentration of heparan sulfate, hyaluronan, chondroitin sulfate and syndecan-1 and if these serum concentrations can be used to identify...

Extraction of Glycosaminoglycans Containing Glucosamine and Chondroitin Sulfate from Chicken Claw Cartilage

Chicken cartilage (claw) is a waste of chicken cuts which are widely available in Indonesia. Cartilage part of chicken claw becomes a potential source of chondroitin sulfate (CS) and glucosamine (GS). This study aims to determine the most optimal extraction methods of CS and GS from cartilage of chicken claw. Various types of extraction methods used in this study are taken from the extraction by using boiling water (2 and 2.5 hours), acetic acid (7 and 17 hours), as well as proteolysis by papain (24 and 48 hours). Parameters observed include chemical characteristics of powdered cartilage of chicken claw as well as CS and GS levels in powdered cartilage of chicken claw extract. The results of this research show that the levels of CS and GS of chicken claw cartilage powder were 2.17% and 13%. Meanwhile, the highest GS level was obtained from the extraction with water treatment for 2.5 hours which was 8.1%. The treatment and duration of extraction will significantly affect the number of GS which was produced. The highest content of CS was obtained from the extraction with the enzyme treatment for 48 hours which was 2.47%. The best treatment is the extraction with water treatment for 2.5 hours which were the extracts with GS levels of 8.1% and 2.03% CS was selected through the analysis of multiple attribute .

RNA Contaminates Glycosaminoglycans Extracted from Cells and Tissues.

Glycosaminoglycans (GAGs) are linear negatively charged polysaccharides and important components of extracellular matrices and cell surface glycan layers such as the endothelial glycocalyx. The GAG family includes sulfated heparin, heparan sulfate (HS), dermatan sulfate (DS), chondroitin sulfate (CS), keratan sulfate, and non-sulfated hyaluronan. Because relative expression of GAGs is dependent on cell-type and niche, isolating GAGs from cell cultures and tissues may provide insight into cell- and tissue-specific GAG structure and functions. In our objective to obtain structural information about the GAGs expressed on a specialized mouse glomerular endothelial cell culture (mGEnC-1) we adapted a recently published GAG isolation protocol, based on cell lysis, proteinase K and DNase I digestion. Analysis of the GAGs contributing to the mGEnC-1 glycocalyx indicated a large HS and a minor CS content on barium acetate gel. However, isolated GAGs appeared resistant to enzymatic digestion by heparinases. We found that these GAG extracts were heavily contaminated with RNA, which co-migrated with HS in barium acetate gel electrophoresis and interfered with 1,9-dimethylmethylene blue (DMMB) assays, resulting in an overestimation of GAG yields. We hypothesized that RNA may be contaminating GAG extracts from other cell cultures and possibly tissue, and therefore investigated potential RNA contaminations in GAG extracts from two additional cell lines, human umbilical vein endothelial cells and retinal pigmental epithelial cells, and mouse kidney, liver, spleen and heart tissue. GAG extracts from all examined cell lines and tissues contained varying amounts of contaminating RNA, which interfered with GAG quantification using DMMB assays and characterization of GAGs by barium acetate gel electrophoresis. We therefore recommend routinely evaluating the RNA content of GAG extracts and propose a robust protocol for GAG isolation that includes an RNA digestion step.

Suppressive Activity of Chondroitin Sulfate on Periostin Production by Synoviocytes from Knee Osteoarthritis In Vitro

Background: Knee osteoarthritis (OA) is well known to be one of the most common joint disease in elderly people and characterized by pain, stiffness and loss of function in articulating joints, which develop slowly and worsen over time. Chondroitin sulfate (CS) and glucosamine hydrochloride are used for the treatment of OA and reported that these substances are favorably modify the clinical symptoms, especially pain, however, the therapeutic mechanisms of these substances are not fully understood. The present study was undertaken to examine the influence of CS on the production of periostin, which is one of the important molecules associated with OA development, by synoviocytes from an OA patient by an in vitro cell culture technique. Methods: Synoviocytes (1 x 105 cell/ml) were stimulated with 10.0 ng/ml IL-13 in the presence of various concentrations of CS. After 48 h, periostin content in culture supernatants was examined by ELISA. We also examined the influence of CS on transcription factor, STAT6, activation and periostin mRNA expression in synoviocytes 12 and 24 h after IL-13 stimulation, respectively. Results: Addition of CS into cell cultures caused the suppression of periostin production from synoviocytes induced by IL-13 stimulation through the inhibition of STAT6 activation and periostin mRNA expression. The minimum concentration of CS that caused significant suppression of periostin production, STAT6 activation and mRNA expression was 10.0 μg/ml. Conclusion: These results strongly suggest that the ability of CS to suppress periostin production from synoviocytes may account, at least in part, for the clinical efficacy of CS on OA.

PROMOTION EFFECT OF CHONDROITIN SULFATE ON PROLIFERATION OF MYOBLASTS

To research the effect of chondroitin sulfate (CS) on the proliferation of myoblasts and the formation of myotube. The myoblasts at passage 5 were used to prepare the cells suspension (1×108 cells/mL), and the experiment was divided into 4 groups based on CS concentration in the medium:group A (0 μg/mL), group B (50 μg/mL), group C (100 μg/mL), and group D (200 μg/mL). The cell morphology and myotube formation were observed by inverted microscope at 4, 5, and 8 days after treatment; MTT was used to detect the cell proliferation at 6 days, and the number of myotube was calculated by HE staining at 8 days. Cells showed spindle shape after adherent, with ovoid nuclei and dense cytoplasm under inverted microscope. When the cell adherent rate was 90%, cells arranged in whorls swirled and showed long fusiform adherent growth; and then nuclei fusion resulted in formation of multincleated myotubes. At 8 days, most myoblasts fused to form myotube in group A, but less myotube was observed in groups B and C, and the least myotube in group D. The absorbance (A) values of groups A, B, C, and D were 0.045 2±0.004 4, 0.540 4±0.096 7, 0.660 9±0.143 4, and 1.069 0±0.039 0 respectively, showing significant difference between other groups (P<0.05) except between groups B and C P>0.05). HE staining observation showed that most myoblasts fused to form myotube in group A, but less myotube in groups B and C, and the least myotube in group D. The number of myotube of groups A, B, C, and D were 222.01±30.02, 193.13±42.46, 170.26±11.96, and 136.88±16.78 respectively, showing no significant difference among groups (F=1.658, P=0.252). CS can significantly promote the proliferation of myoblast, the promotion is the biggest when CS concentration is 200 μg/mL.

Chondroitin sulphate inhibits NF-κB activity induced by interaction of pathogenic and damage associated molecules

To evaluate the anti-inflammatory mechanism of action of Chondroitin Sulphate (CS). THP-1 macrophages were cultured with a range of sizes and concentrations of HA fragments with TLR4 (LPS in a physiologically relevant concentration determined by analyses of sera of a community clinic ascertained knee osteoarthritis (OA) cohort) or TLR2 (heat killed listeria bacteria) agonists and varying concentrations of CS in a physiologically relevant range (10-200μg/ml). We measured IL-1β release, intracellular IL-1β, proIL-1β, caspase-1 and NF-κB activity and DNA binding activity of NF-κB transcription factors from nuclear and cytoplasmic extracts. Serum LPS was significantly associated with radiographic knee joint space narrowing (JSN) (P=0.02) in the OA cohort (n=40). The priming dose of LPS used for these experiments (10ng/ml) was below the lowest serum concentration of the OA cohort (median 47.09, range 14.43-81.36ng/ml). Priming doses of LPS and HA fragments alone did not elicit an inflammatory response. However, primed with LPS, HA fragments produced large dose-dependent increases in IL-1β that were inhibitable by CS. CS did not inhibit caspase-1 activity but in physiologically achievable concentrations, attenuated NF-κB activity induced by either the TLR4 (LPS 1000ng/ml) or TLR2 agonists alone or in combination with HA fragments. LPS induced and CS significantly reduced activity of canonical NF-κB transcription factors, p65, p50, c-Rel and RelB. Subinflammatory concentrations of pathogenic (LPS, listeria) and damage associated (HA) molecules interact to induce macrophage-related inflammation. CS works upstream of the inflammasome by inhibiting activation of NF-κB transcription factors.

The combined application of human adipose derived stem cells and Chondroitinase ABC in treatment of a spinal cord injury model

BackgroundAlthough stem cell therapy has become a major focus as a new option for management of spinal cord injury (SCI), its effectiveness should be promoted. In this study, we investigated the effects of co-administrating human adipose-derived stem cells (hADSCs) and Chondroitinase ABC (ChABC) in a rat model of spinal cord injury. Material and methodshADSCs derived from superficial layer of abdominal adipose tissue were used to treat a contusion-induced SCI. Animals were randomly allocated to five equal groups including sham (only laminectomy), SCI (SCI+vehicle injection), hADSCs (1×10⁶ hADSCs/10μl intra-spinal injection), ChABC (10μl of 100U/ml ChABC intra-spinal injection injection), and hADSCs+ChABC. Basso, Beattie and Bresnahan tests were used to evaluate locomotor function. 8weeks after treatment, cavity size, myelination, cell differentiation (neuron and astrocyte), and chondroitin sulfate amount were analyzed. ResultshADSC transplanted animals, ChABC injected animals (P<0.001), and hADSC+ChABC treated rats (P<0.001) displayed significant motor improvement compared to SCI group. Combination therapy of hADSCs and ChABC led to greater locomotor recovery compared to using hADSCs (P<0.001) or ChABC (P<0.01) alone. Spinal cords in the combined and single therapy groups had cavities filled with myelinated areas and less chondroitin sulfate content in comparison with the control group (P<0.001). hADSCs expressed GFAP, B III tubulin and Map2. ConclusionCombination therapy with ChABC and hADSCs exhibits more significant functional recovery than single therapy using either. This result may be applicable in selection of the best therapeutic strategy for SCI.

Application of Chondroitin Sulfate on Organogenesis of Two &lt;i&gt;Cymbidium&lt;/i&gt; spp. under Different Sources of Lights

The aim of this study was to present chondroitin sulfate as a plant growth regulator and to give an overview about light effects on PLBs (protocorm like bodies) culture of Cymbidium dayanum and Cymbidium finlaysonianum cultured in vitro. Chondroitin sulfate is a sulfated glycosaminoglycan (GAG) composed of a chain of alternating sugars N-acetylgalactosamine and glucuronic acid. It is widely used as a material for food ingredients, cosmetics and medicine. PLBs were cultured on modified MS medium containing different concentration of chondroitin sulfate (0, 0.1, 1 and 10 mg/l), under four sources of lights: conventional white fluorescent tube, red LED, green LED and blue LED. In C. dayanum, 100% PLBs formation rate was observed at 0.1 mg/l chondroitin sulfate with modified MS medium under green LED and 1 mg/l chondroitin sulfate under blue LED; the maximum shoots and roots formation were observed under green LEDs (93% and 80% respectively) when media contained 0.1 mg/l chondroitin sulfate. In C. finlaysonianum, every concentrations of chondroitin sulfate enhanced the growth rate of PLBs when compared to control treatment, under all four sources of lights. The highest values were recorded with 0.1 mg/l chondroitin sulfate which induced 100% PLBs formation under blue LED, while 10 mg/l chondroitin sulfate had induced 100% PLBs formation under green LED. The highest percentage of shoots (73%) was initiated in the medium containing 10 mg/l chondroitin sulfate under green LED. Plant development was strongly influenced by the light quality and plant growth regulator functions as chemical messengers for intercellular communication of plant. The results demonstrated that low concentrations of chondroitin sulfate could promote PLBs, shoots and roots formation of Cymbidium spp. under green and blue LED.

A Study on Development of High Functional Materials Producing Technique using By-products from Skate Processing (1) - Development of Chondroitin Sulfate Materials using Skate Cartilages

For the purpose of reuse the wasted by-products from the skate process to the health functional food or medicinal material, chondroitin sulfate was extracted from the skate cartilage with the method of hydrolysis with protease enzyme, and the contents of chondroitin sulfate and hydrolyzed protein were measured qualitatively and quantitatively. The effects of chondroitin sulfate on body weight or liver weight changes, hepatotoxicity elimination and anti-inflammatory actions were measured from in vivo test with feed-treated mice. From the hydrolytic extraction of skate cartilage with the mixture of 1% alcalase and 1% protease for 4 hours, the extraction yield of chondroitin sulfate was about 32.55%. The content and molecular weight of chondroitin sulfate was 26.63% and ., respectively and the content ratio of chondroitin sulfate to protein was measured to 1 to 2.76 with gel permeation chromatography. For the odor component, trimethylamine decreased about 30% but almost not ammonia from chondroitin sulfate with the treatment of activated carbon. From the feeding chondroitin sulfate to mice, the control effect of body and liver weights decrease was measured, anti-inflammatory action and hepatotoxicity elimination action were also measured. From these results, process operation conditions for manufacturing of chondroitin sulfate were suggested.

Structural characteristics are crucial to the benefits of guar gum in experimental osteoarthritis

Protein-free guar gum (DGG) was oxidized (DGGOX) or sulfated (DGGSU) by insertion of new groups in C-6 (manose) and C-6 (galactose), for DGGOX and DGGSU, respectively. Rats were subjected to anterior cruciate ligament transection (ACLT) of the knee, joint pain recorded using the articular incapacitation test, and the analgesic effect of intraarticular 100μg DGG, DGGOX or DGGSU solutions at days 4–7 was evaluated. Other groups received DGG or saline weekly, from days 7 to 70 and joint damage assessed using histology and biochemistry as the chondroitin sulfate (CS) content of cartilage. The molar mass of CS samples was obtained by comparing their relative electrophoretic mobility to standard CS. DGG but not DGGOX or DGGSU significantly inhibited joint pain. DGG significantly reversed the increase in CS, its reduced electrophoretic mobility, and histological changes following ACLT, as compared to vehicle. Structural integrity accounts for DGG benefits in experimental osteoarthritis.

S&T-51 EXTRACELLULAR MATRIX CHARACTERISTICS OF TUNICA ALBUGINEA IN MEN WITH AND WITHOUT PEYRONIE'S DISEASE

INTRODUCTION AND OBJECTIVES: Peyronie’s disease (PD) is characterized by the formation of fibrous plaques in the tunica albuginea (TA) of the penis, causing pain and penile curvature during erection, with great impact on quality of life of patients. There is a lack of highly effective medical treatments for PD, mainly because it’s pathophysiology is not completely understood. The aim of this study was to compare possible markers related to alterations and remodeling of extracellular matrix which can be involved on the development of the disease in men with and without PD METHODS: Samples of TA of 10 patients with PD were obtained at the moment of surgical correction of penile curvature. Samples of TA of 10 corpses with normal penis at the physical exam were obtained (control group). Tissues were processed and stained with Hematoxilin, submitted to immunohistochemistry (IHQ) with the following primary antibodies: heparanase (HPSE), heparanase2 (HPSE2), metalloproteinases 2 and 9 (MMP2 and MMP9) and TGF-s. IHQ reaction used peroxidase kit LSAB -peroxidase (DakoCytomation , Copenhagen, Dinamarca). Genic expression of heparanase-1 (HPSE), heparanase2 (HPSE2), metalloprotease 9 (MMP9), syndecan 1 (SYN1), interleukin 6 (IL-6), hyaluronic acid sintetase 1 e 2 (HAS1 e HAS2), hyaluronidases 1 e 2 (HYL1 e HYL2), was done by quantitative RTPCR (qRT-PCR). Statystical analyses were done using software Graph PRISM 5 . Project was reviewed and approved by the Research Ethical Committee of Faculdade de Medicina do ABC, (# 242/2010). RESULTS: Histopathological evaluation showed higher number of cells, reduction of apoptosis events, alteration of the distribution of collagen fibers and diminished number of vessels on TA of patients with PD compared to the controls TA. qRT-PCR analysis showed increased concentration of TGF-s and interleucine1 reduction on HSPE-1 expression and decreased concentration of dermatan sulfate and chondroitin sulfate in patients with PD (p<0.05). However, there were no alterations on HPSE2, MMP2 e MMP9 expressions. Those findings suggest a higher remodeling of extracellular matrix in patients with PD. CONCLUSIONS: In comparison with the controls the TA of patients with PD showed lesser number of vessels and apoptosis events; increased concentration of TGF-s and interleukin-1, reduction of heparanase-1 expression and decreased concentration of dermatan sulfate and chondroitin sulfate.