OBJECTIVES: This study aims to investigate the effects and mechanisms of chondroitin sulfate (CS), dermatan sulfate (DS), and heparin (HEP) on chondrogenesis of murine chondrogenic cell line (ATDC5) cells and the maintenance of murine articular cartilage in vitro. METHODS: ATDC5 and articular cartilage tissue explant were cultured in the medium containing different sulfated glycosaminoglycans. Cell proliferation, differentiation, cartilage formation, and mechanism were observed using cell proliferation assay, Alcian blue staining, real-time quantitative polymerase chain reaction (RT-qPCR), and Western blot, respectively. RESULTS: Results showed that HEP and DS primarily activated the bone morphogenetic protein (BMP) signal pathway, while CS primarily activated the protein kinase B (AKT) signal pathway, further promoted ATDC5 cell proliferation and matrix production, and increased Sox9, Col2a1, and Aggrecan expression. CONCLUSIONS This study investigated the differences and mechanisms of different sulfated glycosaminoglycans in chondrogenesis and cartilage homeostasis maintenance. HEP promotes cartilage formation and maintains the normal state of cartilage tissue in vitro, while CS plays a more effective role in the regeneration of damaged cartilage tissue.
Animals ; Mice ; Cartilage/metabolism* ; Cell Differentiation ; Cells, Cultured ; Chondrocytes/metabolism* ; Chondrogenesis/physiology* ; Glycosaminoglycans/pharmacology*

OBJECTIVE: The effects of the staurosporine on contraction of self-assembled constructs and extracellular matrix syntheses of goat temporomandibular joint discs were investigated. METHODS: Goat temporomandibular joint disc cells were isolated and cultured to P3, and 5.5×10⁶ cells were combined with different concentrations of staurosporine (0, 0.1, 1, 10, 100 nmol·L⁻¹) in agarose wells and cultured for one week. The samples were frozen and sectioned. Safranin-O,  Picro-sirius red and immunohistochemical staining were performed to observe the distributions of the extracellular matrix and the expression of alpha-smooth muscle actin (α-SMA). Enzyme linked immunosorbent assay (ELISA) and Blyscan kits were utilized to quan--titatively detect the contents of type Ⅰ collagen (ColⅠ) and glycosaminoglycans (GAGs). RESULTS: Each group of goat temporo-mandibular joint disc cells in the agarose wells were gathered to self-assemble into a disc-shaped base for 4 hours and then to gradually contract into a round shape. The Picro-sirius red staining was strong and indicated collagen distribution. The Safranin-O staining observed GAGs throughout the entire construct. The expression of ColⅠ was strongly posi-tive in the staurosporine groups; however, the expression of α-SMA was weak. ColⅠ and GAGs contents in the stau-rosporine groups were greater than that of the control group, especially in the 10 nmol·L⁻¹ group (P<0.01). CONCLUSIONS Staurosporine has a certain effect on the shrinkage of self-assembled constructs; however, such effect is not prominent. Staurosporine contributes to the construction synthesis of extracellular matrix.
Animals ; Collagen Type I ; Glycosaminoglycans ; Goats ; Staurosporine ; pharmacology ; Temporomandibular Joint ; Temporomandibular Joint Disc ; cytology ; drug effects

Hyaluronic acid (HA) has long been studied in diverse applications. It is a naturally occurring linear polysaccharide in a family of unbranched glycosaminoglycans, which consists of repeating di-saccharide units of N-acetyl-D-glucosamine and D-glucuronic acid. It is almost ubiquitous in humans and other vertebrates, where it participates in many key processes, including cell signaling, tissue regeneration, wound healing, morphogenesis, matrix organization, and pathobiology. HA is biocompatible, biodegradable, muco-adhesive, hygroscopic, and viscoelastic. These unique physico-chemical properties have been exploited for several medicinal purposes, including recent uses in the adjuvant treatment for chronic inflammatory disease and to reduce pain and accelerate healing after third molar intervention. This review focuses on the post-operative effect of HA after third molar intervention along with its various physio-chemical, biochemical, and pharmaco-therapeutic uses.
Acetylglucosamine ; Biocompatible Materials ; Glycosaminoglycans ; Humans ; Hyaluronic Acid ; Molar, Third ; Morphogenesis ; Regeneration ; Vertebrates ; Wound Healing

Mucopolysaccharidosis (MPS) is an inherited metabolic disorder caused by a deficiency in enzymes that participate in the degradation of glycosaminoglycans (GAGs) such as heparin sulfate and dermatan sulfate. Left untreated, patients show progressive mental and physical deterioration due to deposition of GAGs in organs. Death often occurs due to cardiac or respiratory failure before patients reach their early twenties. MPS has several oral and dental manifestations. An enlarged head, short neck, and open mouth associated with a large tongue are major characteristics of MPS patients. Dental complications can be severe, including unerupted dentition, dentigerous cyst-like follicles, malocclusions, condylar defects, and gingival hyperplasia. A 21-year-old female patient with MPS was described in this article, with special emphasis on oral manifestations and dental treatment.
Dental Pulp Cavity ; Dentition ; Dermatan Sulfate ; Female ; Gingival Hyperplasia ; Glycosaminoglycans ; Head ; Heparin ; Humans ; Malocclusion ; Mouth ; Mucopolysaccharidoses ; Neck ; Oral Manifestations ; Respiratory Insufficiency ; Root Canal Therapy ; Tongue ; Young Adult

It is very useful to evaluate the content and 3D distribution of extracellular matrix non-destructively in tissue engineering. This study evaluated the feasibility of using micro-computed tomography (µCT) with Hexabrix to measure quantitatively sulfated glycosaminoglycans (GAGs) of engineered cartilage. Rabbit chondrocytes at passage 2 were used to produce artificial cartilages in polyglycolic acid scaffolds in vitro. Engineered cartilages were incubated with Hexabrix 320 for 20 min and analyzed via µCT scanning. The number of voxels in the 2D and 3D scanning images were counted to estimate the amount of sulfated GAGs. The optimal threshold value for quantification was determined by regression analysis. The 2D µCT images of an engineered cartilage showed positive correlation with the histological image of Safranin-O staining. Quantitative data obtained with the 3D µCT images of 14 engineered cartilages showed strong correlation with sulfated GAGs contents obtained by biochemical analysis (R² = 0.883, p < 0.001). Repeated exposure of engineered cartilages to Hexabrix 320 and µCT scanning did not significantly affect cell viability, total DNA content, or the total content of sulfated GAGs. We conclude that µCT imaging using Hexabrix 320 provides high spatial resolution and sensitivity to assess the content and 3D distribution of sulfated GAGs in engineered cartilages. It is expected to be a valuable tool to evaluate the quality of engineered cartilage for commercial development in the future.
Cartilage* ; Cell Survival ; Chondrocytes ; DNA ; Extracellular Matrix ; Glycosaminoglycans* ; In Vitro Techniques ; Ioxaglic Acid ; Polyglycolic Acid ; Tissue Engineering

Clinical investigations have shown a significant relationship between osteoarthritis (OA) and estrogens levels in menopausalwomen. Therefore, treatment with exogenous estrogens has been shownto decrease the risk ofOA.However, the effect estrogen has not been clearly demonstrated in the chondrocytes using phytoestrogens, which lack the specific side-effects of estrogens, may provide an alternative therapy. This study was designed to examine the possible effects of phytoestrogen (daidzein) on human chondrocyte phenotype and extracellular matrix formation. Phytoestrogens which lack the specific side-effects of estrogens may provide beneficial effect without causing hormone based side effect. Human chondrocytes cells were cultured in 2D (flask) and 3D (PCL-CA scaffold) systems. Daidzein cytotoxic effect was determined by MTT assay. Chondrocyte cellular content of glycosaminoglycans (GAGs), total collagen and chondrogenic gene expression were determined in both culture systems after treatment with daidzein.Daidzein showedtime-dependent and dose-independent effects on chondrocyte bioactivity.Thecompound at low doses showed significant (p<0.05) increase in total collagen andGAGsproduction at similar levels in 2Dand 3Dculture environment. The mRNA levels of Collagen II and Sox9 were increased significantly (p<0.01) after the treatment while the upregulation in COMP expression was statistically insignificant (p>0.05). The expression levels of Fibronectin, Laminin and Integrin b1were significantly increased especially in3Dculture system. This studywas illustrated the potential positive effects of daidzein onmaintenance of human chondrocyte phenotype and extracellular matrix formation suggesting an attractive and viable alternative therapy for OA.
Chondrocytes* ; Collagen ; Estrogens ; Extracellular Matrix ; Fibronectins ; Gene Expression ; Glycosaminoglycans ; Humans* ; Laminin ; Osteoarthritis ; Phenotype* ; Phytoestrogens* ; RNA, Messenger ; Up-Regulation

In this paper we report the differentiating properties of platelet-rich plasma releasates (PRPr) on human chondrocytes within elastomeric polycaprolactone triol–citrate (PCLT–CA) porous scaffold. Human-derived chondrocyte cellular content of glycosaminoglycans (GAGs) and total collagen were determined after seeding into PCLT–CA scaffold enriched with PRPr cells. Immunostaining and real time PCR was applied to evaluate the expression levels of chondrogenic and extracellular gene markers. Seeding of chondrocytes into PCLT–CA scaffold enriched with PRPr showed significant increase in total collagen and GAGs production compared with chondrocytes grown within control scaffold without PRPr cells. The mRNA levels of collagen II and SOX9 increased significantly while the upregulation in Cartilage Oligomeric Matrix Protein (COMP) expression was statistically insignificant. We also report the reduction of the expression levels of collagen I and III in chondrocytes as a consequence of proximity to PRPr cells within the scaffold. Interestingly, the pre-loading of PRPr caused an increase of expression levels of following extracellular matrix (ECM) proteins: fibronectin, laminin and integrin β over the period of 3 days. Overall, our results introduce the PCLT–CA elastomeric scaffold as a new system for cartilage tissue engineering. The method of PRPr cells loading prior to chondrocyte culture could be considered as a potential environment for cartilage tissue engineering as the differentiation and ECM formation is enhanced significantly.
Blood Platelets* ; Cartilage Oligomeric Matrix Protein ; Cartilage* ; Chondrocytes ; Collagen ; Elastomers ; Extracellular Matrix* ; Fibronectins ; Glycosaminoglycans ; Humans* ; Laminin ; Methods ; Phenotype* ; Platelet-Rich Plasma ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; Tissue Engineering ; Up-Regulation

OBJECTIVETo study the effects of pulsed ultrasound (PUS) and pulsed electromagnetic fields (PEMF) on the secretion of extracellular matrix from a culture complex during in vitro chondrogenesis.

METHODSAll the rat bone marrow mesen- chymal stem cell pellets were cultured in achondrogenic medium. Different intensities of PUS (100, 150, and 200 mW · cm⁻²) and PEMF (1, 2, and 5 mT) were applied to the cell pellets for 2 weeks. Group N was cultured without PUS and PEMF stimu- lation as control. The culture medium was collected after 2 weeks of culture. Enzyme-linked immunosorbent assay (ELISA) was used to detect the type of collagen and glycosaminoglycan (GAG) in the culture medium.

RESULTSPUS increased the secreting-type collagen and GAG from cell pellets compared with group N (P < 0.05), whereas there was no difference in different intensities (P > 0.05). PEMF had no significant effect on the secretion of the type of collagen (P > 0.05). A PEMF of 1 mT had no significant effect on the secretion of GAG (P > 0.05). A PEMF 2 and 5 mT decreased the secretion of GAG (P < 0.05).

CONCLUSIONTo prevent the secretary of extracellular matrix may play a role in chondrogenic effect of PEMF.

Animals ; Bone Marrow Cells ; radiation effects ; Cells, Cultured ; Chondrogenesis ; radiation effects ; Electromagnetic Fields ; Extracellular Matrix ; Glycosaminoglycans ; Hematopoietic Stem Cells ; Mesenchymal Stromal Cells ; radiation effects ; Rats ; Ultrasonic Waves

BACKGROUND: There is evidence that glycosaminoglycans (GAGs) are present in the hair shaft within the follicle but there are no studies regarding GAGs isolation and measurement in the human hair shaft over the scalp surface, it means, in the free hair shaft. OBJECTIVE: The purpose of our research was to isolate and measure the total GAGs from human free hair shaft. METHODS: Seventy-five healthy individuals participated in the study, 58 adults, men and women over the age of 50 and 17 children (aged 4~9). GAGs in hair samples, received from the parietal and the occipital areas, were isolated with 4 M guanidine HCl and measured by the uronic acid-carbazole reaction assay. RESULTS: GAGs concentration was significantly higher in the occipital area than in the parietal area, in all study groups. GAG levels from both areas were significantly higher in children than in adults. GAG levels were not associated with gender, hair color or type. CONCLUSION: We report the presence of GAGs in the human free hair shaft and the correlation of hair GAG levels with the scalp area and participants' age.
Adult ; Child ; Female ; Glycosaminoglycans* ; Guanidine ; Hair Color ; Hair* ; Humans* ; Male ; Proteoglycans ; Rabeprazole ; Scalp

OBJECTIVEThis study aims to explore the clinical applicability and relevance of glycosaminoglycan Chemical Exchange Saturation Transfer (gagCEST) for intervertebral disc.

METHODS25 subjects ranging in age from 24 yrs to 74 yrs were enrolled. gagCEST was acquired using a single-slice TSE sequence on a 3T. Saturation used a continuous rectangular RF pulse with B1=0.8 µT and a fixed duration time=1100 ms. Sagittal image was obtained firstly without saturation pulse, and then saturated images were acquired at 52 offsets ranging from ±0.125 to ±7 parts per million (ppm). MR T2 relaxivity map was acquired at the identical location. Six subjects were scanned twice to assess scan-rescan reproducibility.

RESULTSGagCEST intraclass correlation coefficient (ICC) of six subjects was 0.759 for nucleus pulposus (NP) and 0.508 for annulus fibrosus (AF). Bland-Altman plots showed NP had a mean difference of 0.10% (95% limits of agreement: -3.02% to 3.22%); while that of AF was 0.34% (95% limits of agreement: -2.28% to 2.95%). For the 25 subjects, gag CEST in NP decreased as disc degeneration increased, with a similar trend to T2 relaxivity. Gag CEST of AF showed a better correlation with disc degeneration than T2 relaxivity.

CONCLUSIONGagCEST in NP and AF decreased as disc degeneration increased, while gagCEST in AF showed a better correlation than T2 relaxivity.

Adult ; Aged ; Biomarkers ; analysis ; Case-Control Studies ; Female ; Glycosaminoglycans ; chemistry ; metabolism ; Humans ; Intervertebral Disc ; chemistry ; metabolism ; Intervertebral Disc Degeneration ; diagnosis ; metabolism ; Lumbar Vertebrae ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged