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PubMed1979

Cell-free synthesis of hyaluronic acid in Marfan syndrome.

The cell-free synthesis of hyaluronic acid has been demonstrated in extracts of cultured human fibroblasts. Preparations from fibroblasts of normal individuals as well as those from patients with Marfan syndrome incorporate glucuronic acid and N-acetylglucosamine from their UDP derivatives into hyaluronic acid. Extracts from Marfan fibroblasts demonstrate 3 to 10 times more total and specific hyaluronic acid synthetase activity than do preparations from normal fibroblasts. All synthetic activity was found in particulate fractions with the bulk of activity localized in material sedimenting as large membrane fragments. Marfan and normal preparations exhibited similar properties with respect to substrate, cofactor, pH requirements, and heat stability. Neither the Marfan nor normal enzyme systems could be stimulated by exogenous acceptors, nor did either preparation contain a soluble factor which stimulated or inhibited the enzymic activity of the other. The genetic defect in Marfan syndrome appears to result in increased activity of hyaluronic acid synthetase without demonstrable changes in properties of the particulate enzymes involved.

Cell-Free System↗
PubMed2004

Therapeutic effects of hyaluronic acid on osteoarthritis of the knee. A meta-analysis of randomized controlled trials.

BACKGROUND: The magnitude of the therapeutic effects of intra-articular injection of hyaluronic acid on osteoarthritis of the knee is still in question. The aim of this meta-analysis was to elucidate the therapeutic efficacy and safety of intra-articular injection of hyaluronic acid for osteoarthritis of the knee. METHODS: We conducted a meta-analysis of twenty blinded randomized controlled trials that compared the therapeutic effect of intra-articular injection of hyaluronic acid with that of intra-articular injection of a placebo to treat osteoarthritis of the knee. The outcome end points were classified into three categories: pain with activities, pain without activities, and function. The outcome measures of the efficacy of hyaluronic acid were the mean differences in the efficacy scores between the hyaluronic acid and placebo groups. The outcome measure of the safety of hyaluronic acid was the relative risk of adverse events. RESULTS: Intra-articular injection of hyaluronic acid can decrease symptoms of osteoarthritis of the knee. We found significant improvements in pain and functional outcomes with few adverse events. However, there was significant between-study heterogeneity in the estimates of the efficacy of hyaluronic acid. Subgroup analysis and meta-regression analysis showed that lower methodological quality such as a single-blind or single-center design resulted in higher estimates of hyaluronic acid efficacy, that introduction of acetaminophen as an escape analgesic in the trial resulted in lower estimates of hyaluronic acid efficacy, and that patients older than sixty-five years of age and those with the most advanced radiographic stage of osteoarthritis (complete loss of the joint space) were less likely to benefit from intra-articular injection of hyaluronic acid. CONCLUSIONS: This meta-analysis confirmed the therapeutic efficacy and safety of intra-articular injection of hyaluronic acid for the treatment of osteoarthritis of the knee. Additional well-designed randomized controlled trials with high methodological quality are needed to resolve the continued uncertainty about the therapeutic effects of different types of hyaluronic acid products on osteoarthritis of the knee in various clinical situations and patient populations. LEVEL OF EVIDENCE: Therapeutic study, Level II-3b (systematic review; nonhomogeneous Level-I studies). See Instructions to Authors for a complete description of levels of evidence.

Activities of Daily Living↗
PubMed1988

Light and electron microscopic studies on the localization of hyaluronic acid in developing rat cerebellum.

The hyaluronic acid-binding region was prepared by trypsin digestion of chondroitin sulfate proteoglycan aggregate from the Swarm rat chondrosarcoma, and biotinylated in the presence of hyaluronic acid and link protein. After isolation by gel filtration and HPLC in 4 M guanidine HCl, the biotinylated hyaluronic acid-binding region was used, in conjunction with avidin-peroxidase, as a specific probe for the light and electron microscopic localization of hyaluronic acid in developing and mature rat cerebellum. At 1 w postnatal, there is strong staining of extracellular hyaluronic acid in the presumptive white matter, in the internal granule cell layer, and as a dense band at the base of the molecular layer, surrounding the parallel fibers. This staining moves progressively towards the pial surface during the second postnatal week, and extracellular staining remains predominant through postnatal week three. In adult brain, there is no significant extracellular staining of hyaluronic acid, which is most apparent in the granule cell cytoplasm, and intra-axonally in parallel fibers and some myelinated axons. The white matter is also unstained in adult brain, and no staining was seen in Purkinje cell bodies or dendrites at any age. The localization of hyaluronic acid and its developmental changes are very similar to that previously found in immunocytochemical studies of the chondroitin sulfate proteoglycan in nervous tissue (Aquino, D. A., R. U. Margolis, and R. K. Margolis. 1984. J. Cell Biol. 99:1117-1129; Aquino, D. A., R. U. Margolis, and R. K. Margolis. J. Cell Biol. 99:1130-1139), and to recent results from studies using monoclonal antibodies to the hyaluronic acid-binding region and link protein. The presence of brain hyaluronic acid in the form of aggregates with chondroitin sulfate proteoglycans would be consistent with their similar localizations and coordinate developmental changes.

Aggrecans↗
PubMed1982

Quantitation of hyaluronic acid in equine synovia.

An assay for the determination of hyaluronic acid concentration in equine synovia (synovial fluid) was investigated. The method involved the degradation of hyaluronic acid, using a highly specific microbial enzyme. The unsaturated tetra- and hexasaccharide products were oxidized with periodate and caused to react with thiobarbituric acid for form a chromophore which absorbed in the visible range of the spectrum. The method was precise within acceptable limits for synovia and compared favorably with the widely used carbazole method of Bitter and Muir (Anal Biochem 4:330-334, 1962). The between-assay coefficient of variation was 1.7% and the method was found to be linear to 100 microgram of hyaluronic acid. The mean value for hyaluronic acid concentration in synovia of health horses was 127 mg/100 ml.

Animals↗
PubMed2006

Hyaluronic acid predicts hepatic fibrosis in children with hepatic disease.

BACKGROUND: Hyaluronic acid (HA) is removed by the liver via sinusoidal cell adhesion molecules. This is impeded in fibrosis, leading to a rise in serum HA. As a noninvasive marker of fibrosis, HA may obviate the need for liver biopsy. OBJECTIVE: To evaluate HA as a marker of hepatic fibrosis, in unselected children undergoing liver biopsy. METHODS: Ninety-three unselected consecutive children (median age, 7.5 years; range, 0.07-19 years) undergoing a liver biopsy between April 2003 and March 2004 were prospectively recruited. Liver biopsy and fasting HA levels were taken simultaneously. The Ishak score was used to stage fibrosis. Scores of 3 or greater were regarded as significant fibrosis. Hyaluronic acid levels were measured using an enzyme-linked binding protein assay (2002 Corgenix, Inc) (adult reference range, 0-75 ng/mL; pediatric reference range, 0-30 ng/mL). RESULTS: Twenty-three (25%) of 93 biopsies had significant fibrosis, and HA levels in this group were significantly higher than those with mild fibrosis (Ishak score, <3), (median level, 72 ng/mL vs 30 ng/mL; Mann-Whitney U test; P < 0.005). Hyaluronic acid level of 50 ng/mL had a positive predictive value 40% and negative predictive value 86% for significant fibrosis. An HA level 200 ng/mL has a sensitivity of 26% and specificity of 90%. CONCLUSIONS: Hyaluronic Acid is a valid noninvasive predictor of hepatic fibrosis in unselected children with liver disease. An HA level of 200 ng/mL strongly suggests significant fibrosis. Hyaluronic acid level of less than 50 ng/mL accurately identifies those who do not have significant fibrosis.

Adolescent↗
PubMed1990

Elevated level of hyaluronic acid binding protein in diabetic rats.

Hyaluronic acid binding protein (HABP), an extracellular matrix glycoprotein which interacts specifically with hyaluronic acid (HA) has been purified to homogeneity by HA affinity chromatography. Antibody was raised against it and the specificity of antibody towards HABP was confirmed by western blot analysis. A specific and sensitive assay method has been developed adopting solid phase non-competitive double sandwich method of ELISA. This new assay method enabled us to determine the levels of HABP in different tissue extracts of normal, diabetic and reverse diabetic rats. A significant increase in the levels of HABP was observed in diabetic animals, which however attained normal levels with insulin treatment.

Animals↗
PubMed1992

Correlation of hyaluronic acid accumulation and the growth of preneoplastic mammary cells in collagen: a longitudinal study.

Hyaluronic acid accumulation is characteristic of mammary tumor cells, and the amount that accumulates seems to correlate with the degree of malignancy of the producing cells. We have tested directly the relationship between hyaluronic acid accumulation and the replication rate of preneoplastic mammary cells in culture. We used nontumorigenic but immortal CL-S1 mouse mammary cells that were derived from a hyperplastic alveolar nodule. Using a collagen gel culture system, we found clear differences in the growth properties of cells before and after Passages 68 to 70. Late passage cells replicated earlier and faster than early passage cells in collagen and on plastic. The rate of cycling resembled that of tumorigenic mouse mammary cells during the first week of culture. Cells seeded at low densities cycled faster than those seeded at high densities during the second week in culture. Exogenous hyaluronic acid, at 10 to 1000 micrograms/ml, neither enhanced nor inhibited CL-S1 cell growth significantly in collagen, regardless of passage. However, by the third day in collagen, late passage cells produced 7 times more total glycosaminoglycans and 12 times more hyaluronic acid per cell than did early passage cells. Late passage cells also deposited 12 times more labeled hyaluronic acid in the matrix than did early passage cells, on a per-cell basis. After a decline in the deposition of hyaluronic acid in the extracellular matrix, growth ceased. The late passage cells did not grow in soft agar, indicating that they had not become neoplastic spontaneously during passage. However, their accelerated growth rate, coupled with the synthesis and secretion of large amounts of hyaluronic acid into the extracellular matrix, may characterize a distinct step in tumor progression in preneoplastic CL-S1 cells.

Animals↗
PubMed1979

Hyaluronic acid vitreous substitute. A six-year clinical evaluation.

Hyaluronic acid vitreous substitute (HYVISC) was used during 347 operations performed on 294 eyes of 286 patients. It was injected intraocularly in 266 operations (73 scleral bucklings, 175 open-sky vitrectomies, among others) on eyes with complex retinal detachments that were considered inoperable by ordinary surgical techniques. During 81 closed vitrectomies, it was applied extraocularly between the surgical contact lens and the cornea. Hyaluronic acid was found to be well tolerated by human eyes. In eyes that had been given a poor prognosis, scleral buckling with hyaluronic acid tamponade produced reattachment in 16% of the eyes; open-sky vitrectomy, scleral buckling, and hyaluronic acid salvaged 18%. It also appeared helpful in the preservation of corneal epithelial integrity and clarity during closed vitrectomy.

Adolescent↗
PubMed1994

Hyaluronic acid. A review of its pharmacology and use as a surgical aid in ophthalmology, and its therapeutic potential in joint disease and wound healing.

Hyaluronic acid is a naturally occurring polysaccharide with distinct physicochemical properties which underlie its application as a viscoelastic tool in ophthalmological surgery. In cataract surgery the role of hyaluronic acid in facilitating procedures and protecting the corneal endothelium is well established. Some benefit has also been gained with the use of hyaluronic acid in penetrating keratoplasty, trabeculectomy, retinal reattachment and trauma surgery, although its efficacy in these indications is less well-defined in the published literature. In addition to its lubricating and cushioning properties, demonstration of some in vitro anti-inflammatory activity and a possible disease-modifying effect for hyaluronic acid in animals has prompted its investigation as a treatment in osteoarthritis and, to a much lesser extent, in rheumatoid arthritis. Hyaluronic acid 20 mg, as weekly intra-articular injections for 3 to 7 weeks, improved knee pain and joint motion in patients with osteoarthritis. Although this occurred to a greater degree than with placebo in most comparisons, the effects of hyaluronic acid was similar to those of placebo in the largest trial. In the few available comparisons with other agents, hyaluronic acid appeared equivalent to methylprednisolone 40 mg (for 3 weeks) and to a single injection of triamcinolone 40 mg. Hyaluronic acid was distinguished from other therapies by providing a sustained effect after treatment discontinuation. Together with its very good tolerability profile, these properties justify further study of hyaluronic acid in patients with osteoarthritis. Some limited evidence of improvement in patients with rheumatoid arthritis, and a possible healing effect of hyaluronic acid on tympanic membrane perforations, represent additional areas of interest for future investigation. In summary, hyaluronic acid is a well-established adjunct to cataract surgery and may prove to be a promising option in the treatment of patients with osteoarthritis. Its very good tolerability provides further impetus for examination of its potential role in an extended scope of arthritic and ophthalmological indications, and in wound healing.

Adjuvants, Pharmaceutic↗
PubMed1981

Effect of cycloheximide and dexamethasone phosphate on hyaluronic acid synthesis and secretion in cultured human skin fibroblasts.

We have studied some aspects of the regulation of hyaluronic acid synthesis in cultured human skin fibroblasts derived from normal individuals. In these cells 85% of radiolabeled glucosamine is incorporated into two types of macromolecules, hyaluronic acid and glycoproteins, characterized by district partitions between the cell and medium compartments. Treatment of cells with cycloheximide under conditions in which protein and glycoprotein synthesis is inhibited by at least 90% has no effect on hyaluronic acid synthesis. This result is consistent with the hypothesis that protein synthesis is not necessary for the synthesis of hyaluronic acid. Conversely, 5 X 10(-6) M dexamethasone phosphate inhibits hyaluronic acid by 60% but has no effect on protein or glycoprotein or glycoprotein synthesis. Concomitant treatment with dexamethasone phosphate and cycloheximide restores the synthesis of hyaluronic acid to normal values. The dexamethasone phosphate effect on hyaluronic acid synthesis is therefore not a primary induction response since it requires protein synthesis. The secretion of hyaluronic acid and glycoproteins from the cells into the medium is not affected by either of the two drugs. This experimental approach will be useful in the study of the regulation of glycosaminoglycan metabolism in human cells especially with respect to disorders of connective tissue.

Cells, Cultured↗
PubMed1986

Pharmacological effect of hyaluronic acid (HA) on phagocytes: hypothesis for an HA-induced monocyte chemotactic factor for neutrophils.

The connection between hyaluronic acid and phagocyte function is not well documented in the literature. Its action may either inhibit or enhance polymorphonuclear neutrophil (PMN) function, depending on its concentration. Studies were conducted to verify the effect of hyaluronic acid on phagocytes, both directly using hyaluronic acid and indirectly via a mediated route using the medium from a hyaluronic acid monocyte incubation. Determinations were made of phagocytosis, reduction of nitroblue tetrazolium, superoxide production, and chemotaxis before and after incubation with hyaluronic acid. Chemotaxis of PMNs was used to evaluate the chemotactic action of a medium in which monocytes had been incubated with hyaluronic acid. This method resulted in progressive improvement in the chemotactic index. The authors conclude that the monocytes incubated with hyaluronic acid produce a chemotactic factor for neutrophils.

Cell Movement↗
PubMed1988

The role of hyaluronic acid in inflammation and wound healing.

We have previously hypothesized that hyaluronic acid (HA) and fibrin specifically interact and help orchestrate the early stages of the inflammatory response and wound healing (J. Theoretic. Biol. 119, 219, 1986). In the present work we have extended studies to confirm this prediction. Several approaches were used to show that human fibrinogen specifically binds to hyaluronic acid. In addition, this latter glycosaminoglycan greatly stimulated the in vitro formation of fibrin clots induced by thrombin. The presence of hyaluronic acid also altered the structure of the final fibrin gel. Removal of circulating hyaluronic acid in blood is therefore probably vital in order to maintain normal haemostasis. Evidence is presented to suggest that an endocytic receptor recycling process is responsible for the ability of liver endothelial cells to perform this function and remove HA from the blood. Although hyaluronic acid levels are initially low in a blood clot, the proposed wound-healing model explains why and how HA levels increase and the functional and structural significance of this polysaccharide during wound healing.

Animals↗
PubMed1984

Inhibition of oxidative degradation of hyaluronic acid by uric acid.

It has been postulated that glycosaminoglycans in the trabeculum have an influence on aqueous humor drainage. Ascorbate reduces the viscosity of hyaluronic acid, and also increases outflow facility. Our recent observation of high urate concentrations in some glaucomatous eyes led us to study the influence of urate on oxidative degradation of hyaluronic acid by ascorbate. The viscosity of rooster comb hyaluronic acid was reduced slowly by ascorbate. Cupric sulfate accelerated ascorbate oxidation and also enhanced hyaluronic acid degradation. Urate inhibited ascorbate oxidation and prevented the copper catalyzed oxidative degradation of rooster comb hyaluronic acid. The range of urate concentrations used in this study was within the range of urate concentrations observed in glaucomatous eyes. The partially purified umbilical cord hyaluronic acid had lower viscosity than rooster comb hyaluronic acid, and rapidly degraded in the presence of ascorbate. The ascorbate effect on umbilical cord hyaluronic acid was partially prevented by urate.

Animals↗
PubMed1999

Molecular weight independence of the effect of additive hyaluronic acid on the lubricating characteristics in synovial joints with experimental deterioration.

OBJECTIVE: To assess the molecular weight dependence of the lubricating characteristics of additive hyaluronic acid in animal joints with experimental deterioration. DESIGN: An experimental study in rabbits stifles in vitro. BACKGROUND: Hyaluronic acid is used as a drug for osteoarthritis. However, the molecular weight dependence of the effect of hyaluronic acid has been unclear. METHODS: Using a robotic arm under force-control, frictional coefficients were measured in fourteen rabbit stifles. The experimental conditions examined were: (1) intact joint surfaces; (2) after the joint surfaces were washed with physiological saline; (3) after the addition of 1% hyaluronic acid solution with a molecular weight of one million daltons and two million daltons, respectively. RESULTS: The increased frictional coefficient produced by washing with saline was decreased significantly by the addition of hyaluronic acid to the joint surfaces. This effect was observed regardless of the molecular weight of the hyaluronic acid. CONCLUSION: The lubricating characteristics of hyaluronic acid in synovial joints do not change with molecular weight. RELEVANCE: Hyaluronic acid solution is used clinically for the treatment of osteoarthritis. In the present study, the molecular weight dependency of the lubricating characteristics of hyaluronic acid was tested using the friction in synovial joints.

Animals↗
PubMed1976

[Conformational changes of hyaluronic acid in acid medium (author's transl)].

Both vitreous body homogenate and a fibrous product containing hyaluronic acid and collagen isolated from vitreous humor by acetone precipitation were investigated to elucidate the liquefaction of vitreous body by acids. Graphic evaluation of the titration with hydrochlorid acid and viscosity measurements suggest a changed of the hyaluronic acid molecule in the pH-range between 5.2 and 4.1. Between these pH-values the ionization of carboxyl groups is decreased accompanied by conformational changes of the hyaluronic acid molecule. These results are in agreement with the conclusion of other authors that by lowering the pH of hyaluronic acid solutions a random coil to double helix transition of hyaluronic acid occurs.

Animals↗
PubMedDate not supplied

Hyaluronic acid degradation products induce neovascularization and fibroplasia in fetal rabbit wounds.

Scarless fetal wound healing occurs with mild fibroplasia and neovascularization, while the wound is persistently enriched with hyaluronic acid. Conversely, adult wounds are characterized by prominent fibroplasia, neovascularization, and scar formation, and hyaluronic acid is a transient component of the early adult wound matrix. Our group has reported that enzymatic degradation of fetal rabbit wound hyaluronic acid results in marked increases in fibroplasia, collagen deposition, and neovascularization. This altered, adultlike healing response is hypothesized to have resulted from the generation of biologically active hyaluronic acid degradation products. Therefore, this study analyzes the fibrovascular inductive activity of hyaluronic acid degradation products. Fetal rabbit wounds were treated with hyaluronic acid degradation products generated by methods known to produce oligosaccharides with significant angiogenic activity. Implants from wounds treated with either of the control solutions (n = 4 for each control) had identical histologic features characterized by a mild mononuclear cell infiltrate but neither infiltrating fibroblasts nor collagen. In marked contrast, implants from wounds treated with hyaluronic acid degradation products contained infiltrating fibroblasts and collagen intermixed with numerous blood vessels. Quantitation of capillary ingrowth showed a sixfold increase in the neovascular response in wounds treated with hyaluronic acid degradation products compared with either controls (p < 0.05). This study shows that hyaluronic acid degradation products stimulate neovascularization and fibroplasia in fetal wounds. These observations suggest that a balance between hyaluronic acid accumulation and degradation has significant regulatory influence in fetal tissue repair.

Journal Article↗
PubMed1995

In vitro lead cation-hyaluronic acid interaction.

It is the purpose of this investigation to study the in vitro spontaneous interaction lead-hyaluronic acid using the equilibrium dialysis technique. The following working conditions were used: membrane retention capacity starting from 12000 D; solvent was an aqueous buffer solution (pH 7.4); relation of internal-external volume of 0.5 : 25 mL: dialysis time 24 hours; lead concentration in dialyzing solution 4 micrograms/mL (always outside the bag); temperature 4 degrees C; concentrations of hyaluronic acid 0.3, 1.2, 2.1 and 3.0 mg/mL; concentration of plasma proteins 1.5 mg/mL; lead was determined in the internal solution of the bag, and was measured quantitatively by atomic absorption spectrometry. Testing separately hyaluronic acid and plasma proteins both types of marcromolecules turned out to bind lead. As the concentration of hyaluronic acid increases, the amount of lead retained increases as well, although this relation is only approximately linear. At the normal concentrations of plasma proteins and hyaluronic acid in synovial fluid, the proteins solution bound more lead than hyaluronic acid solution 16.2 +/- 2.1 and 5.7 +/- 0.4 micrograms/mL respectively (P < 0.0001). What these data show ist that both, hyaluronic acid and proteins are involved in the in vitro lead retention. The effect of both ligands must be synergistic, because when the two molecules are present, a higher amount of lead is retained compared to the amount bound separately, suggesting that lead can replace calcium by complexation with mucopolysaccharides; this explains the lead accumulation in synovial fluid since this fluid is rich in hyaluronic acid and proteins.

Cations↗