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Hyaluronic acid influences the migration of myoblasts within the avian embryonic wing bud.

Myoblasts migrate in a proximodistal direction within the avian embryonic wing bud during normal limb development. Since the presence and distribution of hyaluronic acid within the wing bud coincide with the time and with the direction of the migration of myoblasts, we microinjected hyaluronic acid into chicken wing buds that had received grafts containing quail myoblasts. It was found that injected hyaluronic acid has a strong positive effect on the migration of myoblasts: it causes a migration of myoblasts in donor-host combinations in which this is normally not the case, and it can cause migration in a proximal direction, a phenomenon not observed during normal development. From this it may be concluded that hyaluronic acid can influence myoblast migration in vivo. A similar effect could be observed after the microinjection of dextran sulfate, a synthetic compound having similar physicochemical properties. Hyaluronic acid, therefore, may play an important role in the control of the migration of myogenic cells in vivo by its physiocochemical properties.

Animals↗

Studies on the affinity of hyaluronic acid binding protein to glycosaminoglycans.

The affinity of hyaluronic acid binding protein (HBP) to different glycosaminoglycans (GAGs) was examined. The purified protein was pretreated with hyaluronic acid (HA), heparin, glucuronic acid and N-Acetyl-glucosamine and was loaded onto Hyaluronate-Sepharose affinity column. The binding of HBP to HA immobilized on sepharose column was specifically blocked only by pretreatment of HBP to HA and the elution of HBP was decreased proportionately with the addition of higher quantity of HBP. The specificity of HBP to HA was confirmed as it did not bind to Heparin-Sepharose or Chondroitin-4-Sulphate-Sepharose columns. The complex of HBP in association with HA was further shown on Sephadex G-200 and 7.5% polyacrylamide gel. All the experimental findings indicate that HBP binds specifically to HA only.

Animals↗

Influences of sulfated glycosaminoglycans on biosynthesis of hyaluronic acid in rabbit knee synovial membrane.

The effect of various sulfated glycosaminoglycans on glycoconjugates syntheses in synovial membranes of rabbit knee joints in culture was investigated by two different approaches. In the first approach, synovial membranes isolated from rabbit knee joints were cultured in the presence of sulfated glycosaminoglycans and [14C]glucosamine. In the second approach, solutions of sulfated glycosaminoglycans were injected into rabbit knee joints and synovial membranes isolated from the joints were cultured in the presence of [14C]glucosamine. The major part of [14C]glucosamine-labeled glycoconjugates associated with the synovial membranes and secreted into culture medium was hyaluronic acid. Of the natural glycosaminoglycans tested, dermatan sulfate gave the maximum stimulation of hyaluronic acid synthesis followed by chondroitin 4- and 6-sulfate. Heparin, heparan sulfate, keratan sulfate, keratan polysulfate, and hyaluronic acid had no significant effect. Of the chemically polysulfated glycosaminoglycans, GAGPS (a persulfated derivative of chondroitin sulfate) gave high stimulation but N-acetylchitosan 3,6-disulfate had no effect. The effect of sulfated glycosaminoglycans on hyaluronic acid synthesis was the same in both experimental approaches. The increase in the amount of secreted hyaluronic acid in culture medium paralleled that in synovial membranes. The results indicate that the galactosamine-containing sulfated glycosaminoglycans have a specific stimulatory effect on hyaluronic acid synthesis. A high degree of sulfation of the molecules appeared to potentiate the stimulatory effect.

Animals↗

HOST-PARASITE RELATIONSHIPS AMONG GROUP A STREPTOCOCCI. I. HYALURONIC ACID PRODUCTION BY VIRULENT AND AVIRULENT STRAINS.

Willoughby, Donald S. (University of Minnesota, Minneapolis), Yael Ginzburg, and Dennis W. Watson. Host-parasite relationships among group A streptococci. I. Hyaluronic acid production by virulent and avirulent strains. J. Bacteriol. 87:1452-1456. 1964.-Intradermal passage of group A streptococcal strains in rabbits is an easy and effective technique for selecting organisms capable of producing increased amounts of hyaluronic acid. Glutamine was required for in vitro hyaluronate production, although the requirement varied with the organism tested. The virulent type 18 cells produced maximal amounts of hyaluronic acid in the presence of 0.01% glutamine compared with 0.1% for the more avirulent type 28; after animal passage, the glutamine requirement for type 28 decreased to 0.01%. Organisms selected by animal passage produced small quantities of hyaluronic acid in the absence of an exogenous source of glutamine. Glucose at a concentration of 1% gave the greatest stimulation for hyaluronate formation. Growth studies indicated similar rates of multiplication for types 18 and 28; yet, the rate of hyaluronate production under these conditions was much greater for the more virulent type 18. Because of these observations, it is suggested that the rate of hyaluronate production is a contributing factor in the virulence of type 18.

Animals↗

The effect of diflunisal on hyaluronic acid production by 'activated' human synovial fibroblasts.

1 Cultured human synovial fibroblasts were 'activated' by leucocytes or poly (I) . poly (C) to overproduce hyaluronic acid. 2 Diflunisal in concentrations of 30, 15 and 10 microgram/ml completely abrogated the overproduction of hyaluronic acid induced by leucocytes. 3 Diflunisal (10 microgram/ml), indomethacin (2 microgram/ml) and aspirin (200 microgram/ml) partially inhibited the overproduction of hyaluronic acid induced by poly (I) . poly (C), although none of them decreased it to control non 'activated' levels. 4 Diflunisal 25 microgram/ml completely abrogated hyaluronic acid overproduction induced by poly (I) . poly (C). 5 This model could be an in vitro indicator for in vivo pharmacological levels of anti-inflammatory drugs.

Anti-Inflammatory Agents↗

Age-related changes in the molecular weight of hyaluronic acid from rat skin.

Age-related changes in the molecular weight of hyaluronic acid from the skin of rats was studied using fetuses, newborn animals and rats 1, 2, 3, 4, 13, 26 and 52 weeks of age. Average values of the molecular weight of hyaluronic acid varied with age in three phases: 39 X 10(4) in fetuses and up to the age of one week, 55 X 10(4) in individuals aged to 26 weeks and 87 X 10(4) in individuals 52 weeks of age. The distribution of molecular weight was quite different between 4 weeks and 52 weeks of age. At 4 weeks of age, the molecular weight of hyaluronic acid was widely distributed, ranging from 18 X 10(4) to 79 X 10(4), while that from 52-week-old individuals was very high with a distribution ranging from 92 X 10(4) to 119 X 10(4).

Aging↗

Simultaneous detection of submicrogram quantities of hyaluronic acid and dermatan sulfate on agarose-gel by sequential staining with toluidine blue and Stains-All.

A new discontinuous agarose-gel electrophoresis in 0.05 M HCl/0.04 M barium acetate combined with the highly sensitive visualization technique using toluidine blue/Stains-All has been developed for the simultaneous assaying of hyaluronic acid (HA) and dermatan sulfate (DS) with a detection limit at submicrogram level greater than other conventional procedures. Furthermore, this procedure also separates and reveals chondroitin sulfate (CS). The densitometric analysis of bands resulted in a linear response between 0.01 and 0.5 microg of glycosaminoglycans (GAGs) with correlation coefficients greater than approximately 0.94. Hyaluronic acid and dermatan sulfate extracted and purified from the abdominal skin of six rats were separated and quantified in comparison with the evaluation made by treatment of chondroitin ABC lyase and separation of Delta-disaccharides from hyaluronic acid (DeltadiHA) and dermatan sulfate/chondroitin sulfate (Deltadi4s and Deltadi6s) by HPLC. The total amount of rat skin polysaccharides (hyaluronic acid and dermatan sulfate) was 1.24+/-0.26 microg/mg of tissue by discontinuous agarose-gel electrophoresis and 1.20+/-0.33 microg/mg by HPLC with hyaluronic acid and dermatan sulfate percentages of 50.32+/-2.38 and 49.66+/-2.53, respectively. The analyses also confirmed that hyaluronic acid and dermatan sulfate are the main rat abdominal skin polysaccharides with chondroitin sulfate present in trace amounts. This new agarose-gel electrophoresis could be particularly useful in the study of the distribution of glycosaminoglycans in the skin from different body sites of animals and normal human subjects and may be of importance in understanding the changes that occur in the skin, especially the metabolism of extracellular matrix constituents, in connective tissue disorders.

Animals↗

Studies in fetal wound healing, VII. Fetal wound healing may be modulated by hyaluronic acid stimulating activity in amniotic fluid.

Fetal wound healing occurs rapidly and without inflammation, fibrosis, or scar formation. It is a process fundamentally different from adult wound healing. The mechanisms that underlie such unique healing properties are unknown. However, hyaluronic acid, a glycosaminoglycan component of the extracellular matrix, is prominent throughout the course of fetal wound healing, and is thought to play a major role in the healing process. Amniotic fluid contains high levels of hyaluronic acid. Amniotic fluid also contains a number of potent growth factors that are critical for fetal development. In this report, a new factor in amniotic fluid that stimulates deposition of hyaluronic acid is described. This activity is measured in an in vitro assay system in which cultured fibrosarcoma cells are used as indicator cells. Amniotic fluid thus provides two separate mechanisms for the deposition of hyaluronic acid. One is by exogenous application directly onto fetal skin wounds; the second is by providing a factor to increase the production of hyaluronic acid endogenously, by stimulating cells around the wound site. The resulting hyaluronic acid-rich area may support the ability of the fetal wound to heal with its unique properties.

Amniotic Fluid↗

Hyaluronic acid in the pulmonary secretions of patients with asthma.

Hyaluronic acid was the only glycosaminoglycan found in the pulmonary secretions of patients with asthma. The compound had a hexuronate/hexosamine molar ratio of about 1:1. Glucosamine constituted over 98% of the hexosamines, the remaining being galactosamine. The compound moved as a single spot with the mobility of standard hyaluronic acid on cellulose acetate electrophoresis, and this spot disappeared after digestion with testicular hyaluronidase. Even after extensive proteolysis and purification, the compound was associated with small amounts of protein, the major amino acids of which were aspartic acid, threonine, serine, glutamic acid, glycine and valine.

Adult↗

Hyaluronic acid deposition in cardiac myxomas: localization using a hyaluronate-specific binding protein.

Myxomas are the most common primary tumors of the heart. These lesions contain a gelatinous, ground-substancelike material which has been described as glycosaminoglycan in nature. Using a newly developed, cartilage-derived hyaluronic acid-binding protein and a modification of the avidin-biotin immunostaining procedure, we demonstrate that hyaluronic acid is contained in the jellylike material of cardiac myxomas.

Adult↗

Increase of ICSI efficiency with hyaluronic acid binding sperm for low aneuploidy frequency in pig.

The present study was designed to evaluate the ability of hyaluronic acid binding sperm (HABS) in increasing the efficiency of intracytoplasmic sperm injection (ICSI) in terms of the production of chromosomally normal porcine embryos. Porcine embryos were produced by in vitro fertilization (IVF), ICSI and ICSI using hyaluronic acid binding sperm (ICSI-HABS). Chromosome aneuploidy in sperm and embryos was evaluated using chromosome 1 submetacentric probe for fluorescence in situ hybridization (FISH) analysis. No significant differences were observed in the blastocysts rates (18.6, 23.6 and 23.8%) and cell numbers (61.8+/-12.5, 55.5+/-7.3 and 59.3+/-9.6) among embryos derived from IVF, ICSI, and ICSI-HABS. However, the frequency of normal diploidy in ICSI-HABS (75.5%) was significantly higher (P<0.05) than that in IVF (57.0%) and ICSI (68.2%). Embryos from ICSI-HABS showed significantly lower chromosome abnormality rate (P<0.05). Both ICSI and IVF embryos showed higher rates of polyploidy, and hence chromosomally abnormal embryos, in comparison to ICSI-HABS embryos. In addition, we investigated the chromosomal complement of porcine spermatozoa by FISH. The rate of chromosome number abnormality in porcine sperm was found to be 6.25% (70/1120). Thus, we conclude that the use of hyaluronic acid binding sperm is superior to morphological sperm selection for ICSI in producing chromosomally normal embryos and increasing the ICSI efficiency by lowering the aneuploidy frequency. Our results indicate that the selection of normal sperm with hyaluronic acid binding assay might help to reduce the early embryonic mortality due to chromosomal aneuploidy thereby increasing the success rate of embryo transfer technology in pigs.

Aneuploidy↗

Hyaluronic acid. An indicator of pathological conditions of human lungs?

Hyaluronic acid was the only glycosaminoglycan found in detectable amounts in the pulmonary secretions of patients with cystic fibrosis. The compound gave a hexuronate/hexosamines molar ratio of approximately 1. Glucosamine represented over 98% of the total hexosamines, the remainder being galactosamine. No hexoses or sulfate could be detected. It moved as a single spot with the mobility of standard hyaluronic acid on cellulose acetate electrophoresis and this spot disappeared after digestion with testicular hyaluronidase. It was associated with trace amounts of protein, the major amino acids of which are aspartic acid, glutamic acid, glycine, and alanine.

Amino Acids↗

Evidence for naturally occurring hyaluronic acid binding protein in rat liver.

Hyaluronic acid binding protein (HBP) was purified homogeneously from normal adult rat liver by hyaluronate-sepharose affinity chromatography. The molecular weight of this protein as determined by gel filtration was found to be 64,000 daltons. This protein HBP appeared as a single band in non-dissociating gel electrophoresis and has a subunit of molecular weight approximately 12,000 as determined by SDS-gel electrophoresis.

Animals↗

Human anti-hyaluronic acid antibodies: is it possible?

BACKGROUND: Although hyaluronan has been acknowledged as being free of species and organ specificity, for 4 years I have encountered a variety of adverse reactions to injectable hyaluronic acid as used in aesthetic medicine. OBJECTIVE: I have tried to prove that some of those side effects may be allergic reactions to the commercial preparations of injectable hyaluronic acid. METHODS: I began with intradermal tests to the reactive patients and to 2 witnesses; then lymphocyte transforming tests were performed at the University of Geneva (Switzerland). Histology was performed on the skin tests and on reactive treated areas of the face of different patients. A serum analysis was then done by Pr. Sainte Laudy of Laboratoire Pasteur--Cerba (France). RESULTS: The skin tests were positive for one or the other or both of the injectable hyaluronic acid preparations used in aesthetic medicine. The different biopsies have shown for some a chronic inflammatory reaction, even 11 months after the treatment or a severe granulamatous reaction to foreign bodies. Serum analysis revealed positive antibodies against Restylane and/or Hylaform and even IgG and E anti-hyaluronic acid. CONCLUSION: Since 1995, I have 8 patients with adverse reactions to injectable hyaluronic acid, which after several tests, may be allergic to those products. Isn't it time to introduce intradermal tests before any injection of this type, as done with injectable bovine collagen?

Animals↗

Double-blind, randomized clinical study comparing hyaluronic acid cream to placebo in patients treated with radiotherapy.

PURPOSE: The effect of hyaluronic acid (Ialugen cream) on acute skin reactions after radiotherapy, was assessed in a randomized, double-blind, placebo-controlled study. MATERIAL AND METHODS: Out of the 152 patients presenting with head and neck, breast or pelvic carcinomas and registered in the study, 134 cases-70 in the Ialugen group (IA) and 64 in the placebo group (PBO)-completed their IA or PBO treatment. At the time of randomisation, these two groups were balanced for sex, age, weight and height. The mean total dose of radiation given during the study was 60.6 +/- 10.9 Gy in the IA group and 64.3 +/- 10.8 Gy in the PBO group (P = 0.47). RESULTS: Acute radio-epithelitis scores were significantly higher in the PBO group than in the IA group, starting from the control at week 3 and throughout the 6 weeks of treatment (P < 0.01 from week 3 to week 7; P < 0.05 at weeks 8 and 10). Likewise, the global efficacy judgement expressed, at the end of treatment, by both the physician and the patient showed a significant difference in favour of Ialugen (P < 0.01 and P < 0.05, respectively). There was no significant difference of tolerance between the IA and PBO treatments (P = 0.18 according to the physician and P = 0.42 from the patient's viewpoint). CONCLUSION: The prophylactic use of a cream with hyaluronic acid is shown to reduce the incidence of high grade radio-epithelitis, suggesting an interesting role of the hyaluronic acid cream as supportive treatment to improve compliance and quality of life in patients undergoing radiation therapy.

Adult↗

Hyaluronic acid for the treatment of knee osteoarthritis: long-term outcomes from a naturalistic primary care experience.

OBJECTIVE: Intraarticular hyaluronic acid is indicated for patients with osteoarthritis of the knee. However, clinical experience, especially efficacy and adverse events, with repeated injection series in the long term are limited. DESIGN: Patients were referred to a large primary care center for management of osteoarthritis of the knee. All were naive to intraarticular hyaluronic acid therapy and met our entry criteria, including resting visual analog scale pain of > 45 mm, radiographic confirmation of unilateral knee grade 1-3 osteoarthritis, and willingness to receive intraarticular therapy. Patients received a three-intraarticular injection series with Suplasyn (10 mg/ml, 2-ml injection) over 3 wks. Patients were instructed to return for consideration of repeat injection series based on their perception of pain restricting daily activity and a resumption of severity similar to their initial presentation. This prospective naturalistic cohort was followed for 6.7 yrs. Patients completed baseline assessment of rest and walking visual analog scale pain (primary efficacy variable), completed a 5-point categorical global satisfaction score, and recorded adverse events and concomitant therapeutic modality use at each study visit. Patients returned for consideration of a repeat injection series based on their perception of symptom severity and were eligible if their resting visual analog scale pain was > 45 mm. The three-injection series and data collection were repeated, and again, patients were given similar instructions regarding consideration of a third injection series. RESULTS: Of 897 referral patients, 537 (mean age, 68 +/- 8 yrs; mean duration of symptoms, 7.4 +/- 4.1 yrs) met our criteria, and only 21 patients did not return for a second injection series. The mean time between first and second series was 27 +/- 7 wks. The change in walking visual analog scale pain was significantly improved from baseline after the first series (81.3%, P < 0.001) and second series (86.7%, P < 0.0001). Similarly, resting visual analog scale pain was significantly decreased from baseline after the first (P < 0.001) and second (P < 0.001) series, and patient satisfaction was significantly improved with each injection series (P < 0.03 and P < 0.01). Very few adverse events were recorded and were limited to local pain and swelling. Use of concomitant therapeutic modalities at presentation for a second injection series included: nonsteroidal anti-inflammatory drugs/cyclooxygenase-2 medications (37%), acetaminophen (31%), oral nutraceuticals (12%), and physical therapy and bracing (12%). CONCLUSIONS: Intraarticular hyaluronic acid injections were highly effective in improving resting and walking pain in patients with osteoarthritis of the knee on a first and a second treatment series. Duration of symptom control was about 6 mos, and the therapy was highly satisfactory to patients and was associated with very few local adverse events and limited use of concomitant therapeutic modalities. These data support the potential role of intraarticular hyaluronic acid as an effective long-term therapeutic option for patients with osteoarthritis of the knee.

Adjuvants, Immunologic↗

Determination of molecular weight of hyaluronic acid in bovine vitreous humour and Healon by high performance gel permeation chromatography and its depolymerization with ascorbic acid.

A method has been developed for the determination of the molecular weight of hyaluronic acid by high performance gel permeation chromatography. The molecular weight distribution was determined using three polymeric columns. Columns were calibrated using Pullulan Polymer molecular weight standards. The average molecular weight (+/- SD) of the hyaluronic acid in bovine vitreous humour and Healon were 1.20 x 10(6) Da (+/- 0.49 x 10(6) and 3.01 x 10(6) Da (+/- 0.14 x 10(6), respectively. Furthermore, the depolymerization of the hyaluronic acid in the presence of ascorbic acid (the concentration found in human eye) was studied. Samples incubated in the presence of atmospheric air showed more depolymerization than those where atmospheric air had been flushed-out with nitrogen.

Animals↗