Hyaluronan Concentration Research Articles

The effect of molecular weight and concentration of hyaluronan on the recovery of the rat sciatic nerve sustaining acute traumatic injury

Acute traumatic peripheral nerve injury remains a significant clinical issue affecting mostly young individuals and their productivity in spite of advances in current medicine. Hyaluronan has been explored in this scenario for its anti-adhesive and high biocompatibility properties for decades. The molecular weight and concentration of the locally applied hyaluronan has been overlooked and not optimized. We used different molecular weights and concentrations of hyaluronan in a rat sciatic nerve crush injury model and found better overall outcomes with high molecular weight (3000 kDa) hyaluronan. The anti-inflammatory effect of the higher molecular weight hyaluronan may have a more favorable effect. We conclude that the optimization of hyaluronan is necessary when incorporating hyaluronan in the engineering of biomaterials for use in acute traumatic peripheral nerve injury.

Endothelial glycocalyx biomarkers increase in patients with infection during Emergency Department treatment

PurposeEndothelial glycocalyx (EG) shedding may promote organ failure in sepsis. This study describes temporal changes in EG biomarkers from Emergency Department (ED) arrival, and associations with clinical characteristics. Materials and MethodsThis prospective observational study included 23 patients with simple infection, 86 with sepsis and 29 healthy controls. Serum EG biomarkers included syndecan-1, syndecan-4 and hyaluronan. Samples were taken on enrolment in the ED (T0), 1 hour (T1), 3 hours (T3) and 12 to 24 hours (T24) later. ResultsSyndecan-1 concentration increased incrementally over time (T0-T24, both patient groups, P < .001) whereas hyaluronan concentration peaked at T3 (T0-T3, sepsis group, P < .001). Hyaluronan was positively associated with cumulative fluid volumes (P < .001) at T0, T1, and T3, independent of illness severity. Both syndecan-1 (OR 1.04, 95% CI 1.01-1.07, P = .017) and hyaluronan (OR 1.83, 95% CI 1.46-2.30, P < .001) were associated with organ failure, independent of age and comorbidity.Syndecan-4 concentration was not different between groups or over time. ConclusionsIn contrast to previous ICU studies, EG biomarkers increased during the first 24 hours of sepsis treatment and were associated with fluid volumes and organ failure. Further investigation is required to determine if interventions delivered in the ED contribute to EG shedding.

Improved quality of porcine embryos cultured with hyaluronan due to the modification of the mitochondrial membrane potential and reactive oxygen species level

Although considerable progress has been made in pig embryo culture systems, the developmental competence and quality of the produced embryos are still lower than their in vivo-derived counterparts. Because hyaluronan (HA) regulates various cellular processes and possesses antioxidant properties, this glycosaminoglycan seems to be a promising supplement in culture media. However, until now, its beneficial influence on in vitro pig embryo development has been debatable. Hence, we aimed to investigate the effect of 0.25 mg/mL, 0.5 mg/mL and 1 mg/mL concentrations of HA on the developmental potential and quality of cultured porcine embryos. We found that 1 mg/mL HA supplementation significantly increased the obtained percentages of cleaved embryos to ∼95%, morulae to ∼87% and blastocysts to ∼77%. At 0.5 mg/mL and 1 mg/mL HA concentrations, we observed a significantly improved blastocyst quality, expressed as the total number of cells per blastocyst, number of cells in the inner cell mass, number of TUNEL-positive nuclei per blastocyst, the TUNEL index and the blastocyst diameter. Because the inner mitochondrial membrane potential (ΔΨm) and reactive oxygen species (ROS) level are important for proper embryo development, for the first time, we measured these two parameters in cultured embryos at various HA concentrations and during their development up to the expanded blastocyst stage. For blastocysts cultured with 1 mg/mL HA, the ΔΨm and ROS level were ∼1.6 and 2.7 times lower, respectively, than those of the control blastocysts. Both ΔΨm and the ROS level were increased in parallel during in vitro embryo development with and without HA, but this increase was less pronounced in the presence of HA. Hence, our quantitative data unequivocally show that supplementation of NCSU-23 culture medium with 1 mg/mL HA improves the developmental potential and quality of pig embryos. This effect results from a significant decrease in the ROS level induced by the HA-dependent ΔΨm reduction.

Controlling adsorption of albumin with hyaluronan on silica surfaces and sulfonated latex particles

Polysaccharides are known to modify binding of proteins at interfaces and this paper describes studies of these interactions and how they are modified by pH. Specifically, the adsorption of human serum albumin on to polystyrene latex and to silica is described, focusing on how this is affected by hyaluronan. Experiments were designed to test how such binding might be modified under relevant physiological conditions. Changes in adsorption of albumin alone and the co-adsorption of albumin and hyaluronan are driven by electrostatic interactions. Multilayer binding is found to be regulated by the pH of the solution and the molecular mass and concentration of hyaluronan. Highest adsorption was observed at pH below 4.8 and for low molecular mass hyaluronan (≤150kDa) at concentrations above 2mgml−1. On silica with grafted hyaluronan, albumin absorption is reversed by changes in solvent pH due to their strong electrostatic attraction. Albumin physisorbed on silica surfaces is also rinsed away with dilute hyaluronan solution at pH 4.8. The results demonstrate that the protein adsorption can be controlled both by changes of pH and by interaction with other biological macromolecules.

The effect of vasopressin on the expression of genes of key enzymes of the interstitial hyaluronan turnover and concentration ability in WAG rat kidneys

In mammals arginine-vasopressin (AVP) is a major hormone involved in the regulation of renal water reabsorption, acting via the increase of the osmotic permeability of the epithelium of the collecting duct. The AVP-induced intracellular events include, as a core step, the trafficking of the vesicles containing the water channels, aquaporin-2, to the apical plasma membrane of the collecting duct’s principal cells. The interstitium of the renal inner medulla contains abundant linear negatively charged glycosaminoglycan, hyaluronan (HA), which affects the water flow between structures of the concentrating mechanism, depending on its polymeric state. Using real-time RT-PCR, we tested the assumption that the renal hyaluronan may be involved in the long-term vasopressin effect on water reabsorption. The expression of the genes encoding hyaluronan synthase-2 (Has2), hyaluronidase-1, and hyaluronidase-2 (Hyal1 and Hyal2) in the kidneys of Wistar Albino Glaxo (WAG) was studied. The Has2 mRNA content was the highest in the papilla of the kidneys of the hydrated rats. The V2 receptor-selective vasopressin analog dDAVP (100 μg/kg of body weight, twice a day intraperitoneally for 2 days) induced a considerable decrease in the Has2 mRNA content in the papilla with less pronounced changes in the cortex. In contrast to Has2, the dDAVP treatment caused a significant increase in the Hyal1 and Hyal2 mRNA content in the renal papilla. There was a good fit between the Hyal1 and Hyal2 transcriptional levels and changes in hyaluronidase activity in the renal tissue. It was suggested that vasopressin is able to inhibit the synthesis of hyaluronan and concomitantly promotes its degradation in the renal papilla interstitium, thereby facilitating water flow between the elements of the renal countercurrent system. The implications for this effect are discussed in the context of the data in the literature.

Research on the efficacy of different concentrations of sodium hyaluronate for the treatments of ocular surface acid burns

Objective To observe the efficacy of different concentrations of sodium hyaluronate for the treatments of ocular surface acid burns. Methods The records of 90 eyes of 82 patients, which suffered from Ⅰ-Ⅱ degree ocular acid burn were analyzed restrospectively.All cases treated with different concentrations of sodium hyaluronate eye drops and were divided into two groups randomly.The concentration of sodium hyaluronate was 0.1% in group A (49 eyes) and 0.3% in group B (41 eyes). And the tear film break up time and repair time of corneal epithelium were compared between two groups. Results The tear film break up time after surgery was (3.1±1.8)s in group A and (5.4±1.9)s in group B. The repair time of corneal epithelium in group A and group B were (4.0±1.6)d and(2.6±1.2)d respectively.The difference in the corneal epithelium repair time and tear film break up time were statistically significant(P=0.001). Conclusion High concentrations of sodium hyaluronate eye drops (0.3%) are better than the low concentration of the same eyedrop in promoting cornea and conjunctiva repair. Key words: Acid burn, ocular surface; Repair time, epithelium, cornea; Eye drops, sodium hyaluronate

Effect of hyaluronic acid initial concentration on cross-linking efficiency of hyaluronic acid - based hydrogels used in biomedical and cosmetic applications.

This work was aimed to explore the potential effect of hyaluronic acid (HA) initial concentration (7.0 - 14.0 % w/v) on cross-linking efficiency of HA hydrogels cross-linked with 1,4-butanediol diglycidyl ether (BDDE). The results revealed that the hydrogel prepared at 10.0 % HA concentration exhibited a slower degradation rate, a lower swelling ability and more regular porosity than those prepared at either lower or higher HA concentration. After four days incubating with hyaluronidase, the content of NAG (N-acetyl glucosamine) remaining in the 10.0 HA hydrogel was 25.1±1.9 % with respect to the total NAG content found in the original mass. In contrast, the hydrogels prepared at 7.0 % and 14.0 % HA concentration showed a less remaining content of NAG equaled to approximately 15.9±5.4 % and 19.5±2.6 % respectively. On the other hand, the swelling ability of tested hydrogels was steadily decreased with the increase of HA initial concentration until the 10.0 % HA hydrogel and then showed an opposite trend. Based on this finding, the 10.0 % HA hydrogel exhibited the lowest swelling ratio which was observed at 129±3.2 g/g in distilled water and at 116±2.4 g/g in phosphate buffer saline (PBS). The SEM images showed various morphologies within the entire range of tested hydrogels. However, the hydrogel prepared at 10.0 % HA concentration was more homogenous and appeared with narrower pore-size distribution ranged in diameter from less than 50 μm to approximately 300 μm. Finally, the effect of HA initial concentration was investigated by FTIR which confirmed that the 10.0 % HA hydrogel was subject to a greater loss of (- OH) at 3343 cm-1 than other hydrogels except the 11.0 % HA hydrogel. This phenomenon was probably attributed to the formation of pendants that allowed the 11.0 % HA hydrogel to appear with a lower peak intensity than the 10.0 % HA hydrogel in the FTIR spectra. In conclusion, the HA initial concentration plays a crucial role in determining the cross-linking efficiency of HA hydrogels cross-linked with BDDE.

The Macula Flava of the Human Vocal Fold as a Stem Cell Microenvironment.

1. There is growing evidence to suggest that the cells in the maculae flavae are tissue stem cells of the human vocal fold and maculae flavae are a candidate for a stem cell niche. 2. The latest research shows that the cells in the human maculae flavae are involved in the metabolism of extracellular matrices that are essential for the viscoelasticity in the human vocal fold mucosa as a vibrating tissue, and considered to be important cells in the growth, development, and aging of the human vocal fold mucosa. 3. The cells in the human maculae flavae possess proteins of all three germ layers, indicating they are undifferentiated and have the ability of multipotency. 4. The cell division in the human adult maculae flavae is reflective of asymmetric self-renewal and cultured cells form a colony-forming unit. Therefore, the phenomenon gives rise to the strong possibility that the cells in the human maculae flavae are tissue stem cells. 5. Recent research suggests that the cells in the human maculae flavae arise from the differentiation of bone marrow cells via peripheral circulation. 6. The hyaluronan concentration in the maculae flavae is high and contains cells which possess hyaluronan receptors, indicating that the maculae flavae are hyaluronan-rich matrix, which is required for a stem cell niche. 7. A proper microenvironment in the maculae flavae of the human vocal fold mucosa is necessary to be effective as a stem cell niche maintaining the stemness of the contained tissue stem cells.

Structural behaviour of sodium hyaluronate in concentrated oppositely charged surfactant solutions.

This work discusses the polyelectrolyte sodium hyaluronate (HA) and its polyelectrolyte/surfactant complexes (PESCs) with tetradecyltrimethylammonium bromide (TTAB) in the semi-dilute regime of HA and at high concentrations of TTAB. The structure and flow properties in the surfactant excess region were studied by light scattering and small angle neutron scattering (SANS) as well as by rheology. The unique behaviour of HA to maintain its high viscosity was observed even at very high TTAB concentrations of 496 mM and this effect was systematically studied in the concentration range from 1 to 25 mM HA. From the data, it could be concluded that: (1) extended rod-like structures of the PESCs prevent molecular dissolution of HA by TTAB. (2) HA and TTAB micelles interact rather weakly as seen by a low fraction of bound micelles. (3) At very high TTAB concentrations a decompaction of PESCs (fractal dimension Df going from 2.0 to 1.2) occurs with increasing HA concentration but (4) both the entanglement of HA and the structure of the micelles are not affected.

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...

Anti-tumor and anti-invasion effects of a combination of 4-methylumbelliferone and ionizing radiation in human fibrosarcoma cells.

Hyaluronan (HA) is a major component of the extracellular matrix that is synthesized in excess in cancer tissues. 4-methylumbelliferone (MU) inhibits the synthesis of HA and is closely related to the invasion and metastasis of cancer. However, the effects of MU in conjunction with cancer radiotherapy remain unknown. The present study assessed the anti-tumor and anti-invasion effects of the concomitant use of ionizing radiation (IR) and 100 µM MU on human fibrosarcoma HT1080 cells. Cell viability and cellular invasion potency assays were performed. There was a greater decrease in the viability of cells cultured with a combination of 2 Gy IR and MU compared with untreated control cells. In addition, cell cycle distribution analysis demonstrated that a higher proportion of these cells were in the sub-G1 phase and higher fractions of annexin-V positive, propidium iodide positive cells (i.e., apoptotic cells) were observed. HA concentration in the 2 Gy irradiated culture was similar to that in the non-irradiated control culture, however, it significantly decreased following the administration of both MU alone and 2 Gy IR with MU. Furthermore, treatment with 2 Gy IR and MU resulted in a significant decrease in the invasion rate and matrix metalloproteinase (MMP)-2 and MPP-9 expression. Taken together, these results suggest that the administration of MU with 2 Gy IR is effective at reducing HA production, cell invasion and the metastatic potential of cancer cells.

Increased concentration of hyaluronan in tears after soaking contact lenses in Biotrue multipurpose solution.

PurposeThis study was conducted to determine 1) the concentration of hyaluronan (HA) in the tear films of contact lens (CL) wearers versus non-CL wearers and 2) whether HA sorbed from Biotrue, an HA-containing multipurpose solution (MPS), onto senofilcon A lenses affects the concentration of HA in tears after 2 hours of wear.Patients and methodsTears of habitual CL wearers and non-CL wearers were collected on Schirmer strips at baseline and after 2 hours of wear of senofilcon A CLs that had first been either rinsed with Sensitive Eyes Saline or soaked in Biotrue MPS for 14 hours. HA concentrations were measured by enzyme-linked immunosorbent assay (ELISA) and adjusted for sample volumes.ResultsNo difference in baseline concentrations of HA in tears was found between CL wearers and non-CL wearers (P=0.07), nor between males and females (P=0.06). However, age was significantly negatively associated with HA concentration (P<0.01), and mostly, CL wear contributed to a significant association (P<0.01). Among saline-rinsed CL wearers, no change in HA concentration in tears was observed after 2 hours of wear (P=0.38). By contrast, a significant increase in HA concentration was observed in the tears from eyes that had worn CLs soaked in Biotrue MPS when compared to baseline (P=0.01) or to saline-rinsed control (P=0.03).Conclusion1) In this study population, no difference in baseline concentration of HA was observed between CL wearers and non-CL wearers, and 2) after 2 hours of wear of senofilcon A lenses that were soaked in Biotrue MPS, HA concentrations in the tear films of CL wearers increased.

The effect of the intracervical administration of FSH or LH on the levels of hyaluronan, COX2, and COX2 mRNA in the cervix of the nonpregnant ewe

During the periovulatory period, the cervix relaxes in response to changes in circulating concentrations of reproductive hormones. The present study investigated the role of gonadotrophins in cervical function by examining the expression of cyclooxygenase-2 (COX2) and COX2 mRNA and the concentration of hyaluronan (HA) in the cervix, after intracervical treatment with either FSH or LH. Eighteen ewes were assigned to four groups. They were then treated with commercial intravaginal progestagen sponges and eCG to synchronize their estrous cycles. Intracervical treatments were given 24 hours after removal of the sponges as follows: group 1: FSH, 2 mg; group 2: LH, 2 mg; group 3: vehicle; and group 4: control. Cervices were collected 54 hours after sponge removal and then divided into three regions. The expression of COX2 and COX2 mRNA was determined by immunohistochemistry and in situ hybridization and those of HA by ELISA. The levels of expression of COX2, COX2 mRNA, and HA were compared in six tissue layers (luminal epithelium, subepithelial stroma, circular, longitudinal and transverse muscle, and serosa) and in three cervical regions (vaginal, mid, and uterine). The results showed that both FSH and LH significantly increased the levels the COX2 mRNA and COX2 in the cervix, but the effects of the gonadotrophins were selective. The effects of both FSH and LH were most evident at the vaginal end of the cervix and least at the uterine end of the cervix. Furthermore, their effects were confined to the stroma and smooth muscle layers of the cervix in the case of FSH and to smooth muscle only in the case of LH. Neither FSH nor LH affected the concentration of HA in the cervix although FSH but not LH reduced the concentration of HA in cervical mucus. These findings suggest that the gonadotrophins regulate the expression of COX2 in the cervix and that they may have a role facilitating relaxation of the cervix during estrus in the ewe.

Effect of vasopressin on the expression of genes for key enzymes of interstitial hyaluronan turnover and concentration ability in WAG rat kidneys

In mammals, arginine-vasopressin (AVP) is a major hormone involved in the regulation of renal water reabsorption, acting via an increase in the osmotic permeability of the collecting duct epithelium. The AVP-induced intracellular events include, as an essential step, the trafficking of the vesicles containing the water channels, aquaporin-2, to the apical plasma membrane of the collecting duct principal cells. The interstitium of the renal inner medulla contains abundant linear negatively charged glycosaminoglycan hyaluronan (HA), which affects the water flow depending on their polymeric state. Using real-time RT-PCR, we tested the assumption that the renal hyaluronan may be involved in the longterm vasopressin effect on water reabsorption. The expression of the genes encoding hyaluronan synthase-2 (Has2) and hyaluronidase-1, 2 (Hyal1, Hyal2) in the kidneys of Wistar Albino Glaxo (WAG) was studied. Has2 mRNA content was the highest in the kidney papilla of the hydrated rats. The V2 receptor-selective vasopressin analog dDAVP (100 μg/kg bw, ip, twice a day for 2 days) induced a considerable decrease in Has2 mRNA content in the papilla with less pronounced changes in the cortex. In contrast to Has2, dDAVP treatment caused a significant increase in Hyal1 and Hyal2 mRNA content in the renal papilla. There was a good fit between Hyal1 and Hyal2 transcriptional level and changes in hyaluronidase activity in the renal tissue. It was suggested that vasopressin is able to inhibit the synthesis of hyaluronan and concomitantly promotes its degradation in the renal papilla interstitium, thereby facilitating water flow between elements of the renal countercurrent system. The implications for this effect are discussed in the context of the literature data.

Decreased hyaluronan concentration during primary rhegmatogenous retinal detachment.

To evaluate the concentration and molecular weight of hyaluronan (HA) polysaccharides as well as hyaluronidase activity in patients with rhegmatogenous retinal detachment (RRD). Twenty vitreous samples from 20 patients with RRD and 19 samples from 19 patients with idiopathic epiretinal membrane, macular hole, or vitreomacular traction syndrome were collected during surgical management with pars plana vitrectomy. The molecular weight of various HA fragments was assessed using agarose gel electrophoresis. Enzyme-linked immunosorbent assay was employed for the measurement of HA (in μg/mL). Hyaluronidase activity was evaluated using substrate (HA) sodium dodecyl sulfate polyacrylamide gel electrophoresis. Agarose gel electrophoresis showed that the vitreous of the control group contained HA of high molecular mass, in contrast with the patient group. Mean HA concentration in the patient group was 50.96 μg/mL and differed significantly from that of the control group, which was 271.81 μg/mL (p&lt;0.0005). Hyaluronidase activity was significantly higher in the vitreous of patients with RRD (p = 0.037). The vitreous of patients with RRD is characterized by decreased HA concentration compared to controls of the same age and sex and shows higher hyaluronidase catalytic activity. Hyaluronan degradation could be associated with specific vitreous alterations that potentially contribute to retinal break formation and consequently detachment.

First in vivo detection and characterization of hyaluronan-coated extracellular vesicles in human synovial fluid.

Extracellular vesicles (EVs) function in intercellular signaling by transporting different membrane and cytosolic molecules, including hyaluronan (HA) and its synthesis machinery. As both EVs and HA are abundant in synovial fluid, we hypothesized that HA synthesized in synovial membrane would be carried on the surface of EVs. Synovial fluid (n = 15) and membrane samples (n = 5) were obtained from knee surgery patients. HA concentrations were analyzed in synovial fluid and HA and its synthesis machinery were examined with histochemical stainings in synovial membrane. To assess the size distribution of EVs in synovial fluid and to visualize HA on EVs, nanoparticle tracking analysis (NTA), confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM) were utilized. The average HA concentration in synovial fluid was 2.0 ± 0.21 mg/ml without significant differences between the patients with trauma/diagnostic arthroscopy and primary or post-traumatic osteoarthritis. Positive stainings of HA synthases (HAS1-3), HA and its receptor CD44 in synovial cells indicated active HA secretion in synovial membrane. According to NTA, EVs were abundant in synovial fluid and their main populations were ≤300 nm in diameter after differential centrifugation. There were no significant differences in the EV counts between the patients with primary or post-traumatic osteoarthritis. TEM verified that HA-positive particles detected by CLSM were lipid membrane vesicles surrounded by a HA coat. Our results provide the first in vivo evidence that human synovial fluid contains HA-positive EVs, one source of which presumably is the long HAS-positive protrusions of synovial fibroblasts. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1960-1968, 2016.

Interaction between Cationic Micelles and Hyaluronan

Micelles are particles of colloidal dimension and it is known that in an aqueous phase can solubilize water insoluble molecules (hydrophobic biologically active substances) in their inner core. However surfactants may have adverse effects on biological structures. Association of micelles with suitable bioacceptable molecules (e.g. hyaluronan) may lead to enhance their biocompatibility, decrease the nanoparticles cytotoxicity and also to target to the specific receptors in a human body. In this study, results of the research of electrostatic coating of cationic micelle with hyaluronan are reported. The cationic micelle/hyaluronan complexes were studied using turbidimetry and dynamic light scattering method and were evaluated to determine their structure, average size and zeta potential as a function of the amount of the both component in the system. Turbidimetric titration was chosen as indicator of the loss of intensity of transmitted light due to the scattering effect of particles associated from hyaluronan and surfactant in it. Dynamic light scattering measurement provided information about size distribution and zeta potential during mixing of component. Pilot solubilization experiments were performed to evaluation of stability and solubilization ability of the complexes. Results of turbidimetry revealed that aggregates formation (turbidity increasing) depends especially on hyaluronan concentration while surfactant concentration (above critical micelle concentration) affects interaction insignificantly. Dynamic light scattering experiments indicate that hyaluronan molecular weight has insignificant effect on isoelectric point of the systems. It was found that solubilization ability of the complexes is influenced by molecular weight of hyaluronan.

Sevoflurane mitigates shedding of hyaluronan from the coronary endothelium, also during ischemia/reperfusion: an ex vivo animal study.

Glycosaminoglycan hyaluronan (HA), a major constituent of the endothelial glycocalyx, helps to maintain vascular integrity. Preconditioning the heart with volatile anesthetic agents protects against ischemia/reperfusion injury. We investigated a possible protective effect of sevoflurane on the glycocalyx, especially on HA. The effect of pre-ischemic treatment with sevoflurane (15 minutes at 2% vol/vol gas) on shedding of HA was evaluated in 28 isolated, beating guinea pig hearts, subjected to warm ischemia (20 minutes at 37°C) followed by reperfusion (40 minutes), half with and half without preconditioning by sevoflurane. HA concentration was measured in the coronary effluent. Over the last 20 minutes of reperfusion hydroxyethyl starch (1 g%) was continuously infused and the epicardial transudate collected over the last 5 minutes for measuring the colloid extravasation. Additional hearts were fixed by perfusion after the end of reperfusion for immunohistology and electron microscopy. Sevoflurane did not significantly affect post-ischemic oxidative stress, but strongly inhibited shedding of HA during the whole period, surprisingly even prior to ischemia. Immunohistology demonstrated that heparan sulfates and SDC1 of the glycocalyx were also preserved by sevoflurane. Electron microscopy revealed shedding of glycocalyx caused by ischemia and a mostly intact glycocalyx in hearts exposed to sevoflurane. Coronary vascular permeability of the colloid hydroxyethyl starch was significantly decreased by sevoflurane vs the control. We conclude that application of sevoflurane preserves the coronary endothelial glycocalyx, especially HA, sustaining the vascular barrier against ischemic damage. This may explain beneficial effects associated with clinical use of volatile anesthetics against ischemia/reperfusion injury.

Effects of estradiol on reduction of osteoarthritis in rabbits through effect on matrix metalloproteinase proteins.

Osteoarthritis (OA), as a known degenerative joint disease, is the most common form of arthritis. In this study, we aimed to elucidate unclear pathogenesis of OA. Rabbit models of OA were established by the transection of the anterior cruciate ligament. Rabbits were randomly divided into three equal groups: the experimental group (OA modeling, treated with estradiol), the control group (OA modeling, treated with normal saline) and the normal group (without OA modeling). The glycosaminoglycan (GAG) and hyaluronan (HA) content of knee joint were collected and assayed. In addition, gene expression of matrix metalloproteinase (MMP)-1, MMP-13 and tissue inhibitor of metalloproteinase (TIMP)-1 were evaluated by real-time PCR and Western blot analysis. Animal models were developed successfully. GAG and HA concentrations were significantly increased in the experimental and the normal group compared with the control group (P<0.05 and P<0.01, respectively). Significant increase of GAG level in 6, 9 and 12 week-samples were found in the experimental group compared with the control group (P<0.01). The expression level of MMP-1 and MMP-13 in the experimental group were lower than the control group (P<0.01), but still higher than those of the normal group (P<0.01). TIMP-1 expression level was found to be higher in the experimental group than that of the control and normal group (P<0.01). The results suggested the possible role of estradiol in the pathological process of OA via its effect on the MMPs. The results also implied the effect of estradiol intervention on OA.

Respective roles of hyaluronidases 1 and 2 in endogenous hyaluronan turnover.

We studied the physiologic roles of the hyaluronidase (HYAL) 1 and HYAL2 in hyaluronan (HA) turnover. HA was localized and quantified using HA binding proteins in various tissues of Hyal1(-/-) and Hyal2(-/-) mice (knockout mice) as well as control mice. HA MW was determined using gel filtration chromatography. HA endocytosis in liver nonparenchymal cells (NPCs) was quantified in vivo Both Hyal1 and Hyal2 knockout mice showed HA accumulation in peripheral tissues without changes in HA MW distribution. HYAL2 deficiency induced buildup of very high MW (>3.10(6) Da) HA in lymph and serum with severe lymph node distortion. The lack of HYAL2 also impaired high MW HA endocytosis by liver NPCs. HYAL1 deficiency led to a moderate increase in serum HA concentration without changes in HA MW distribution and to HA overload of liver NPCs. Wild-type C57BL/6 mice served as controls. In HA injection experiments, saline-injected mice served as additional controls. We conclude that: 1) HYAL1 and HYAL2 are both needed for tissue HA catabolism; 2) HYAL2 is required for high MW HA clearance in lymph nodes and plasma and for HA endocytosis by liver NPCs; and 3) the main role of HYAL1 is HA degradation within liver NPCs.-Bourguignon, V., Flamion, B. Respective roles of hyaluronidases 1 and 2 in endogenous hyaluronan turnover.