Protection of corneal endothelium from irrigation damage:A comparison of sodium hyaluronate and hydroxypropylmethyl-cellulose

Purpose: To compare the endothelial protection of sodium hyaluronate and hydroxypropylmethylcellulose against endothelial damage induced by irrigation. Methods: An in vitro assay with freshly excised porcine eyes was developed using the Janus green photometry technique. Irrigation and aspiration technique was standardised. Forty pairs of porcine eyes were used. One randomly chosen eye was filled with sodium hyaluronate (SH) and the other with hydroxypropylmethylcellulose (HPMC). Irrigation and aspiration was carried out with balanced salt solution for 5 min. Twenty additional pairs of porcine eyes served as controls. Student's t-test was used for statistical analysis. Results: Both viscoelastic agents protected the endothelium as compared with controls. The endothelial protection, determined with the Janus green photometric technique, was significantly greater with HPMC than with SH. Conclusions: Viscoelastic agents are effective in protecting the endothelium from irrigation damage in porcine eyes in vitro. HPMC provided greater protection than SH in this particular model.

Effects of sodium hyaluronate and triamcinolone acetonide on proteoglycan metabolism in equine articular chondrocytes treated with interleukin-1

The objective of this study is to determine if the effects of a high molecular weight sodium hyaluronate (HA) alone or in combination with triamcinolone acetate (TA) can mitigate chondrocyte proteoglycan catabolism caused by interleukin-1 (IL-1) administration. Chondrocytes were collected from fetlock joints of ten horses euthanized for reasons unrelated to joint disease. Chondrocyte pellets were treated with media (negative control); media containing IL-1 only (positive control); or media containing IL-1 with HA only (0.5 or 2.0 mg/mL), TA only (0.06 or 0.6 mg/mL), or HA (0.5 or 2.0 mg/mL) and TA (0.06 or 0.6 mg/mL) in combination. Chondrocyte pellets were assayed for newly synthesized GAG, total GAG content, total DNA content, and mRNA levels of collagen type II, aggrecan, and COX-2. The high concentration of HA (2.0 mg/mL) increased GAG synthesis while the high concentration of TA (0.6 mg/mL) decreased loss of GAG into the media. Both the high concentration of HA and TA increased the total GAG content within the pellet. There was no change in pellet DNA content with either treatment. TA reduced COX-2 mRNA levels as well as aggrecan and collagen type II expression. Treatment with HA had no effect on mRNA levels of COX-2, aggrecan or collagen type II. These results indicate that the high concentration of HA or TA alone or in combination will mitigate effects of IL-1 administration on proteoglycan catabolism of equine articular chondrocytes.

Uso de viscoelásticos na facoemulsificação em cães portadores de catarata: efeitos sobre a pressão intraocular, a morfologia das células endoteliais e a espessura corneana

Avaliaram-se as células endoteliais, a espessura corneana e a pressão intraocular (PIO) de cães portadores de catarata madura, empregando-se viscoelástico à base de hialuronato de sódio 3% e sulfato de condroitina 4% e hidroxipropilmetilcelulose 2%, utilizando-se 20 cães, distribuídos entre os dois grupos dos viscoelásticos. A técnica cirúrgica adotada foi a da facoemulsificação bimanual. As avaliações tonométricas foram efetuadas antes e após o ato cirúrgico, aos 1, 7, 14, 21, 28 e 60 dias de pós-operatório, e a microscopia especular, antes e após 7, 28 e 60 dias. Não houve diferença estatística entre os grupos quanto à PIO, com exceção aos 14 dias, em que se observou maior PIO com o uso de hialuronato de sódio 3% e sulfato de condroitina 4%. Não houve diferença entre os grupos quanto aos parâmetros relacionados ao endotélio, com diminuição discreta da densidade celular endotelial e aumento da área celular com a utilização de hidroxipropilmetilcelulose 2%. A utilização de ambos os dispositivos viscoelásticos analisados é recomendada para o procedimento de facoemulsificação em cães.

Light microscopic evaluation and scanning electron microscopic analysis of horse eyes following deep anterior lamellar keratectomy

Objective To describe the technique of deep anterior lamellar keratoplasty (DALK) with Descemet's membrane (DM) exposure in horse eyes. Also, to compare the efficacy and safety of viscodissection and big-bubble techniques for DALK. Animals studied Thirty-four ex vivo horse eyes. Procedure Deep anterior lamellar keratoplasty was performed in 34 ex vivo horse eyes. Two groups (Group V - viscodissection - 2% sodium hyaluronate; Group A - air - big-bubble) of 17 eyes were studied. Other than the substance used, the surgical technique was similar for both groups. Nonperforated eyes were submitted for light microscopic histologic evaluation and scanning electron microscopic (SEM) analysis. Results Group V - Perforations occurred in 18% of the eyes during surgery. Light microscopy revealed exposure of DM in 28% of the eyes with mean thickness of the remaining stroma being 70.4μm. Group A - Perforations occurred in 42% of the eyes. Light microscopy revealed exposure of DM in 60% of the eyes with mean thickness of the remaining stroma being 23.3μm. No significant differences in safety, efficacy and thickness of the remaining stroma (including all eyes or excluding those with DM exposure) were observed. SEM of the surgical site revealed a more even surface in those eyes with DM exposure compared to eyes with thicker remaining stroma in both groups. Conclusions We describe two DALK techniques (viscodissection and big-bubble) for use in horses. No significant differences in safety, efficacy and thickness of the remaining stroma were observed. However, a nonsignificant trend toward the big-bubble technique being more efficacious but less safe was observed. © 2012 American College of Veterinary Ophthalmologists.

Análise comparativa de dois viscoelásticos dispersivos na facoemulsificação em cães portadores de catarata

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Uso de hialuronidato de sódio para tratamento de deslocamento anterior de disco articular com travamento bucal

Temporomandibular joint disorders causing limitation of mouth opening and pain may be related to the restriction of slipping or forward displacement of the mandibular condyle. Temporomandibluar arthrocentesis is the lavage of the joint and is regarded as a simple method that produces good results in patients with these symptoms. This article reports a case of disc displacement and closed lock in a patient who reported limited mouth opening and pain as the main symptoms. The treatment proposed was arthrocentesis and lavage, combining a corticosteroid with a sodium hyaluronate solution. It is concluded that arthrocentesis is a simple, efficient and barely invasive method for particular types of temporomandibular joint disorders and should be considered before opting for more invasive surgical procedures.

Properties of an injectable low modulus PMMA bone cement for osteoporotic bone

The use of polymethylmethacrylate (PMMA) cement to reinforce fragile or broken vertebral bodies (vertebroplasty) leads to extensive bone stiffening. Fractures in the adjacent vertebrae may be the consequence of this procedure. PMMA with a reduced Young's modulus may be more suitable. The goal of this study was to produce and characterize stiffness adapted PMMA bone cements. Porous PMMA bone cements were produced by combining PMMA with various volume fractions of an aqueous sodium hyaluronate solution. Porosity, Young's modulus, yield strength, polymerization temperature, setting time, viscosity, injectability, and monomer release of those porous cements were investigated. Samples presented pores with diameters in the range of 25-260 microm and porosity up to 56%. Young's modulus and yield strength decreased from 930 to 50 MPa and from 39 to 1.3 MPa between 0 and 56% porosity, respectively. The polymerization temperature decreased from 68 degrees C (0%, regular cement) to 41 degrees C for cement having 30% aqueous fraction. Setting time decreased from 1020 s (0%, regular cement) to 720 s for the 30% composition. Viscosity of the 30% composition (145 Pa s) was higher than the ones received from regular cement and the 45% composition (100-125 Pa s). The monomer release was in the range of 4-10 mg/mL for all porosities; showing no higher release for the porous materials. The generation of pores using an aqueous gel seems to be a promising method to make the PMMA cement more compliant and lower its mechanical properties to values close to those of cancellous bone.

Particle engineering for the production of respirable dry powder formulations

Hyaluronan (HA) plays an important role in lung pathophysiology. For this reason it has attracted great attention both as active ingredient and as excipient in treating lung diseases by direct pulmonary HA administration. The aim was the production of highly respirable and flowable HA powders either as a potential carrier for drug delivery or for being delivered directly by inhalation. Engineered sodium hyaluronate powders were produced by spray-drying technique. All the spray-dried powders were characterised in terms of particle size distribution, drug content, morphology and in vitro respirability. HA was successfully formulated with salbutamol sulphate in combination with leucine and highlighted remarkable aerodynamic performance (emitted dose equal to 83 % and FPF % equal to 97.1%). Moreover, HA colloidal solutions were designed and they were spray-dried. In order to improve particle aerodynamic characteristics, different types of excipients were investigated. In particular, stearylamine (5% w/w) allowed to obtain the best performance throughout the experimental set. Finally, in vitro biocompatibility was carried out by MTT assay and High Content Analysis for selected dry powder formulations and starting materials. The assays demonstrated the same outcome by confirming the HA biocompatibility and by producing the same rank of toxicity for the surfactants. The general conclusion of the project is that formulation containing HA and stearyl alcohol represents the best performing formulation.

A double blind, randomized, multicenter, parallel group study of the effectiveness and tolerance of intraarticular hyaluronan in osteoarthritis of the knee

Objective. To investigate the efficacy and tolerability of a course of 5 injections of hyaluronan (HA) given at intervals of one week in patients with symptomatic, mild to moderate osteoarthritis (OA) of the knee. Methods: A double blind, randomized, parallel group, multicenter (17 centers), saline vehicle-controlled study was conducted over 18 weeks. Patients received either 25 mg (2.5 ml) HA in a phosphate buffered solution or 2.5 ml vehicle containing only the buffer by intraarticular injection. Five injections were given at one week intervals and the patients were followed for a further 13 weeks. The Western Ontario McMaster (WOMAC) OA instrument was used as the primary efficacy variable and repeated measures analysis of covariance was used to compare the 2 treatments over Weeks 6, 10, 14, and 18. Results. Of 240 patients randomized for inclusion in the study, 223 were evaluable for the modified intention to treat analysis. The active treatment and control groups were comparable for demographic details, OA history, and previous treatments. Scores for the pain and stiffness subscales of the WOMAC were modestly but significantly lower in the HA-treated group overall (Weeks 6 to 18; p < 0.05) and the statistically significant difference from the control was not apparent until after the series of injections was complete. The physical function subscale did not reach statistical significance (p = 0.064). Tolerability of the procedure was good and there were no serious adverse events that were considered to have a possible causal relationship with the study treatment. Conclusion. Intraarticular HA treatment was significantly more effective than saline vehicle in mild to moderate OA of the knee for the 13 week postinjection period of the study.

Viscosupplementation for the treatment of osteoarthritis of the knee (Review)

Background Osteoarthritis (OA) is the most prevalent chronic joint disorder worldwide and is associated with significant pain and disability. Objectives To assess the effects of viscosupplementation in the treatment of OA of the knee. The products were hyaluronan and hylan derivatives (Adant, Arthrum H, Artz (Artzal, Supartz), BioHy (Arthrease, Euflexxa, Nuflexxa), Durolane, Fermathron, Go-On, Hyalgan, Hylan G-F 20 (Synvisc Hylan G-F 20), Hyruan, NRD-101 (Suvenyl), Orthovisc, Ostenil, Replasyn, SLM-10, Suplasyn, Synject and Zeel compositum). Search strategy MEDLINE (up to January (week 1) 2006 for update), EMBASE, PREMEDLINE, Current Contents up to July 2003, and the Cochrane Central Register of Controlled Trials (CENTRAL) were searched. Specialised journals and reference lists of identified randomised controlled trials (RCTs) and pertinent review articles up to December 2005 were handsearched. Selection criteria RCTs of viscosupplementation for the treatment of people with a diagnosis of OA of the knee were eligible. Single and double-blinded studies, placebo-based and comparative studies were eligible. At least one of the four OMERACT III core set outcome measures had to be reported (Bellamy 1997). Data collection and analysis Each trial was assessed independently by two reviewers for its methodological quality using a validated tool. All data were extracted by one reviewer and verified by a second reviewer. Continuous outcome measures were analysed as weighted mean differences (WMD) with 95% confidence intervals (CI). However, where different scales were used to measure the same outcome, standardized mean differences (SMD) were used. Dichotomous outcomes were analyzed by relative risk (RR). Main results Seventy-six trials with a median quality score of 3 (range 1 to 5) were identified. Follow-up periods varied between day of last injection and eighteen months. Forty trials included comparisons of hyaluronan/hylan and placebo (saline or arthrocentesis), ten trials included comparisons of intra-articular (IA) corticosteroids, six trials included comparisons of nonsteroidal anti-inflammatory drugs (NSAIDs), three trials included comparisons of physical therapy, two trials included comparisons of exercise, two trials included comparisons of arthroscopy, two trials included comparisons of conventional treatment, and fifteen trials included comparisons of other hyaluronans/hylan. The pooled analyses of the effects of viscosupplements against 'placebo' controls generally supported the efficacy of this class of intervention. In these same analyses, differential efficacy effects were observed for different products on different variables and at different timepoints. Of note is the 5 to 13 week post injection period which showed a percent improvement from baseline of 28 to 54% for pain and 9 to 32% for function. In general, comparable efficacy was noted against NSAIDs and longer-term benefits were noted in comparisons against IA corticosteroids. In general, few adverse events were reported in the hyaluronan/hylan trials included in these analyses. Authors' conclusions Based on the aforementioned analyses, viscosupplementation is an effective treatment for OA of the knee with beneficial effects: on pain, function and patient global assessment; and at different post injection periods but especially at the 5 to 13 week post injection period. It is of note that the magnitude of the clinical effect, as expressed by the WMD and standardised mean difference (SMD) from the RevMan 4.2 output, is different for different products, comparisons, timepoints, variables and trial designs. However, there are few randomised head-to-head comparisons of different viscosupplements and readers should be cautious, therefore, in drawing conclusions regarding the relative value of different products. The clinical effect for some products, against placebo, on some variables at some timepoints is in the moderate to large effect-size range. Readers should refer to relevant tables to review specific detail given the heterogeneity in effects across the product class and some discrepancies observed between the RevMan 4.2 analyses and the original publications. Overall, the analyses performed are positive for the HA class and particularly positive for some products with respect to certain variables and timepoints, such as pain on weight bearing at 5 to 13 weeks postinjection. In general, sample-size restrictions preclude any definitive comment on the safety of the HA class of products; however, within the constraints of the trial designs employed no major safety issues were detected. In some analyses viscosupplements were comparable in efficacy to systemic forms of active intervention, with more local reactions but fewer systemic adverse events. In other analyses HA products had more prolonged effects than IA corticosteroids. Overall, the aforementioned analyses support the use of the HA class of products in the treatment of knee OA.

Microbial hyaluronic acid production

Hyaluronic acid (HA) is a commercially valuable medical biopolymer increasingly produced through microbial fermentation. Viscosity limits product yield and the focus of research and development has been on improving the key quality parameters, purity and molecular weight. Traditional strain and process optimisation has yielded significant improvements, but appears to have reached a limit. Metabolic engineering is providing new opportunities and HA produced in a heterologous host is about to enter the market. In order to realise the full potential of metabolic engineering, however, greater understanding of the mechanisms underlying chain termination is required.

Sodium 2-nitrocinnamate dihydrate: a one-dimensional hydrogen-bonded coordination polymer

The title compound catena-poly[aqua-mu3-2-nitrocinnamato], [Na(C9H6NO4)(H2O)2]n, the sodium salt of trans-2-nitrocinnamic acid, is a one-dimensional coordination polymer based on six-coordinate octahedral NaO6 centres comprising three facially-related monodentate carboxylate O-atom donors from separate ligands (all bridging)[Na-O, 2.4370(13)-2.5046(13)A] and three water molecules (two bridging, one monodentate) [Na-O, 2.3782(13)-2.4404(17)A]. The structure is also stabilized by intra-chain water-O-H...O(carboxylate) and O-H...O(nitro) hydrogen bonds.

Fluorescence spectrometric study on the interactions of isoprocarb and sodium 2-isopropylphenate with bovine serum albumin

The binding interaction of the pesticide Isoprocarb and its degradation product, sodium 2-isopropylphenate, with bovine serum albumin (BSA) was studied by spectrofluorimetry under simulated physiological conditions. Both Isoprocarb and sodium 2-isopropylphenate quenched the intrinsic fluorescence of BSA. This quenching proceeded via a static mechanism. The thermodynamic parameters (ΔH°, ΔS° and ΔG°) obtained from the fluorescence data measured at two different temperatures showed that the binding of Isoprocarb to BSA involved hydrogen bonds and that of sodium 2-isopropylphenate to BSA involved hydrophobic and electrostatic interactions. Synchronous fluorescence spectroscopy of the interaction of BSA with either Isoprocarb or sodium 2-isopropylphenate showed that the molecular structure of the BSA was changed significantly, which is consistent with the known toxicity of the pesticide, i.e., the protein is denatured. The sodium 2-isopropylphenate, was estimated to be about 4–5 times more toxic than its parent, Isoprocarb. Synchronous fluorescence spectroscopy and the resolution of the three-way excitation–emission fluorescence spectra by the PARAFAC method extracted the relative concentration profiles of BSA, Isoprocab and sodium 2-isopropylphenate as a function of the added sodium 2-isopropylphenate. These profiles showed that the degradation product, sodium 2-isopropylphenate, displaced the pesticide in a competitive reaction with the BSA protein.