Decreased Sulfate Content and Zeta Potential Distinguish Glycosaminoglycans of the Extracellular Matrix of Osteoarthritis Cartilage.

We aimed to determine the characteristics that distinguish glycosaminoglycans (GAGs) from osteoarthritis (OA) and normal cartilage and from men and women. Cartilage samples from 30 patients subjected to total joint arthroplasty secondary to OA or fracture (control) were evaluated, and the GAG content (µg/mg dry cartilage) after proteolysis was determined by densitometry, using agarose-gel electrophoresis. Relative percentages of carbon (C), nitrogen (N), and sulfur (S) in GAGs were determined by elemental microanalysis, as well as the zeta potential. Seventeen samples (56.6%) were from patients >70 years old, with 20 (66.6%) from women, and most [20 (66.6%)] were from the hip. The GAG content was similar regardless of patients being >/≤ 70 years old with 96.5 ± 63.5 and 78.5 ± 38.5 µg/mg (P = 0.1917), respectively. GAG content was higher in women as compared to men, with 89.5 ± 34.3 and 51.8 ± 13.3 µg/mg, respectively (P = 0.0022), as well as in OA than fracture samples, with 98.4 ± 63.5 and 63.6 ± 19.6 µg/mg, respectively (P = 0.0355). The GAG extracted from the cartilage of patients >70 years old had increase in N, and there were no gender differences regarding GAG elemental analysis. GAG from OA had a highly significant (P = 0.0005) decrease in S% (1.79% ± 0.25%), as compared to fracture samples (2.3% ± 0.19%), with an associated and significant (P = 0.0001) reduction of the zeta potential in the OA group. This is the first report of a reduced S content in GAG from OA patients, which is associated with a reduced zeta potential.

Galactomannan of Delonix regia seeds reduces nociception and morphological damage in the rat model of osteoarthritis induced by sodium monoiodoacetate.

This study investigated the effects of the protein-free galactomannan obtained from Delonix regia seeds (GM-DR) in an experimental osteoarthritis (OA) model. GM-DR was obtained from water-homogenized endosperms by collection of the supernatant and precipitation with ethanol. The remaining proteins in the galactomannan were removed by alkaline hydrolysis. Weight average molar mass (Mw) of the galactomannan was estimated in 5.8 × 105 g mol-1, presenting mannose:galactose ratio of 2.39:1. Rats received sodium monoiodoacetate (OA groups, 1 mg/25 µL) or saline (sham group) in the right tibio-tarsal joint. GM-DR (30-300 µg) was administered by intra-articular route at days 14 and 21 after OA induction. Hypernociception was evaluated daily by the measurement of the mechanical threshold required to cause joint flexion and paw withdrawal reflex. The 56-day animal groups were euthanized for joint histopahological analysis using the OARSI score system. Lower doses of GM-DR (30 and 100 µg) promoted antinociception from day 15 until the endpoint at day 56. Joint damage was reduced by GM-DR administration (100 µg) in OA-subjected animals, compared to the vehicle-treated OA group (5.9 ± 1.8 vs 19.0 ± 1.8, respectively, p < 0.05). Conclusion: Both antinociception and damage reduction suggest that Delonix regia galactomannan is a promising approach for osteoarthritis therapy.

Increase in molar mass distinguishes chondroitin sulfate from osteoarthritis and normal extracellular cartilage matrix.

PURPOSE: To determine alterations of chondroitin sulfate (CS) that reflect cartilage damage in an experimental osteoarthritis (OA) model as well as in human OA samples. MATERIALS AND METHODS: Rats were subjected to anterior cruciate ligament transection (ACLT; OA) or a sham procedure and sacrificed 14, 28, or 70 days after ACLT for histopathology and analysis of extracted CS. Cartilage samples from 14 patients undergoing hip or shoulder arthroplasty secondary to OA or fracture (control) were subjected to the same protocol. The CS content (µg/mg dry cartilage) after proteolysis was determined by densitometry, using agarose-gel electrophoresis. Molar mass (MM) and peak MM of CS were determined using high-performance size-exclusion chromatography (HPSEC). RESULTS: OA and sham joints at 70 d had 24 [22-24] and 3 [1-6] median histopathology scores, respectively (p < 0.001). Relative CS content (77.7 ± 8.3 µg/mg) was significantly increased in OA samples 70 d after ACLT, as compared to sham (53.5 ± 10.0 µg/mg). Peak MM of CS was higher in OA than in sham samples (P < 0.05). Similarly, CS content and peak MM were higher in cartilage from human OA patients, as compared to fracture samples, reproducing experimental data. CONCLUSION: Cartilage matrix from experimental and human OA samples has increased in the relative CS content. Increase in the peak MM distinguishes CS of the extracellular matrix of OA from normal cartilage.

Bacterial cellulose aerogels: Influence of oxidation and silanization on mechanical and absorption properties.

Biodegradable aerogels may help to develop eco-friendly technological pathways to increase the efficiency of chemical processes. In the present work, we describe the preparation of a novel bacterial cellulose aerogel oxidized by TEMPO, nanofibrillated in a blender, and silanized with methyltrimethoxysilane, resulting in four different types of aerogel. The aerogels produced from the double-functionalized cellulose suspension (BCOXNS) were compared to other non-oxidized (BCN and BCNS) and non-silanized (BCOXN) aerogels All aerogels were very light (density 10-14 kg.m-3) and very porous (porosity >99 %). The aerogels of BCOXNS showed better mechanical properties (tension of 13.0 kPa, modulus of elasticity of 39.4 kPa) and hydrophobicity, and could absorb organic solvents of different polarities. The BCOXNS could be recycled at least 7 times after absorbing organic solvents while retaining an absorption capacity of 83 %. This material can be used as a standard for the further development of aerogels based on bacterial biopolymers.

Galactomannan from the seeds of Caesalpinia pulcherrima prevents indomethacin-induced gastrointestinal damage via neutrophil migration.

Galactomannans are neutral polysaccharides isolated from the endosperm of some Leguminosae seeds. They consist of a (1 → 4) linked ß-mannopyranosyl backbone partially substituted at O-6 with α-d-galactopyranosyl side groups. C. pulcherrima have anti-inflammatory and muco-adhesive proprieties. Acute gastritis is an inflammatory disease triggered by use of non-steroidal anti-inflammatory drugs. We investigated the gastroprotective effect of galactomannan obtained from the seeds of Caesalpinia pulcherrima L. (GM-CP) in acute gastritis model induced by indomethacin. Gastritis was induced with indomethacin (30 mg/kg, P.·O.) in female Swiss mice. Animal groups (n = 7) were pretreated with saline-dissolved GM-CP (3 mg/kg, 10 mg/kg, 30 mg/kg, P.O.) or vehicle 1 h before gastritis induction. Mice were euthanized seven hours after the induction. The stomach and blood samples were collected for analysis. At 10 mg/kg, GP-CP reduced the extension of macroscopic lesion and the loss of superficial cells by alleviating inflammatory symptoms (neutrophil infiltration, migration and adhesion of mesenteric leukocytes, production of TNF-α and thiobarbituric acid reactive species (TBARS) and helping to maintain mucin labeling of the tissue. Thus, the findings of the study suggest that GM-CP exhibits gastroprotective effects.

Acetylated cashew gum-based nanoparticles for the incorporation of alkaloid epiisopiloturine.

The natural alkaloid epiisopiloturine has recently become the focus of study for various medicinal properties, particularly for its anti-inflammatory and antischistosomal effect. The incorporation of active molecules in natural polymeric matrices has garnered increasing interest during recent decades. A new derivative of cashew gum successfully obtained by gum acetylation has shown great potential as a carrier in controlled drug release systems. In this work, epiisopiloturine was encapsulated in acetylated cashew gum nanoparticles in order to increase solubility and allow slow release, whereas the morphology results were supported by computer simulations. The particles were produced under a variety of conditions, and thoroughly characterized using light scattering and microscopic techniques. The particles were spherical and highly stable in solution, and showed drug incorporation at high levels, up to 55% efficiency. Using a dialysis-based in vitro assay, these particles were shown to release the drug via a Fickian diffusion mechanism, leading to gradual drug release over approximately 6 h. These nanoparticles show potential for the use as drug delivery system, while studies on their potential anti-inflammatory action, as well as toxicity and efficacy assays would need to be performed in the future to confirm their suitability as drug delivery candidates.

Bacterial cellulose nanocrystals produced under different hydrolysis conditions: Properties and morphological features.

Bacterial cellulose (BC) is a polymer with interesting physical properties owing to the regular and uniform structure of its nanofibers, which are formed by amorphous (disordered) and crystalline (ordered) regions. Through hydrolysis with strong acids, it is possible to transform BC into a stable suspension of cellulose nanocrystals, adding new functionality to the material. The aim of this work was to evaluate the effects of inorganic acids on the production of BC nanocrystals (BCNCs). Acid hydrolysis was performed using different H2SO4 concentrations and reaction times, and combined hydrolysis with H2SO4 and HCl was also investigated. The obtained cellulose nanostructures were needle-like with lengths ranging between 622 and 1322nm, and diameters ranging between 33.7 and 44.3nm. The nanocrystals had a crystallinity index higher than native BC, and all BCNC suspensions exhibited zeta potential moduli greater than 30mV, indicating good colloidal stability. The mixture of acids resulted in improved thermal stability without decreased crystallinity.

Acetylated cashew gum-based nanoparticles for transdermal delivery of diclofenac diethyl amine.

Nanoprecipitation and dialysis methods were employed to obtain nanoparticles (NPs) of acetylated cashew gum (ACG). NPs synthesized by dialysis showed greater average size compared to those synthesized by nanoprecipitation, but they presented improved stability and yield. NPs were loaded with diclofenac diethylamine and the efficiency of the drug incorporation was over 60% for both methods, for an ACG:NP a weight ratio of 10:1. The cytotoxicity assay demonstrated that the NPs had no significant effect on the cell viability, verifying their biocompatibility. The release profile for the diclofenac diethylamine associated with the ACG-NPs showed a more controlled release compared to the free drug and a Fickian diffusion mechanism was observed. Transdermal permeation reached 90% penetration of the drug.

Clinical relevance of trans 1,4-polyisoprene aging degradation on the longevity of root canal treatment.

This in vivo study investigated the time of degradation of root filling material (trans 1,4-polyisoprene) retrieved from endodontically treated teeth and correlated the occurrence of degradation with the longevity of endodontics. Thirty-six root-filled teeth with different filling times (2 to 30 years) and with and without periapical lesions were selected. All teeth presented clinical indication for root canal retreatment. The association among filling time, presence of periapical lesion and root filling material degradation was investigated. Root filling samples were retrieved from the root canals using a Hedströ m file without solvent. The trans 1,4-polyisoprene was isolated by root filling solubilization in chloroform followed by filtration and centrifugation. GPC and FT-IR were the analytical techniques utilized. Degradation of trans 1,4-polyisoprene occurred with time, as a slow process. It is an oxidative process, and production of carboxyl and hydroxyl groups in the residual polymer were observed. Statistically significant decrease of molar mass was observed after 5 (p=0.0001) and 15 (p=0.01) years in teeth with and without periapical lesion, respectively. Bacteria participated in polymer degradation. Gutta-percha aging was proven an important factor for the long-term success of endodontic treatment. The findings of the present study showed that, after 15 years, polymer weight loss may decrease the capacity of the filling mass to seal the root canal space and prevent re-infection, thus compromising significantly the longevity of root canal therapy.

Brazilian gutta-percha points. Part II: thermal properties.

This study was undertaken to explore the effect of heating on gutta-percha, analyzing the occurrence of endothermic peaks corresponding to the transformation that occurs in the crystalline structure of the polymer during thermal manipulation. This study also sought to determine the temperature at which these peaks occur, causing a transformation from the beta- to the alpha-form, and from the alpha- to the amorphous phase. Eight nonstandardized gutta-percha points commercially available in Brazil (Konne, Tanari, Endopoint, Odous, Dentsply 0.04, Dentsply 0.06, Dentsply TP, Dentsply FM) and pure gutta-percha (control) were analysed using differential scanning calorimetry (DSC) and thermogravimetry analysis (TGA). The transition temperatures were determined and analysed. With the exception of Dentsply 0.04 and Dentsply 0.06, the majority of the products showed thermal behaviour typical of beta-gutta-percha, with two endothermic peaks, exhibiting two crystalline transformations upon heating from ambient temperature to 130 degrees. Upon cooling and reheating, few samples presented two endothermic peaks. It was concluded that heating dental gutta-percha to 130 degrees C causes changes to its chemical structure which permanently alter its physical properties.

Brazilian gutta-percha points. Part II: thermal properties

This study was undertaken to explore the effect of heating on gutta-percha, analyzing the occurrence of endothermic peaks corresponding to the transformation that occurs in the crystalline structure of the polymer during thermal manipulation. This study also seeked to determine the temperature at which these peaks occur, causing a transformation from the beta- to the alpha-form, and from the alpha- to the amorphous phase. Eight nonstandardized gutta-percha points commercially available in Brazil (Konne, Tanari, Endopoint, Odous, Dentsply 0.04, Dentsply 0.06, Dentsply TP, Dentsply FM) and pure gutta-percha (control) were analysed using differential scanning calorimetry (DSC) and thermogravimetry analysis (TGA). The transition temperatures were determined and analysed. With the exception of Dentsply 0.04 and Dentsply 0.06, the majority of the products showed thermal behaviour typical of beta-gutta-percha, with two endothermic peaks, exhibiting two crystalline transformations upon heating from ambient temperature to 130°. Upon cooling and reheating, few samples presented two endothermic peaks. It was concluded that heating dental gutta-percha to 130°C causes changes to its chemical structure which permanently alter its physical properties.

Este estudo teve como objetivo analisar, através da Calorimetria Diferencial de Varredura (DSC) e Análise Termogravimétrica (TGA), os efeitos do aquecimento sobre o polímero guta-percha, bem como explorar a ocorrência de picos endotérmicos, os quais correspondem às transformações cristalinas do polímero guta-percha, o que é traduzido em transições de fases (fase beta para alfa e fase alfa para amorfa). Foram utilizadas 8 marcas comerciais de cones de guta-percha não-estandardizados disponíveis no mercado brasileiro (Konne, Tanari, Endopoint, Odous, Dentsply 0.04, Dentsply 0.06, Dentsply TP, Dentsply FM), além da guta-percha pura (controle). As temperaturas de transição foram determinadas e analisadas. Com exceção das amostras Dentsply 0.04 e Dentsply 0.06, todas as demais apresentaram duas transformações cristalinas de fase quando submetidas ao aquecimento da temperatura ambiente até 130°C, comportamento típico de guta-percha em fase beta. Ao serem resfriadas e reaquecidas, poucas amostras apresentaram dois picos endotérmicos. É possível concluir que o aquecimento a 130°C causa danos na estrutura química do polímero guta-percha, o qual altera de forma definitiva suas propriedades físicas.

Analgesic activity of a polysaccharide in experimental osteoarthritis in rats.

Viscosupplementation efficacy has been related to the high molecular weight of hyaluronic acid-like compounds, as well as to gel formulation. We evaluated the effect of a galactomannan polysaccharide derived from Guar gum (GG) in joint pain in an osteoarthritis (OA) model. Wistar rats (six animals/group) were subjected to anterior cruciate ligament transection (ACLT-OA group). The OA group was compared to a false-operated group (sham). Joint pain was recorded daily, using the articular incapacitation test, until 7 days after ACLT. Solutions or gel preparations of GG (100 microg) or Hylan G-F 20 (100 microg), used as a comparator, were given intraarticularly (i.a.) at day 4 after ACLT. Controls received saline i.a. The OA group had significantly increased joint pain as compared to sham (P<0.001). GG, either as a gel or solution, significantly inhibited joint pain similar to the inhibition achieved with Hylan G-F20. This is the first demonstration that a galactomannan derived from GG reduces joint pain in experimental OA. This analgesia is independent of the colloidal state. We propose that the analgesic benefit of viscosupplementation may be due to an intrinsic carbohydrate-mediated mechanism rather than to the rheologic properties of the material.

Clinical relevance of trans 1.4-polyisoprene aging degradation on the longevity of root canal treatment

This in vivo study investigated the time of degradation of root filling material (trans 1,4-polyisoprene) retrieved from endodontically treated teeth and correlated the occurrence of degradation with the longevity of endodontics. Thirty-six root-filled teeth with different filling times (2 to 30 years) and with and without periapical lesions were selected. All teeth presented clinical indication for root canal retreatment. The association among filling time, presence of periapical lesion and root filling material degradation was investigated. Root filling samples were retrieved from the root canals using a Hedströ m file without solvent. The trans 1,4-polyisoprene was isolated by root filling solubilization in chloroform followed by filtration and centrifugation. GPC and FT-IR were the analytical techniques utilized. Degradation of trans 1,4-polyisoprene occurred with time, as a slow process. It is an oxidative process, and production of carboxyl and hydroxyl groups in the residual polymer were observed. Statistically significant decrease of molar mass was observed after 5 (p=0.0001) and 15 (p=0.01) years in teeth with and without periapical lesion, respectively. Bacteria participated in polymer degradation. Gutta-percha aging was proven an important factor for the long-term success of endodontic treatment. The findings of the present study showed that, after 15 years, polymer weight loss may decrease the capacity of the filling mass to seal the root canal space and prevent re-infection, thus compromising significantly the longevity of root canal therapy.

Este estudo in vivo avaliou a degradação do material obturador e a influência deste fator na longevidade do tratamento endodôntico. Foram selecionados 36 pacientes (3-30 anos) com canais tratados endodonticamente, com e sem lesões periapicais, e indicação de retratamento endodôntico. Foi investigada a associação entre o tempo de tratamento, presença de lesão periapical e a degradação do material obturador. O material obturador foi removido com uma lima Hedströ em sem uso de solvente. O polímero trans 1,4- poliisopreno foi isolado do material obturador através de solubilização em clorofórmio, seguido de filtragem e centrifugação. GPC e FT-IR foram os métodos analíticos utilizados. A degradação do trans 1,4- poliisopreno foi observada com o tempo, sendo um processo lento e oxidativo, com formação de grupos carboxílicos e hidroxilas no polímero residual. Após 5 (p=0,0001) e 15 (p=0,01) anos, em dentes com e sem lesões periapicais, respectivamente, houve decréscimos significantes na massa molar do material obturador. A infecção bacteriana participa no processo de degradação do polímero. O envelhecimento da guta-percha é um fator que influencia o sucesso a longo prazo do tratamento endodôntico. Após 15 anos, a longevidade do tratamento pode ser significantemente afetada pela redução da capacidade de selamento causada pela perda de massa molar do polímero, permitindo a reinfecção do sistema de canais radiculares.

Brazilian gutta-percha points. Part I: chemical composition and X-ray diffraction analysis.

Eight nonstandardized gutta-percha points commercially available in Brazil (Konne, Tanari, Endopoint, Odous, Dentsply 0.04, Dentsply 0.06, Dentsply TP and Dentsply FM) were analysed chemically and by X-ray diffraction, and their chemical compositions were compared. The organic fraction (gutta-percha polymer and wax/resin) of the gutta-percha points was separated from the inorganic fraction (ZnO and BaSO4) by dissolving them in chloroform. The gutta-percha polymer was precipitated with acetone. The inorganic fraction was analysed by elemental microanalysis. Energy-dispersive X-ray microanalysis (EDX) and X-ray diffraction were employed to identify the chemical elements and compounds (barium sulfate and zinc oxide). The barium sulfate content was calculated based on the percentage of sulfur found in the elemental microanalysis. All analyses were repeated three times. The means and standard deviations of the percentage by weight of gutta-percha in the points were: Konne (17.6 +/- 0.30), Tanari (15.2 +/- 0.30), Endopoint (16.7 +/- 0.23), Odous (18.8 +/- 0.20), Dentsply 0.04 (15.7 +/- 0.17), Dentsply 0.06 (16.6 +/- 0.17), Dentsply TP (21.6 +/- 0.15) and Dentsply FM (16.3 +/- 0.23). The means and standard deviations of the zinc oxide content were: Konne (79.9 +/- 0.10), Tanari (81.9 +/- 0.07), Endopoint (81.3 +/- 0.40), Odous (79.7 +/- 0.26), Dentsply 0.04 (77.9 +/- 0.03), Dentsply 0.06 (78.2 +/- 0.07), Dentsply TP (69.8 +/- 0.19) and Dentsply FM (72.6 +/- 0.70). The method utilized was appropriate to quantify gutta-percha, wax/resin, zinc oxide and barium sulfate. Cone brands without barium sulfate were found. An unusual high wax/resin percentage was detected in Dentsply FM (p = 0.0003). Dentsply TP showed the highest gutta-percha percentage.

Brazilian gutta-percha points: Part I: chemical composition and X-ray diffraction analysis / Cones nacionais de guta-percha. Parte I: composição química e análise por difração de raios X

Oito marcas de cones de guta-percha disponíveis no mercado brasileiro (Konne, Tanari, Endopoint, Odous, Dentsply 0.04, Dentsply 0.06, Dentsply TP e Dentsply FM) foram analisadas quimicamente e por difração de raios X; e suas composições, comparadas. A porção orgânica (guta-percha e ceras/resinas) foi separada da porção inorgânica (ZnO e BaSO4) através da dissolução em clorofórmio. O polímero guta-percha foi precipitado com adição de acetona. A fração inorgânica foi analisada por microanálise elementar. Microanálise por energia dispersiva de raios X (EDX) e difração por raios X foram utilizadas para identificar os elementos e compostos (BaSO4 e ZnO). A quantidade de sulfato de bário foi calculada através da porcentagem de enxofre detectada na microanálise elementar. Todas as análises foram executadas em triplicata. As médias e os desvios padrões das porcentagens em peso de guta-percha nas diferentes marcas analisadas foram: Konne (17,6 ± 0,30), Tanari (15,2 ± 0,30), Endopoint (16,7 ± 0,23), Odous (18,8 ± 0,20), Dentsply 0.04 (15,7 ± 0,17), Dentsply 0.06 (16,6 ± 0,17), Dentsply TP (21,6 ± 0,15) e Dentsply FM (16,3 ± 0,23). As médias e os desvios padrões das quantidades de óxido de zinco encontrados foram: Konne (79,9 ± 0,10), Tanari (81,9 ± 0,07), Endopoint (81,3 ± 0,40), Odous (79,7 ± 0,26), Dentsply 0.04 (77,9 ± 0,03), Dentsply 0.06 (78,2 ± 0,07), Dentsply TP (69,8 ± 0,19) e Dentsply FM (72,6 ± 0,70). O método utilizado foi apropriado para quantificação dos componentes guta-percha, ceras/resinas, óxido de zinco e sulfato de bário. Foram encontrados cones que não apresentaram sulfato de bário na composição. Foi detectada uma concentração exagerada de ceras/resinas no cone Dentsply FM (p = 0,0003). O cone Dentsply TP apresentou a maior porcentagem de guta-percha.