Induction of M2 Macrophages Prevents Bone Loss in Murine Periodontitis Models.

Periodontitis is characterized by the progressive destruction of tooth-supporting alveolar bone, which is mainly caused by chronic inflammation in response to persistent bacterial insult. It has recently become clear that the pathogenesis of periodontitis is associated with a high ratio of proinflammatory M1 (classically activated) macrophages to anti-inflammatory M2 (alternatively activated). To decrease the inflammatory activity, we locally delivered the C-C motif chemokine ligand 2 (CCL2) using controlled-release microparticles (MPs). CCL2 is known to promote chemotaxis of M0 or M2 phenotype macrophages to the inflamed site and induce M2 phenotype polarization locally. Our in vitro data showed that CCL2 increased the number of M2 phenotype macrophages, decreased TNF-α secretion, and enhanced chemotaxis of RAW264.7 cells toward CCL2 MPs. Moreover, we induced periodontal disease in 2 animal models through inoculation of Porphyromonas gingivalis and ligature around the murine molar. Micro-computed tomography analysis showed significant reduction of alveolar bone loss in the CCL2 MP treatment group when compared with a blank MP group and a no-treatment periodontitis group in both models. Immunohistologic analysis showed a significant increase in the M2 phenotype subset and a decrease in the M1 phenotype subset in the CCL2 MP group of the P. gingivalis-induced model. Also, in both models, tartrate-resistant acidic phosphatase staining showed significantly fewer numbers of osteoclasts in the CCL2 MP group in alveolar bone area. Moreover, quantitative polymerase chain reaction results showed a significant increase in IL-1RA (interleukin 1 receptor antagonist) mRNA expression and a decrease in RANKL (receptor activator of nuclear factor kappa-Β ligand) mRNA expression in the CCL2 MP group in the ligature model. In summary, manipulation of endogenous M2 phenotype macrophages with CCL2 MPs decreased the M1 phenotype:M2 phenotype ratio and prevented alveolar bone loss in mouse periodontitis models. The delivery of CCL2 MPs provides a novel approach to treat periodontal disease.

Permeability hysterisis of limestone during isotropic compression.

The evolution of permeability hysterisis in Indiana Limestone during application of isotropic confining pressures up to 60 MPa was measured by conducting one-dimensional constant flow rate tests. These tests were carried out either during monotonic application of the confining pressure or during loading-partial unloading cycles. Irreversible permeability changes occurred during both monotonic and repeated incremental compression of the limestone. Mathematical relationships are developed for describing the evolution of path-dependent permeability during isotropic compression.

Dermatan sulfates of normal and scarred fascia.

We evaluated the composition of dermatan sulfates (DS) derived from 23 samples of normal and 23 samples of scarred fascia lata. We analyzed the molecular weight of intact DS chains and the length of chain regions comprising: (1) clusters of L-iduronate-containing disaccharides ("iduronic sections"); (2) clusters of D-glucuronate-containing disaccharides ("glucuronic sections"); and (3) copolymeric sections with both types of disaccharides. A portion of scarred fascia DS chains demonstrated higher molecular weight compared with those from normal tissue. Most disaccharides of DS chains derived from both fascia types form copolymeric segments - heterogeneous in size - with alternatively distributed single disaccharides with glucuronic residues and mainly single ones with iduronate. Only a small number of disaccharides form "glucuronic sections" of heterogeneous size or short "iduronic sections". However, the scarred fascia DS chains demonstrate an increased content of shorter "glucuronic sections" and shorter, often oversulfated, copolymeric segments. It seems that in normal fascia, the DS chain type with a single, long copolymeric region and a single, shorter "glucuronic section" is predominant, while in scarred tissue an increase in multidomain DS chain content may occur.

An accumulation of proteoglycans in scarred fascia.

A little is known about proteoglycan (PG) changes, occuring in the course of scarring of tissues another than skin. The aim of present study was biochemical characterization of glycosaminoglycans (GAGs) and proteoglycans (PGs) of normal and scarred fascia. Samples of normal fascia lata were taken at autopsy from 23 individuals and samples of scarred fascia lata were removed from 23 patients at reoperations for femoral fracture. The obtained tissues were divided into two samples: first of them was submitted to GAG isolation and the second one to PG isolation. GAGs were extracted by extensive papain digestion followed by the fractionation using cetylpyridinium chloride. In order to qualitative and quantitative characterization GAGs were submitted to electrophoresis on cellulose acetate before and after treatment with enzymes, specifically depolymerizing some kinds of GAGs. PGs were extracted using 4 M guanidine HCl followed by purification by forming complexes with Alcian blue. PGs were submitted to gel permeation chromatography on Sepharose 4B. In order to obtain core proteins PGs were depolymerized with chondroitinase ABC. The purified PGs and their core proteins were separated with sodium dodecyl sulphate/polyacrylamide gel electrophoresis (SDS/PAGE). It was found that total GAGs content was significantly elevated in scarred fascia. Both types of fascia contained chondroitin-, dermatan- and heparan sulphates and hyaluronic acid. Dermatan sulphates (DS) were the predominant GAGs of normal and scarred fascia. The contents of all GAG types were increased in scarred fascia. Both types of fascia contained two kinds of dermatan sulphate proteoglycans (DSPGs); first being similar to biglycan and the second one similar to decorin, as it was judged by molecular weight of their native molecules and core proteins as well as type of GAG components. Densitometric analysis showed that decorin is a predominant DSPG in both fascia types, but in scarred tissue the ratio of biglycan to decorin is considerably higher. Moreover, in scarred fascia a large chondroitin sulphate proteoglycan (CSPG) was also observed. The obtained results have shown that the scar formation is accompanied by quantitative and qualitative alterations in GAGs/PGs resembling those observed in hypertrophic skin scars. The biochemical modification of the scarred fascia lata may partly explain the clinically manifested damage to biomechanical properties of this tissue.

[Therapeutic efficacy of low-dose alpha interferon therapy in liver cirrhosis associated with HBV]. / Skutecznosc terapeutyczna niskich dawek interferonu alfa w wyrównanej marskosci watroby zwiazanej z zakazeniem HBV.

The aim of this study was the assessment the efficacy and safety of therapy with interferon alpha (Intron A) administered s.c. 3 MU x 3/week for 12 weeks for patients with HBV related liver cirrhosis (Child's class A). Fifteen patients completed therapy and 12 months follow-up. At the end of follow-up sustained response to the therapy, defined by clearance of HBV-DNA, normalization of ALAT activity in serum and improvement in the liver histology was achieved in 46.6% of treated patients. Moreover, among few patients from group of nonresponders (patients without sustained clearance of HBV-DNA) decrease of HBV-DNA level, ALAT activity in serum and improvement in the liver histology were observed. Adverse effects of IFN alpha therapy were typical, but in any case were no necessity terminate the therapy.

Glycosaminoglycans of human serum and their alterations in diabetes mellitus.

Human serum contains several glycosaminoglycans (GAGs), mainly chondroitin sulphates and significantly less of heparan sulphate + heparin and dermatan sulphate. The non-insulin-dependent diabetes mellitus (with vascular complications) was associated with a significant increase in total serum GAG concentration, mainly of chondroitin sulphates and dermatan sulphate, with a simultaneous decrease in heparan sulphate + heparin level. These alterations were much more evident in patients with poor metabolic control. Hyaluronic acid (undetectable in healthy subjects and in patients with good metabolic control) appeared only in trace amounts in poorly controlled diabetic individuals. The obtained data allow to conclude that the diabetes mellitus-associated disturbances in tissue GAG metabolism lead to significant alterations in serum GAG composition.

[Plasma fibronectin in chronic liver disease--marker of fibrosis?]. / Fibronektyna osoczowa w przewleklych chorobach watroby--znacznik wlóknienia?

The determination of fibronectin (FN) concentration in plasma has been performed in the group of 77 patients (60-with various chronic liver diseases, 6-with AIDS IVc, 11-healthy patients). The purpose of this study was: evaluation of the value of plasma FN determination in assessment the degree of liver fibrosis and the degree of liver damage. The obtained results were compared with routine biochemical tests and histopathological picture of liver sections. Among patients with liver diseases, we observed that plasma FN concentration was significantly lower only in the group with decompensated liver cirrhosis, in relation to control group. Non significant lower values of FN was observed in the group of patients with chronic hepatitis, as well as non significant higher ones in the group with cholestasis and fibrosis. It has been concluded that determination of plasma FN concentration has not any importance in evaluation of degree of liver fibrosis and its only one from many functional liver tests.

Influence of chronic mercury poisoning upon the connective tissue in rats. I. Effect of mercuric chloride on glycosaminoglycan levels in tissues, serum and urine.

Rats were intoxicated with mercuric chloride (1mg/kg b.w.) daily, for 12 weeks. A decrease in total glycosaminoglycan content was shown in the skin, the lungs, the liver and the heart muscle. These changes were accompanied by a slight alteration of the glycosaminoglycan pattern, a decrease in hyaluronic acid in the skin, the lungs and the heart muscle and an enhancement of heparan sulphate level in the kidneys. In serum of mercury-intoxicated rats, an increase in total glycosaminoglycan levels was shown. This enhancement was caused by elevation of almost all fractions. Urine output of glycosaminoglycans was higher in mercury-treated animals as compared to the controls.

[Experimental evaluation of toxicity of selected mixtures used in metallurgy]. / Doswiadczalna ocena toksycznosci wybranych mieszanin uzywanych w przemysle hutniczym.

The investigation is aimed at evaluation of the effects of copper-coating and bonderizing solutions and oil--emulsion used in technological processes in the metallurgic industry upon hematological and biochemical changes in rats' peripheral blood. The studies were performed on male Wistar rats. The solutions were carried onto hairy and depilated skin. Acute toxicity was evaluated in two-weeks' experiment and chronic toxicity in three--months' experiment. The copper-coating solution was found to be most hazardous, whereas the oil emulsion-the least hazardous. The copper-coating and bonderizing solutions, when chronically applied, result in anaemia and changes in the percentage picture of leucocytes, and also increased blood concentration of uric acid, creatinine and hydroxyproline. No significant changes in concentrations of the remaining biochemical indices were found. The results point to synergistic effects of oil-emulsion and copper-coating solution.

Effect of irradiation on glycosaminoglycans content in rat tissue.

Glycosaminoglycan (GAGs) fractions were determined in tissues (skin, liver, lungs, aortic wall) and blood serum of rats irradiated with a single dose of 500 R. An increase of total GAGs as well as changes in the fractions were found in the tissues and urine of exposed rats.

Glycosaminoglycan content in tissues and body fluids of rats treated with atherogenic diet.

An increase of total glycosaminoglycan content in aortic wall and liver as well as changes in the concentration of glycosaminoglycan fractions in aorta, skin, liver, and blood serum were found in white rats fed with atherogenic diet. Urinary excretion of glycosaminoglycans was increased in experimental animals.