Clinical application of autologous fibroblast cell culture in gingival recession treatment.

BACKGROUND AND OBJECTIVE: Gingival recession is defined as soft and hard tissue displacement resulting in root surface exposure. The optimal outcome of gingival recession treatment is complete, predictable and long-lasting root coverage with a significant level of tissue regeneration. Tissue engineering, which applies active regeneration principles, presents the contemporary treatment approach in the restitution and regeneration of lost tissues. The objective of the present study was to evaluate and compare the clinical results of application of an autologous fibroblast cell culture (AFCC) on a collagen matrix and a connective tissue graft (CTG) placed under a coronally advanced flap (CAF), in the treatment of single and multiple gingival recessions. MATERIAL AND METHODS: Eighteen patients from the Department of Periodontology, School of Dentistry, University of Belgrade, were randomly enrolled in this study. Inclusion criteria were the bilateral presence of Miller Class I or II single or multiple maxillary gingival recessions. A split-mouth design was used in the study. The experimental group was treated with AFCC on a collagen scaffold, which was placed under a CAF. The control group received a combination of CTG and CAF. Clinical parameters such as gingival recession coverage, keratinized tissue width, clinical attachment level and gingival index were recorded at baseline and at 12 mo postoperatively. The oral hygiene level was assessed by plaque index evaluation. Postoperative healing was evaluated through the healing index, recorded 1, 2 and 3 wk postoperatively. The final esthetic outcome was assessed using the mean root coverage esthetic score (RES). RESULTS: Statistically significant improvement of all parameters assessed was found compared with baseline. A statistically significant difference between groups was observed only in keratinized tissue width. Greater keratinized tissue width is still obtained with the use of CTG. Regarding the tissue-healing results, no statistically significant difference was achieved. The RES results were similar for both groups. CONCLUSIONS: Within the limitations of the present study, both procedures proved to be efficient in gingival recession treatment. AFCC, as a novel tissue-engineering concept and living cell-based therapy, proved to be a reliable and successful treatment concept.

Recurrent hemolytic uremic syndrome with hypocomplementemia and intestinal lymphangiectasia.

A 23-year old male patient had eight distinct episodes of hemolytic uremic syndrome (HUS) between 8.5 and 15 years of age, five of them accompanied by hypocomplementemia. In the further course, severe hypertension, renal insufficiency as well as protein-losing enteropathy due to intestinal lymphangiectasia developed, whilst hypocomplementemia persisted. The association of recurrent HUS with hypocomplementemia and intestinal lymphangiectasia may represent a new association within a subgroup of the atypical HUS.

[Contamination of IgG preparation for intravenous use with IgA. I. Development of ELISA tests for determination of low concentrations of IgAc, IgA1, IgA2, IgG and IgM]. / Kontaminacija imunoglobulinima A preparata imunoglobulina G3A intravensku upotrebu. I) Razvoj postupaka enzimskog imunoeseja za merenje niskih koncentracija imunoglobulina A1, A2, G i M.

Very low amounts of immunoglobulin contaminants present in preparations of immunoglobulin G for intravenous use (IVIG) require sophysticated procedures for detection of such low concentrations in milieu of comparatively very high level of IgG. Standard RID and nephelometry procedures are not adequate for these purposes. In process of removal of IgA contamination from IVIG, which is currently under development in Blood Transfusion Institute, Belgrade, it is necessary to have senzitive and very specific procedures for determination of and follow up of very low concentrations of IgA remauning in IVIG. We have established a highly senzitive and specific ELISA precedures for determination of very low concentrations of total IgA (IgAc), IgA1, IgA2, IgG and IgM. Commercial standards of IgAc, IgA1, IgA2, IgG and IgM were bound to PVC plates coated with corresponding monoclonal antibodies anti IgAc-IgA1, anti-IgG and anti-IgM. Resulting standard curves have shown high correlation coeficients with limited range of detection (two orders of magnitude in nanogram range). Very high specificity of ELISA tests were obtained due to the highly specific monoclonal antibodies used. Very high specificity and very low detection level of developed tests are advantageous compared to standard procedures for Ig concentration determination.

[Contamination of IgG preparation for intravenous use with IgA. II. Removal of IgA1 from IgG preparation using affinity chomatography on lectin jacalin columns]. / Kontaminacija imunoglobulinima A preparata imunoglobulina G3A intravensku upotrebu. II) Uklanjenje imunoglobulina A1 iz preparata imunoglobulina G pomocu afinitetne hromatografije na koloni lektina zakalina.

Administration of preparations of IgG for intravenous use (IVIG) in individuals with high anti-IgA antibody levels in circulation is frequently accompained with anafilactoid reactions. Thus, there is a requirement for preparations of IVIG with very low levels of IgA. Reduction of IgA concentration in IgG preparations is generally difficult to achieve, since physical and chemical characteristics, on which proceses for Ig purification are based on, are similar for these two Igs. In this work the ability of lectin jacalin to selectively bind IgA1 subclass was exploited for removal of IgA1 from IVIG currently under development in Blood Transfusion Institute of Republic Serbia, Belgrade. Concentrations of Igs in this preparation of IVIG were measured by ELISA tests developed for these purposes. The results have shown 280 times decrease of IgA1 content without changes of IgG concentration. Therefore, affinity chomatography on jacalin column is a suitable method for removal of IgA1 subclass from IVIG. Because of a favourable pH and ionic strength conditions used in process, this procedure has no effect on native state of IgG.

Postcholecystectomy diarrhea from villous atrophy of the terminal ileum.

We report a 60-year-old man with intermittent diarrhea for 15 years which was worsened by cholecystectomy. The results of routine laboratory tests, x-ray studies of the gut, and endoscopy were normal. Fecal fat excretion was normal, but total bile acid excretion was increased in the alkaline feces. The retention of 75SeHCAT was low. Jejunal mucosal biopsy showed normal mucosa. Ileal biopsy was performed at ileoscopy some 10 to 15 cm proximal to the ileocecal valve. Scanning-electron and light microscopy showed a transitional zone between villous atrophy in the distal and normal mucosa in the proximal part of the biopsy specimen. The response of diarrhea to cholestyramine treatment was unequivocal, suggesting bile acid malabsorption. The increased enterohepatic cycling of bile acids after cholecystectomy may be responsible for a decrease in their ileal uptake. Very often, as in our patient, the diarrhea precedes cholecystectomy and is considerably increased by it. Cholecystectomy may unmask a previously unrecognized bile acid transport defect. In our patient, this defect might have been caused by the villous atrophy of the terminal ileum.

Complement activation in stored platelet concentrates.

Activation of platelets during preparation and/or storage of platelet concentrates in plastic containers at room temperature has recently been recognized. Many different biologic causes of this activation have been postulated. Activated complement, as a multi-enzyme system, is one of the possible sources of molecules leading to platelet activation. To detect complement activation, functional complement activity and the generation of complement-derived ligands were investigated in platelet concentrate supernatant plasma during 5 days of storage at room temperature. Hemolytic tests for functional classical and alternative pathway activity were used, as was the kinetic test for complement-mediated inhibition of immune complex precipitation. The presence of C3 activation products (C3, C3c, C3dg) was investigated in plasma by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting procedures and on platelets by immunofluorescence. Activation of complement was evident during storage, and C3c and C3d fragments were clearly demonstrated in plasma. The amount of C3d fragments on platelets gradually rose during the first 3 days of storage. At the end of 5 days of storage, the platelets became C3d negative. There are two possible mechanisms of C3d disappearance--shedding and/or further degradation of C3d fragments. Those results indicated that complement activation and the generation of complement-dependent ligand-receptor interaction may be mechanisms for platelet activation in concentrates stored at room temperature.

[The terminal ileum and chronic diarrhea: endoscopy, histology and parasitology]. / Terminalni ileum i hronicni proli: endoskopija, histologija i parazitologija.

In 53 patients with chronic diarrhea ileoscopy was done following colonoscopy. Beside the microscopic examination, terminal ileum biopsies and mucosal smears were also performed. Endoscopy of the terminal ileum was abnormal in eight patients (15.1%); biopsy itself was diagnostic in 22 patients (41.5%): primary bile acid malabsorption with mucosal atrophy and reduced retention of 75ScHCAT (10), mucosal atrophy after cholecystectomy (4), Crohn's disease (6), backwash ileitis in ulcerative colitis (1), and postirradiation ileitis (1). Biopsies were normal but mucosal smear indicated the cause of diarrhea in a further 10 patients: giardiasis was found in 7, and candidiasis in 3 patients. All in all, endoscopy, biopsy and mucosal smear of terminal ileum showed a sensitivity of 58.5%. In 38 patients in whom laboratory, roentgenologic and endoscopic investigation failed to establish the etiology of diarrhea, the sensitivity of ileoscopy itself was 0%, of ileoscopy with biopsy 36.8% and ileoscopy with biopsy and mucosal smear 47.4%. We conclude that endoscopy, biopsy and mucosal smear of the terminal ileum are indicated in the investigation of patients with chronic diarrhea.

Primary bile acid malabsorption. Histologic and immunologic study in three patients.

Three patients are presented with a history of chronic watery diarrhea due to bile acid malabsorption, proved by the tauro-23[75Se]selena-25 homocholic acid test and an unequivocal response to cholestyramine therapy. Fecal fat tests, Schilling tests, and barium studies of the small intestine and colon were all normal. Jejunal biopsies were normal but multiple biopsies of the terminal ileum, performed by retrograde ileoscopy, showed uniform crypt hyperplastic villous atrophy and features of colonic metaplasia with increased mononuclear infiltration of the lamina propria. All 3 patients demonstrated evidence of abnormal immune function, including the presence of serum autoantibodies, circulating immune complexes, and hypocomplementemia. One patient had Sjögren's syndrome. This disorder, which might be immunologically mediated, should be called primary bile acid malabsorption and should be distinguished from other ileal disorders.