Host Connective Tissue Research Articles

Characterization of an experimental model to determine streptococcal M protein–induced autoimmune cardiac and neurobehavioral abnormalities

Group A streptococcal (GAS) infection is associated with a spectrum of autoimmune diseases including acute rheumatic fever/rheumatic heart disease (ARF/RHD) and neurobehavioral abnormalities. Antibodies against GAS M proteins cross‐react with host tissue proteins in the heart and brain leading to the symptomatology observed in ARF/RHD. As throat carriage of Streptococcus dysgalactiae subspecies equisimilis (SDSE) has been reported to be relatively high in some ARF/RHD endemic regions compared with GAS, and both SDSE and GAS express coiled‐coil surface protein called M protein, we hypothesized that streptococci other than GAS can also associated with ARF/RHD and neurobehavioral abnormalities. Neurobehavioral assessments and electrocardiography were performed on Lewis rats before and after exposure to recombinant GAS and SDSE M proteins. Histological assessments were performed to confirm inflammatory changes in cardiac and neuronal tissues. ELISA and Western blot analysis were performed to determine the cross‐reactivity of antibodies with host connective, cardiac and neuronal tissue proteins. Lewis rats injected with M proteins either from GAS or SDSE developed significant cardiac functional and neurobehavioral abnormalities in comparison to control rats injected with phosphate‐buffered saline. Antibodies against GAS and SDSE M proteins cross‐reacted with cardiac, connective and neuronal proteins. Serum from rats injected with streptococcal antigens showed higher immunoglobulin G binding to the striatum and cortex of the brain. Cardiac and neurobehavioral abnormalities observed in our experimental model were comparable to the cardinal symptoms observed in patients with ARF/RHD. Here for the first time, we demonstrate in an experimental model that M proteins from different streptococcal species could initiate and drive the autoimmune‐mediated cardiac tissue damage and neurobehavioral abnormalities.

Reconciling VEGF With VPF: The Importance of Increased Vascular Permeability for Stroma Formation in Tumors, Healing Wounds, and Chronic Inflammation.

It is widely believed that vascular endothelial growth factor (VEGF) induces angiogenesis by its direct mitogenic and motogenic actions on vascular endothelial cells. However, these activities are only detected when endothelial cells are cultured at very low (0.1%) serum concentrations and would not be expected to take place at the much higher serum levels found in angiogenic sites in vivo. This conundrum can be resolved by recalling VEGF’s original function, that of an extremely potent vascular permeability factor (VPF). In vivo VPF/VEGF increases microvascular permeability such that whole plasma leaks into the tissues where it undergoes clotting by tissue factor that is expressed on tumor and host connective tissue cells to deposit fibrin and generate serum. By providing tissue support and by reprogramming the gene expression patterns of cells locally, fibrin and serum can together account for the formation of vascular connective tissue stroma. In sum, by increasing vascular permeability, VPF/VEGF triggers the “wound healing response,” setting in motion a fundamental pathophysiological process that induces the mature stroma that is found not only in healing wounds but also in solid tumors and chronic inflammatory diseases. Once initiated by increased vascular permeability, this response may be difficult to impede, perhaps contributing to the limited success of anti-VEGF therapies in treating cancer.

First molecular characterization of hypodermin genes of Hypoderma bovis and serodiagnosis of bovine hypodermosis with recombinant hypodermin C antigen and a synthetic peptide containing its linear B-cell epitope

Hypodermins A (HA), B (HB), and C (HC) of warble flies are modulatory antigens involved in host inflammation and immune responses during migration of the warble fly larvae through host connective tissues. In the current study, molecular characteristics of the genes encoding HA, HB, and HC were revealed from cDNA constructs of third-instar larvae of Hypoderma bovis. The open reading frame (ORF) of each hypodermin gene was amplified with modified gene-specific primers, and the resulting PCR products were cloned into pGEM-T Easy Vector to produce recombinant plasmids (rHA, rHB, and rHC). The ORF sequences of rHA, rHB, and rHC genes are 705 bp, 771 bp, and 783 bp long and encode proteins of 234, 256, and 263 amino acids with predicted sizes of 25.74 kDa, 27.79 kDa, and 28.51 kDa, respectively. The rHC gene was subcloned into the pET 100/D-TOPO Expression Vector, and the recombinant HC was purified using affinity chromatography. Western blotting indicated that rHC was recognized by the sera of cattle naturally infested with H. bovis. The rHC and a synthetic peptide (sHC) containing its linear B cell-specific epitope were evaluated as serological markers in indirect ELISA (iELISA) for the diagnosis of bovine hypodermosis. Both sHC and rHC iELISAs had sensitivity values equal to or higher than 90 % and specificity values of 100 %. A total of 200 serum samples from cattle in the Central Anatolia Region of Turkey were also analyzed by rHC and sHC-iELISAs to reveal the seroprevalence of bovine hypodermosis. The results of both iELISAs were consistent with one another and revealed a hypodermosis prevalence of 62 %. Our study provides the first data on molecular characterization of hypodermin genes of H. bovis and indicates the efficacy of recombinant antigen and peptide-based iELISA for serodiagnosis of bovine hypodermosis.

Proteolytic cleavage of HLA class II by human neutrophil elastase in pneumococcal pneumonia

Bacterial and viral respiratory infections can initiate acute lung injury and acute respiratory distress syndrome. Neutrophils and their granule enzymes, including neutrophil elastase, are key mediators of the pathophysiology of acute respiratory failure. Although intracellular neutrophil elastase functions as a host defensive factor against pathogens, its leakage into airway spaces induces degradation of host connective tissue components. This leakage disrupts host innate immune responses via proteolytic cleavage of Toll-like receptors and cytokines. Here, we investigated whether neutrophils possess proteases that cleave adaptive immune molecules. We found that expression of the human leukocyte antigen (HLA) class II molecule HLA-DP β1 was decreased in THP-1-derived macrophages treated with supernatants from dead neutrophils. This decreased HLA-DP β1 expression was counteracted by treatment with neutrophil elastase inhibitor, suggesting proteolytic cleavage of HLA-DP β1 by neutrophil elastase. SDS-PAGE showed that neutrophil elastase cleaved recombinant HLA-DP α1, -DP β1, -DQ α1, -DQ β1, -DR α, and -DR β1. Neutrophil elastase also cleaved HLA-DP β1 on extracellular vesicles isolated from macrophages without triggering morphological changes. Thus, leakage of neutrophil elastase may disrupt innate immune responses, antigen presentation, and T cell activation. Additionally, inhibition of neutrophil elastase is a potential therapeutic option for treating bacterial and viral pneumonia.

Allomyrina dichotoma larval extract attenuates intestinal barrier disruption by altering inflammatory response and tight junction proteins in lipopolysaccharide-induced Caco-2 cells

Intestinal epithelial cells form a barrier between the intestinal lumen and host connective tissues and play an important role in maintaining intestinal nutrient homeostasis. This study investigated effects of Allomyrina dichotoma (rhinoceros beetle) larval extract (ADLE) on the intestinal barrier damage and explored mechanisms for reversing intestinal barrier dysfunction in lipopolysaccharide (LPS)-stimulated Caco-2, human intestinal epithelial cells. LPS reduced intestinal epithelial barrier function by increasing transepithelial electrical resistance, and this effect was significantly attenuated by ADLE treatment. ADLE also significantly countered the inhibition of tight junction-related protein expression in both LPS-induced Caco-2 cells and intestine from HFD-induced mice. Moreover, ADLE significantly decreased expression and production of inflammatory factors, such as iNOS, cox-2, nitric oxide, and cytokines induced by LPS stimulus. Reduction in phosphorylation of adenosine monophosphate-activated protein kinase was averted by ADLE treatment in LPS treated INS-1 cells. Finally, reactive oxygen stress level was decreased and ATP production was increased by ADLE treatment. ADLE appears to display gut health-promoting effects by reducing inflammation and inducing tight junction proteins in Caco-2 cells. Therefore, ADLE might be useful for preventing or treating intestine cell damage in inflammatory bowel disease.

Fabrication of ultrahigh-molecular-weight polyethylene porous implant for bone application

Abstract Porous implants play a crucial role in allowing ingrowth of host connective tissue and thereby help in keeping the implant in its place. With the aim of mimicking the microstructure of natural extracellular matrix, ultrahigh-molecular-weight polyethylene (UHMWPE) porous samples with a desirable pore size distribution were developed by combining thermally induced phase separation and salt leaching techniques. The porous UHMWPE samples consisted of a nanofibrous UHMWPE matrix with a fibre diameter smaller than 500 nm, highly interconnected, with a controllable pore diameter from nanoscale to 300 µm. Moreover, a porous UHMWPE sample was also developed as a continuous and homogeneous coating onto the UHMWPE dense sample. The dense/porous UHMWPE sample supported human foetal osteoblast 1.19 cell line proliferation and differentiation, indicating the potential of porous UHMWPE with a desirable pore size distribution for bone application. An osseointegration model in the sheep revealed substantial bone formation within the pore layer at 12 weeks via SEM evaluation. Ingrown bone was more closely opposed to the pore wall when compared to the dense UHMWPE control. These results indicate that dense/porous UHMWPE could provide improved osseointegration while maintaining the structural integrity necessary for load-bearing orthopaedic application.

Obesity and periodontal disease: A review.

Periodontal diseases usually refer to inflammatory disorders that are caused by pathogenic bacteria in the subgingival biofilm in association with impaired host immune response and connective tissue breakdown. The bacterial challenge exacerbates the cytokine production by the gingival epithelium, resulting in an uncontrolled inflammation that leads to tooth loss in adults from different populations. The prevalence of these diseases increases with aging, longer retention of teeth, and increased incidence of obesity and diabetes among the population. The prevalence demonstrates an increasing trend and a correlation with numerous comorbidities. Hence, as a family physician one should have the in-depth knowledge regarding the relationship between obesity and periodontitis to create awareness among people to provide primary care. Thus, it is relevant to develop new methods capable of detecting these diseases in the early stages and following up on their progression.

Role of Salivary Interleukin 1 in Chronic Periodontitis: A Review

Periodontal diseases are a group of inflammatory disorders that are caused by pathogenic bacteria in the sub gingival biofilm leading to association with impaired host immune response and connective tissue breakdown. There has been substantial evidence for the central role played by cytokine networks in periodontal diseases of which interleukin 1 has a significant role in periodontal disease pathogenesis. Studies of the immunopathogenesis of periodontitis and analysis of mediators in saliva have allowed the identification of many potentially useful biomarkers.[1] This current review focuses on the role of salivary interleukin 1 level in chronic periodontitis

Cavisoma magnum (Cavisomidae), a unique Pacific acanthocephalan redescribed from an unusual host, Mugil cephalus (Mugilidae), in the Arabian Gulf, with notes on histopathology and metal analysis.

Cavisoma magnum (Southwell, 1927) Van Cleave, 1931 was originally described from a sea bass, Serranus sp. and spotted surgeonfish, Ctenochaetus strigosus (Perciformes) off Sri Lanka before its more recent redescription from milkfish in the Philippines in 1995. These reports were based on only light infections of their host fishes. Of the few flathead grey mullets, Mugil cephalus (Mugilidae), that we examined in the Arabian Gulf, one fish was infected with 1,450 worms. One milkfish, Chanos chanos (Chanidae), from the same location in the Arabian Gulf, was also heavily infected with specimens of C. magnum. The descriptions of this unique large worm are revised and for the first time, we provide SEM images, new systematic observations, metal analysis of hooks showing extremely high levels of sulfur, and histopathology in the mullet intestinal tissue. Adjustments and corrections of previous descriptive accounts are made. The histopathology studies show extensive damage to the host intestinal tissue including epithelial necrosis, hemorrhaging and worm encapsulation. There is an extensive amount of host connective tissue surrounding the worm. Results of x-ray analysis displayed high levels of sulfur in proboscis hooks, especially at the tips and edges of these attachment structures.

Regional differentiation of the cuticular surface structure in the mesoparasitic copepod Cardiodectes shini (Siphonostomatoida: Pennellidae) on a pygmy goby

Cardiodectes shini is a mesoparasitic copepod found on the heads of pygmy gobies. The copepod inserts its cephalothorax with antennary processes into the host tissues, while the trunk and egg sacs remain outside the host. The ultrastructure of the epicuticle surface differs among the antennary processes, cephalothorax, and trunk. In the antennary process, the epicuticle appears fuzzy and is less electron-dense than other parts of the body. This loose cuticle structure may be related to the absorption of nutrients in the host hemolymph. The cephalothorax and trunk have an electron-dense epicuticle: there is an array of minute protuberances on the epicuticle of the cephalothorax, whereas the trunk cuticle has no protuberances. This array of protuberances might be involved in suppression of the host immune response, because the cephalothorax has direct contact with the host connective tissues and similar structures are found on other parasitic copepods inhabiting host tissue.

Morphological and ultrastructural aspects of Myxobolus niger n. sp. (Myxozoa) gill parasite of Corydoras melini (Siluriformes: Callichthyidae) from Brazilian Amazon

Myxobolus niger n. sp. (Myxozoa) is described in the connective tissue of the serosa layer of the gill arch of Corydoras melini (Callichthyidae) captured from the Negro River, Amazonas State, Brazil. The prevalence of the parasite was 20% and the range intensity was 1–2 cysts per fish. The plasmodia were white and spherical to ellipsoidal, measuring 175μm in diameter and were surrounded by a well-defined capsule of host connective tissue, with distinct delicate and interlaced collagen fibers. The myxospores body was ellipsoidal in frontal view and biconvex in sutural view. Spore dimensions were 11.3±0.4μm in length, 6.8±0.2μm in width and 4.1±0.2μm in thickness. The valves were symmetrical and smooth. The two polar capsules were elongated as pyriform and equal in size, measure 5.0±0.3μm in length and 2.0±0.1μm in width. The polar capsule had six to seven polar filament turns. Some aberrant spores were round in shape and had three polar capsules. The sporoplasm was binucleated and contained moderated number of sporoplasmosomes. The development of the plasmodia was asynchronic, with mature and immature spores. The plasmodium had moderated pynocitic channels. There were no projections, no invaginations and no microvilli in the plasmodial wall. This study is the first description of Myxobolus species in the fish of the Callichthyidae family.

Multiple gingival recession coverage with an allogeneic biostatic fascia lata graft using the tunnel technique—A histological assessment

BackgroundAutogenous connective tissue graft (CTG) that can be safely harvested from the palatal mucosa is limited. Often a multi-stage surgical procedure is needed to cover multiple gingival recessions (MGR). To address this problem, efforts are being made to explore substitutes suitable in size to ensure surgical treatment in a single visit.The objective of the present study was the histological evaluation of tissue in the recipient site after augmentation with a hydrated biostatic Fascia Lata Allograft (FLA) in conjunction with MGR coverage at different healing stages. Material and methodsTwelve patients needing bilateral multiple gingival recession coverage participated in this study. On the test side, the tunnel technique with FLA was used, while CTG, harvested from the palatal mucosa, was used to cover MGR on the control side. Histological assessment was performed 3, 6, 9 and 12 months after augmentation. ResultsFLA was well tolerated by the host tissue. During all investigation periods histological images of all patients in the test side revealed a slow process of incorporation of the material grafted in the host connective tissue, showing a colonization of the graft with host fibroblasts and formation of new blood vessels. After 12 months, the graft had fully remodeled into connective tissue of the host gingiva. ConclusionApart from the limitations of the present study, we conclude that the FLA may serve as a substitute for autogenous CTG harvested from the palatal mucosa and can be applied as a technique for covering MGR in a single visit.

In Vivo Repair of Rat Annulus Fibrosus Defects using High-Density Collagen Gels

Introduction Although a discectomy successfully relieves the neurological symptoms of a herniated intervertebral disc (IVD), it does not treat the underlying degenerative process; the annular defect remains unrepaired. Persistent annular defect is associated with an increased risk of recurrent herniation and progressive degenerative changes to the IVD. It may also be the primary cause of chronic low back pain following discectomy. To date, there is no established method for repairing annular defects in vivo. The presented study aims to determine whether injectable high-density collagen (HDC) gels can reduce further disc herniation and inhibit degenerative changes in a needle-punctured rat-tail model, and whether riboflavin (RF) cross-linking of injected collagen influences the repair process. Materials and Methods A total of 31 athymic rats were punctured with an 18-gauge needle in C3/4 of the caudal spine. They were divided into four groups: group 1 punctured and injected with HDC cross-linked with 0.5 mM (n = 6) or 0.75 mM (n = 7) RF; group 2 punctured and injected with noncross-linked collagen (n = 6); group 3 punctured and untreated (n = 8); group 4 punctured and injected with FITC-labeled cross-linked collagen (n = 4). Degenerative changes to punctured discs, NP size, and NP hydration were analyzed using X-ray, MRI), and histology. Functionality of repaired AF tissue was measured by mechanical tests, by comparing the hydraulic permeability of treated discs to that of healthy discs. Results After 5 weeks, untreated discs showed signs of terminal degenerative changes on MRI and histological sections ( Fig. 1 ). No NP tissue remained in the disc space. In contrast, discs treated with RF cross-linked collagen gels retained 63% of NP and 80% of disc height at 5 weeks, and showed minimal degenerative changes on histological section. Interestingly, a series of in vivo imaging showed that after 5 weeks, discs treated with cross-linked HDC demonstrated consistent improvement in both volume and hydration over time, while untreated discs and discs treated with noncross-linked HDC, lost volume, and water content. Injected collagen was seen to form a zipper-like adhesion to the host AF and connective tissue after 1 week and by 5 weeks a fibrous cap that persisted until 18 weeks. NP hydration and IVD functionality of treated discs were found to be similar to those of adjacent healthy discs at 18 weeks. [Figure: see text] Conclusion Injection of high-density collagen gel cross-linked with RF can repair annular defects, prevent the degenerative cascade, and maintain the functionality of IVDs in a rat-tail spine. Further studies will be performed to confirm the ability of HDC gels to repair annular defects in a large animal model.

Comparative analysis of gingival crevicular fluid β-glucuronidase levels in health, chronic gingivitis and chronic periodontitis.

Background and Objectives:Current methods available for periodontal disease diagnosis are seriously deficient in terms of accuracy, in the ability to predict ongoing or future disease activity and indeed in determining whether previously diseased sites are in an arrested phase or still active. One area that is receiving a great deal of attention is the biochemical investigation of gingival crevicular fluid (GCF). β-glucuronidase (βG) is one of the enzymes found in GCF that is involved in degradation of the ground substance and fibrillar components of host connective tissue. GCF βG activity might be a good indicator or predictor of periodontal disease activity. This study was conducted to estimate and compare the GCF βG levels in patients with healthy periodontium, chronic gingivitis, and chronic periodontitis.Methodology:Subjects were classified into three groups of 20 patients each; healthy individuals, chronic gingivitis, and chronic periodontitis. After recording the plaque index, gingival index and probing pocket depth, 1 μL GCF was collected by placing a calibrated microcapillary pipette extracrevicularly and transferred to sterile plastic vials containing 350 μL of normal saline with 1% bovine serum albumin. Analysis of βG was done by spectrophotometryResults:βG levels in GCF were significantly higher in chronic periodontitis group (mean value - 2.04743), followed by chronic gingivitis group (mean - 1.11510) and healthy group (0.53643).Conclusion:Increased βG levels were observed in patients with increased periodontal destruction, hence GCF βG levels can be used as biochemical marker for periodontal disease activity.

Soft tissue wound healing at teeth, dental implants and the edentulous ridge when using barrier membranes, growth and differentiation factors and soft tissue substitutes

To review the biological processes of wound healing following periodontal and periimplant plastic surgery when different technologies are used in a) the coverage of root and implant dehiscences, b) the augmentation of keratinized tissue (KT) and c) the augmentation of soft tissue volume. An electronic search from The National Library of Medicine (MEDLINE-PubMed) was performed: English articles with research focus in oral soft tissue regeneration, providing histological outcomes, either from animal experimental studies or human biopsy material were included. Barrier membranes, enamel matrix derivatives, growth factors, allogeneic and xenogeneic soft tissue substitutes have been used in soft tissue regeneration demonstrating different degrees of regeneration. In root coverage, these technologies were able to improve new attachment, although none has shown complete regeneration. In KT augmentation, tissue-engineered allogenic products and xenogeneic collagen matrixes demonstrated integration within the host connective tissue and promotion of keratinization. In soft tissue augmentation and peri-implant plastic surgery there are no histological data currently available. Soft tissue substitutes, growth differentiation factors demonstrated promising histological results in terms of soft tissue regeneration and keratinization, whereas there is a need for further studies to prove their added value in soft tissue augmentation.

Use of Curcumin in Periodontal Inflammation

Periodontitis is a bacterial initiated but host modulated chronic infection that leads to destruction of the connective tissue supporting the teeth. Immune and inflammatory response directed against specific bacteria and its products become responsible for the local periodontal tissue loss in susceptible persons. Non-surgical therapy has been the mainstay of periodontal management with mechanical removal of plaque being the predominant method. However some individuals non responsive with only mechanical therapy benefit from supplementation with antimicrobial therapy. The use of adjunctive antimicrobial therapy has been plagued by problems of microbial resistance of local and gut flora. The identification of modified tetracycline made a paradigm shift in host modulation wherein the inflammatory pathway of host connective tissue destruction was altered without affecting the microbial profile. Systemic and locally delivered curcumin shows potential for having similar anti-inflammatory properties. Here we review the anti-inflammatory properties of curcumin and its various forms in modulatory host response as a potential therapy in periodontal diseases.

Updated morphology, histopathology and molecular phylogeny of Myxobolus hearti, cardiac myxosporea in gibel carp, Carassius gibelio (Bloch)

The original description of Myxobolus hearti is supplemented with new data on spore morphology, histopathology and molecular phylogeny. Myxobolus hearti are found in the heart ventricle of the gibel carp, Carassius gibelio (Bloch), where they form whitish oval or irregularly shaped plasmodia. Mature spores are oval or shortly ellipsoidal in frontal view, lemon-shaped in sutural view and eye-shaped in apical view. The spores are 14.12 ± 0.35 (13.6-15) μm long (mean ± SD), 11.85 ± 0.34 ± 0.36 (11-12) μm wide and 7.32 ± 0.36 (7-8) μm thick. The two polar capsules are equal in size, 6.11 ± 0.29 (6-7) μm long and 3.89 ± 0.31(3-4) μm wide, and are long pyriform in shape. Polar filaments have six or seven coils situated perpendicular to the longitudinal axis of the polar capsules. Histopathology indicates that the plasmodia are encased by the host connective tissue, and no inflammatory responses are found in the heart ventricles. Phylogenetic analysis based on the 18S small-subunit ribosomal DNA sequences indicates that M. hearti is, genetically, most similar to Henneguya doneci, a gill-infecting species.

Acellular Porcine Urinary Bladder Matrix for the Treatment of an Avulsive Scalp Wound in Irradiated Tissue on a Chronically Immunocompromised Patient: A Case Presentation

Biological materials composed of extracellular matrix (ECM) harvested from a wide variety of tissues and organs are widely used as viable alternative options for tissue reconstruction. A new product, MatriStem by ACell, has shown promise in regards to regeneration of healthy functional tissue as opposed to scar formation. MatriStem is comprised of an acellular extracellular matrix scaffold comprised of porcine urinary bladder matrix (UBM). Its unique bimodal surface characteristics consist of an intact basement membrane and an organized connective tissue bed comprised of urinary bladder lamina propia. The basement membrane is conducive to epithelial and endothelial cell attachment, proliferation and differentiation, while the lamina propia allows for integration of the product into the wound bed and host connective tissues, and supports vascular ingrowth. In addition, the processing of the MatriStem product allows for retention of multiple growth factors including vascular endothelial growth factor (VEGF), transforming growth factor beta (TGFβ), platelet derived growth factor (PDGF), bone morphogenic protein 4 (BMP4), and basic fibroblast growth factor (BFGF) to assist in regeneration and healing. Recent studies have demonstrated antimicrobial properties of UBM-ECM to both S. aureus and E. colias well as a predominance of macrophage phenotype 2 (M2) in the early postoperative period which has been shown to correlate with less scar formation and more healthy functional tissue regeneration with increased chemotactic activity.

Characterization and expression of cathepsin B2 in Fasciola gigantica

Fasciola gigantica cathepsin B belongs to a family of cysteine proteases which is involved in invasion of host tissues. In this study, the recombinant cathepsin B2 (rFgCatB2), synthesized in Pichia pastoris, showed enzymatic activity on a fluorometric substrate Z-Phe-Arg-AMC and gelatin. Furthermore, this recombinant enzyme could degrade IgG and type I collagen. Mouse antiserum against rFgCatB2 reacted with the native FgCatB2 in whole body (WB) extracts of metacercariae (MET), newly excysted juveniles (NEJ) and 2week-old juveniles, but not in 3, 4week-old juveniles and adult flukes. Immunolocalization showed the presence of cathepsin B2 only in the caecal epithelium of MET, NEJ and 2week-old juveniles. Co-localization of FgCatB2 and a prominent antigen of NEJ, FgCatB3, revealed that these proteins were expressed at the same regions in the caecal epithelium. Anti-rFgCatB2 showed no cross reaction with the other parasites’ antigens by Western blotting. These findings suggest that CatB2 is expressed only in early stages of the parasite and may be involved in digestion of host connective tissues and evasion of the host immune system during their penetration and migration. Thus, CatB2 could be considered as an immunodiagnostic and vaccine candidate for fasciolosis.

Clinical and histological healing of a new collagen matrix in combination with the coronally advanced flap for the treatment of Miller class-I recession defects: an experimental study in the minipig

To describe the histological and clinical outcomes of the use of a xenogeneic collagen matrix (CM) in combination with the coronally advanced flap (CAF) in the treatment of localized Miller class-I gingival recessions. Gingival recession defects were surgically created on 12 minipigs. The defects were randomly treated with either the CAF procedure and the interposition of a CM (test) or the CAF alone (control). Clinical and histological outcomes at 1, 4 and 12 weeks were evaluated. Histometrically, in the test group, there was a shorter junctional epithelial dimension [2.26 (SD 0.23) mm] compared with the control [2.79 (SD 0.77) mm]. On the contrary, the amount of newly formed cementum was larger in the test group [1.08 (SD 0.41) mm] than in the control group [0.75 (SD 0.25) mm], although the differences were not statistically significant. Both techniques rendered similar clinical outcomes, achieving complete root coverage at the end of the study. Nevertheless, the CM graft attained more tissue regeneration, characterized by a shorter epithelium and a larger new cementum formation. The use of a xenogeneic CM resulted in the incorporation of the xenograft within the adjacent host connective tissues in the absence of significant inflammation.