Formalin-induced experimental sclerosing cholangitis in the rat.

The few reported cases of sclerosing cholangitis following removal of an echinococcus cyst are thought to be a consequence of the chemical action of formalin used for sterilization of the residual cavity. The aim of this study was to assess this hypothesis. We injected 0.15ml of 2% buffered formalin solution into the central hepatic lobe of five rats, after a midline laparotomy. At 6, 12, 18 and 24 weeks after formalin injection all rats were reoperated upon and a sample of hepatic parenchyma from both the central and the left hepatic lobe was obtained for microscopic evaluation. Our findings, dilatation of portal tracts and bile canaliculi, thickening of the pericanalicular cytoplasm, portal and periportal inflammatory cell infiltration and fibrosis and enlargement of the perisinusoidal space of Disse, suggest that 2% formalin solution leads to the development of essential phenomena of cholestasis and sclerosing cholangitis in the rat, so thus it should be avoided in liver hydatid disease surgery.

Ultrastructural study of bronchial epithelium in chronic respiratory diseases.

The fine structure of bronchial epithelium in thirty-six patients, thirty-one men and five women, suffering from chronic obstructive pneumonopathy or bronchial carcinoma was studied. No remarkable alterations were found with electron microscopy, in most non-smokers in contrast to the smokers who presented destruction of the epithelial cells and loss of the cilia or many pathological cilia with an abnormal microtubular configuration and irregular orientation. The severity, however, of the alterations was not related to the severity of smoking and to the presence of bronchial cancer.

Ultrastructure of the rat hippocampus after isobaric respirative hyperoxia.

After exposing rats to an environment of isobaric hyperoxia, the ultrastructural alterations of the hippocampus were studied. No major alterations were found in the nerve cells. Of importance was the moderate osmiophilia and the spindle-like transformation of the mitochondria. Vacuolated synapses and neuraxons were found, containing amorphous material. Astrocytic perivascular end feet were found vacuolated in many places. Many endothelial cells of the capillaries presented high osmiophilia, which sometimes prevented structural details. Quantitatively, the findings were proportionally related to the time of exposure in the pure oxygen atmosphere (24, 48 and 65 hours).

Hypoxic pretreatment protects against neuronal damage of the rat hippocampus induced by severe hypoxia.

The present study investigates whether under conditions of successive hypoxic exposures pretreatment with mild (15% O(2)) or moderate (10% O(2)) hypoxia, protects hippocampal neurones against damage induced by severe (3% O(2)) hypoxia. The ultrastructural findings were also correlated with regional superoxide dismutase (SOD) activity changes. In unpretreated rats severe hypoxia induced ultrastructural changes consistent with the aspects of delayed neuronal death (DND). However, in preexposed animals hippocampal damage was attenuated in an inversely proportional way with the severity of the hypoxic pretreatment. The ultrastructural hypoxic tolerance findings were also closely related to increased regional SOD activity levels. Thus the activation of the endogenous antioxidant defense by hypoxic preconditioning, protects against hippocampal damage induced by severe hypoxia. The eventual contribution of increased endogenous adenosine and/or reduced excitotoxicity to induce hypoxic tolerance is discussed.

Dentinogenic activity of allogenic plasma fibronectin on dog dental pulp.

The response of ectomesenchymal cells of dog dental pulp to implantation of Millipore filters supplemented with bovine plasma fibronectin was evaluated after observation periods of one or four weeks. Two concentrations of plasma fibronectin were used (0.2 and 1 mg/mL). Experiments also included implants treated with control solutions (PBS or 1 mg/mL of dog albumin). Formation of a layer of elongated, polarized cells was demonstrated in direct contact with the implants treated with 1 mg/mL of plasma fibronectin solution, after one week post-operatively. Microfilamentous organization and orientation of rough endoplasmic reticulum was observed mainly in the supranuclear zone of the polarized cells. Implants treated with the same solution were consistently surrounded by a thick layer of dentinal matrix after four weeks of their exposure to pulp sites. Implants treated with control solutions or with the low concentration of fibronectin never showed any sign of cell polarization and matrix synthesis. These data provide evidence that the pulp cells can express their odontoblastic phenotype in response to a surface containing concentrated fibronectin (even allogenic), without the need of other molecules as exogenous inductive factors.

Short-term dentinogenic response of dog dental pulp tissue after its induction by demineralized or native dentine, or predentine.

The events initiating the expression of odontoblastic potential by pulpal ectomesenchymal cells were investigated by exposing the pulp to demineralized, native and unmineralized autogenous dentine. The pulp responses to implants were histologically evaluated 3, 7 and 10 days postoperatively, while the surface structure of the newly mineralized matrices was examined 12 and 28 days after implantation. Differentiation of odontoblast-like cells in close proximity to the implanted matrix was consistently demonstrated after exposure to predentine. Scattered columnal cells undergoing polarization, characterized ultrastructurally by the orientation of their rough endoplasmic reticulum, were also found in direct contact with the demineralized dentine. However, in response to demineralized implants, groups of differentiated odontoblast-like cells were clearly seen only in association with a zone of matrix secreted in a polar, predentine-like pattern, indicating an asynchronous inductive influence of this type of implant on pulp cells. Further, the response of pulp cells to native dentine was characterized by the elaboration of a two-layered matrix (a fibrous and a polarly deposited matrix) before initiation of secondary dentinogenesis. Scanning electron microscopy of the newly deposited matrices revealed differences between the indirect matrix synthesis, observed in short-term response to implants of demineralized or native dentine, and the specific, dentinogenic function of the odontoblast-like cells. These observations indicate that the dentine-induced dentinogenesis is initiated by two mechanisms--direct induction of odontoblast-like cells as well as indirect matrix synthesis, which further controls cell polarization. Immobilization of the cells on implanted matrix seems to be the critical requirement for direct expression of the odontoblastic phenotype.

Effects of recombinant basic fibroblast growth factor, insulin-like growth factor-II and transforming growth factor-beta 1 on dog dental pulp cells in vivo.

The effects of recombinant basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF)-II and transforming growth factor (TGF)-beta 1 on dental pulp cells were investigated by light and transmission electron microscopy after their implantation for 1 and 3 weeks at central sites of mechanically exposed pulps in dog molar and canine teeth. The implants were Millipore filters that have been soaked with solutions containing 100 or 500 ng/ml of bFGF or IGF-II or 100 ng/ml of TGF-beta 1. Control filters were soaked with dog albumin. No changes in cell organization or matrix synthesis were seen after implantation of control filters. Groups of columnar, polarized cells with numerous mitochondria and Golgi elements or elongated cells unassociated with any matrix deposition were demonstrated after 1 or 3 weeks, respectively, in close proximity to the filters that had been soaked with bFGF solution; at a distance from these implants enhanced formation of an osteotypic matrix was seen beneath the exposure site. No particular response was found in close proximity to the filters that had been soaked with IGF-II solution after 1 or 3 weeks implantation but thick zones of osteodentine were found beneath the exposure site and at adjacent circumferential dentine sites. Numerous elongated, polarized cells with long cytoplasmic extensions invading the filter pores were consistently seen after 1 week in close proximity to the filters that had been soaked with TGF-beta 1 solution. After 3 weeks implantation of these filters, deposition of a tubular matrix surrounding the implants was seen in association with the highly elongated odontoblast-like cells, while enhancement of circumferential dentine formation was also found at adjacent peripheral sites. These experiments demonstrate that TGF-beta 1 when implanted for short term periods at central pulp sites exerted dentine-specific effects, inducing differentiation of odontoblast-like cells and stimulating primary odontoblasts. Implantation of bFGF and IGF-II did not result in reparative dentine formation, but did stimulate osteotypical matrix deposition at a distance from the implants.

Induction of odontoblast-like cell differentiation in dog dental pulps after in vivo implantation of dentine matrix components.

The effects of dentine extracellular matrix components on dental mesenchymal cells were studied by light and transmission electron microscopy after their implantation at central sites of mechanically exposed pulps in dog molar teeth. The implants were Millipore filters that had been soaked with solutions containing 30 or 300 micrograms/ml of an EDTA-soluble fraction of rabbit incisor dentine. Control filters were soaked with dog albumin or phosphate buffered saline. Columnar, polarized cells were consistently seen after 8 days in close proximity to the filters coated with both concentrations of dentine matrix components. Characteristic features of these polarized cells included widened cisternae of the rough endoplasmic reticulum, a rich microfilamentous network in the long cytoplasmic extensions invading the filter pores and numerous cytoplasmic bodies. These cells also showed evidence of functional as well as cytological differentiation. Polarized processing of secretory granules could be observed after 8 days' implantation, and also the presence of matrix vesicles and deposition of a fine, collagenous matrix into the filters apically to the distal end of the cytoplasmic processes. After 24 days' implantation, secretion of a tubular matrix could be consistently seen in association with the odontoblast-like cells. No changes in cell organization or matrix synthesis were seen after implantation of control filters. These studies demonstrate that bioactive components present in the EDTA-soluble dentine matrix fraction are able to directly induce cell polarization and apical secretion of tubular matrix when implanted in contact with dental pulp cells at sites remote from the odontoblast layer.

Inability of calcium hydroxide to induce reparative dentinogenesis at non-peripheral sites of dog dental pulp.

The ability of 2 calcium hydroxide-containing materials to induce initiation of reparative dentinogenesis was tested at sites remote from the dentinogenically-active regions of the pulp periphery. Pieces of the cements, Dycal and Life, were implanted in central parenchymal sites of dog dental pulps for periods of 6, 14 and 42 days, respectively. Similar pieces were placed in peripheral capping sites as controls. The responses were analyzed by light and transmission electron microscopy. Induction of tubular dentin matrix lined with elongated and polarized odontoblast-like cells was only seen at peripheral capping sites. In response to the centrally implanted cements, only atubular hard tissue with lining fibroblast-like cells was deposited.

The dentinogenic effect of mineral trioxide aggregate (MTA) in short-term capping experiments.

AIM: The objective of the present experiment was to study the early pulpal cell response and the onset of reparative dentine formation after capping application of MTA in mechanically exposed pulps. METHODOLOGY: Thirty-three teeth from three dogs, 12-18 months of age were mechanically exposed via class V cavities. Light pressure was applied to control haemorrhage. ProRoot MTA (Dentsply Simfra, Paris) was placed at the exposure site and light pressure was applied with a wet cotton pellet. The cavities were restored with amalgam and the pulpal tissue reactions were assessed by light and electron microscopy (transmission and scanning) after healing intervals of 1, 2 or 3 weeks. RESULTS: A homogenous zone of crystalline structures was initially found along the pulp-MTA interface, whilst pulpal cells showing changes in their cytological and functional state were arranged in close proximity to the crystals. Deposition of hard tissue of osteotypic form was found in all teeth in direct contact with the capping material and the associated crystalline structures. Formation of reparative dentine (tubular matrix formation in a polar predentine-like pattern by elongated polarized cells) was consistently related to a firm osteodentinal zone. CONCLUSIONS: The present experiments indicate that MTA is an effective pulp-capping material, able to stimulate reparative dentine formation by the stereotypic defensive mechanism of early pulpal wound healing.

Congestive gastropathy and antral varices: is there an association?

Gastric mucosal lesions are frequently observed during endoscopy in cirrhotics, but only recently has attention been paid to the gastric mucosal vessels themselves as the cause of such lesions. The presence - observed during endoscopy - of areas of erythema outlined by a yellow mosaic-like network, and the existence - ascertained by histology - of focal ectasia of mucosal vessels are termed "congestive gastropathy". In a prospective investigation of this entity, gastric mucosal biopsies have been obtained in 20 portal hypertensive patients with or without signs of congestive gastropathy at their initial sclerotherapy session. During the progress of the study two - Child's C - patients (having negative endoscopic and microscopic findings of congestive gastropathy) exhibited severe lesions of congestive gastropathy at the 2nd and 3rd session, respectively, and further biopsies were therefore obtained. Histologic examination after eosin-hematoxylin and Masson's trichrome staining revealed dilated mucosal vessels. At the 3rd and the 5th sclerotherapy session, respectively, - their esophageal varices having been obliterated - antral varices were prominent. We ascribe the endoscopic changes observed in the gastric mucosa to the alteration of gastric vascularity due to blood flow redistribution, but we pose the question as to whether there is any association between these progressive mucosal alterations and the formation of antral varices.

Ultrastructural study of rabbit sciatic nerve regeneration following experimental excisional transection and autograft reconstruction.

This experimental study presents the ultrastructure of regenerating sciatic nerve of the rabbit, after transection and immediate end to end anastomosis, using perineural fascicular nerve autograft, in a sterile environment. Twenty-four hours, 1, 2, and 6 weeks after the anastomosis, the treated sciatic nerves were exposed and three segments were excised and studied. The first at the region of the graft and the others from the proximal and distal stump of the nerve, in the vicinity of the graft suture. The sections taken from the proximal part showed that the nerve structure was identical with the control. Degeneration and regeneration of nerve fibres were observed on the sections taken from the region of the grafts and from the distal parts. Macrophagic activity appeared mainly one week after the operation. Fibroblastic invasion started 24 hours after operation. A moderate amount of collagen fibres was gradually formed. The fibres were disposed in parallel with the neuraxon. Schwann cells were slightly affected initially but consequently they fully recovered and showed signs of extra-activity of the cytoplasm organelles, e.g., enlargement of the granular endoplasmic reticulum cisternae. The present study showed that the bridging of experimental gaps of rabbit's sciatic nerve, by means of autograft and by use of perineural suturing, was successful. The regenerating nerve fibres were growing through the graft towards the distal part of the nerve. In this process Schwann cells and fibroblastic activity play a key role, which is most favourably influenced by using the technique described in this paper.