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Biomedical subjects

J A Gilles

Publications and source records attributed to J A Gilles.

8 recordsLinked to original sources

Inflammation-associated lysyl oxidase protein expression in vivo, and modulation by FGF-2 plus IGF-1.

Lysyl oxidase is the extracellular enzyme that catalyzes oxidative deamination of peptidyl-lysine residues in elastin precursors, and lysine and hydroxylysine residues in collagen precursors to form peptidyl-aldehydes. These aldehydes then spontaneously condense to crosslink collagen and elastin and thereby allow the formation of a mature and functional extracellular matrix. In the present study, cryosections made from aseptic immune-induced periapical lesions experimentally generated in laboratory rats were examined by immunohistochemistry to investigate whether lysyl oxidase protein expression is altered in inflamed oral tissues. Periapical lesions are experimentally induced endodontic lesions of tooth roots. In addition, the effect of administration of a mixture of fibroblast growth factor (FGF)-2 and insulin-like growth factor (IGF)-1 into these lesions on lysyl oxidase expression was determined. Lysyl oxidase expression was found to be increased in non-mineralized connective tissue adjacent to inflamed lesions. Morphometric analyses indicated that maximum lysyl oxidase expression occurred at a discrete distance from the lesion not exceeding 350 microm from the inflammatory cells. Staining was associated with mesenchymal cells with a fibroblastic morphology. No lysyl oxidase staining was found near teeth where no lesion was induced. Application of a mixture of FGF-2 and IGF-1 resulted in a further twofold increase in lysyl oxidase expression. These results provide a new in vivo model to study lysyl oxidase regulation, and suggest that inflammatory cells may control lysyl oxidase expression in oral tissues, possibly by a mechanism involving secretion of cytokines and other factors, probably contributing to the regulation of extracellular matrix accumulation.

Animals↗

Oral bone loss is increased in ovariectomized rats.

Alveolar bone loss associated with periodontal disease occurs frequently in postmenopausal females, the same group that is predisposed to osteoporosis. To determine if the estrogen-deficient state enhances oral bone loss, we studied ovariectomized rats administered the potent bone-resorbing cytokine interleukin-1 or the periodontal pathogen Campylobacter rectus lipopolysaccharide (LPS). Distal root canals of first mandibular molars were instrumented with endodontic files, and bone resorbing factors were deposited and sealed into the root canal. Radiographs of periapical bone loss were evaluated using computer assisted image analysis to determine lesion size. Both interleukin-1 and C. rectus LPS caused a significant increase in lesion area in both ovariectomized and normal rats when compared with controls and a significant increase in ovariectomized animals compared to nonovariectomized animals receiving LPS. Using this endodontic model, we have demonstrated that estrogen deficiency results in increased oral bone loss in rats.

Alveolar Bone Loss↗

Monocyte chemoattractant protein-1 induces monocyte recruitment that is associated with an increase in numbers of osteoblasts.

Monocyte chemoattractant protein-1 (MCP-1) is a member of the chemokine family of cytokines. The principal function of MCP-1 is thought to be the stimulation of monocyte recruitment. Monocyte products are potential regulators of bone cell activity. Growth factors produced by monocytes may stimulate bone formation, while cytokines such as IL-1 and IL-6 can induce bone resorption. To determine whether MCP-1 enhances recruitment of monocytes during bone healing, studies were carried out in which MCP-1 was applied to osseous sites in vivo. Changes in monocyte number were determined by immunohistochemistry using the antibody ED-1 specific for peripheral monocytic cells. The effect of MCP-1 on osteoblast number was determined by counting the number of alkaline phosphatase positive cells in close proximity to bone. For comparison, osteoblast number was also determined following stimulation with platelet-derived growth factor (PDGF)-BB plus IGF-1 in vivo. Results indicate that MCP-1 stimulated a large increase in monocyte recruitment compared to vehicle alone. An increase in monocytes induced by MCP-1 was associated with an increase in the number of osteoblasts lining the bone surface, although not to the same magnitude as a positive control, PDGF-BB, and IGF-1. These results indicate that MCP-1 induces the recruitment of monocytes to bone and suggest that the recruitment is associated with an increase in osteoblast number. This is likely to occur via indirect mechanisms, because MCP-1 did not directly enhance DNA synthesis in osteoblastic cells in vitro. Thus, activated mononuclear phagocytes may play an important role in osseous wound healing by stimulating proliferation of osteoblastic cells, presumably through the elaboration of growth factors.

Alkaline Phosphatase↗

Development of an in vitro culture system for the study of osteoclast activity and function.

Resorption is a difficult process to study in vivo because of the complex organization of mineralized tissue. The objective of this study was to develop an in vitro culture system for studying osteoclast attachment and activity. Transparent, microporous substrata were prepared using radio frequency-sputtering techniques to apply a thin layer of hydroxyapatite on polycarbonate filters previously coated with Type I collagen. Dentinoclasts isolated from the root surface of rat molar teeth undergoing inflammatory resorption and osteoclasts isolated from the endosteal surfaces of rat long bones were incubated on the experimental substrata. The culture system proved successful, supporting osteoclast viability and activity as demonstrated by cellular attachment and spreading seen by scanning electron microscopy. Activity was further demonstrated by monitoring acid production using a fluorescent probe, acridine orange. Isolated osteoclasts or dentinoclasts, when combined with synthetically prepared substrata, were observed to respond by mechanisms comparable to those that occur in vivo. Therefore, the system developed permits basic observations of osteoclasts in an in vitro system.

Animals↗

Anti-inflammatory effects of dexamethasone on periapical tissues following endodontic overinstrumentation.

An animal model was developed in which the anti-inflammatory effects of dexamethasone could be examined histologically in periapical tissues following endodontic overinstrumentation. Mandibular first molars containing vital or partially necrotic pulp tissue were instrumented beyond the apical foramen in rats. Fifty microliters of sterile saline or dexamethasone sodium phosphate (0.4 mg/ml) were administered in a blind manner by supraperiosteal infiltration into the buccal vestibule. The rats were killed after 6, 24, and 48 h. Block sections of the mandibular molars were processed for histological examination and stained with hematoxylin and eosin. The number of polymorphonuclear neutrophils present in the periapical tissues was counted in a blind manner and statistical analysis of the results was performed by two-way analysis of variance. Following endodontic overinstrumentation, local infiltration of dexamethasone produced a significant anti-inflammatory effect on the periapical tissues of teeth with vital or partially necrotic pulp tissue.

Analysis of Variance↗

Comparison of the Canal Master endodontic instrument and K-type files for enlargement of curved root canals.

Twenty mandibular molars with 40 mesial curved root canals were instrumented with a circumferential technique using K files and the Canal Master instrumentation technique by junior dental students whose only endodontic experience had been completion of a preclinical endodontic course. Preoperative and postoperative radiographs were taken facially and mesially with instruments in place. The canals were evaluated radiographically for transportation 1 mm and 4 mm from the apex. The mesial roots were cross-sectioned at the same level. The sections were evaluated for roundness of the canal preparation. It was concluded that the Canal Master instrumentation technique transported the root canal less and produced rounder preparations than the K file circumferential technique. There appears to be a greater tendency for breakage with the Canal Master.

Dental Pulp Cavity↗

Dimensional stability of temporary restoratives.

The effects of thermal cycling on the dimensional stability of temporary restoratives were studied. Stability appeared to be affected significantly by water content. On thermal cycling, Cavit showed less linear dimensional change than two cements containing zinc oxide-eugenol and gutta-percha temporary stopping.

Dental Restoration, Temporary↗