Norris L. O'Dell

The Effects of Dietary Ferric Iron and Iron Deprivation on the Bacterial Composition of the Mouse Intestine

The influence of dietary ferric iron on the intestinal microbiota of mice was investigated with a view to promoting benign lactic acid bacteria (which have minimal iron requirements) in order to enhance colonization-resistance potential. Three groups of eight mice received a diet differing only in iron content, for a period of 12 weeks. Dietary iron deprivation resulted in overall increased small intestinal bacterial populations, including lactic acid bacteria, but these differences were generally not significant (p > 0.05). With the exception of coliforms, all examined bacterial groups (anaerobes, micro-aerophiles, lactobacilli, and enterococci) were significantly (p < 0.05) elevated in the colons of iron-deprived mice. The relatively low numbers of total anaerobes in the colons of iron-replete and iron-overloaded mice suggested that, as well as promotion of bacteria under iron-deprived condition, provision of ferric iron suppressed bacteria, probably by oxidation of normally reduced environments.

Morphological Alterations in the Elastic Fibers of the Rabbit Craniomandibular Joint following Experimentally Induced Anterior Disk Displacement

Elastic fibers are important components of the connective tissue that attaches the articular disk of the craniomandibular joint (CMJ) to the skull and mandible. Biopsies of the articular disk proper and bilaminar zone (BZ) tissues from patients with anterior disk displacement (ADD) have shown previously that there is a marked loss of elastic fibers. In the present study, the effects of inducing ADD on the elastic fibers in the rabbit CMJ disk proper, BZ and condylar cartilage were investigated. The right CMJ was exposed surgically and the discal attachments were severed except for the BZ attachments. Then, the disk was displaced anteriorly and sutured to the zygomatic arch. The CMJs were removed after 1, 2 or 6 weeks and processed for histochemical demonstration of elastic fibers. The results showed osteoarthritic changes following ADD, and a significant decrease in the number of the elastic fibers in the disk proper and BZ. The remaining elastic fibers were abnormal in their appearance and orientation. In addition, ADD led to the appearance of fine elastic fibers among the chondrocytes in the hyaline cartilage of the condyle that were not present in the cartilage of the control condyle. We conclude that induced ADD can lead to a significant loss of elastic fibers in the articular disk, and result in the appearance of elastic fibers within the cartilage of the mandibular condyle.

Quantitative concentration profiling of nickel in tissues around metal implants: a new biomedical application of laser ablation sector field ICP-MS

Laser-ablation sector-field (high resolution) inductively coupled plasma mass spectrometry (LA-HR-ICP-MS) has been used for in situ determination and spatial elemental profiling of nickel concentrations in tissues that have been exposed to nickel wire. Nickel has a number of adverse biological effects that have made the use of nickel (or any other metal) in biomedical implants controversial. Yet, information about the distribution of nickel in tissues around nickel-containing implants is scarce. This study examines the diffusion of nickel with time and the spatial distribution of nickel around nickel-containing implants in vivo. Pure nickel wires were implanted subcutaneously into rats for seven days and the tissues were analyzed for nickel content and degree of inflammation away from the implants using 24Mg and 60Ni isotopes. Data were obtained by ablation with Nd:YAG laser operating in the UV region (266 nm and 213 nm) and element analysis with a high resolution ICP-MS. A 50 ppm glass standard (NIST-612) was also analyzed for the same isotopes. Quantification was performed by assuming a uniform nominal magnesium concentration value of 97 µg g−1 in untreated tissue and using 24Mg intensity for internal calibration. The precision (RSD%) of measurements for 24Mg for the NIST-612 Glass standard was within 3.8% to 4.6% and for the tissue samples was within 3.2% to 4.5%. The precision of analysis for 60Ni for the NIST-612 Glass standard was 5.4%. There was a significant penetration of nickel ions into tissues exposed to nickel wire implants. The concentration of nickel reached values as high as 60 µg g−1 near the implants, falling exponentially to undetectable levels 3–4 mm from the implants. The study showed that the laser ablation technique was well suited for analysis of soft tissues for metal ion content. This technique also allowed metal concentration spatial profiling as a function of time.

Morphological and biochemical evidence for elastic fibres in the Syrian hamster temporomandibular joint disc.

Elastic fibres are considered to be important for the normal biomechanical functions of the TMJ. The objective here was to correlate morphological evidence for the presence of elastic fibres in discal tissues with biochemical evidence for elastin. For light microscopy, the joints were removed en bloc, processed for paraffin embedding, sectioned and stained with resorcin-fuchsin. For biochemical study, a radioimmunoassay for desmosine was used to estimate the amount of elastin in excised articular discs. The histological preparations showed that numerous elastic fibres were present in various areas of the disc and in some of the discal attachments to surrounding bone. Radioimmunoassay also indicated that elastin was present in these tissues. Therefore, the biochemical findings support the morphological in suggesting that elastic fibres are present in the articular disc of the hamster TMJ.

Distribution of Putative Elastic Fibers in Rabbit Temporomandibular Joint Tissues

This study describes the general distribution of putative elastic fibers in the connective tissues that comprise the articular disk and some of the adnexal tissues of the rabbit temporomandibular joint. Joints were removed en bloc and processed for light-microscopic study. The fibroelastic tissues of the bilaminar zone of the articular disk and the various attachments of the disk to the mandibular condyle contained numerous elastic fibers. Since these morphologic data indicate the presence of many elastic fibers, we suggest that the rabbit temporomandibular joint may serve as a model system to study the functional consequences of selectively altering the quality or quantity of elastic fibers in these tissues.

Musculoskeletal Arrangements for Lateral Mandibular Movements in the Rabbit and Rat: Electromyographic and Other Analyses

Electromyography using wire microelectrodes and multichannel recording of the masseter and medial pterygoid muscles demonstrated that in the rabbit, the ipsilateral medial pterygoid muscle actively opposes lateral mandibular movement. In the rat, the ipsilateral medial pterygoid muscle and contralateral masseter muscle oppose the movement. The results correlate with the musculoskeletal arrangements of the two animals.

Morphological study of lectin binding in the rabbit temporomandibular joint disc.

SummaryGlycoconjugates of the extracellular matrix are important for the normal mechanical functions of connective tissue structures such as the temporomandibular joint disc. Since lectins are known to bind to sugar residues with high affinity, a variety of lectins were used to study the presence and distribution of glycoconjugates in the temporomandibular joint disc. Discs were removed from 6 to 8-month-old rabbits and either sectioned in a cryostat and processed for light microscopy or fixed in 2% glutaraldehyde and processed for electron microscopy. The frozen sections were incubated with fluorescein- or peroxidaseconjugated lectin solutions. Ultrathin sections mounted on grids were incubated with lectins combined with a colloidal gold marker system for electron microscopical study. Our results indicate thatCanavalia ensiformis agglutinin (ConA) showed little or no binding to the discal tissue.Triticum vulgaris agglutinin (WGA) andMacluras pomifera (MPA) were bound strongly to both the synovium and the extracellular matrix and WGA also bound to the territorial matrix of chondrocyte-like cells.Glycine max andArachis hypogoea agglutinins (SBA and PNA), were localized in the synovium and extracellular matrix but to a lesser degree than WGA and MPA. WGA, MPA,Griffonia simplicifolia II andUlex europaeus were bound by discal fibroblasts. WGA was also localized in lysosomes of synovial A-cells (macrophages). The electron microscopical studies with lectins and colloidal gold marker systems indicated that some areas of the disc may be fibrocartilagenous as had been suggested by earlier immunohistochemical studies using monoclonal antibodies to characteristic glycosaminoglycans (GAGs) in cartilage.

In vitro Effects of Elastase on Rabbit Craniomandibular Joint Articular Disks

This in vitro study correlates morphologic and radioimmunoassay (RIA) findings on the effects of elastase on the elastic fibers that are found in the rabbit craniomandibular joint (CMJ) articular disk. Articular disks were removed from rabbit CMJs at necropsy, and cut sagittally into two pieces which were incubated in 0.3 ml of phosphate-buffered saline containing either 0, 12.5, 25 or 50 units of porcine pancreatic elastase for either 1, 3 or 24 h. The quantitative RIA findings correlated well with the qualitative light-microscopic observations in that both methods showed a reduction in the amounts of elastin in the CMJ disks following enzyme treatment. However, the morphologic appearance of most of the elastase-treated disks suggested that the destruction of the elastic fibers was more extensive than was suggested by the results of the RIA which indicated that some elastin remained in the tissues of the disks even when the highest enzyme level and longest incubation period were combined. The results of this study also support the interpretation that the resorcin-fuchsin-stained fibers in the rabbit CMJ disk are elastic fibers.

Histochemical demonstration of monoaminergic and cholinesterase-positive nerve fibres in regenerating rat submandibular gland autografts.

SummaryA cholinesterase localization method and a monoamine histofluorescence technique were used to locate nerve fibres in regenerating rat submandibular gland autografts. Experimental rats had a portion of one submandibular gland excised and cut into small fragments which were autografted immediately into the middle one-thrid of the tongue. Control rats had a portion of one submandibular gland removed and discarded, and their tongues were sham-operated. Seven to ten weeks later, the rats were killed and the tongues were removed, frozen and sectioned in a cryostat. A light microscopical study of the tongue sections subjected to the cholinesterase technique showed that the submandibular gland autografts contained many nerve fibres that exhibited cholinesterase activity. These cholinesterase-positive nerve fibres were distributed throughout the autografts. The fibres were associated with the numerous duct-like structures and the less numerous acini. In addition, ultraviolet illumination of tongue sections after treatment with a glyoxylic acid mixture revealed histofluorescent monoaminergic nerves within the autografts. These fibres were less prominent than the cholinesterase-positive fibres and appeared to run primarily along blood vessels within the autografts. The results suggest that autonomic nerves are present within regenerating submandibular gland autografts.

Effects of in vivo single and multiple isoproterenol injections on subsequently explanted submandibular glands

The present study reports on the effects of in vivo single and multiple isoproterenol (ISP) injections on the proliferative response of subsequently explanted mouse submandibular glands. Explants were cultured in Waymouths medium supplemented with 10% fetal bovine serum, and were harvested after 1,2,4 or 6 days in culture. 24 h prior to harvest, the explants were fed medium containing [3H] thymidine. At the time of harvest, the explants were fixed in situ and processed for light-microscopic radioautography. The number of labeled nuclei and the total number of nuclei were counted, and labeling indices were calculated for the control and ISP-treated explant sectons. The results show that the ISP-treated explants had more cells actively synthesizing DNA than did the corresponding control explants after 1 day in culture. In addition, after 2 days in culture, the explants from mice that had received a single dose of ISP incorporated more [3H] thymidine than the corresponding control explants. These results suggest that the in vivo ISP stimulus initiated a hyperplastic response in vivo that was completed during the 1st or 2nd day in vitro.