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PubMed1992

Cutaneous vascular anatomy of the thoracic region of the dorsum and its role in flap design in the rat.

The cutaneous vascular anatomy of the proximal one-half of the dorsum (the thoracic region) and its role in flap design was studied in the rat. The investigation included anatomical dissection, ink injection into the axial artery, and flap harvesting in live animals. Anatomical dissection and india ink injection of the thoracic region revealed that the skin derives its principal blood supply from the lateral thoracic artery. The cutaneous vascular territory of the lateral thoracic artery was defined as follows: the medial border, along the midline of the back from the level of the inferior angle of the scapula to the lower border of the last rib; the lateral border, anterior axillary line corresponding with the length of the medial border; the cephalic border, a line joining the medial and lateral border at the level of the inferior angle of the scapula; and the caudal border, a line corresponding to the lower border of the last rib. Bilateral vascular pedicle island cutaneous flaps were harvested in living rats based on the vascular territory demarcated by India ink injection. All flaps survived; hence, this flap is reliable, with consistent vascularity, and is easy to harvest, and therefore, can be used as a vascular pedicle experimental model to study flap hemodynamics.

Animals↗
PubMed2001

Development of a device for the delivery of agents to bone during distraction osteogenesis.

Various agents have been theoretically and experimentally implicated as mediators of distraction osteogenesis (DO). The purpose of this study was to develop a vehicle for the potential delivery of these factors to the region of the distraction site in an attempt to manipulate this biologic process. Three adult mongrel dogs (12 months old) had oblique osteotomies performed bilaterally through the gonial regions. In group I, the external distracter was affixed to the right hemimandible of two dogs (n = 2 hemimandibles) with cannulated pins (external diameter = 1.5 mm; lumen diameter = 1.0 mm; length = 60 mm), whereas the distracter on the left was affixed with standard, noncannulated pins of the same dimensions. In group II, cannulated pins were used to affix the external distracter to both hemimandibles (n = 2 hemimandibles) of a dog. The devices were activated after a 5-day latency period and were lengthened at a rate of 1 mm/day for 20 days. During the distraction period, 0.1 ml/d of sterile india ink was injected into the cannulated pins, after which the sterile stylet was replaced. The activation protocol was followed by 28 days of fixation (consolidation period). The hemimandibles from group I underwent removal of soft tissues, acetone fixation, and gross examination/photography, whereas the hemimandibles from group II were prepared for histologic evaluation (whole mount, hematoxylin and eosin staining). All dogs survived to the end of the study and demonstrated successful DO without evidence of complications. Hemimandibles in group I displayed evidence of india ink on both the lingual and buccal cortex around the cannulated pin site, in the regenerate and on the neocortices of the distracted segment. Hemimandibles of group II showed histologic evidence of the india ink being deposited densely around the cannulated pin site and extending in a radial fashion around the pin site into the regenerate. This study demonstrates for the first time a vehicle device for the delivery of an inert dye to the regenerate site during distraction osteogenesis. This vehicle offers the potential of delivery of various factors implicated in distraction osteogenesis (i.e., mitogens) in an attempt to alter this process and also substances (i.e., chemotherapy, antibiotics, etc.) for use in the treatment of various osteopathies.

Animals↗
PubMed1983

The intracerebral microcirculation of the rat in hemorrhagic shock.

The intracerebral microcirculation of the isocortex was studied in unanesthetized rats under hemorrhagic shock. To observe the microvessels, three markers were separately injected intravenously during the shock period: (a) Evans blue for fluorescence microscopic visualization of the vessels. (b) India ink for gross and light microscopic evidence of retention of carbon. (c) Horseradish peroxidase (HRP) for light and electron microscopic study. Lack of spontaneous recovery from shock was associated with: (a) 55-65% blood loss, a low blood pressure (30-40 mm Hg), and a dramatic increase in pulse rate; (b) marked Evans blue fluorescence along the vessels; (c) no retention of India ink in the microcirculation; (d) peroxidase activity on the luminal surface of the endothelium. Absence of India ink in the microcirculation of the isocortex during the shock period, as shown by light and electron microscopy, suggests that there is sufficient cerebral blood flow to clear the carbon particles from the blood stream and that there are no openings greater than 30 nm in the endothelial layer allowing seepage of carbon particles through or between endothelial cells. Vascular Evans blue fluorescence and peroxidase activity were both demonstrated on the luminal surface of the endothelial cells, by light microscopy, indicating that these markers are abnormally retained. Ultrastructural demonstration of increased HRP uptake and adherence onto the endothelial cells confirms these observations. These results show that regional endothelial alterations occur in this model of hemorrhagic shock.

Anesthesia, Inhalation↗
PubMed1987

Surgical skin-marking techniques.

Surgical skin-marking inks and dyes are in everyday use for designing and planning incisions in plastic and reconstructive surgery. We have traced the historical development of surgical skin-marking techniques from ancient times to the present. The biochemical characteristics of the commonly used marking agents are discussed. A three-part experiment utilizing a pig model was carried out to test the tissue inflammatory response to the various dyes and inks when used intradermally as tattoos, the persistence of such tattoos, and the ease of skin erasure for each of eight stains. Methylene blue and gentian violet are recommended as the best all-purpose marking agents. The use of proprietary inks is discouraged.

Animals↗
PubMed1997

The contribution of the SMAS to the blood supply in the lateral face lift flap.

The purpose of this study was to compare the arterial blood supply of the lateral face lift flap when the flap is elevated either above or below the SMAS. The transverse facial artery supplies blood to a large portion of the lateral face lift flap. This vessel passes through the superficial musculoaponeurotic system (SMAS) in its course toward the flap. The transverse facial artery is at risk for transection during both elevation of the skin flap and elevation of the SMAS. Nine fresh cadavers underwent a single-plane rhytidectomy either superficial to the SMAS or in a sub-SMAS plane. In half the subjects, the transverse facial artery perforator was identified prior to dissection using surface landmarks and preserved. In the other half of dissected specimens, this perforator was transected by undermining through the transverse facial artery perforation site (near the zygomatic ligament). This transection is typically performed during a face lift. The facial artery and transverse facial artery were selectively injected with ink, and cutaneous staining was recorded. It was found that the pattern and total area of ink staining occurred in an all-or-none fashion depending on whether the transverse facial artery had been preserved or transected. Inclusion or exclusion of the SMAS layer in the dissected lateral face lift flap had no effect on the ink injection pattern, suggesting that the SMAS plays little or no role in lateral face lift flap viability.

Cadaver↗
PubMed1979

The lymphatic drainage of the rat prostate and its status as an immunologically privileged site.

Recent suggestions that the rat prostate is an alymphatic, immunologically privileged site stimulated further investigation of its status using a variety of techniques. Fixation of certain organs by vascular perfusion of glutaraldehyde followed by plastic embedding avoids the distortion and stromal reorganization that often obscures evidence of lymphatic vessels in conventional histological preparations. When applied to the rat prostate, this technique revealed small lymphatic vessels at irregular intervals throughout the sparse stroma. Ink injected into the prostate drained from the organ into iliac lymph nodes within 3 to 4 hr. Enlargement of iliac lymph nodes within 1 week after injection of parental strain lymphoid cells into the prostate of F1 rats confirmed the drainage pattern. Ink and lymphoid cell injections into the bladder wall yielded comparable results, although ink reached iliac nodes sooner than it did in prostate injections. Immunological privilege was investigated by determining survival of skin allografts implanted in the prostate. Skin-into-prostate grafts bearing either major or minor histocompatibility antigens were rejected within a few days of similar orthotopic grafts. The inability of the rat prostate to allow significantly prolonged allograft survival as well as its demonstrated lymphatic drainage argues against an immunologically privileged status.

Animals↗
PubMed2007

Tracing of gastric reflux into the middle ear in a mongolian gerbil model.

OBJECTIVES: The purpose of this study was to trace induced gastric reflux and to examine whether it reaches the middle ear in a Mongolian gerbil model. BACKGROUND: Otitis media with effusion is the most frequent middle ear disease in childhood. Gastroesophageal reflux is suspected to be a possible factor in its pathogenesis. METHODS: Seventeen Mongolian gerbils were assigned to three groups: the control (phosphate-buffered saline application to the lower esophageal sphincter) and two experimental groups (Aquo-Trinitrosan [Merck, Darmstadt, Germany] application to the lower esophageal sphincter, low gastric pressure and Aquo-Trinitrosan application, higher gastric pressure). We injected Chinese ink into the stomach to trace the path of a potential gastroesophageal reflux in all three groups. The traces of ink were investigated by ear and larynx endoscopy and histology. RESULTS: There were no signs of gastroesophageal reflux based on the data obtained from the control group. In animals with traceable laryngeal reflux, the ink was also shown to advance through the eustachian tube and reach the middle ear. In addition, we found that when reflux reaches the middle ear on one side, it also reaches the contralateral middle ear in most cases. CONCLUSION: Gastroesophageal reflux induced by relaxation of the lower esophageal sphincter was shown to reach the middle ear in our Mongolian gerbil model. These results support recent hypotheses linking gastroesophageal reflux to the development of otitis media with effusion.

Animals↗
PubMed1992

Quantitative analysis of the fine vascular anatomy of articular ligaments.

An image analysis technique has been developed to quantitatively describe the fine vascular patterns observed in ligament tissue. The longitudinal orientational distribution and total vessel volume of India-ink-perfused blood vessel segments in normal and healing ligaments were determined. The methods involved special vascular preparation of adult rabbit knee medial collateral ligaments (MCL) by India-ink perfusion. Black and white microscope images of ink-perfused tissue sections were subjected to a thresholding procedure to binarize digitized ligament images, which were then skeletonized and analyzed for directional distribution based on the least-squares technique. Analysis of medial collateral ligaments in New Zealand White rabbits using this method has shown that scarred tissue is more vascular and has a more chaotic angular distribution of blood-vessel segments than normal ligament tissue.

Algorithms↗
PubMed1998

Suppressive effect of the angiogenesis inhibitor TNP-470 on the development of carcinogen-induced hepatic nodules in rats.

Tumor metastasis can be prevented by inhibiting angiogenesis. In the present study, we have demonstrated that the angiogenesis inhibitor TNP-470 also suppresses the development of primary hepatic nodules. Hepatocarcinogenesis was performed by the feeding of 2-acetylaminofluorene to hepatectomized rats during 8-14 weeks of age. Predominantly arterial-to-portal circulation and sinusoidal capillarization were determined by the staining of nodules with arterially infused ink and immunostaining for factor VIII-related antigen, respectively. Intraperitoneal administration of 30 mg/kg b.w. of TNP-470 twice a week significantly reduced the number of hepatic nodules. Among the nodules, hyperplastic nodules stained with ink, atypical hyperplastic nodules and hepatocellular carcinoma, all of which possess structurally altered sinusoidal endothelial cells or capillary-type endothelial cells, were dramatically decreased in number. Suppression was observed equally in nodules of all sizes. TNP-470 was more effective when administered during 8-20 weeks than during 14-26 weeks. In contrast, ink-non-stained hyperplastic nodules, which have normal sinusoidal endothelial cells, were not affected at all. The present results indicate that TNP-470 suppresses the development of primary hepatic nodules whose microvessels are capillaries or transitional forms from sinusoids to capillaries.

2-Acetylaminofluorene↗
PubMed1995

Reliability of reduced air pressure methods used to assess the apical seal.

The reliability of two commonly used methods of sealability testing, dye leakage under normal atmospheric pressure and dye leakage in a partial vacuum, was tested using a repeated studies design. Forty extracted teeth were instrumented and obturated using lateral condensation of gutta-percha and Sealapex sealer. One-half of the teeth were placed in India ink under normal atmospheric pressure and the other half underwent air evacuation before immersion in the ink. After 24 h the teeth were removed from the ink and rendered transparent for linear leakage analysis. A second study, repeated within the exact parameters of the first, was performed approximately 1 month later. Mann-Whitney U tests indicated that there was no significant difference (P>0.05) between the air and vacuum groups or within each group (air or vacuum) between trials, indicating that the methodology is repeatable and that application of reduced air pressure did not enhance reliability. A large majority of the teeth placed under partial vacuum showed evidence of vacuum-induced artefacts that may negate the value of vacuum as a research tool.

Air Pressure↗
PubMed1994

Periodontal ligament injection in the dog primary dentition: spread of local anaesthetic solution.

The spread of local anaesthetic solution administered with a pressure syringe has not been studied as extensively in young animals having primary or mixed dentitions as in adult animals. The purpose of this investigation was to study the distribution of local anaesthetic solution injected into the periodontal ligament of young dogs. India ink was added to carpules containing 2% lidocaine and 1:100,000 epinephrine, and the spread of solution was examined macroscopically and microscopically. Injections were made with a pressure syringe (Ligmaject) at 58 sites mesial and distal to primary teeth in five dogs aged 3-9 months. Three dogs were killed within 12 hours, the other two after 5 days. Spread of the ink was studied in non-decalcified slabs, in three-dimensional cleared specimens, and in histological sections. The solution usually reached the alveolar bone crest, seeped under the periosteum and alongside vascular channels into bone marrow, reaching natural cavities such as the crypts of tooth buds and the mandibular canal. The ink did not penetrate into the enamel organ or contact the permanent tooth buds. The solution appeared to spread along the path of least resistance, governed by the intricacies of anatomical structures and fascial planes. Therefore the risk of mechanical damage to permanent tooth germs appears to be minimal.

Anesthesia, Dental↗
PubMed1993

Encapsulation of streptococci isolated from bovine milk.

Capsule expression was assessed by India ink staining in 95 streptococcal and enterococcal isolates representing 8 species. Organisms were evaluated after primary culture on blood agar and after storage in 10% skim milk at -80 degrees C. Seventeen of 95 primary culture samples were positive for an unstained halo (capsule) by India ink. Only 3 isolates were positive for capsule after being maintained for an extended period of time. To confirm results of India ink tests, an unencapsulated and encapsulated strain of each species was observed by electron microscopy. Cells of encapsulated strains were surrounded by thread-like fibers indicative of capsule, whereas cells of unencapsulated strains had a smooth surface. The importance of capsule in the pathogenesis of streptococcal mastitis is unknown. Studies on capsule expression by streptococci may facilitate investigations on the pathogenesis of streptococcal mastitis.

Animals↗
PubMed1995

The Q-switched Alexandrite laser's effects on tattoos in guinea pigs and harvested human skin.

BACKGROUND: Developmental work in tattoo removal is currently carried out in animal models whose skin has different optical and mechanical properties from human skin. OBJECTIVE: To observe laser effects on tattoos in a new human skin model and that of in vivo guinea pig skin. METHODS: A comparison of the efficacy of the Q-switched Alexandrite laser (750 nm, 100-120 nanoseconds) in tattooed harvested human skin and guinea pig skin was performed. Visual assessments as well as histologic and electron microscopy evaluation before and after treatment are reported. RESULTS: Mild to moderate clearing of black tattoo pigment, as judged by visual assessment, was found after one laser treatment in both the in vitro human skin model and guinea pig skin. Blue and green ink showed similar results to black ink in human skin whereas red and yellow ink did not respond in our model. CONCLUSION: Possible mechanisms of tattoo lightening and the usefulness of the in vitro human skin model are discussed.

Animals↗
PubMed1985

Studies on the vascular drainage system of the hamster buccal pouch.

Because some investigators feel that the hamster buccal pouch is an immune privileged site with no lymphatic drainage, this study was done to demonstrate and further elucidate a drainage system from the pouch to the cervical nodes. Eight hamsters were injected with india ink, submucosally in the right pouch. Dissections at one or 2 days revealed ink in macrophages in the right submandibular nodes at 2 days. Histologically, the ink was noted within macrophages which aggregated around blood vessels. No lymph vessels were noted in the pouch. Six hamsters were injected with fluorochrome-labelled latex microspheres. At 2 and at 4 days the microspheres were noted in both the pouches and the ipsilateral submandibular nodes. The results indicate there is a delayed lymphatic drainage from cheek pouches to the submandibular nodes mediated by macrophages.

Animals↗
PubMed1995

Sealing ability of potential retrograde root filling materials.

The sealing ability of two potential retrograde root filling materials, a light-cured glass ionomer cement (Vitrebond) and a reinforced zinc oxide-eugenol cement (Kalzinol) was compared with that of amalgam using three methods of assessment: bacterial leakage, confocal microscopy and Indian ink leakage. the root canals of 80 extracted human single-rooted teeth were prepared. All the teeth were apicected, retrograde cavities were prepared and then divided into four equal groups of 20 teeth. The teeth were sterilised by autoclaving and the retrograde cavities filled with the test materials. In the control group, retrograde cavities in 10 teeth were left unfilled while cavities in another 10 teeth were sealed with cyanoacrylate cement. The teeth were first subjected to a bacterial leakage test using Enterococcus faecalis as a leakage marker. After which, the adaptation of the retrograde root fillings was assessed using a confocal optical microscope. Finally, the teeth were processed for the Indian ink leakage test. Bacterial leakage occurred in more teeth filled with amalgam compared with both Vitrebond and Kalzinol (P<0.001), between which there were no differences. With confocal microscopy, the size of the marginal gap was largest with amalgam and smallest with Vitrebond; all differences were statistically significant (<0.001). Finally, there was more Indian ink penetration with amalgam compared with both Vitrebond and Kalzinol (P<0.001) but there were no differences between Vitrebond and Kalzinol. Although there were individual differences within samples, overall, the three methods of assessment produced similar results. The sealing ability of Vitrebond and Kalzinol was similar and both materials were better than amalgam.

Carbon↗
PubMed2004

Cryptococcus neoformans CAP59 (or Cap59p) is involved in the extracellular trafficking of capsular glucuronoxylomannan.

Several genes are essential for Cryptococcus neoformans capsule synthesis, but their functions are unknown. We examined the localization of glucuronoxylomannan (GXM) in strain B-3501 and in cap59 mutants B-4131 and C536. Wild-type strain B-3501 showed a visible capsule by India ink staining and immunofluorescence with anticapsular monoclonal antibodies (MAbs) 12A1 and 18B7. B-4131, a mutant containing a missense mutation in CAP59, showed no capsule by India ink staining but revealed the presence of capsular polysaccharide on the cell surface by immunofluorescence. The cap59 gene deletion mutant (C536), however, did not show a capsule by either India ink staining or immunofluorescence. Analysis of cell lysates for GXM by enzyme-linked immunosorbent assay revealed GXM in C536 samples. Furthermore, the epitopes recognized by MAbs 12A1, 2D10, 13F1, and 18B7 were each detected in the cytoplasm of all strains by immunogold electron microscopy, although there were differences in location consistent with differences in epitope synthesis and/or transport. In addition, the cells of B-3501 and B-4131, but not those of the cap59 deletant, assimilated raffinose or urea. Hence, the missense mutation of CAP59 in B-4131 partially hampered the trafficking of GXM but allowed the secretion of enzymes involved in hydrolysis of raffinose or urea. Furthermore, the cell diameter and volume for strain C536 are higher than those for strain B-3501 or B-4131 and may suggest the accumulation of cellular material in the cytoplasm. Our results suggest that CAP59 is involved in capsule synthesis by participating in the process of GXM (polysaccharide) export.

Biological Transport↗
PubMed1992

Evidence that Candida albicans binds via a unique adhesion system on phagocytic cells in the marginal zone of the mouse spleen.

We recently demonstrated by using an ex vivo adhesion assay that Candida albicans yeast cells exhibit a unique binding affinity for the marginal zone of the spleen. This binding event provides a working model for studying mechanisms of organ dissemination of the fungus from the blood. By using the ex vivo assays reported here, we showed by bright-field and electron microscopic techniques that mouse spleen marginal zone cells capable of ingesting India ink particles are also involved in yeast cell attachment. During splenic clearance of yeast cells from the circulation in vivo, C. albicans is also associated exclusively with marginal zone cells capable of ingesting India ink. The ability to ingest the ink particles is not necessarily related to yeast cell adherence, because the fungal cells did not bind to phagocytic cells in the splenic red pulp. In fact, the marginal zone phagocytic cells appear to have a unique binding system, because yeast cells also did not bind to phagocytes in other tissues, such as the thymus and peritoneum, or to seven different myeloid cell lines. In addition, antibodies to a number of well-characterized murine adhesion molecules, such as leukocyte integrins, LECAM-1, and CD44, had no effect on binding. On the basis of these results, we propose that splenic marginal zone phagocytes express a novel adhesion system that involves either a unique adhesion molecule or previously described adhesion molecules with unique binding activities.

Adhesiveness↗
PubMed1992

Heterogeneity of rabbit aortic endothelial cells, with special reference to phagocytosis.

Heterogeneity of aortic endothelial cells with regard to phagocytotic ability was examined by injecting India ink into normal rabbits. Light and electron microscopic analyses revealed that particles of India ink were phagocytosed in the endothelial cells, which in turn were localized at the distal side of the orifice of aortic branches, especially those of brachiocephalic, left clavicular, and dorsal intercostal arteries. No remarkable differences were found ultrastructurally between phagocytosing and nonphagocytosing endothelial cells. Ingested India ink particles were present within phagosomes of the endothelial cells for several hours after injection; the particles eventually accumulated in the subendothelial space twenty-four hours after injection. These results indicate that an active transport system of large molecules via the phagocytotic processes is present in endothelial cells located at the distal sides of the orifice of aortic branches. These regions are known to develop initial atherosclerotic lesions in hypercholesterolemic animals. Thus, a possible correlation between phagocytotic ability of endothelial cells and development of atherosclerosis is suggested.

Animals↗