India Ink Injection Research Articles

Laparoscopic colonic surgery: The unseen lesion

SummaryPre-operative contrast radiology should be used to confirm the location of colonoscopically diagnosed lesions due to the inability to directly palpate the colon during laparoscopic assisted colectomies. If the lesion is small, then injection of India ink should be used to facilitate localization at laparoscopy.

Endoscopic India ink injection: a method for preparation, sterilization, and administration

India ink tattooing of the colon has been used to mark lesions for subsequent surgical identificationl -3 and to identify polypectomy sites to facilitate followUp.4 In animal studies, endoscopic injection of dyes in the colon has been shown to be an effective and welltolerated method for mucosal marking.5 Unfortunately, several side effects have been attributed to the injection of India ink. Reports include influenza infection in mice pre-treated with India ink,6 phlegmonous gastritis after injection into the stomach,7 fat necrosis,s inflammatory pseudotumor (which appeared to consist of inflammatory cells and fibroblast proliferation),S and abscess.9 Possible causes of these complications may be an inflammatory reaction to the ink (carbon particles) or diluent (either via a direct irritant effect ofthe undiluted ink or secondary to a hypersensitivity reaction), or inadequate sterilization. For example, in the case of phlegmonous gastritis7 and the cases of infection in mice,5 the ink was not sterilized. In the report of pseudotumor,s the ink (Higgins, Faber-Castell, Lewisberg, Tenn.), comprising about 7 % carbon pigment, 5% propylene glycol, and smaller concentrations of shellac, ammonium hydroxide, and surfactant, was used in a very concentrated form (approximately equal parts India ink and bacteriostatic water). Anyone ofthe multiple diluents or the carbon pigments may have caused this inflammatory response. In addition, post-autoclave contamination of the ink may have occurred. Similar reasons might account for the case offat necrosis,s in which the type of ink was unknown but was injected in concentrated form (approximately equal parts India ink and bacteriostatic water). The same possible causes could account for the abscess9 found incidentally during operative resection for tumor. The India ink used was Pelikan brand (Pelikan, Hanover, Germany), containing ethylene glycol, sodium tetraborate decahydrate, and ammonia in 1:10 dilution with sterile water. This for-

Observations on experimental flow-through venous flaps

The aim of this work was to compare the survival of an arteriovenous island flap with the survival of an island flap with a flow-through venous supply. Our experimental studies were performed on 95 Wistar rats randomised into six groups: Group 1: Indian ink injection of flow-through venous flaps with capillary network; Group 2: control group deprived of vascularisation; Group 3: control group with arteriovenous supply; Group 4: flow-through venous flaps of group 1; Group 5: epigastric flow-through venous flaps with a main venous trunk; Group 6: histological examination of flow-through venous flaps. The survival of flaps was monitored by direct examination, histological examination, capillaroscopy, and laser Doppler. Three out of 50 flow-through venous flaps survived. There was a statistically significant difference in the delay of clinical necrosis between the composite non vascularised free grafts (2.8 ± 1.2 days) and the flow-through venous flaps (4.1 ± 1.3 to 4.9 ± 1.1 days depending on the type of flap). A 20% decreased venous blood flow was observed in the flow-through venous flaps.

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.

An experimental study on direct revascularization of bronchial circulation by microvascular anastomosis

The effects of direct revascularization of the bronchial artery after bronchoplasty were estimated by laser Doppler velocimetry and india ink injection in dogs. Bronchoplastic surgery at the right main bronchus was performed in all dogs, and the bronchial artery was reconstructed using the internal thoracic artery in the reconstruction group. The mucosal blood flow was measured at the distal side of the anastomosis. India ink was injected into the aorta in the nonreconstruction group and into the internal thoracic artery in the reconstruction group. The peripheral blood flow had diminished immediately after surgeries to 59% of the baseline value and took 14 days to recover to the baseline value in the nonreconstruction group. However, in the reconstruction group, the blood flow recovered at once to 78% of the baseline value and had returned to that value in 5 days. Statistically significant differences were noted between the groups from just after operation to day 7. India ink data confirmed these findings. In the nonreconstruction group, no ink was observed in the peripheral bronchial vessels on day 3; it was noted in part of the vessels on day 7 and in most on day 14. On the other hand, a relatively large number of vessels were stained just after operation in the reconstruction group. Thus reconstruction of the bronchial artery by means of the anastomosis with the internal thoracic artery can be said to be a useful and effective method for preventing airway ischemia.

Anatomic and clinical considerations of an internal mammary artery harvest.

In an effort to understand the perceived correlation of internal mammary artery harvesting and wound healing difficulties in the inferior margins of the sternotomy incision, we showed the cutaneous vascular perfusion in the sternal and xiphoid areas by India ink injection studies in cadavers. With these studies, we demonstrated an inherent paucity of nutrient supply to the inferior sternum and xiphoid area. The classic internal mammary artery harvest further compromises the blood supply to these areas. We believe that limiting the most inferior dissection of the internal mammary artery and not including the distal bifurcation leaves intact the lateral musculophrenic nutrient supply to the inferior sternum and xiphoid area and to the ipsilateral abdominal rectus muscle. These guidelines will help to prevent ischemic complications of this area and may aid in reconstruction. If the bifurcation is harvested, we believe that the removal of the avascular xiphoid cartilage at the time of the initial bypass procedure may eliminate this as a potential septic focus.

Pericranial healing and the temporalis myo-osseous flap in the rabbit model.

The purpose of this animal study was to determine the rate of revascularization of a temporalis myo-osseous (TMO) flap after pericranial elevation. In 24 rabbits, the right pericranium was raised in entirety through a bicoronal flap at the first operation. The pericranium was then reapproximated in situ. The pericranium was allowed to heal for 1 to 28 days before the second operation. At the second operation, through the same bicoronal flap, right and left temporalis myo-osseous flaps were raised. The left temporalis myo-osseous flap served as a control. Revascularization and viability of the temporalis myo-osseous flaps were studied by using technetium bone scans, india ink injection studies, and histologic study. Results demonstrated that 4 days following pericranial elevation, the temporalis myo-osseous flap is viable and revascularized by the pericranium. Immediate bone scanning and india ink injection showed patent pericranial circulation to the osseous portion of the temporalis myo-osseous flap at 4 days. Histologic study confirmed the viability of the temporalis myo-osseous flap. In conclusion, after pericranial elevation, pericranial healing and revascularization were complete at 4 days. This allowed a viable temporalis myo-osseous flap to be raised successfully at this time.

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.

The Blood Supply of the Human Distal Radioulnar Joint and the Microvasculature of Its Articular Disk

The arterial blood supply of the distal radioulnar joint was investigated in 35 upper extremities taken from 22 fresh cadavers (11 newborns and 11 adults using the India ink injection and tissue-clearing techniques according to Spalteholz). Microvasculature of the articular disk of the distal radioulnar joint was also performed in 35 articular disks taken from 22 fresh human cadavers, 11 newborn and 11 adults using the same technique. It was found that the general blood supply to the joint is received mainly from the palmar and dorsal branches of the anterior interosseous artery. These branches, after dividing at the proximal border of the pronator quadratus, arborize in a fanlike fashion around the joint and their small ramifications penetrate and vascularize the capsule and the articular disk from the palmar, dorsal, and medial sides. The terminal branches of the anterior interosseous artery reinforced by the posterior interosseous artery and a small branch of the ulnar artery give the direct peridiscal vessels to the palmar, medial, and dorsal margins of the articular disk, which arborize and anastomose with one another and form the terminal capillary networks that end at the peripheral segments of the disk in a series of terminal capillary loops, leaving the inner segments devoid of blood vessels. The posterior interosseous artery anastomoses at the distal part of the forearm with one of the terminal rami of the dorsal branch of the anterior interosseous artery and, in that way, contribute to the vascularization of the dorsal capsule of the distal radioulnar joint. The ulnar artery gives off a small branch that anastomoses with one of the terminal ramifications of the palmar branch of the anterior interosseous artery and contributes to the formation of a small arterial arch on the anteromedial side of the distal ulna, supplying the anteromedial capsule and the basistyloid area of the ulna. Both ulnar and radial arteries contribute to the vascularity of the joint through the collateral network of the palmar and dorsal carpal arches. In the articular disk, the major central portion of the disk is avascular and only its peripheral, palmar, medial, and dorsal margins are vascularized. The proportion of vascularized zone to avascular zone depends on the age of the subject and, in newborns, is approximately 33%. In adults, only 25% of the peripheral segments are vascularized.(ABSTRACT TRUNCATED AT 400 WORDS)

A new experimental model: the vascular pedicle cutaneous flap over the dorsal aspect (flank and hip) of the rat

Cutaneous vascular anatomy of the flank and the hip was studied in the rat. Investigation was done by anatomical dissection, ink injection to the axial artery and flap harvesting in living rats. Anatomical dissection and India ink injection revealed that the rat's flank and hip skin derives its blood supply from the iliac branch of the iliolumbar artery, which originates from the lateral aspect of the abdominal aorta. The vascular territory of the iliac branch of the iliolumbar artery was defined as follows: the medial border—along the midline of the back from the 12th rib to the proximal part of the tail; 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 12th rib; and the caudal border—the posterior lateral line of the hip. On the basis of the vascular territory demarcated by India ink injection, bilateral vascular pedicle island cutaneous flaps were harvested in living rats. Results showed total survival of all flaps. This flap is easily made, and it could be used as a reliable model for flap research.

Prefabrication of free flaps using the omentum in rats.

The omentum used as a vascular carrier is a new experimental model for prefabrication of free flaps in the rat. A 2.5 x 4 cm patch of omentum with gastroepiploic vessels and its rich vascular arcades was transferred under a bipedicled 2.5 x 6 cm right abdominal panniculocutaneous flap. At subsequent time intervals (postoperative days 0, 3, 5, 7, and 14), the skin pedicles were divided and the skin flap was raised as a composite island flap vascularized only by the underlying omental patch. The composite flap was then sutured back in place. Prefabricated flaps examined postoperatively demonstrated a dye fluorescence index (DFI) of 4.60% +/- 2.82% and 1.67% +/- 4.08% flap survival (FS) in the day 0 group; in the day 3 group, 9.83% +/- 1.47% of DFI and 15.00% +/- 10.35% of FS; in the day 5 group, 28.39% +/- 5.25% of DFI and 86.0% +/- 12.74% of FS (P less than 0.05); in the day 7 group, 38.19% +/- 7.52% of DFI and 98.13% +/- 3.72% of FS (P less than 0.05); and, in the day 14 group, 49.55% +/- 6.70% of DFI and 100% of FS (P less than 0.05). India-ink injection and histologic examination confirmed revascularization of the overlying skin by day 5.

Lymphatic drainage of the prostate gland in canines.

Lymphography of the penis was performed in 3 canines, and direct intraprostatic injection of India ink was carried out in 8 canines to visualize lymphatic drainage of the prostate gland. In penile lymphography, the contrast medium drained in 2 directions: (1) lymphatic drainage into the external and common iliac lymph nodes through the superficial inguinal lymph nodes; (2) lymphatic drainage into the presacral lymph nodes along the urethra and posterior wall of the pelvis. Lymphatics of the prostate gland drained in 3 directions: (1) lymphatics from the prostate gland mainly drained along the prostatic and internal iliac vessels into the internal and common iliac lymph nodes, (2) lymphatics from the dorsal region near the urinary bladder drained along the ureter into the common iliac lymph nodes, and (3) lymphatics from the apex of the prostate gland drained along the posterior wall of the pelvis into the presacral lymph nodes. Prostate cancer developed at the apex is estimated to preferentially metastasize to the presacral lymph nodes, and drug injection into the penile lymphatics is considered to be a good route to treat metastases of lymph nodes in the pelvis.

On the morphology of the terminal microvasculature during endochondral ossification in rats

The investigation was carried out on the proximal tibia metaphysis by means of India ink-injection, reconstruction, corrosion casting combined with scanning electron microscopy, and transmission electron microscopy of thin sections. The terminal vasculature during endochondral ossification proved to be a labyrinth of interconnected sinusoidal complexes with bulbous buds invading the chondrocyte cavities of the growth plate. This system of vessels was canalized throughout. A special feature was the existence of endothelial pockets. The formation of the terminal microvasculature as a labyrinthine cavity during endochondral ossification, together with the still incompletely developed bone could be a weak point for compression injuries.

The blood vasculature of the gastrointestinal tract in chinook, Oncorhynchus tshawytscha (Walbaum), and coho, O. kisutch (Walbaum), salmon

The venous and arterial vasculature of the chinook and coho salmon gastrointestinal tract were examined using corrosion casts and India ink injection techniques. Observations derived from 28 individuals of various sizes and of both sexes were used to construct simplified venous and arterial plans. Examination of the blood vasculature revealed the presence of a variety of anastomoses hitherto undescribed in teleosts.

The Fasciovascular Pedicle for Revascularization of Other Tissues

A fasciovascular pedicle based on the epigastric vessels was developed in a rat model to determine if it could be used as a "universal carrier" to revascularize a new composite flap. The effects of time course, carrier size, and flap ischemia on the revascularization process were studied. A 2.5 x 4-cm or 1 x 4-cm fascial patch pedicled on the vessels was transferred under bipedicled 2.5 x 4-, 6-, or 8-cm abdominal panniculocutaneous flaps. At different time intervals, the flap was raised as an island flap connected only by it vascular bundle and then sutured back in place. The skin perfusion by dermofluorometry and flap survival were both markedly increased on day 5 (p less than 0.001). The wide carrier had a 93% survival area, whereas the narrow carrier had only 71%. The wide carrier induced relatively faster and better revascularization (p less than 0.05). Moderate ischemia promoted revascularization (p less than 0.01). An india ink injection study and histological examination provided visual evidence of revascularization. This fasciovascular pedicle is a promising model for prefabrication of complex new composite flaps and for studying the process of revascularization between the layers. Based on these findings and further investigations, a thin, prefabricated abdominal free flap was successfully transferred for facial resurfacing in humans.

The microvascular anatomy of the canine stomach: A comparison between the body and the antrum

To investigate whether there is a difference in the microvascular architecture between the body and the antrum of the canine stomach, these two locations were compared with respect to microsphere entrapment and the microvascular architecture and diameter in relation to histology by corrosion casting and by intraarterial injection of india ink. There was 63% shunting of 9-micron microspheres in the antrum, but none in the body. Corrosion cast and Indian ink studies showed that in the body there was a single microvascular network of capillaries that appeared to originate from the arterioles in the submucosa and were in close apposition to the epithelial cells of the gastric glands. The diameter of these capillaries was 8.6 +/- 0.2 microns. In contrast, there were two distinct capillary networks in the antrum: a basal and a superficial. The capillaries of the basal network of the antrum originated from the arterioles at the level of the muscularis mucosa and drained into the capillaries of the superficial mucosa. The capillaries of the superficial network had a significantly larger diameter (10.8 +/- 0.4 microns) than those of the basal network (7.3 +/- 0.2 microns). In many instances the capillaries of the superficial network originated directly from the ascending arterioles passing through the basal network. These direct arteriocapillary connections may have permitted the shunting of 9-microns spheres in the antrum.

Role of interfragmentary strain in fracture healing: Ovine model of a healing osteotomy

It has been hypothesized that the histological pattern of fracture healing is controlled at least in part by the local mechanical strains in the interfragmentary region. To test this "interfragmentary strain hypothesis," we applied cyclic bending deformations to tibial osteotomies in 11 sheep. An instrumented flexible plate spanning a 1-mm osteotomy gap was deformed to create a gradient of tissue elongation from 10% under the plate to 100% at the opposite cortex. The cyclic deformations were applied three times per minute, 24 h per day, for 1-5 weeks. However, as a result of tissue differentiation, the bone-plate complex increased in stiffness with healing time, resulting in a marked reduction of the gap deformation at approximately 4 weeks. Fracture healing was evaluated using vascular injection of India ink and conventional histology. A nonlinear three-dimensional finite element model of the interfragmentary tissue at the initial stage of healing was used to predict the complex tissue strains. The ingrowth of vascularized soft tissue into the interfragmentary gap, as well as the subsequent differentiation of this tissue, occurred earlier and to a greater degree in regions of lower strain. In contrast, the proliferation of callus tissue was greatest at the periosteal and endosteal surfaces of the cortex opposite the plate. Direct comparison of the finite element predictions with the histology demonstrated that the spatial distribution of bone resorption at the fracture fragment ends directly corresponded to the locations of elevated tissue strain and stress. However, there was no consistent numerical relationship between the magnitude of these local peak strains and the corresponding volume of cortical bone resorption over the bone cross section.

Topographie des Collecteurs Lymphatiques Mammaires de la Chienne

SummaryOn the topography of the lymph drainage from the mammary glands of the bitchThe topography of the lymph drainage was examined by in vivo injections of india ink into the mammary parenchyma of 73 female dogs. Only one gland was injected at a time on each side. Only the cranial abdominal mammary gland had direct drainage to the axillary as well as to the superficial inguinal nodes. The principal routes of drainage differ for each gland. The glands on each side are completely independent as regards their lymph drainage. The cranial thoracic mammary glands on each side had their own collecting duct which led to the sternal lymph nodes. The results of this study suggest to develop a new model for the surgical extirpation of mammary tumors in the bitch.

Urologic Function After Experimental Cauda Equina Compression Cystometrograms Versus Cortical-Evoked Potentials

Twenty female beagle dogs underwent an L6-7 laminectomy and six dogs each had 25, 50 or 75% constriction of the cauda equina and 2 control dogs had laminectomy only. Cystometrograms were performed pre- and post-operatively and three months after constriction. Cortical evoked potentials were monitored pre- and post-operatively and monthly for three months. After three months of constriction, the cauda equina of these dogs in each group was examined histologically and vascular circulation was examined by latex and India ink injection (Spalteholz technique). The control dogs had normal CMGs and CEPs. Twenty-five percent constriction caused no CMG changes and mild CEP changes. Fifty percent constriction caused no statistically significant CMG changes, major CEP changes and venous congestion of the nerve roots and dorsal root ganglia. Seventy-five percent constriction produced severe CMG changes with detrusor areflexia, increased bladder capacity and clinical incontinence. CEPs also had marked deterioration. Vascular analysis revealed severe arterial narrowing at the level of constriction and venous congestion of the nerve roots and dorsal root ganglia. Blockage of axoplasmic flow and nerve root atrophy was seen in all dogs with 75% constriction. Cortical evoked potentials were the most sensitive predictor of neural compression. CMGs were not sensitive until severe compression was achieved. Bladder dysfunction, i.e., detrusor areflexia, appears to occur with blockage of axoplasmic flow and early sensory changes occur with neurovenous congestion.

Vascularization and related distribution of leucocytes in carp, Cyprinus carpio L., head kidney

The distribution of lymphoid cells in the carp head kidney was investigated in relation to the vascular system. Blood vessels in the head kidney were histologically identified into arteries, sinusoids and two types of veins: renal veins and portal, which were distinguished by India ink injection into the caudal vein. By histological and histoplanimetrical observations it was found that the head kidney contained a number of lymphoid cells, which mainly aggregate around the connections between the portal veins and sinusoids, and that the cellular density of the aggregations was higher than in the thymus.Pigment‐containing cell clusters were also observed around these connections. This arrangement of the blood vessels suggests that it is one of the structures able to trap foreign materials, and the occurrence of the lymphoid clusters around the portal veins is a phylogenetic sign of the morphological division between granulopoietic and lymphatic tissues in the carp head kidney.