Regression and persistence: remodelling in a tissue engineered axial vascular assembly

In later stages of vasculoangiogenesis a vascular network is going through a metamorphosis for optimal perfusion and economy of energy. In this study we make a quantitative approach to phenomena of remodelling in a bioartificial neovascular network and suggest variance of calibre as a parameter of neovascular maturation. For this study, 18 male Lewis rats were subjected to the AV loop operation in combination with a hard porous biogenic matrix and an isolation chamber. The animals were allocated into three groups for different explantation intervals set to 2, 4 and 8 weeks, respectively. Collective attributes like vascular density, percent fractional area and variance of calibre were evaluated for a predefined region of interest (ROI). Late morphogenesis was evaluated by means of scanning electron microscopy. After the fourth week the absolute number of vessels within the ROI decreased (P < 0.03) whereas, on the contrary, the fractional area of all segments increased (P < 0.02). The variance in calibre was significantly increased in the 8‐week group (P < 0.05). Lymphatic growth after week 4, early pericyte migration as well as intussusceptive angiogenesis were identified immunohistologically. Phenomena of remodelling were evaluated quantitatively in a neovascular network and variance could be proposed as a parameter of net vascular maturation.     

Microvascularization of the thyroid glands in larval and adult Xenopus laevis‐ histomorphology and scanning electron microscopy of vascular corrosion casts

This study demonstrates gross arterial supply, venous drainage and microvascular patterns of larval and adult thyroid glands in the African Clawed Frog, Xenopus laevis by scanning electron microscopy of vascular corrosion casts and light microscopy of stained serial tissue sections. Results confirm published findings gained by microscopical dissections with respect to gross arterial supply. However, in adult frogs one rather than two thyroid veins are found. This study reveals for the first time that bilaterally located thyroid glands in premetamorphosis have immature capillary networks, lack a clear hierarchy of blood vessels, and show many signs of sprouting angiogenesis. During metamorphic climax, blood vessels gain a clear hierarchy and capillaries form closed networks around thyroid follicles. From climax onwards, non‐sprouting angiogenesis (intussusceptive microvascular growth) becomes the prevailing mode of angiogenesis intensifying follicle capillarization. Due to narrow interfollicular spaces, thyroidal arterioles remain superficial while draining venules are located interfollicularly. In contrast with the mammalian thyroid gland where most thyroid follicles have their own capillary beds, most thyroid follicles in Xenopus share their capillary beds with neighbouring follicles. Consequently, the concept of individual morphological and functional angiofollicular units applicable to adult mammalian and human thyroid glands is not applicable to larval and adult amphibian thyroid glands.     

Vascularization of the brains of the Atlantic and Pacific hagfishes, Myxine glutinosa and Eptatretus stouti: a scanning electron microscope study of vascular corrosion casts

The microvascularization of the brains of the hagfishes, Myxine glutinosa L. and Eptatretus stouti, were studied by scanning electron microscopy (SEM) of microvascular corrosion casts. Sections of these casts were used to determine the vascular territories of defined brain areas. Histological serial sections (10 microm) of the brains served for correlation of findings. Analysis of the microvascular casts of both species revealed that the blood supply to and from these brains arose ventrally and dorsally, respectively. Neither species possesses an arterial circle (Circulus Willisi) and both have similar microvascular patterns. The only difference between Myxine and Eptatretus was that the posterior cerebral artery in Myxine divides into mesencephalic and rhombencephalic branches, and in Eptatretus a third branch, termed telencephalic branch, arises from the posterior cerebral artery. 3D-morphometry revealed that luminal diameters of: 1) intracerebral arteries and arterioles range from 35.11 +/- 5.66 microm (mean +/- SEM) in the hypothalamus to 92.69 +/- 14.48 microm in the thalamus; 2) capillaries range from 17.8 +/- 0.44 microm in the olfactory bulb to 21.70 +/- 0.87 microm in the basal ganglia; and 3) intracerebral venules and veins range from 49.38 +/- 4.17 microm in the hypothalamus to 75.58 +/- 6.59 microm in the rhombencephalon. Interbranching distances of arteries and arterioles range from 179.19 +/- 11.32 microm in the optic tectum to 235.19 +/- 94.64 microm in the hypothalamus. Capillaries range from 91.07 +/- 6.22 microm in the hypothalamus to 116.15 +/- 9.45 microm in the thalamus, and venules and veins range from 137.30 +/- 18.11 microm in the hypothalamus to 189.83 +/- 17.47 microm in the optic tectum. Intervascular distances range from 70.58 +/- 3.58 microm in the olfactory bulb to 89.52 +/- 5.74 microm in the optic tectum. Branching angles of arteries and arterioles range from 38.39 +/- 10.9 degrees in the olfactory bulb to 100.73 +/- 9.4 degrees in the optic tectum, and the branching angles of capillaries range from 74.40 +/- 5.42 degrees in the optic tectum to 90.24 +/- 4.66 degrees in the olfactory bulb. Finally, the branching angles of the venules and veins range from 67.84 +/- 6.83 degrees in the tegmentum of the mesencephalon to 92.30 +/- 6.35 degrees in the optic tectum.     

Microvascularization of the spleen in larval and adult Xenopus laevis: Histomorphology and scanning electron microscopy of vascular corrosion casts

Microvascular anatomy and histomorphology of larval and adult spleens of the Clawed Toad, Xenopus laevis were studied by light microscopy of paraplast embedded serial tissue sections and scanning electron microscopy (SEM) of vascular corrosion casts (VCCs). Histology showed i) that white and red pulp are present at the onset of metamorphic climax (stage 57) and ii) that splenic vessels penetrated deeply into the splenic parenchyma at the height of metamorphic climax (stage 64). Scanning electron microscopy of VCCs demonstrated gross arterial supply and venous drainage, splenic microvascular patterns as well as the structure of the interstitial (extravasal) spaces representing the “open circulation routes.” These spaces identified themselves as interconnected resin masses of two distinct forms, namely “broccoli‐shaped” forms and highly interconnected small resin structures. Arterial and venous trees were clearly identified, as were transitions from capillaries to interstitial spaces and from interstitial spaces to pulp venules. Venous sinuses were not diagnosed (nonsinusal spleen). The splenic circulation in Xenopus laevis is “open.” It is hypothesized that red blood cells circulate via splenic artery, central arteries, penicillar arteries, and red pulp capillaries primarily via “broccoli‐shaped” interstitial spaces, pulp venules and veins into subcapsular veins to splenic veins while lymphocytes circulate also via the interstitial spaces represented by the highly interconnected small resin structures in vascular corrosion casts. In physiological terms, the former most likely represent the fast route for blood circulation, while the latter represent the slow route. J. Morphol. 277:1559–1569, 2016. © 2016 Wiley Periodicals, Inc.     

Microvascular anatomy of the brain of the adult pipid frog,Xenopus laevis (Daudin): A scanning electron microscopic study of vascular corrosion casts

To demonstrate the 3D microvascular anatomy of the brain of the model organism Xenopus laevis Daudin scanning electron microscopy of vascular corrosion casts was correlated with light microscopy of stained 7 µm thick serial tissues sections. Results showed that supplying arteries descended from the leptomeningeal surface without remarkable branchings straight to the subventricular zone where they branched and capillarized. Capillaries showed few H‐ and/or Y‐shaped anastomoses during their centrifugal course toward the leptomeningeal surface where they drained into cerebral venules and veins. Apart from the accessory olfactory bulb and the vestibule‐cochlear nucleus where capillaries were densely packed, capillaries formed a wide‐meshed 3D network throughout the brain parenchyma and thus contrasted to urodelian brains where hairpin‐shaped capillaries descend from the leptomeningeal vessels into varying depths of the brain parenchyma. In about two‐third of specimens, a closed arterial circle of Willis was found at the base of the brain. If this circle in Xenopus might serve the same two functions as in men is briefly discussed. Choroid plexuses of third and fourth ventricle were found to have a high venous, but a low arterial inflow via one small choroidal artery only. Findings are compared with previous studies on the vascularization of the anuran brain and discrepancies in respect to presence or absence of particular arteries and/or veins in Ranids, Bufonids, and Pipids studied so far are discussed with particular emphasis on the techniques used in the various studies published so far.     

Vascularization of the telencephalic choroid plexus of a ganoid fish [Acipenser ruthenus (L.)

The vascularization of the telencephalic choroid plexus of the sterlet Acipenser ruthenus, a ganoid fish, was examined by vascular corrosion casting and by light and transmission electron microscopy. The arterial supply is from the dorsal mesencephalic artery via: 1) the ventral choroidal arteries (left and right); 2) the dorsal choroidal arteries (left and right); 3) the caudal choroidal arteries (left and right); 4) the ventral arteries of the dorsal sac; and, from the olfactory arteries, via 5) the rostral choroidal arteries. The venous drainage is mainly through a single main choroidal vein that can take various courses either directly to the anterior cardinal vein or via the middle cerebral vein to the anterior cardinal vein. To a lesser extent, the plexus is drained via the lateral telencephalic veins and the ventral vein of the dorsal sac to the middle cerebral vein. By angioarchitecture and form, the plexus can be subdivided into five distinct parts: the surface network, the median folds, the large lateral folds, the small lateral folds, and the area common to the bottom of the dorsal sac and the telencephalic plexus. Diameters of terminal vessels as measured from vascular corrosion casts and from paraplast, semithin, and ultrathin sections were never less than 10 micron. It is suggested that the different areas in one plexus may have different functions with respect to secretion and absorption of cerebrospinal fluid.     

Vascularization of the pars distalis of the hypophysis in the toad,Bufo bufo (L.) (Amphibia,Anura)

Summary The vascularization of the pars distalis of the hypophysis of the toad, Bufo bufo (L.), was studied by the traditional method of injecting a mixture of India-ink and gelatine into the circulatory system of the head via the arteria carotis communis. Further, methyl-methacrylate corrosion casts of the brains were made; the hypothalamo-adenohypophysial region of these corrosion casts was studied with the scanning electron microscope. The results showed that the portal vessels which arise from the median eminence do not supply distinct areas in the pars distalis as is supposed by the point-to-point-hypothesis. The portal vessels enter the ventro-median region of the pars distalis and branch off into a three-dimensional network of the secondary capillary plexus of the pars distalis. The plexus is made up mostly by four- to six-sided meshes. This angioarchitecture guarantees an optimal supply of the glandular cells of the pars distalis with nutritional factors and releasing hormones, on the one hand, and facilitates the removal of the hormones which are released by these cells, on the other hand. The venous drainage of the pars distalis is exerted mainly by two large veins, which bilaterally leave the dorso-lateral region (venous pole) of the pars distalis and by a few small veins, which drain into the wide, sinus-like vessel, which curves around the dorso-caudal region of the pars distalis and joins bilaterally the vena hypophysea transversa.This work was supported by the Fonds zur Förderung der wissenschaftlichen Forschung (Projekt Nr. 2183/N 39) and by the Stiftungs- und Förderungsgesellschaft der Paris-Lodron-Universität SalzburgThe authors wish to thank Mag. Ursula Albrecht for excellent technical assistance and Mr. Gerhard Sulzer for photographic work     

Vascularization of the pars intermedia of the hypophysis in the toad,Bufo bufo (L.) (Amphibia,Anura)

The vascularization of the pars intermedia of the hypophysis of the toad, Bufo bufo (L.) was studied by injection of a mixture of India-ink and gelatine into the circulatory system of the head via the arteria carotis communis. Further methyl-methacrylate corrosion casts of brains were made and the hypophysial region of the corrosion casts was examined with the scanning electron microscope. The results showed that the vascularization of the pars intermedia of the toad hypophysis consists of a single-layered vascular network, which is located on the ventral surface of the pars intermedia. The network is formed by capillaries, which primarily run caudally in a fan-like manner and which show only a few cross-connections. In the rostral region of the pars intermedia this network lies rather superficially, while in the caudal region it slightly penetrates the parenchyma. The vascular network originates from vessels of the neural stalk and from wide capillaries of the rostro-ventral region of the neurointermediate junction. The venous drainage of the pars intermedia is exerted by veins, which leave the caudal region and drain into the veins leaving the venous pole of the pars distalis. The flat, wide meshed vascular net on the ventral side of the pars intermedia, demonstrated in this study, fits into the concept that the pars intermedia of the anuran hypophysis is under the control of nerve fibers coming from the hypothalamus.