The dynamics of morphofunctional changes in the microcirculation of the pelvic extremity after modeling the chief stages of its replantation]

Under study was the effect of autotransplantation in its "pure form" upon the morpho-functional reconstruction and structural mechanisms of adaptation of the blood and lymphatic links of the microcirculatory bed of extremities during early postoperation period up to 10 days. The pathophysiological state of the extremity sufficiently close to its autotransplantation was obtained by means of circular transection of soft tissues of the medial third of the femur together with the nerves and deep collecting lymphatic vessels. It was found that after modeling the main stages of replantation in the fascia and periosteum of the operated extremity there developed a spasm of the arteriolar link and dilatation of the venular and lymphatic links of the microcirculatory bed. The areas of leukocytic infiltration with the phenomena of diapedesis and microhemorrhages were revealed along the course of postcapillaries and venules in the paravasal connective tissue. The amount of functioning arteriole-venular anastomoses was increased. Against the background of pronounced oedema of soft tissues of the operated extremity the venous pressure increased and the rate of the capillary bloodflow in the skin and muscles decreased. The above changes tend to be reduced by the 10th day after modelling the main stages of replantation of the extremity.     

Contribution of Subcutaneous Connective Tissues to the Epithelialization and Cyst Formation by the Skin Transplanted Subcutaneously

Skins and hollow organs have been shown to form epithelialized cysts when transplanted into subcutaneous tissue of a recipient animal, expanding their surface areas. This system seems to offer a good potential for regenerating organs. We investigated the functional and structural contribution of epithelia and connective tissue compartments in this regeneration system with two experimental systems.Key Words: skin, epithelialization, transplantation, GFP, epidermis, cystDispase-separated epidermis often forms epithelialized cysts when combined with dermal connective tissue whereas dispase-separated epidermis alone does not form cysts or epithelialize, indicating the functional importance of the dermal connective tissue in the regeneration process.When GFP rats were used as donors for the skin, the donor-derived tissue was composed of whole epidermis and parts of the connective tissue cells and blood vessels under the newly epithelialized portion of the cyst wall. Small capillaries of granulation tissues were shown to be of recipient origin, but some large vessels were of donor origin. These results showed the significant functional and structural contribution of dermal connective tissue in the regeneration of the skin in subdermal transplant.     

Contribution of Subcutaneous Connective Tissues to the Epithelialization and Cyst Formation by the Skin Transplanted Subcutaneously

Skins and hollow organs have been shown to form epithelialized cysts when transplanted into subcutaneous tissue of a recipient animal, expanding their surface areas. This system seems to offer a good potential for regenerating organs. We investigated the functional and structural contribution of epithelia and connective tissue compartments in this regeneration system with two experimental systems.

Dispase-separated epidermis often forms epithelialized cysts when combined with dermal connective tissue whereas dispase-separated epidermis alone does not form cysts or epithelialize, indicating the functional importance of the dermal connective tissue in the regeneration process.

When GFP rats were used as donors for the skin, the donor-derived tissue was composed of whole epidermis and parts of the connective tissue cells and blood vessels under the newly epithelialized portion of the cyst wall. Small capillaries of granulation tissues were shown to be of recipient origin, but some large vessels were of donor origin. These results showed the significant functional and structural contribution of dermal connective tissue in the regeneration of the skin in subdermal transplant.     

Morphofunctional changes in the hemo- and lymphomicrocirculatory bed of the uterus and its regional lymph nodes in venous congestion during pregnancy

Interrelations of the blood and lymphatic systems of the uterus have been examined in rats at pregnancy complicated with the ++phlebo-occlusive syndrome. Blood stream impediment in the caudal vena cava results in increasing diameter of the arterioles, capillaries and venules of the endo- and myometrium. Certain disturbances of blood circulation in the uterus at the ++phlebo-cclusive syndrome in the pregnancy animals, as a rule, reflect in the uterine lymph outflow; this is demonstrated as dilatation and deformity of the lymphatic vessels and capillaries, appearance of protrusions of the lymphatic vessels wall. There is a definite co-ordination in the reaction to the caudal vena cava occlusion in the intra-, ++extra-organic uterine lymphatic bed and in its regional lymph nodes, manifested as the following morphological signs: decreasing relative volume, that the lymphoid tissue occupies and increasing relative volume of the cortical and medullary intermediate sinuses.     

An anatomic study of the septocutaneous vessels of the leg

The vascular anatomy of the skin and fascia of the leg were studied in 20 cadaver legs that were injected and dissected under magnification to identify the origin, course, and distribution of vessels from the subfascial level to the skin. In addition to the longitudinally oriented fasciocutaneous arteries and the musculocutaneous perforators, the study demonstrated a third and important system of blood supply: the septocutaneous vessels. These vessels arise directly from the posterior tibial, anterior tibial, and peroneal arteries, run along the intermuscular septum, pierce the crural fascia, and ramify radially in the subcutaneous tissue superficial to the fascia. Longitudinally oriented anastomotic arcades are formed along the leg between branches of adjacent septocutaneous vessels. Each septocutaneous vessel has one or two venae comitantes. Selected methylene blue injections of the septocutaneous vessels revealed rich staining of the superficial surface of the fascia, the subcutaneous tissue, and distinct longitudinally oriented skin territories. There was no injection of dye in the deep surface of the fascia. It is felt that the septocutaneous vessels constitute an important source of skin circulation in the leg and form the basis for various fasciocutaneous flaps that have useful clinical applications.     

Electrical Impedance of Acupuncture Meridians: The Relevance of Subcutaneous Collagenous Bands

Background

The scientific basis for acupuncture meridians is unknown. Past studies have suggested that acupuncture meridians are physiologically characterized by low electrical impedance and anatomically associated with connective tissue planes. We are interested in seeing whether acupuncture meridians are associated with lower electrical impedance and whether ultrasound-derived measures – specifically echogenic collagenous bands - can account for these impedance differences.

Methods/Results

In 28 healthy subjects, we assessed electrical impedance of skin and underlying subcutaneous connective tissue using a four needle-electrode approach. The impedances were obtained at 10 kHz and 100 kHz frequencies and at three body sites - upper arm (Large Intestine meridian), thigh (Liver), and lower leg (Bladder). Meridian locations were determined by acupuncturists. Ultrasound images were obtained to characterize the anatomical features at each measured site. We found significantly reduced electrical impedance at the Large Intestine meridian compared to adjacent control for both frequencies. No significant decrease in impedance was found at the Liver or Bladder meridian. Greater subcutaneous echogenic densities were significantly associated with reduced impedances in both within-site (meridian vs. adjacent control) and between-site (arm vs. thigh vs. lower leg) analyses. This relationship remained significant in multivariable analyses which also accounted for gender, needle penetration depth, subcutaneous layer thickness, and other ultrasound-derived measures.

Conclusion/Significance

Collagenous bands, represented by increased ultrasound echogenicity, are significantly associated with lower electrical impedance and may account for reduced impedances previously reported at acupuncture meridians. This finding may provide important insights into the nature of acupuncture meridians and the relevance of collagen in bioelectrical measurements.     

The influence of differing connective tissue substrates on the maintenance of adult stratified squamous epithelia

Summary The interaction between adult stratified squamous epithelium and its supporting connective tissue possibly involves both permissive and directive influences. To examine the effect of vitality and specificity of connective tissue on the maintenance of epithelial structure and histo-differentiation, specimens of skin and oral mucosa from various regions of adult mice were separated using either EDTA or trypsin. Prior to transplantation, the epithelium was recombined with either inverted homologous connective tissue or with connective tissue that had been killed either by heating or repeated freeze-thawing. Epithelial sheets were also transplanted onto the graft bed alone or in combination with striated muscle or tendon.Normal patterns of cytodifferentiation were maintained when the epithelium was recombined with inverted or frozen-thawed subepithelial connective tissue but there was a loss of spatial organization on the frozen-thawed connective tissue. In contrast, heat-killed or trypsin-treated frozen-thawed subepithelial connective tissue and non-dermal connective tissue failed to maintain a viable epithelium. These observations suggest that subepithelial connective tissues (dermis, lamina propria) but not deep connective tissues facilitate epithelial proliferation and histodifferentiation.Supported by NIH/NIDR RO1 DEO5190     

Trabecular barriers (FRENULAE) of the extracapsular clefts (38563).

Ferrocyanide was perfused through the vascular system of the isolated legs of 10 dogs. Ferric ion was then injected intramuscularly (not intravascularly) into the leg. The injected materials spread along the trabeculae of the tissues, rupturing the sheets of fibrous connective tissue (frenulae) inpeded the movement of materials in the trabeculae. The barriers consist of sheets of connective tissue which insert on adjacent fibrous capsules. The capsules enclose each cluster of muscle, glandular or other cells. These barriers appear to function in conjunction with the lymphatic drainage system.     

Scanning electron microscope study of the liver microcirculation

During the experimental investigation performed in dogs and rats, by means of scanning electron microscopy of corrosive anatomical preparations, the spatial organization of all parts of the hepatic vascular bed (arterial, venous and lymphatic) has been studied, specific features of their components construction have been described. Within the limits of one hepatic lobule the number of vessels included in the portal vein system exceeds that of the arterial ones, originating from the proper hepatic artery system. In every part of the vascular bed the gradient of the form, orientation and pronouncement of the nuclei-containing zones in endotheliocytes and myocytes has been established. Various appliances participating in the blood and lymph stream regulation in different parts of the vascular bed have been revealed. As initial elements of the lymph bed, closed digital or loop-like capillaries should be regarded, they localize in the organ's connective tissue framework. Around the portal and hepatic veins and their branches, as well as around the biliary ducts, well developed plexuses of the lymphatic and blood capillaries and vessels localize, they are the main draining pathways of the organ. The degree of development and pronouncement of these plexuses depends on the lumen size in the formation they accompany.     

The subcutaneous vascular system (lower extremity): studies on micro-preparations]

At the level of fascial penetration the cutaneous arteries of the lower extremity are constantly accompanied by 1 or 2 communicating veins of varying diameter. Usually, they penetrate the fascia in rows via the muscular interstitium. There they give up branches to the fascial vascular network, to pre- and subfascial areolar tissue and possibly to the muscle origins on the fascia, or to the intermuscular septa. The texture of the fascia itself determines the structure of fascial openings for the cutaneous vessels. In the subcutaneous tissue the arteries are accompanied by 1 or 2 veins (Vv. comitantes). Numerous arterio-arterial and intervenous anastomoses form a subcutaneous network of vascular bundles. Two anastomosing venous systems can be distinguished in the subcutis: The small Vv. comitantes are fed primarily by the subcutaneous adipose lobes, and end in communicating veins or flow into the subcutaneous veins (epifascial veins). These large subcutaneous veins on the other hand are responsible for the actual outflow from the venous network of the cutis. They form the saphenous system or empty into larger communicating veins. Between the subcutaneous arterioles and accompanying venules there are numerous capillary webs. In addition direct capillaries and looped as well as meandering or knotted arterio-venous shunts are found. The subcutaneous vascular bundles are fixed by a connective tissue. There are often typical capillary meshes within arterial sheat, oriented like a rope-ladder. They undergo prenatal development. The subcutaneous (epifascial) veins are surrounded by areolar tissue on the cutaneous as well as on the fascial side. Fibers from the accompanying connective tissue criss-cross into the adventitia and thereby anchor the veins in a movable fashion. Typical vascular patches with plane capillary networks characterize the areolar tissue around the subcutaneous veins, which is differentiating at the 2nd half of the fetal period. Within are also capillary loops and convoluted arterio-venous shunts.     

Tissue basophils in thermal damage to the skin in an experiment (morphometric analysis and statistical modelling)

By means of quantitative morphological analysis tissue basophils (TB) (mast cells) in the derm and in the subcutaneous loose connective tissue have been studied at various degree of thermal lesion of the skin. Average diameter, amount of the cells per 0.1 mm2 of the slice area and distance between them serve as criteria for estimation of the TB functional activity. Certain differences have been revealed in the TB state at a weak and severe thermal lesion of the skin. At a threshold thermal action, metabolic processes in the TB become active. Burns of the skin of the II and IIIa degree result in an increased discharge of the granular component from cytoplasm and often in death of the cells. By means of multiple linear regression equations certain dependence of the degree of the thermal skin lesion on the state of the TB population in various dermal layers and in the subcutaneous loose connective tissue is stated.     

Microscopic and ultrastructural study of the lymphatic system in the human parotid gland

The localization and fine structure of the lymphatic system vessels are examined in the human parotid gland. A network of lymphatic capillaries extends in the intralobular connective septa around the striated ducts. These lymphatics converge in collectors frequently bordering the excretory ducts. On the contrary, no lymphatics are present next to the intercalated ducts and adhenomers. Ultrastructurally, the lymphatic capillaries are characterized by a very thin endothelial wall and by slightly complicated intercellular adhesions. Open junctions are also present. The presence of numerous lymphatic capillaries bordering the striated ducts and their blood microvasculature is discussed in relation to the functional activities of the striated ducts in the modification of the saliva.     

Structure of the excretory ducts, intraorganic lymphatic vessels, tissue canals and intercellular space of various organs (data of scanning electron microscopy injection replicas)

Architectonics of the biliary canaliculi and intrahepatic ducts systems, as well as intraorganic urinary pathways in white rats have been investigated by means of scanning electron microscopy of injection replica. Acinar structure of the intralobular part of the biliary bed has been proved. Anastomoses between the biliary canaliculi of the neighbouring lobules have been demonstrated. A useful method for obtaining injection replica of the intraorganic lymphatic vessels is filling of the ductal system of the parenchymatous organs with solid resins (methylmethacrylate+) under a high (nonphysiological) pressure. Casts of periportal and paravenous hepatic lymphatic vessels have been obtained. An ability of methylmethacrylate to replicate intercellular and connective tissue spaces is verified. Casts of the perisinusoid spaces (Disse) are obtained for the first time.     

The reversed fasciosubcutaneous flap in the leg

A reversed fasciosubcutaneous tissue flap in the leg is described. This distally based flap is vascularized by the perforating cutaneous branches of the peroneal and tibialis posterior arteries. It must carry all its subcutaneous tissue. A study on the vascularization of the subcutaneous tissue reveals the predominance of the vascular network in this layer with regard to the dermal or fascial plane. The dermal vascular network at the donor site is sufficient to let the skin survive without its underlying subcutaneous vascular support. The flap can reach the malleolar and heel region. The advantages of this technique are (1) easy dissection, (2) preservation of the major vascular pedicles of the lower limb, (3) skin preservation at the donor site, thus preserving the shape of the limb, and (4) versatility (it is supple and can adapt to every surface, and it can be grafted on the deep or the superficial side). The addition of this technique to the armamentarium of the reconstructive surgeon has proved to be very useful in repairing soft-tissue defects in the lower limb. Often it can replace the classical fasciocutaneous flap or even a free flap.     

Light and electron microscope observations of the lymphatic drainage units of the peritoneal cavity of rodents

Fluid, particles, and cells are taken up from the peritoneal cavity by lymphatic drainage units, which, in the mouse and rat, are located along the peritoneal surface of the muscular portion of the diaphragm. The drainage units are composed of three specifically differentiated components: a lymphatic lacuna, a covering of lacunar mesothelium, and intervening submesothelial connective tissue. The units are drained by connecting lymphatic vessels that cross the diaphragm to empty into collecting lymphatic vessels running along the pleural surface of the diaphragm. The collecting lymphatics empty into parasternal lymphatic trunks. In this report, we briefly review critical features of the drainage apparatus and describe new observations, summarized below, about their structure. Around the rim of stomata, the mesothelial openings that lead into the lymphatic lacunae, plasma membranes of lacunar mesothelial cells and of lacunar endothelial cells abut but are not linked to one another by recognizable junctional specializations. Lacunar endothelial cells often extend valve-like processes that bridge the distal end of the channel beneath the stoma. The configuration of the endothelial processes may be complex. Occasionally, processes from fibroblasts in the submesothelial connective tissue adjacent to stomata make contact with the interstitial surface of lacunar endothelial cells. A discontinuous elastic layer in the submesothelial connective tissue spans the roof of each lacuna. Connecting and collecting lymphatics, which drain lymphatic lacunae, possess endothelial valves. Possible functions for each of these newly described structural features are discussed.     

Development of the Mouse Dermal Adipose Layer Occurs Independently of Subcutaneous Adipose Tissue and Is Marked by Restricted Early Expression of FABP4

The laboratory mouse is a key animal model for studies of adipose biology, metabolism and disease, yet the developmental changes that occur in tissues and cells that become the adipose layer in mouse skin have received little attention. Moreover, the terminology around this adipose body is often confusing, as frequently no distinction is made between adipose tissue within the skin, and so called subcutaneous fat. Here adipocyte development in mouse dorsal skin was investigated from before birth to the end of the first hair follicle growth cycle. Using Oil Red O staining, immunohistochemistry, quantitative RT-PCR and TUNEL staining we confirmed previous observations of a close spatio-temporal link between hair follicle development and the process of adipogenesis. However, unlike previous studies, we observed that the skin adipose layer was created from cells within the lower dermis. By day 16 of embryonic development (e16) the lower dermis was demarcated from the upper dermal layer, and commitment to adipogenesis in the lower dermis was signalled by expression of FABP4, a marker of adipocyte differentiation. In mature mice the skin adipose layer is separated from underlying subcutaneous adipose tissue by the panniculus carnosus. We observed that the skin adipose tissue did not combine or intermix with subcutaneous adipose tissue at any developmental time point. By transplanting skin isolated from e14.5 mice (prior to the start of adipogenesis), under the kidney capsule of adult mice, we showed that skin adipose tissue develops independently and without influence from subcutaneous depots. This study has reinforced the developmental link between hair follicles and skin adipocyte biology. We argue that because skin adipocytes develop from cells within the dermis and independently from subcutaneous adipose tissue, that it is accurately termed dermal adipose tissue and that, in laboratory mice at least, it represents a separate adipose depot.     

Lymphatic endothelial progenitor cells contribute to de novo lymphangiogenesis in human renal transplants

De novo lymphangiogenesis influences the course of different human diseases as diverse as chronic renal transplant rejection and tumor metastasis. The cellular mechanisms of lymphangiogenesis in human diseases are currently unknown, and could involve division of local preexisting endothelial cells or incorporation of circulating progenitors. We analyzed renal tissues of individuals with gender-mismatched transplants who had transplant rejection and high rates of overall lymphatic endothelial proliferation as well as massive chronic inflammation. Donor-derived cells were detected by in situ hybridization of the Y chromosome. We compared these tissues with biopsies of essentially normal skin and intestine, and two rare carcinomas with low rates of lymphatic endothelial proliferation that were derived from individuals with gender-mismatched bone marrow transplants. Here, we provide evidence for the participation of recipient-derived lymphatic progenitor cells in renal transplants. In contrast, lymphatic vessels of normal tissues and those around post-transplant carcinomas did not incorporate donor-derived progenitors. This indicates a stepwise mechanism of inflammation-associated de novo lymphangiogenesis, implying that potential lymphatic progenitor cells derive from the circulation, transmigrate through the connective tissue stroma, presumably in the form of macrophages, and finally incorporate into the growing lymphatic vessel.     

Use of a calf prosthesis and tissue expansion in aesthetic reconstruction of the leg.

We present a technique for reconstruction of the legs in patients with soft-tissue loss and formation of large scars with retraction of this tissue in the pretibial region. In such patients, a subcutaneous tissue expander is placed in the region adjacent to the scar tissue. With expansion, we obtained sufficient skin for use in the reconstruction, and the resulting asymmetry in leg diameter was compensated for by means of one or two calf prostheses, depending on the patient.