The human lung is born with a fraction of the adult complement of alveoli. The postnatal stages of human lung development comprise an alveolar stage, a stage of microvascular maturation, and very likely a stage of late alveolarization. The characteristic structural features of the alveolar stage are well known; they are very alike in human and rat lungs. The bases for alveolar formation are represented by immature interairspace walls with two capillary layers with a central sheet of connective tissue. Interalveolar septa are formed by folding up of one of the two capillary layers. In the alveolar stage, alveolar formation occurs rapidly and is typically very conspicuous in both species; it has therefore been termed ‘bulk alveolarization’. During and after alveolarization the septa with double capillary networks are restructured to the mature form with a single network. This happens in the stage of microvascular maturation. After these steps the lung proceeds to a phase of growth during which capillary growth by intussusception plays an important role in supporting gas exchange. In view of reports that alveoli are added after the stage of microvascular maturation, the question arises whether the present concept of alveolar formation needs revision. On the basis of morphological and experimental findings we can state that mature lungs contain all the features needed for ‘late alveolarization’ by the classical septation process. Because of the high plasticity of the lung tissues, late alveolarization or some forms of compensatory alveolar formation may be considered for the human lung.

In order to investigate the postnatal growth of the gas exchange apparatus, the lungs of rats aged 1, 4, 7, 10, 13, and 21 days were fixed by intratracheal instillation of glutaraldehyde. The analysis and interpretation of the morphological changes observed by light and electron microscopy were based on the results of previous morphometric and autoradiographic studies performed on the same material.The newborn rat has no alveoli, but breathes with smooth walled air channels and saccules, which correspond to the prospective alveolar ducts and alveolar sacs, respectively. The bulk of alveoli are formed between days 4 and 13 by a rapid outgrowth of secondary septa from the primary septa present at birth. The arrangement of elastic fibers during this period suggests that these may play a role in septa1 outgrowth. Based on ultrastructural observations a model is described for the capillarisation of the secondary septa. Some evidence is given that alveoli may also be formed by outpouchings in the walls of terminal bronchioles.Primary and secondary septa have initially an immature appearance. They both show an apparently double capillary network, whereas the mature interalveolar septum is just wide enough to accommodate a single capillary. Possible mechanisms for this structural transformation which occurs within three weeks after birth are discussed.

The postnatal growth of the lung was quantitatively investigated in rats aged 1 , 4 , 7, 10, 13, 21,44 and 131 days by light and electron microscopic morphometry .Lung volume (V,) increased first directly with body weight (W). After day 10 V, followed the function WO.?O. Based on the quantitative findings the postnatal lung growth could be divided into three phases.

This review shall familiarize the reader with the various aspects of intussusceptive angiogenesis (IA). The basic event in IA is the formation of transvascular tissue pillars. Depending on location, timing, and frequency of pillar emergence, the IA process has different outcomes. In capillaries, a primary IA function is to expand the capillary bed in size and complexity (intussusceptive microvascular growth). It represents an alternative to capillary sprouting. Highly ordered pillar formation in a developing capillary network leads to the formation of vascular trees (intussusceptive arborization). In small arteries and veins, pillar formation at the vessels' branching angles leads either to remodeling of the branching geometry or even to vascular pruning (intussusceptive branching remodeling). It appears essential that future angiogenic research considers always both phenomena, sprouting and intussusception. Vascularization of tissues, organs, and tumors rely heavily on both mechanisms; neglecting one or the other would obscure our understanding of the angiogenesis process. Developmental Dynamics 231:474 -488, 2004.

Intussusception (growth within itself) is an alternative to the sprouting mode of angiogenesis. The protrusion of opposing microvascular walls into the capillary lumen creates a contact zone between endothelial cells. The endothelial bilayer is perforated, intercellular contacts are reorganized, and a transluminal pillar with an interstitial core is formed, which is soon invaded by myofibroblasts and pericytes leading to its rapid enlargement by the deposition of collagen fibrils. Intussusception has been implicated in three processes of vascular growth and remodeling. (1) Intussusceptive microvascular growth permits rapid expansion of the capillary plexus, furnishing a large endothelial surface for metabolic exchange. (2) Intussusceptive arborization causes changes in the size, position, and form of preferentially perfused capillary segments, creating a hierarchical tree. (3) Intussusceptive branching remodeling (IBR) leads to modification of the branching geometry of supplying vessels, optimizing pre- and postcapillary flow properties. IBR can also lead to the removal of branches by pruning in response to changes in metabolic needs. None of the three modes requires the immediate proliferation of endothelial cells but rather the rearrangement and plastic remodeling of existing ones. Intussusception appears to be triggered immediately after the formation of the primitive capillary plexus by vasculogenesis or sprouting. The advantage of this mechanism of growth over sprouting is that blood vessels are generated more rapidly in an energetically and metabolically more economic manner, as extensive cell proliferation, basement membrane degradation, and invasion of the surrounding tissue are not required; the capillaries thereby formed are less leaky. This process occurs without disrupting organ function. Improvements in our understanding of the process should enable the development of novel pro- and anti-angiogenic therapeutic treatments.

The theory of bifurcating vascular systems predicts vessel diameters that are related to optimality criteria like minimization of pumping energy or of building material. However, mechanisms for producing the postulated optimality have not been described so far, and quantitative data on bifurcation diameters during development are scarce. We used an embryonic vascular bed that rapidly grows and adapts to changing hemodynamic conditions, the chicken chorioallantoic membrane (CAM), and correlated vascular cast and tissue section morphology with in vivo time-lapse video monitoring. The bifurcation exponent ⌬ and associated parameters were quantitatively assessed in arterial and venous microvessels ranging in diameter from 30 to 100 m. We observed emergence of optimality by means of intussusception, i.e., formation of transvascular tissue pillars. In addition to intussusceptive microvascular growth (IMG ‫؍‬ expansion of capillary networks) and intussusceptive arborization (IAR ‫؍‬ formation of feeding vessels from capillaries) the observed intussusception at bifurcations represents a third variant of nonsprouting angiogenesis. We call it intussusceptive branching remodeling (IBR). IBR occurred in vessels of considerable diameter by means of two alternative mechanisms: either through pillars arising close to a bifurcation, which increased in girth until they merged with the connective tissue in the bifurcation angle; or through pillars arising at some distance from the bifurcation point, which then expanded by formation of ingrowing tissue folds until they became connected to the tissue of the bifurcation angle. Morphologic evidence suggests that IBR is a wide-spread phenomenon, taking place also in lung, intestinal, kidney, eye, etc., vasculature. Irrespective of the mode followed, IBR led to a branching pattern close to the predicted optimum, ⌬ ‫؍‬ 3.0. Significant differences were observed between ⌬ at arterial bifurcations (2.70 to 2.90) and ⌬ at venous bifurcations (2.93 to 3.75). IBR, by means of eccentric pillar formation and fusion, was also involved in vascular pruning. Experimental changes in CAM hemodynamics (by locally increasing blood flow) induced onset of IBR within less than 1 hr. Our study provides morphologic and quantitative evidence that a similar cellular machinery is used for all three variants of vascular intussusception, IMG, IAR, and IBR. It thus provides a mechanism of efficiently generating complex blood transport systems from limited genetic information. Differential quantitative outcome of IBR in arteries and veins, and the experimental induction of IBR strongly suggest that hemodynamic factors can instruct embryonic vascular remodeling toward optimality.

Abstract-Although vascular endothelial growth factor (VEGF) has been described as a potent angiogenic stimulus, its application in therapy remains difficult: blood vessels formed by exposure to VEGF tend to be malformed and leaky. In nature, the principal form of VEGF possesses a binding site for ECM components that maintain it in the immobilized state until released by local cellular enzymatic activity. In this study, we present an engineered variant form of VEGF, ␣ 2 PI 1-8 -VEGF 121 , that mimics this concept of matrix-binding and cell-mediated release by local cell-associated enzymatic activity, working in the surgically-relevant biological matrix fibrin. We show that matrix-conjugated ␣ 2 PI 1-8 -VEGF 121 is protected from clearance, contrary to native VEGF 121 mixed into fibrin, which was completely released as a passive diffusive burst. Grafting studies on the embryonic chicken chorioallantoic membrane (CAM) and in adult mice were performed to assess and compare the quantity and quality of neovasculature induced in response to fibrin implants formulated with matrix-bound

The life of a human lung can be subdivided into five distinct phases: embryonic, pseudoglandular, canalicular, saccular, and alveolar. The embryonic period, during which the lung primordium is laid down as a diverticulum of the foregut, lasts for about seven weeks. From the 5th to the 17th week the lung looks much like a tubulo-acinar gland, with epithelial tubes sprouting and branching into the surrounding mesenchyme. In the last week of this pseudoglandular stage the prospective conductive airways have been formed, and the acinar limits can be recognized. The events of the subsequent canalicular phase (17th-26th week) can be summarized as the widening of the peripheral tubules, the differentiation of the cuboidal epithelium into type I and type II cells, the formation of the first thin air-blood barriers, and the start of surfactant production. During the saccular stage, which follows and lasts until birth, the growth of the pulmonary parenchyma, the thinning of the connective tissue between the airspaces, and the further maturation of the surfactant system are the most important steps towards life. At birth, although already functional, the lung is structurally still in an immature condition, because alveoli, the gas exchange units of the adult lung, are practically missing. The airspaces present are smooth-walled transitory ducts and saccules with primitive type septa that are thick and contain a double capillary network. During the first 1-3 years of postnatal life, alveoli are formed through a septation process that greatly increases the gas exchange surface area. The primitive septa with their capillaries undergo a complete remodeling, gaining the mature slender morphology found in the adult lung.