Quantitative analysis of the thickness of human skin and subcutaneous tissue following controlled expansion with a silicone implant

Histologic quantitation of the thickness of human tissues that were expanded using silicone expanders showed that the epidermis underwent significant thickening after 5 weeks to 5 months of expansion. The dermis and subcutaneous tissue, on the other hand, were significantly thinner after expansion. Capsules were formed in all 19 patients. The capsule was significantly thickest after 2 to 2.5 months of expansion. Expanded tissues 2 years after cessation of expansion had the same thickness as control tissues and had no remnant fibrous capsule.     

Spatial distributions of expansion rate, cell division rate and cell size in maize leaves: a synthesis of the effects of soil water status, evaporative demand and temperature

The spatial distributions of leaf expansion rate, cell division rate and cell size was examined under contrasting soil water conditions, evaporative demands and temperatures in a series of experiments carried out in either constant or naturally fluctuating conditions. They were examined in the epidermis and all leaf tissues. (1) Meristem temperature affected relative elongation rate by a constant ratio at all positions in the leaf. If expressed per unit thermal time, the distribution of relative expansion rate was independent of temperature and was similar in all experiments with low evaporative demand and no water deficit. This provides a reference distribution, characteristic of the studied genotype, to which any distribution in stressed plants can be compared. (2) Evaporative demand and soil water deficit affected independently the distribution of relative elongation rate and had near-additive effects. For a given stress, a nearly constant difference was observed, at all positions of the leaf, between the relative elongation rates of stressed plants and those of control plants. This caused a reduction in the length of the zone with tissue elongation. (3) Methods for calculating cell division rate in the epidermis and in all leaf tissues are proposed and discussed. In control plants, the zone with cell division was 30 mm and 60 mm long in the epidermis and in whole tissues, respectively. Both this length and relative division rate were reduced by soil water deficit. The size of epidermal and of mesophyll cells was nearly unaffected in the leaf zone with both cell division and tissue expansion, suggesting that water deficit affects tissue expansion rate and cell division rate to the same extent. Conversely, cell size of epidermis and mesophyll were reduced by water deficit in mature parts of the leaf.     

Keratinocytes Propagated in Serum-Free,Feeder-Free Culture Conditions Fail to Form Stratified Epidermis in a Reconstituted Skin Model

Primary human epidermal stem cells isolated from skin tissues and subsequently expanded in tissue culture are used for human therapeutic use to reconstitute skin on patients and to generate artificial skin in culture for academic and commercial research. Classically, epidermal cells, known as keratinocytes, required fibroblast feeder support and serum-containing media for serial propagation. In alignment with global efforts to remove potential animal contaminants, many serum-free, feeder-free culture methods have been developed that support derivation and growth of these cells in 2-dimensional culture. Here we show that keratinocytes grown continually in serum-free and feeder-free conditions were unable to form into a stratified, mature epidermis in a skin equivalent model. This is not due to loss of cell potential as keratinocytes propagated in serum-free, feeder-free conditions retain their ability to form stratified epidermis when re-introduced to classic serum-containing media. Extracellular calcium supplementation failed to improve epidermis development. In contrast, the addition of serum to commercial, growth media developed for serum-free expansion of keratinocytes facilitated 3-dimensional stratification in our skin equivalent model. Moreover, the addition of heat-inactivated serum improved the epidermis structure and thickness, suggesting that serum contains factors that both aid and inhibit stratification.     

Growth coordination and the shoot epidermis

Cell-cell communication is essential for growth and development of multicellular organisms. In higher plants, the shoot organs are derived from three clonally distinct cell layers present in the meristem. The role of the outermost L1 cell layer and its derived epidermis in coordinating growth of the inner-cell layers has long been debated. This question has been revisited recently using molecular tools to manipulate cell cycle progression or cell expansion, specifically in the epidermis. These studies conclude that cells in the epidermis both promote and restrict growth of the entire shoot by sending growth signals - either physical or chemical - to the inner layers.     

Specialized extraembryonic cells connect embryonic and extraembryonic epidermis in response to Dpp during dorsal closure in Drosophila

Dorsal closure in Drosophila embryogenesis involves expansion of the dorsal epidermis, followed by closure of the opposite epidermal edges. This process is driven by contractile force generated by an extraembryonic epithelium covering the yolk syncytium known as the amnioserosa. The secreted signaling molecule Dpp is expressed in the leading edge of the dorsal epidermis and is essential for dorsal closure. We found that the outermost row of amnioserosa cells (termed pAS) maintains a tight basolateral cell-cell adhesion interface with the leading edge of dorsal epidermis throughout the dorsal closure process. pAS was subject to altered cell motility in response to Dpp emanating from the dorsal epidermis, and this response was essential for dorsal closure. alphaPS3 and betaPS integrin subunits accumulated in the interface between pAS and dorsal epidermis, and were both required for dorsal closure. Looking at alphaPS3, type I Dpp receptor, and JNK mutants, we found that pAS cell motility was altered and pAS and dorsal epidermis adhesion failed under the mechanical stress of dorsal closure, suggesting that a Dpp-mediated mechanism connects the squamous pAS to the columnar dorsal epidermis to form a single coherent epithelial layer.     

Cells of the Upper and Lower Epidermis of Barley (Hordeum vulgare L.) Leaves Exhibit Distinct Patterns of Vacuolar Solutes

Vacuolar saps were extracted from individual, anatomically uniform cells of the upper (adaxial) and lower (abaxial) epidermis of the third leaf of barley (Hordeum vulgare L.) using a modified pressure probe. Saps (volume 80-200 pL) were sampled at various times between 3 d before and 7 d after full-leaf expansion and were analyzed for their osmolality and their concentrations of NO3-, malate, CI-, K+, and Ca2+. The osmolalities of upper and lower epidermis both increased with time but were similar to each other. In young leaves, K+ and Ca2+ were evenly distributed between the two epidermal layers, but as the leaf aged, the upper epidermis accumulated high (40-100 mM) Ca2+, whereas cells of the lower epidermis accumulated K+ instead. Nitrate concentration was 100 to 150 mM higher in the upper than in the lower epidermis, whereas CI- was 50 to 120 mM higher in the lower epidermis. These differences did not depend on the leaf developmental stage. The uneven distribution of epidermal NO3- and CI- was maintainedover a wide range of epidermal sap concentrations of these ions and was not affected by NO3- or CI- starvation or by an increase in the light intensity from 120 to 400 [mu]mol m-2 s-1. However, the latter did cause a decrease in epidermal NO3- and the appearance and accumulation of epidermal malate, particularly in the upper epidermis. The physiological implications of the results for solute storage in leaves and for the pathways of ion distribution to the epidermis are discussed.     

Morphometric evaluation of collagen fibril dimensions in expanded human breast skin

A great innovation in plastic surgery in recent years has been skin expansion, which has provided the discipline with new possibilities for skin reconstruction. At present, little is known about the biology of skin expansion although it is clear that cell proliferation occurs both in the epidermis and the dermis. During previous morphological investigations of skin under expansion we recorded a number of signs comparable with those seen in wound healing. In the present study, the collagen fibril diameter of of skin before and under expansion has been recorded using an IBAS computer-based morphometric system. Preferentially, we have studied the papillary dermis where the most conspicuous morphological events occur.     

Glutamate transporter Slc1a3 mediates inter‐niche stem cell activation during skin growth

Tissues contain distinct stem cell niches, but whether cell turnover is coordinated between niches during growth is unknown. Here, we report that in mouse skin, hair growth is accompanied by sebaceous gland and interfollicular epidermis expansion. During hair growth, cells in the bulge and outer root sheath temporarily upregulate the glutamate transporter SLC1A3, and the number of SLC1A3⁺ basal cells in interfollicular epidermis and sebaceous gland increases. Fate mapping of SLC1A3⁺ cells in mice revealed transient expression in proliferating stem/progenitor cells in all three niches. Deletion of slc1a3 delays hair follicle anagen entry, uncouples interfollicular epidermis and sebaceous gland expansion from the hair cycle, and leads to reduced fur density in aged mice, indicating a role of SLC1A3 in stem/progenitor cell activation. Modulation of metabotropic glutamate receptor 5 activity mimics the effects of SLC1A3 deletion or inhibition. These data reveal that stem/progenitor cell activation is synchronized over distinct niches during growth and identify SLC1A3 as a general marker and effector of activated epithelial stem/progenitor cells throughout the skin.     

Cell expansion in the epidermis: microtubules, cellulose orientation and wall loosening enzymes

Two models of isolated epidermis were used to demonstrate that the net orientation of cellulose microfibrils in the cell wall is related to mechanical properties of the tissue, and can be used as an indicator for wall anisotropy. In the developing plant epidermis, cells expand in one or two directions in the plane of the plant surface. In epidermis cells actively expanding in one direction (elongation), the orientation of cortical microtubules closely matches the net cellulose orientation. In epidermis cells expanding in two directions, the orientation of the parallel microtubules does not coincide with the net cellulose orientation in the adjacent cell wall. The orientation of cortical microtubules is thus not always a reliable indicator of wall characteristics. In both types of epidermis, a high rate of expansion correlates with a high activity of xyloglucan endotransglycosylase (XET), as determinedin situ. This high activity alone cannot explain unidirectional wall expansion.     

Transient proliferation of proanthocyanidin-accumulating cells on the epidermal apex contributes to highly aluminum-resistant root elongation in camphor tree

Aluminum (Al) is a harmful element that rapidly inhibits the elongation of plant roots in acidic soils. The release of organic anions explains Al resistance in annual crops, but the mechanisms that are responsible for superior Al resistance in some woody plants remain unclear. We examined cell properties at the surface layer of the root apex in the camphor tree (Cinnamomum camphora) to understand its high Al resistance mechanism. Exposure to 500 μm Al for 8 d, more than 20-fold higher concentration and longer duration than what soybean (Glycine max) can tolerate, only reduced root elongation in the camphor tree to 64% of the control despite the slight induction of citrate release. In addition, Al content in the root apices was maintained at low levels. Histochemical profiling revealed that proanthocyanidin (PA)-accumulating cells were present at the adjacent outer layer of epidermis cells at the root apex, having distinctive zones for cell division and the early phase of cell expansion. Then the PA cells were gradually detached off the root, leaving thin debris behind, and the root surface was replaced with the elongating epidermis cells at the 3- to 4-mm region behind the tip. Al did not affect the proliferation of PA cells or epidermis cells, except for the delay in the start of expansion and the accelerated detachment of the former. In soybean roots, the innermost lateral root cap cells were absent in both PA accumulation and active cell division and failed to protect the epidermal cell expansion at 25 μm Al. These results suggest that transient proliferation and detachment of PA cells may facilitate the expansion of epidermis cells away from Al during root elongation in camphor tree.     

Regulation of tomato fruit growth by epidermal cell wall enzymes

Water relations of tomato fruit and the epidermal and pericarp activities of the putative cell wall loosening and tightening enzymes Xyloglucan endotransglycosylase (XET) and peroxidase were investigated, to determine whether tomato fruit growth is principally regulated in the epidermis or pericarp. Analysis of the fruit water relations and observation of the pattern of expansion of tomato fruit slices in vitro , has shown that the pericarp exerts tissue pressure on the epidermis in tomato fruit, suggesting that the rate of growth of tomato fruit is determined by the physical properties of the epidermal cell walls. The epidermal activities of XET and peroxidase were assayed throughout fruit development. Temporal changes in these enzyme activities were found to correspond well with putative cell wall loosening and stiffening during fruit development. XET activity was found to be proportional to the relative expansion rate of the fruit until growth ceased, and a peroxidase activity weakly bound to the epidermal cell wall appeared shortly before cessation of fruit expansion. No equivalent peroxidase activity was detected in pericarp tissue of any age. It is therefore plausible that the expansion of tomato fruit is regulated by the combined action of these enzyme activities in the fruit epidermis.     

Control of Auxin-induced Stem Elongathn by the Epidermis

Segments of the 4th and 5th internodes of light-grown pea seedlings were used for the study of control of stem elongation. With 5th internodes, at low turgor as well as at water saturation auxin primarily appeared to cause a change in cell wall properties of the epidermis but it showed little effect on expansion af the inner tissue. This was confirmed by comparison of expansion between peeled and unpeeled segments, split tests and by measurements of stress-relaxation properties of the epidermal cell wall. Segments with the central part re-moved elongated well in response to auxin, but the isolated epidermis showed neither auxin-induced elongation nor cell wall loosening. A fungal β-1,3-glucanase appeared, at least partly, to have a similar effect as that of auxin on elongation, by changing cell wall properties of the epidermal cell wall. Peeled segments of 4th internodes expanded very little and auxin had little effect on their epidermal cell wall properties.     

Tissue expansion: dividend or loan?

Epidermal mitotic activity during tissue expansion has been assessed in the guinea pig using tritiated thymidine and other confirmatory techniques. Implant inflation results in a threefold elevation of epidermal mitotic activity within 24 hours, followed by a gradual return to normal baseline over 2 to 5 days. Implant deflation, conversely, causes a transient decrease in epidermal mitotic activity. Neither of these phenomena has been previously described. It confirms previous animal studies which illustrate the highly responsive nature of the epidermis to physical stimuli and supports the view that tissue expansion is a highly useful manipulation of normal physiologic processes.     

Interactions of Recombinant Mouse Erythrocyte Transglutaminase with Membrane Skeletal Proteins

In this study, we chose a differentiation-competent rat epidermal keratinocyte (REK) cell line to examine the role of Cx26 and disease-linked Cx26 mutants in organotypic epidermal differentiation. First, we generated stable REK cell lines expressing three skin disease-linked mutants (G59A, D66H and R75W). Second, we used an RNAi approach to knock down the expression of Cx26 in REKs. Interestingly, the three-dimensional (3D) architecture of the organotypic epidermis altered the intracellular spatial distribution of the mutants in comparison to 2D cultured REKs, highlighting the importance of using organotypic cultures. Unexpectedly, the presence of disease-linked mutants or the overexpression of wild-type Cx26 had little effect on the differentiation of the organotypic epidermis as determined by the architecture of the epidermis, expression of molecular markers indicative of epidermis differentiation (keratin 10, keratin 14, involucrin, loricrin) and stratification/cornification of the epidermis. Likewise, organotypic epidermis continued to differentiate normally upon Cx26 knockdown. While Cx26 has been reported to be upregulated during wound healing, no reduction in wound closure was observed in 2D REK cultures that expressed loss-of-function, dominant Cx26 mutants. In conclusion, we demonstrate that gain or loss of Cx26 function does not disrupt organotypic epidermal differentiation and offer insights into why patients harboring Cx26 mutations do not frequently present with more severe disease that encompasses thin skin. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

An accelerated approach to tissue expansion for breast reconstruction: experience with intraoperative and rapid postoperative expansion in 370 reconstructions

Breast reconstruction with tissue expansion is a well-established technique that offers satisfactory aesthetic results with minimal patient morbidity. The traditional period of expansion, however, continues to be a significant source of patient inconvenience and dissatisfaction. The objective of this study was to develop and evaluate a protocol for rapid tissue expansion. A total of 370 breast reconstructions in 314 patients who underwent rapid tissue expansion were retrospectively reviewed. Contraindications to rapid expansion were considered to be previous radiation, mastectomy skin flaps of questionable viability, and an excessively tight skin envelope. All expanders were placed submuscularly and filled to 40 to 50 percent of tissue expander volume. Office expansion was undertaken within 10 to 14 days after the operation and continued on a weekly basis. Each expansion was limited by patient tolerance up to a maximal pressure of 40 mm of water or a volume of 120 cm3. Expansion was considered complete once the expanded breast was 30 to 50 percent larger than the contralateral breast. If required, postoperative chemotherapy was given during the expansion period. Mean patient age was 48 years (range, 23 to 73 years). Two hundred fifty-eight patients had unilateral reconstructions. Three hundred two patients had immediate reconstruction. Mean tissue expander size was 583 cm3 (SD, 108 cm3). Mean intraoperative expansion was 271 cm3, or 46 percent (SD, 9 percent) of the tissue expander size. The first expansion was started 12 days (SD, 3 days) after the operation. The mean volume of each expansion was 88 cm3 (SD, 23 cm3). Expansion was completed in 4.7 office visits (SD, one visit). Mean final expander volume was 672 cm3 (SD, 144 cm3). The expanders were overexpanded by 15.3 percent (SD, 8.4 percent). The mean time between expander placement and the final expansion was 6.6 weeks (SD, 3 weeks). The overall complication rate was 4 percent. Ten patients developed cellulitis, five patients had hematomas requiring drainage, and one expander became exposed. A total of eight expanders were removed: four for cellulitis, one for a hematoma, one because of locally recurrent disease, one because of expander exposure, and one at the patient's request for no medical reason. Intraoperative and rapid postoperative tissue expansion is a safe and reliable technique that offers a significant improvement over conventional techniques. In this accelerated protocol, expansion may be completed in less than 7 weeks. The result is decreased patient morbidity and delays in adjuvant therapy at no detriment to the final surgical outcome.     

Development of a Superficial Meristem During Somatic Embryogenesis from Immature Cotyledons of Soybean (Glycine max L.)

Initial events were studied in the development of an embryogenicmeristem during somatic embryogenesis from in vitro culturedimmature cotyledons of soybean. The presence of 2,4-D in theculture medium led to the formation of a superficial embryogenictissue associated with the abaxial epidermis of 3 mm cotyledons.Additionally, 2,4-D initiated rapid non-morphogenic periclinaldivision in the parenchyma tissues of the cotyledon. Consequentinternal expansion disrupted and eventually ruptured the apparentlyquiescent adaxial epidermis. The profound difference in thein vitro response between abaxial and adaxial epidermes is discussedin relation to their relative roles in nutrient transport duringseed development in vivo. Somatic embryogenesis, transfer cells, Glycine max     

Xyloglucan endotransglucosylase activity loosens a plant cell wall

BACKGROUND AND AIMS: Plant cells undergo cell expansion when a temporary imbalance between the hydraulic pressure of the vacuole and the extensibility of the cell wall makes the cell volume increase dramatically. The primary cell walls of most seed plants consist of cellulose microfibrils tethered mainly by xyloglucans and embedded in a highly hydrated pectin matrix. During cell expansion the wall stress is decreased by the highly controlled rearrangement of the load-bearing tethers in the wall so that the microfibrils can move relative to each other. Here the effect was studied of a purified recombinant xyloglucan endotransglucosylase/hydrolase (XTH) on the extension of isolated cell walls. METHODS: The epidermis of growing onion (Allium cepa) bulb scales is a one-cell-thick model tissue that is structurally and mechanically highly anisotropic. In constant load experiments, the effect of purified recombinant XTH proteins of Selaginella kraussiana on the extension of isolated onion epidermis was recorded. KEY RESULTS: Fluorescent xyloglucan endotransglucosylase (XET) assays demonstrate that exogeneous XTH can act on isolated onion epidermis cell walls. Furthermore, cell wall extension was significantly increased upon addition of XTH to the isolated epidermis, but only transverse to the net orientation of cellulose microfibrils. CONCLUSIONS: The results provide evidence that XTHs can act as cell wall-loosening enzymes.     

Development of dendritic epidermal T cells with a skewed diversity of gamma delta TCRs in V delta 1-deficient mice

One of the most intriguing features of gammadelta T cells that reside in murine epithelia is the association of a specific Vgamma/Vdelta usage with each epithelial tissue. Dendritic epidermal T cells (DETCs) in the murine epidermis, are predominantly derived from the "first wave" Vgamma5+ fetal thymocytes and overwhelmingly express the canonical Vgamma5/Vdelta1-TCRs lacking junctional diversity. Targeted disruption of the Vdelta1 gene resulted in a markedly impaired development of Vgamma5+ fetal thymocytes as precursors of DETCs; however, gammadeltaTCR+ DETCs with a typical dendritic morphology were observed in Vdelta1-/- mice and their cell densities in the epidermis were slightly lower than those in Vdelta1+/- epidermis. Moreover, the Vdelta1-deficient DETCs were functionally competent in their ability to up-regulate cytokines and keratinocyte growth factor-expression in response to keratinocytes. Vgamma5+ DETCs were predominant in the Vdelta1-/- epidermis, though Vgamma5- gammadeltaTCR+ DETCs were also detected. The Vgamma5+ DETCs showed a typical dendritic shape, gammadeltaTCR(high), and age-associated expansion in epidermis as observed in conventional DETCs of normal mice, whereas the Vgamma5- gammadeltaTCR+ DETCs showed a less dendritic shape, gammadeltaTCR(low), and no expansion in the epidermis, consistent with their immaturity. These results suggest that optimal DETC development does not require a particular Vgamma/Vdelta-chain usage but requires expression of a limited diversity of gammadeltaTCRs, which allow DETC precursors to mature and expand within the epidermal microenvironment.     

Patterns of cell division and expansion in developing petals of Petunia hybrida

The definition of the patterns of cell division and expansion in plant development is of fundamental importance in understanding the mechanics of morphogenesis. By studying cell division and expansion patterns, we have assembled a developmental map of Petunia hybrida petals. Cycling cells were labelled with in situ markers of the cell cycle, whereas cell expansion was followed by assessing cell size in representative regions of developing petals. The outlined cell division and expansion patterns were related to organ asymmetry. Initially, cell divisions are uniformly distributed throughout the petal and decline gradually, starting from the basal part, to form a striking gradient of acropetal polarity. Cell areas, in contrast, increased first in the basal portion and then gradually towards the petal tip. This growth strategy highlighted a cell size control model based on cell-cycle departure time. The dorso-ventral asymmetry can be explained in terms of differential regulation of cell expansion. Cells of the abaxial epidermis enlarged earlier to a higher final extent than those of the adaxial epidermis. Epidermal appendage differentiation contributed to the remaining asymmetry. On the whole our study provides a sound basis for mutant analyses and to investigate the impact of specific (environmental) factors on petal growth.     

Fossil leaves of Populus from the Middle Miocene of Yunnan,SW China

Today southern Yunnan, SW China, has a tropical or subtropical climate and seasonal rainforests. In the past, some temperate elements were also present. In this paper, a new species of Populus is reported from the Middle Miocene deposits in Zhenyuan. Its leaves are ovate or ovate-suborbicular, with serrate margins. They have a shallowly cordate to cuneate base without glands, short acuminate apex, and salicoid teeth with spherical glands. The veins are glabrous but unicellular hair bases occur on the lower epidermis of the lamina. Stomata are confined to the lower epidermis. The presence of Populus in the Middle Miocene of the region indicates an expansion of the genus into low-latitude Asia in the late Cenozoic and a more complicated history of vegetational change in southern Yunnan than has so far been assumed.