Subepidermal Connective Tissue Research Articles

In Vivo Tracking of Systemically Administered Allogeneic Bone Marrow Mesenchymal Stem Cells in Normal Rats through Bioluminescence Imaging.

Recently, mesenchymal stem cells (MSCs) are increasingly used as a panacea for multiple types of disease short of effective treatment. Dozens of clinical trials published demonstrated strikingly positive therapeutic effects of MSCs. However, as a specific agent, little research has focused on the dynamic distribution of MSCs after in vivo administration. In this study, we track systemically transplanted allogeneic bone marrow mesenchymal stem cells (BMSCs) in normal rats through bioluminescence imaging (BLI) in real time. Ex vivo organ imaging, immunohistochemistry (IHC), and RT-PCR were conducted to verify the histological distribution of BMSCs. Our results showed that BMSCs home to the dorsal skin apart from the lungs and kidneys after tail vein injection and could not be detected 14 days later. Allogeneic BMSCs mainly appeared not at the parenchymatous organs but at the subepidermal connective tissue and adipose tissue in healthy rats. There were no significant MSCs-related adverse effects except for transient decrease in neutrophils. These findings will provide experimental evidences for a better understanding of the biocharacteristics of BMSCs.

Short term effects of iontophoresis on the structure of the guinea pig tympanic membrane.

Guinea pig tympanic membrane was anesthetized iontophoretically. The animals were sacrificed between 2 h and 4 days after the anesthesia and the tympanic membrane was studied histologically by light microscopy and transmission electron microscopy. Edema in the sub-epidermal connective tissue layer and detachment of the basement membrane from the basal cell layer were observed 2 h after iontophoresis. Eight and 12 h later, the epidermal layer between the handle of the malleus and the bony annulus peeled and retracted towards the bony annulus, leaving only hornified outer epidermis on the fibrous layer. Four days after iontophoresis, the retracted margin of the epidermal layer became greatly thickened and evidence of hyperplasia and migration was observed. This study has demonstrated that iontophoresis of the tympanic membrane produces remarkable histological changes. However, no perforation was observed.

Efficacy of Marigold Extract-Loaded Formulations Against UV-induced Oxidative Stress

The present study investigated the potential use of topical formulations containing marigold extract (ME) (Calendula officinalis extract) against ultraviolet (UV)B irradiation-induced skin damage. The physical and functional stabilities, as well as the skin penetration capacity, of the different topical formulations developed were evaluated. In addition, the in vivo capacity to prevent/treat the UVB irradiation-induced skin damage, in hairless mice, of the formulation with better skin penetration capacity was investigated. All of the formulations were physically and functionally stable. The gel formulation [Formulation 3 (F3)] was the most effective for the topical delivery of ME, which was detected as 0.21 μg/cm(2) of narcissin and as 0.07 μg/cm(2) of the rutin in the viable epidermis. This formulation was able to maintain glutathione reduced levels close to those of nonirradiated animals, but did not affect the gelatinase-9 and myeloperoxidase activities increased by exposure to UVB irradiation. In addition, F3 reduced the histological skin changes induced by UVB irradiation that appear as modifications of collagen fibrils. Therefore, the photoprotective effect in hairless mice achieved with the topical application of ME in gel formulation is most likely associated with a possible improvement in the collagen synthesis in the subepidermal connective tissue.

TRPV1 expression in acupuncture points: Response to electroacupuncture stimulation

The present study was to examine the distribution of transient receptor potential vanilloid type-1 (TRPV1) receptor immunoreactivity in the acupuncture points (acupoint), and determine the influences of electroacupuncture (EA) stimulation on TRPV1 expression. EA stimulation of BL 40 was conducted in two sessions of 20 min separated by an 80 min interval in anesthetized rats. Sections of skin containing BL 40, and its non-meridian control were examined by immunolabeling with antibodies directed against TRPV1. Without EA, the number of subepidermal nerve fibers expressing TRPV1 was higher in the acupoint than in non-acupoint control skin ( p < 0.01). The subepidermal nerve fibers showed the co-localization of TRPV1 with peripherine, a marker for the C-fibers and A-δ fibers. The expression of TRPV1 in nerve fibers is significantly increased by EA stimulation in acupoints ( p < 0.01). However the upregulation in the non acupoint meridian and the non-meridian control skin was short of statistical significance. Double immunostaining of TRPV1 and neuronal nitric oxide synthase (nNOS) revealed their co-localization in both the subepidermal nerve fibers and in the dermal connective tissue cells. These results show that a high expression of TRPV1 endowed with nNOS in subepidermal nerve fibers exists in the acupoints and the expression is increased by EA. We conclude that the higher expression of TRPV1 in the subepidermal nerve fibers and its upregulation after EA stimulation may play a key role in mediating the transduction of EA signals to the CNS, and its expression in the subepidermal connective tissue cells may play a role in conducting the local effect of the EA.

Immunohistochemical study of subepidermal connective of molluscan integument.

Sections of integument from gastropod, bivalve and cephalopod species were studied immunohistochemically to determine reactivity to antibody against the type I-like collagen from Sepia cartilage and antibodies against components of the extracellular matrix (ECM) of vertebrate connective tissue: type I, III, IV, V, and VI collagens, laminin, nidogen and heparan sulphate. All samples exhibited similar reactivities to the antibodies, although differences in the intensity and localization of the immunostaining were found that were clearly correlated with between-species differences in integumental ultrastructure. These findings indicate that the composition of the integumental ECM is similar in the three classes of molluscs examined and that several types of collagen are present. However molluscan subepidermal connective tissue differs from the ECM of vertebrate dermis: molluscan integumental ECM contains collagens similar to type I, V and VI collagens but has no type III-similar collagen. Furthermore molecules similar to the type IV collagen, laminin, nidogen and heparan sulphate of vertebrates were present ubiquitously in molluscan basement membrane, confirming the statement that the structure and composition of basement membrane have remained constant throughout the evolution of all animal phyla.

Radiogenic Lymphangiogenesis in the Skin

The time course of microvascular changes in the environment of irradiated tumors was studied in a standardized human protocol. Eighty skin biopsies from 40 patients with previously treated primary breast cancer were taken from irradiated skin and corresponding contralateral unirradiated control areas 2 to 8 weeks, 11 to 14 months, or 17+ months after radiotherapy (skin equivalent dose 30 to 40 Gy). Twenty-two biopsies of 11 melanoma patients who had undergone lymph node dissection were used for unirradiated control. We found an increase of total podoplanin(+) lymphatic microvessel density resulting mainly from a duplication of the density of smallest lymphatic vessels (diameter <10 microm) in the samples taken 1 year after radiation. Our findings implicate radiogenic lymphangiogenesis during the 1st year after therapy. The numbers of CD68(+) and vascular endothelial growth factor-C(+) cells were highly elevated in irradiated skin in the samples taken 2 to 8 weeks after radiotherapy. Thus, our results indicate that vascular endothelial growth factor-C expression by invading macrophages could be a pathogenetic route of induction of radiogenic lymphangiogenesis.

Horizontal Square Buried Sutures in a Two-Layered Fashion Enable Direct Primary Closure for Small Circular Wounds without Dog-Ears on the Face

Dog-ears often lead to lengthening of an excision, and it is desirable to avoid them. Facial skin, including the subepidermal connective tissue, is flexible and can be used advantageously to minimize dog-ears using a novel buried suture technique. After removing a round lesion, the first horizontal square buried suture (HSBS) was deeply placed parallel to the longitudinal direction of the defect beneath superficial fascia. After the first HSBS was tied, the defect became fusiform but was still largely open. The second HSBS was also placed parallel to the longitudinal direction of the defect but in more superficial fascia and using smaller horizontal buried loops than those of the first deep suture. After the second HSBS in the middle of the dermis was tied, the wound was almost closed without dog-ears. Consequently, few skin sutures were required to finish the operation. Using this technique, a small circular or oval defect on the face up to 1 cm in diameter can be closed without any additional excision of the skin and without creating dog-ears.

Horizontal Square Buried Sutures in a Two-Layered Fashion Enable Direct Primary Closure for Small Circular Wounds without Dog-Ears on the Face

background Dog-ears often lead to lengthening of an excision, and it is desirable to avoid them. objective Facial skin, including the subepidermal connective tissue, is flexible and can be used advantageously to minimize dog-ears using a novel buried suture technique. methods After removing a round lesion, the first horizontal square buried suture (HSBS) was deeply placed parallel to the longitudinal direction of the defect beneath superficial fascia. After the first HSBS was tied, the defect became fusiform but was still largely open. The second HSBS was also placed parallel to the longitudinal direction of the defect but in more superficial fascia and using smaller horizontal buried loops than those of the first deep suture. After the second HSBS in the middle of the dermis was tied, the wound was almost closed without dog-ears. Consequently, few skin sutures were required to finish the operation. conclusions Using this technique, a small circular or oval defect on the face up to 1 cm in diameter can be closed without any additional excision of the skin and without creating dog-ears.

An ultrastructural study of connective tissue in mollusc integument III: Cephalopoda

We studied structure and ultrastructure of the subepidermal connective tissue (SEC) of the integument of three cephalopods ( Sepia officinalis, Octopus vulgaris and Loligo pealii). In all species, three distinct regions of the SEC were recognised: (a) an outer zone (OZ) that included the dermal–epidermal junction, and consisted of a thin layer of connective tissue containing muscles, (b) an extensive middle zone (MZ) containing a compact network of collagen fibres and numerous cells, (c) an inner zone (IZ) of loose connective tissue that merged with muscular fascia. This arrangement differs from that in bivalves and gastropods and recalls vertebrate integument. The dermal–epidermal junction of cephalopods differed from that of bivalves, gastropods and mammals in that the epidermal cells did not possess hemidesmosomes, and their intermediate filaments terminated directly in the plasmamembrane. The thick (120–500 nm) basal membrane (BM) had a superficial zone containing a regular array of granules; a lamina densa composed of a compact network of small filaments and granules; and an IZ distinguished by expansions of granular material protruding into underlying structures. Collagen fibres contained fibroblast-derived cytoplasmic thread, running through their centres and were surrounded by granular material that joins them to adjacent fibres. The collagen fibrils were of medium diameter (30–80 nm) had the typical ultrastructure of fibrillar collagens, and were surrounded by abundant interfibrillar material. The hypodermis was loose, with a network of small bundles of collagen fibrils. Cephalopod integument appears to represent a major evolutionary step distinguishing this class of molluscs.

Immunohistochemical localization of nNOS and VIP in the mantle integument of the mussel, Mytilus galloprovincialis

The phylogeny and functional roles of many bioactive compounds in the invertebrate integument are still unclear. In order to deal with this issue, we performed an immunohistochemical investigation of the integument of the mussel, Mytilus galloprovincialis, to demonstrate the presence of nNOS- and VIP-positive nerve fibers in subepidermal connective tissue of the mantle. Positive nerve cell bodies were detected in this tissue as well as in cortex of sperm follicles, and adjacent to maturating oocytes and spermatocytes located in the thickness of the mantle. These results indicate involvement of a local inhibitory non-adrenergic-non-cholinergic (NANC) regulatory mechanism of epidermal functions, such as mucous secretion and ciliary beating. At the gonadic level, this mechanism probably regulates the cycle of maturation and release of the gametes in both sexes.

An ultrastructural study of connective tissue in mollusc integument: II. Gastropoda

We studied the ultrastructure of the subepidermal connective tissue (SEC) in different zones of the integument in terrestrial, marine and freshwater gastropods (eight species). In all cases, the SEC was a layer of loose connective tissue between the basal membrane (BM) of the epidermis and the connective tissue of the deeper muscle layers. It was of monotonous structure and not differentiated into layers such as are found in mammalian dermis. The extracellular matrix (ECM) consisted of a network of collagen fibrils of variable diameter, with abundant anchoring devices and proteoglycans. In six species, variables quantities of haemocyanin were present within haemocoelic sinuses present in the SEC. The thickness and density of the BM varied from species to species, as well as within species in the various zones of integument. The ultrastructure of the lamina densa (LD) was indistinguishable from that of BM in bivalves and similar to that in mammals, although basotubules and double pegs were absent. An irregularly spaced lamina lucida was usuallypresent and was often shot thorough with filaments and small protrusions of the LD that connected with epithelial plasma membrane or with hemidesmosomes. A lamina fibroreticularis wasnot present. LD protrusions characterize the connection between BM and the ECM of SEC. In theterrestrial gastropods, a spongy matrix with ultrastructure closely similar to LD occupied large tracts of the SEC. In the mantle region of Arion rufus, the integumental SEC contained large cavities filled with spherical concretions, probably representing rudiments of a shell. In the mantle where theintegument contained abundant muscle fibres, the BM was thick and directly connected to the ECMof the SEC which consisted of compact laminae of collagen fibrils with abundant anchoring devices. Along the edge of the foot of Patella ulyssiponensis, the SEC contained a layer of paramyosinic muscle fibres adhering to the epidermis. No differences or gradations in integumental SEC structure could be related to the phylogenetic position of the species examined.

An ultrastructural study of connective tissue in mollusc integument: I. Bivalvia

The ultrastructure of the subepidermal connective tissue (SEC) in different areas of the integument of the bivalves Callista chione, Pecten jacobaeus, Mytilus galloprovincialis and Ostrea edulis was studied by transmission electron microscopy. The main organisation of the SEC was broadly similar in all species: the SEC was connected to the epidermis by a basement membrane and merged directly with the deeper connective tissue surrounding muscles. The SEC was not differentiated into layers like the papillary and reticular dermis of mammals, however, the architecture, thickness and shape of the basement membrane varied from species to species, as well as within species (in the foot, central or marginal zones of the mantle). The ultrastructure of the lamina densa was broadly similar to that in mammals: although basotubules and double pegs were absent, proteoglycans and rod-like units homologous to ‘double tracks’ were always abundant. A zone similar to the lamina lucida was irregularly present and was shot thorough with small protrusions of the lamina densa that connected with the epithelial hemidesmosomes or focal adhesions. Nevertheless zones were observed where the lamina densa fuse directly to the epithelial plasmamembrane. This variability of connection may be related to the various types of epidermal cell. A lamina fibroreticularis was not recognized since anchoring fibrils and microfibrils were not present; lamina densa protrusions into the extracellular matrix (ECM) of SEC characterize the connection between basement membrane and SEC. Collagen fibrils were small and of constant diameter and were never organised into fibres. Anchoring devices – similar to the anchoring plaques of mammalian dermis – were abundant and scattered between SEC collagen fibrils. The orange-pink pigmentation of C. chione seems due to electron-dense granules embedded within the connective ECM.

Pituitary adenylate cyclase activating polypeptide (PACAP) in the rat mammary gland.

Pituitary adenylate cyclase activating polypeptide (PACAP), a member of the vasoactive intestinal polypeptide (VIP) family of peptides, is present in the brain and in neuronal elements of a number of peripheral organs. Since no information on PACAP in the mammary gland exists, we have investigated, by radioimmunoassay and immunohistochemistry, the occurrence and distribution of PACAP immunoreactivity in the mammary gland of lactating and non-lactating rats. A specific monoclonal mouse anti-PACAP antibody'has been used to show that the peptide is located in nerve fibres associated with bundles of circular and longitudinal smooth muscle surrounding the lactiferous duct of the nipple. PACAP-immunoreactive nerve fibres and nerve bundles are present in the subepidermal connective tissue of the nipple and in the mammary parenchyma, some of the fibres being in close contact with blood vessels. Occasionally, a few delicate varicose fibres are associated with secretory alveoli and lactiferous ducts. The majority of PACAP-positive nerve fibres are, however, located in the glabrous skin of the nipple and the hairy skin adjacent to the nipple forming a subepithelial plexus from which delicate varicose nerve fibres enter the overlying epithelium. Double immunostaining for PACAP and a marker for sensory neurons, calcitonin gene-related peptide, has disclosed that the two peptides are almost completely co-localized. A minor population of the PACAP-immunoreactive nerve fibres shows co-existence with VIP. Although no obvious changes at the immunohistochemical level could be observed during pregnancy or lactation, elevated concentrations of immunoreactive PACAP-38 in mammary extracts have been found during lactation. Our data suggest that PACAP is involved in the nervous control of mammary gland function, probably in the transmission of suckling stimuli.

The role of the composition and gas-transporting function of the blood in the fluctuations of the electrical properties of the epidermis and subepidermal tissues

Changes in the composition and gas-transporting function of the blood contribute to the formation of the total electrical impedance of the epidermis, subepidermal connective tissue, and muscles. It is concluded that time fluctuations of the impedance result from the summated influence of the factors characterizing the modulations of the tissue oxygen consumption.

Wound contraction and scar synthesis during development of the amphibian Rana catesbeiana

Certain characteristics of skin wound healing in the North American bullfrog, Rana catesbeiana, were studied grossly and histologically at various stages of development. The contraction kinetics of excisional skin wounds were monitored for 50 days in animals from four larval developmental stages (tadpoles) and from adults (frogs). The percentage of original wound area which remained after contraction had stopped in the four larval groups decreasing steadily from 59.2%+/- 6.8% for the least developed larvae to 9.9%+/- 2.3% for the most developed ones. Wounds in adults contracted at a much lower rate than for larvae and the percentage of original wound area remaining after 50 days was less than 10%. The morphologic features characterizing the intact dermis and epidermis outside the wound bed in the larvae were also observed in qualitatively similar detail inside the wound bed. In the adults, the subepidermal connective tissue inside the wound bed was distinctly different from the physiologic dermis outside and was classified as amphibian scar. We conclude that, during larval development, wound contraction increasingly displaced skin regeneration as a mechanism for wound closure. Adults closed skin wounds by a combination of contraction and synthesis of scar tissue. It is hypothesized that the mechanism for synthesis of scar in this amphibian is established during metamorphosis from larva to adult.

Histochemical study of cutaneous mucins in hypothyroid dogs.

A dermal mucinosis, visualized as dermal alcianophilic material, is occasionally present in canine hypothyroidism (myxedema). Various histochemical reactions (alcian blue/periodic acid-Schiff [PAS], alcian blue at pH 2.6, alcian blue at pH 1.0, critical electrolytical concentrations with and without dimethylsulfoxide, differential hydrolysis by hyaluronidases) were performed on skin biopsies from six dogs (four females and two males ranging from 8 to 13 years) affected by hypothyroidism, all of them presenting dermal mucinosis in hematoxylin and eosin-stained sections. In these dogs, the only polysaccharidic compound involved in the dermal mucinosis was hyaluronic acid. In this study, hyaluronic acid dermal deposits of hypothyroid dogs were significantly different from those of controls in subepidermal connective tissue and loose reticular connective tissue but not in periadnexal zones. We recommend the combined alcian blue/PAS reaction as a routine technique to assess dermal mucinosis in hypothyroid dogs.

Microscopic anatomy and histochemistry of the crayfish parasite Psorospermium haeckeli

DAO Diseases of Aquatic Organisms Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsSpecials DAO 21:79-90 (1995) - doi:10.3354/dao021079 Microscopic anatomy and histochemistry of the crayfish parasite Psorospermium haeckeli Vogt G, Rug M ABSTRACT: Psorospermium haeckeli, a common parasite of the European noble crayfish Astacus astacus, occurs as an egg-shaped spore of ca 100 x 60 x 60 um, particularly within the collagenous layer of the thoracic arteries and the surrounding subepidermal connective tissue. Smaller numbers are also found within the connective tissues of virtually all organs. The morphology and histology of these spores was investigated by microscopic techniques and the composition of their major structural components was identified by histochemistry. Each spore is enveloped by a layer of connective tissue which is assumed to derive from the host. It varies in thickness and composition among the infected organs. The spore proper consists of a 3-layered shell enclosing many globules. The outer shell layer is composed of solid, collagen- and elastin-negative, proteinaceous plates separated by sutures. The medial shell layer appears less compact and includes glycoproteins or neutral mucopolysaccharides. The inner layer is composed of 2 membranous structures including glycoproteins, glycolipids, and collagen- and elastin-like proteins. The interior of the spore contains lipid globules of variable size, intensely staining nonlipid globules with crystalline and vacuolar inclusions, and 2 closely associated nuclei. In most of the crayfish investigated some of the spores were encapsulated by haemocytes and melanized. KEY WORDS: Psorospermium haeckeli . Parasite . Astacus astacus . Crayfish . Microscopic anatomy . Histochemistry Full text in pdf format NextExport citation RSS - Facebook - Tweet - linkedIn Cited by Published in DAO Vol. 21, No. 2. Publication date: March 30, 1995 Print ISSN:0177-5103; Online ISSN:1616-1580 Copyright © 1995 Inter-Research.

Direct application of basic fibroblast growth factor improves tympanic membrane perforation healing

Topical application of basic fibroblast growth factor (b-FGF) on tympanic membrane (TM) perforations was studied in guinea pigs. One-millimeter simple round TM perforations or 2-mm TM perforations with medially flapped borders were performed. Either b-FGF or placebo was instilled in each ear on the day of surgery and daily thereafter. Treatment was applied either directly to the perforation or to a Gelfoam pledget over the defect. When no scaffolding material was interposed, b-FGF induced a faster healing response characterized by a hyperplastic but linear subepidermal connective tissue reaction compared to the control. When Gelfoam was interposed as a scaffold, a voluminous scar protruding into the middle ear cavity and involving the ossicles was observed in both b-FGF and control animals. Gelfoam-induced scars did not decrease after long-term observation, therefore discouraging its use.

Sialomucin in middle ear cholesteatoma perimatrix.

Mucosubstance histochemistry of human middle ear cholesteatoma revealed that sialomucins are abundant and sulfomucins present in small amounts in the glandlike structures of the cholesteatoma perimatrix. Based on the study, various glycoproteins were injected into the dermis of the external ear canal and infiltrated into the tympanic membranes of guinea pigs. Tympanic membranes were obtained 7 days later and light- and electron-microscopically studied. Injection of a sialomucin from bovine submaxillary gland resulted in marked proliferation of epidermis and degeneration of the lamina propria. Asialomucin prepared from sialomucin by hydrolysis produced mild thickening of the epidermis but the lamina propria was not degenerated. Fetuin and bovine serum albumin did not cause proliferation of the epidermis. Cholesterol granuloma formed in some of the specimens injected with sialomucin. The presence of sialomucin in cholesteatoma perimatrix and these experimental studies using tympanic membranes of guinea pigs suggests that sialomucins participate in the proliferation of epidermis and degeneration of subepidermal connective tissue in human middle ear cholesteatoma.

Development of the Hair in the Rat: In vivo and in Transplanted Tissue

Differentiation of the hair was examined in the head skin of rat embryos aged from days 12.5 to 18.5 of gestation. In 12.5-day-old embryos, the skin consisted of single layered epidermis and underlying loose connective tissue. On day 17.5 of gestation, the epidermis appeared to have a two or three layered epithelium, and the subepidermal connective tissue gave rise to an overlying dense tissue, into which blood capillaries arose from the deeper layer. The epidermis showed partial thickenings, where basal cells were arranged perpendicular to the surface of the epidermis. Beneath the epidermal thickenings, a number of connective tissue cells appeared to have accumulated and occasionally were accompanied by blood capillaries. In sequential development of the skin, the thickenings protruded as cords to the underlying dermal tissue and formed hair follicles covered with a connective tissue sheath. Skin obtained from 12.5-day-old embryos was transplanted singly or with cerebral cortex into the third ventricle of adult male rats. Thirty days after the transplantation, hairs and sebaceous glands had developed from the grafted skin. Hair growth tended to be more prominent in the cotransplanted skin grafts. However, no nerve fibers immunoreactive for substance P or calcitonin gene-related peptide (CGRP) were found in the grafted skin, although these fibers appeared in the dermis at the end of gestation in vivo. These results showed that the skin of 12.5-day-old embryos was able to induce hair growth in the third ventricle without accompanying peripheral neuronal fibers or blood capillaries, although the effects of connective tissue cells could not be ruled out.