A Novel Composite Biomaterial Made of Jellyfish and Porcine Collagens Accelerates Dermal Wound Healing by Enhancing Reepithelization and Granulation Tissue Formation in Mice.

Background and Objective: Impaired dermal wound healing represents a major medical issue in today's aging populations. Granulation tissue formation in the dermis and reepithelization of the epidermis are both important and necessary for proper wound healing. Although a number of artificial dermal grafts have been used to treat full-thickness dermal loss in humans, they do not induce reepithelization of the wound, requiring subsequent epithelial transplantation. In the present study, we sought a novel biomaterial that accelerates the wound healing process. Approach: We prepared a composite biomaterial made of jellyfish and porcine collagens and developed a hybrid-type dermal graft that composed of the upper layer film and the lower layer sponge made of this composite biomaterial. Its effect on dermal wound healing was examined using a full-thickness excisional wound model. Structural properties of the dermal graft and histological features of the regenerating skin tissue were characterized by electron microscopic observation and immunohistological examination, respectively. Results: The composite biomaterial film stimulated migration of keratinocytes, leading to prompt reepithelization. The regenerating epithelium consisted of two distinct cell populations: keratin 5-positive basal keratinocytes and more differentiated cells expressing tight junction proteins such as claudin-1 and occludin. At the same time, the sponge made of the composite biomaterial possessed a significantly enlarged intrinsic space and enhanced infiltration of inflammatory cells and fibroblasts, accelerating granulation tissue formation. Innovation: This newly developed composite biomaterial may serve as a dermal graft that accelerates wound healing in various pathological conditions. Conclusion: We have developed a novel dermal graft composed of jellyfish and porcine collagens that remarkably accelerates the wound healing process.

The effects of LXR agonist T0901317 and LXR antagonist GSK2033 on morphogenesis and lipid properties in full thickness skin models.

Full thickness models (FTMs) are 3D-cultured human skin models that mimic many aspects of native human skin (NHS). However, their stratum corneum (SC) lipid composition differs from NHS causing a reduced skin barrier. The most pronounced differences in lipid composition are a reduction in lipid chain length and increased monounsaturated lipids. The liver-X-receptor (LXR) activates the monounsaturated lipid synthesis via stearoyl-CoA desaturase-1 (SCD-1). Therefore, the aim was to improve the SC lipid synthesis of FTMs by LXR deactivation. This was achieved by supplementing culture medium with LXR antagonist GSK2033. LXR agonist T0901317 was added for comparison. Subsequently, epidermal morphogenesis, lipid composition, lipid organization and the barrier functionality of these FTMs were assessed. We demonstrate that LXR deactivation resulted in a lipid composition with increased overall chain lengths and reduced levels of monounsaturation, whereas LXR activation increased the amount of monounsaturated lipids and led to a reduction in the overall chain length. However, these changes did not affect the barrier functionality. In conclusion, LXR deactivation led to the development of FTMs with improved lipid properties, which mimic the lipid composition of NHS more closely. These novel findings may contribute to design interventions to normalize SC lipid composition of atopic dermatitis patients.

Annurca Apple Nutraceutical Formulation Enhances Keratin Expression in a Human Model of Skin and Promotes Hair Growth and Tropism in a Randomized Clinical Trial.

Several pharmaceutical products have been formulated over the past decades for the treatment of male and female alopecia, and pattern baldness, but relatively few metadata on their efficacy have been published. For these reasons, the pharmaceutical and medical attention has recently focused on the discovery of new and safer remedies. Particularly, great interest has been attracted by oligomeric procyanidin bioactivity, able to promote hair epithelial cell growth as well as to induce the anagen phase. Specifically, the procyanidin B2, a dimeric derivative extracted from apples, has demonstrated to be one of the most effective and safest natural compounds in promoting hair growth, both in vitro and in humans by topical applications. By evaluating the polyphenolic content of different apple varieties, we have recently found in the apple fruits of cv Annurca (AFA), native to Southern Italy, one of the highest contents of oligomeric procyanidins, and, specifically, of procyanidin B2. Thus, in the present work we explored the in vitro bioactivity of AFA polyphenolic extract as a nutraceutical formulation, named AppleMets (AMS), highlighting its effects on the cellular keratin expression in a human experimental model of adult skin. Successively, testing the effects of AMS on hair growth and tropism in healthy subjects, we observed significant results in terms of increased hair growth, density, and keratin content, already after 2 months. This study proves for the first time the impact of apple procyanidin B2 on keratin biosynthesis in vitro, and highlights its effect as a nutraceutical on human hair growth and tropism.

In vitro screening for estrogenic endocrine disrupting compounds using Mozambique tilapia and sea bass scales.

A wide range of estrogenic endocrine disruptors (EDCs) are accumulating in the environment and may disrupt the physiology of aquatic organisms. The effects of EDCs on fish have mainly been assessed using reproductive endpoints and in vivo animal experiments. We used a simple non-invasive assay to evaluate the impact of estrogens and EDCs on sea bass (Dicentrarchus labrax) and tilapia (Oreochromis mossambicus) scales. These were exposed to estradiol (E2), two phytoestrogens and six anthropogenic estrogenic/anti-estrogenic EDCs and activities of enzymes related to mineralized tissue turnover (TRAP, tartrate-resistant acid phosphatase and ALP, alkaline phosphatase) were measured. Semi-quantitative RT-PCR detected the expression of both membrane and nuclear estrogen receptors in the scales of both species, confirming scales as a target for E2 and EDCs through different mechanisms. Changes in TRAP or ALP activities after 30minute and 24h exposure were detected in sea bass and tilapia scales treated with E2 and three EDCs, although compound-, time- and dose-specific responses were observed for the two species. These results support again that the mineralized tissue turnover of fish is regulated by estrogens and reveals that the scales are a mineralized estrogen-responsive tissue that may be affected by some EDCs. The significance of these effects for whole animal physiology needs to be further explored. The in vitro fish scale bioassay is a promising non-invasive screening tool for E2 and EDCs effects, although the low sensitivity of TRAP/ALP quantification limits their utility and indicates that alternative endpoints are required.

Fabrication of core-shell nanofibers for controlled delivery of bromelain and salvianolic acid B for skin regeneration in wound therapeutics.

The physiological and pathological complexity of the wound healing process makes it more challenging to design an ideal tissue regeneration scaffold. Precise scaffolding with high drug loading efficiency, efficient intracellular efficacy for therapeutic delivery, minimal nonspecific cellular and blood protein binding, and maximum biocompatibility forms the basis for an ideal delivery system. This paper describes a combinational multiphasic delivery system, where biomolecules are delivered through the fabrication of coaxial electrospinning of different biocompatible polymers. The ratio and specificity of polymers for specific biofunction are optimized and the delivery system is completely characterized with reference to the mechanical property and structural integrity of bromelain (debridement enzyme) and salvianolic acid B (pro-angiogenesis and re-epithelialization). The in vitro release profile illustrated the sustained release of debriding protease and bioactive component in a timely fashion. The fabricated scaffold showed angiogenic potential through in vitro migration of endothelial cells and increased new capillaries from the existing blood vessel in response to an in ovo chicken chorioallantoic membrane assay. In addition, in vivo studies confirm the efficacy of the fabricated scaffold. Our results therefore open up a new avenue for designing a bioactive combinational multiphasic delivery system to enhance wound healing.

[Specificities in children wound healing]. / Particularités de la cicatrisation de l'enfant.

Children have specific characteristics of wound healing. The aim of this study was to describe the specific clinical characteristics of wounds healing in children and to present the current knowledge on the specific mechanisms with regard to infant age. The tissue insult or injury in fetus can heal without scar, mainly due to reduced granulation tissue associated to diminished or even no inflammatory phase, modified extracellular matrix such as the concentration of hyaluronic acid in amniotic liquid, expression and arrangement of collagen and tenascin. Thickness of children skin is a serious negative factor in case of trauma, whereas poor co-morbidities and efficient growth tissue mechanisms are beneficial to good evolution, even in cases of extensive damage and loss of tissue. The subsequent tissue mechanical forces, wound healing during childhood, spanning from the age of 2 until the end of puberty, is associated with more hypertrophic scars, both in duration and in intensity. Consequently, unnecessary surgery has to be avoided during this period when possible, and children with abnormal or pathologic wound healing should benefit from complementary treatments (hydration, massage, brace, silicone, hydrotherapy…), which represent efficient factors to minimize tissue scarring. After wound healing, the growth body rate can be responsible for specific complications, such as contractures, alopecia, and scar intussusceptions. Its evolutionary character implies the need of an attentive follow-up until adult age. Psychologic repercussions, as a consequence of pathologic scars, must be prevented and investigated by the surgeon.

Changes in tissue lipid and fatty acid composition of farmed rainbow trout in response to dietary camelina oil as a replacement of fish oil.

Camelina oil (CO) replaced 50 and 100 % of fish oil (FO) in diets for farmed rainbow trout (initial weight 44 ± 3 g fish(-1)). The oilseed is particularly unique due to its high lipid content (40 %) and high amount of 18:3n-3 (α-linolenic acid, ALA) (30 %). Replacing 100 % of fish oil with camelina oil did not negatively affect growth of rainbow trout after a 12-week feeding trial (FO = 168 ± 32 g fish(-1); CO = 184 ± 35 g fish(-1)). Lipid and fatty acid profiles of muscle, viscera and skin were significantly affected by the addition of CO after 12 weeks of feeding. However, final 22:6n-3 [docosahexaenoic acid (DHA)] and 20:5n-3 [eicosapentaenoic acid (EPA)] amounts (563 mg) in a 75 g fillet (1 serving) were enough to satisfy daily DHA and EPA requirements (250 mg) set by the World Health Organization. Other health benefits include lower SFA and higher MUFA in filets fed CO versus FO. Compound-specific stable isotope analysis (CSIA) confirmed that the δ(13)C isotopic signature of DHA in CO fed trout shifted significantly compared to DHA in FO fed trout. The shift in DHA δ(13)C indicates mixing of a terrestrial isotopic signature compared to the isotopic signature of DHA in fish oil-fed tissue. These results suggest that ~27 % of DHA was synthesized from the terrestrial and isotopically lighter ALA in the CO diet rather than incorporation of DHA from fish meal in the CO diet. This was the first study to use CSIA in a feeding experiment to demonstrate synthesis of DHA in fish.

Vitamin D and aging.

Age-related changes affect vitamin D metabolism and increase the requirement for vitamin D in the elderly. Also there is an age related decrease in calcium absorption and a higher calcium intake is needed. Increasing calcium from dietary sources may be better than supplements, and requires increasing the intake of dairy products or other and calcium-fortified foods. Evidence suggests that vitamin D and calcium nutrition can be improved in the elderly by increasing the vitamin D intake to 800 IU daily together with a total calcium intake of 1000 mg daily. This combination is a simple, inexpensive strategy that can reduce fractures in institutionalized individuals by 30%.

Effect of sex and dietary organic zinc on growth performance, carcass traits, tissue mineral content, and blood parameters of broiler chickens.

Zinc (Zn) is an essential mineral for animal development and function. A study was carried out to evaluate the effect of sex and dietary organic zinc (OZ) on growth performance, carcass traits, tissue mineral content, and blood parameters of broiler chickens. A total of 240 1-day-old male and 240 female broiler chicks (Cobb × Cobb) were assigned to two dietary levels of OZ (2 × 2 factorial) with six replicates per treatment (20 birds/replicate pen). The OZ supplementation levels were 0 and 25 ppm. Results showed that OZ supplementation did not affect the growth performance of male and female broilers, but the males showed significantly better (P < 0.05) growth performance than females did. Similarly, OZ supplementation did not affect the thickness of both the back and thigh skin of male and female broilers; however, males had thicker skin than females. Dietary OZ supplementation did not affect collagen contents in the skin and meat samples. Male broilers had higher skin collagen contents than females, but no sex difference was found in meat collagen contents. OZ supplementation did not affect the shear force values of skin and meat samples. Male broilers had higher shear force values of back skin than females, but not in the meat samples. Dietary OZ supplementation increased (P < 0.05) the thigh meat Zn content in both sexes. The plasma Ca content was significantly (P < 0.05) increased by dietary OZ supplementation; however, other blood parameters were not affected by dietary OZ supplementation. Males had higher plasma glucose and cholesterol content than females. It is concluded that dietary OZ supplementation at the level of 25 ppm does not affect the growth performance and skin quality of broiler chickens but increases the Zn content in thigh meat and Ca content in plasma of broiler chickens. Male broilers had better growth performance and skin quality than females.

Effect of iodine source and dose on growth and iodine content in tissue and plasma thyroid hormones in fattening pigs.

PURPOSE: The aim of the present feeding trial with iodine was to assess pigs' growth performance and carcass characteristics, the iodine accumulation in tissues, and their influences on the thyroid hormones in plasma. METHODS: Eighty pigs (33-115 kg body weight) were allotted to 5 dietary treatments: a control group (150 µg I/kg), two potassium iodide [KI] groups (4,000 and 10,000 µg I/kg), and two potassium iodate [KIO3] groups (4,000 and 10,000 µg I/kg). Iodine concentration was determined in thyroid gland, liver, kidney, muscle, fat, and skin by ICP-MS. Furthermore, thyroxine (T4) and triiodothyronine (T3) in plasma were evaluated. RESULTS: High dietary iodine tended to have a negative effect on younger animals' growth (average daily gain, ADG). However, during the entire growth period, the growth performance and carcass characteristics were not influenced by iodine dosages or sources. Irrespective of iodine source, higher iodine doses of diets affected higher iodine stores in all tested tissues except for abdominal fat. Thus, iodine supplementation with 10,000 µg I/kg feed significantly increased iodine content in thyroid gland (+122%), liver (+260%), kidney (+522%), muscle (+131%), and skin (+321%) compared to the control group. However, there was no significance of thyroid hormones in plasma. CONCLUSIONS: As a result, pork and fat of pigs showed only low iodine accumulation even in the high-iodine groups. Thus, there should be no risk of an iodine excess in human nutrition and animal health, and the EU-upper level for iodine in pig feed can be maintained.

Design and in vivo evaluation of a molecularly defined acellular skin construct: reduction of early contraction and increase in early blood vessel formation.

Skin substitutes are of great benefit in the treatment of patients with full thickness wounds, but there is a need for improvement with respect to wound closure with minimal contraction, early vascularisation, and elastin formation. In this study we designed and developed an acellular double-layered skin construct, using matrix molecules and growth factors to target specific biological processes. The epidermal layer was prepared using type I collagen, heparin and fibroblast growth factor 7 (FGF7), while the porous dermal layer was prepared using type I collagen, solubilised elastin, dermatan sulfate, heparin, fibroblast growth factor 2 (FGF2) and vascular endothelial growth factor (VEGF). The construct was biochemically and morphologically characterised and evaluated in vivo using a rat full thickness wound model. The results were compared with the commercial skin substitute IntegraDRT and untreated wounds. The double-layered construct was prepared according to the design specifications. The epidermal layer was about 40 µm thick, containing 9% heparin and 0.2 µg FGF7 mg per layer, localised at the periphery. The dermal layer was 2.5 mm thick, had rounded pores and contained 10% dermatan sulfate+heparin, and 0.7 µg FGF2+VEGF mg per layer. The double-layered skin construct was implanted in a skin defect and on day 7, 14, 28 and 112 the (remaining) wound area was photographed, excised and (immuno) histologically evaluated. The double-layered skin construct showed more cell influx, significantly less contraction and increased blood vessel formation at early time points in comparison with IntegraDRT and/or the untreated wound. On day 14 the double-layered skin construct also had the fewest myofibroblasts present. On day 112 the double-layered skin construct contained more elastic fibres than IntegraDRT and the untreated wound. Structures resembling hair follicles and sebaceous glands were found in the double-layered skin construct and the untreated wound, but hardly any were found in IntegraDRT. The results provide new opportunities for the application of acellular skin constructs in the treatment of surgical wounds.

Selenoproteins are essential for proper keratinocyte function and skin development.

Dietary selenium is known to protect skin against UV-induced damage and cancer and its topical application improves skin surface parameters in humans, while selenium deficiency compromises protective antioxidant enzymes in skin. Furthermore, skin and hair abnormalities in humans and rodents may be caused by selenium deficiency, which are overcome by dietary selenium supplementation. Most important biological functions of selenium are attributed to selenoproteins, proteins containing selenium in the form of the amino acid, selenocysteine (Sec). Sec insertion into proteins depends on Sec tRNA; thus, knocking out the Sec tRNA gene (Trsp) ablates selenoprotein expression. We generated mice with targeted removal of selenoproteins in keratin 14 (K14) expressing cells and their differentiated descendents. The knockout progeny had a runt phenotype, developed skin abnormalities and experienced premature death. Lack of selenoproteins in epidermal cells led to the development of hyperplastic epidermis and aberrant hair follicle morphogenesis, accompanied by progressive alopecia after birth. Further analyses revealed that selenoproteins are essential antioxidants in skin and unveiled their role in keratinocyte growth and viability. This study links severe selenoprotein deficiency to abnormalities in skin and hair and provides genetic evidence for the role of these proteins in keratinocyte function and cutaneous development.

Transitional change in rat fetal cell proliferation in response to ghrelin and des-acyl ghrelin during the last stage of pregnancy.

Expression of mRNA for the ghrelin receptor, GHS-R1a, was detected in various peripheral and central tissues of fetal rats, including skin, bone, heart, liver, gut, brain and spinal cord, on embryonic day (ED)15 and ED17. However, its expression in skin, bone, heart and liver, but not in gut, brain and spinal cord, became relatively weak on ED19 and disappeared after birth (ND2). Ghrelin and des-acyl ghrelin facilitated the proliferation of cultured fetal (ED17, 19), but not neonatal (ND2), skin cells. On the other hand, with regard to cells from the spinal cord and hypothalamus, the proliferative effect of ghrelin continued after birth, whereas the effect of des-acyl ghrelin on neurogenesis in these tissues was lost at the ED19 fetal and ND2 neonatal stages. Immunohistochemistry revealed that the cells in the hypothalamus induced to proliferate by ghrelin at the ND2 stage were positive for nestin and glial fibrillary acidic protein. These results suggest that in the period immediately prior to, and after birth, rat fetal cells showing proliferation in response to ghrelin and des-acyl ghrelin are at a transitional stage characterized by alteration of the expression of GHS-R1a and an undefined des-acyl ghrelin receptor, their responsiveness varying among different tissues.

Ferrous ferric chloride stimulates the skin cell function and hair growth in mice.

Ferrous ferric chloride (FFC) is a distinct form of aqueous iron composed of a complex of ferrous chloride and ferric chloride that participates in both oxidation and reduction reactions. The author's previous study showed that the supplementation of culture medium with FFC stimulated the proliferation and differentiation of keratinocytes and melanocytes in newborn mice. FFC also stimulated the proliferation of cultured human keratinocytes, fibroblasts, and melanocytes. However, it is not known whether FFC stimulates the proliferation and differentiation of mammalian skin cells as well as hair growth in vivo. To answer this question, FFC-containing skin lotions (FFC Super Essence Plain and Moisture Type, Akatsuka Co.) were painted on the dorsal skin of newborn C57BL/10JHir (B10) mice and tested for their proliferation- and differentiation-stimulating effects on keratinocytes, fibroblasts, and melanocytes as well as for their stimulating effects on the hair growth. This treatment stimulated the proliferation and differentiation of epidermal keratinocytes, dermal fibroblasts, and epidermal and dermal melanocytes in the skin as well as hair growth. From 2 to 3 weeks after birth B10 mice generally lose their hairs except those on the head at the telogen stage of the hair growth cycle due to the expression of the alopecia. The treatment with FFC lotions markedly inhibited the alopecia hair-loss. These results suggest that FFC can stimulate the proliferation and differentiation of keratinocytes, fibroblasts, and melanocytes in the skin as well as the hair growth, and, in addition, can inhibit the alopecia hair-loss.

[Indications for sunflower oil concentrate in the treatment of atopic dermatitis]. / Indicación del destilado oleico de girasol en el tratamiento de la dermatitis atópica.

The treatment of atopic dermatitis, as other diseases that present a sensible skin, includes a series of therapeutic measures initiating with the general cares of the skin and application of elements that allow to preserve the functionality through relipidization and the inhibition of some components of the inflammation. This article includes a series of concepts that justify the use of sunflower oil concentrated like a weapon of forward edge in the treatment of these morbidities.

Development of a keratinocyte-based screening model for antipsoriatic drugs using green fluorescent protein under the control of an endogenous promoter.

Inflamed epidermis (psoriasis, wound healing, ultraviolet-irradiated skin) harbors keratinocytes that are hyperproliferative and display an abnormal differentiation program. A distinct feature of this so-called regenerative maturation pathway is the expression of proteins such as the cytokeratins CK6, CK16, and CK17 and the antiinflammatory protein SKALP/elafin. These proteins are absent in normal skin but highly induced in lesional psoriatic skin. Expression of these genes can be used as a surrogate marker for psoriasis in drug-screening procedures of large compound libraries. The aim of this study was to develop a keratinocyte cell line that contained a reporter gene under the control of a psoriasis-associated endogenous promoter and demonstrate its use in an assay suitable for screening. We generated a stably transfected keratinocyte cell line that expresses enhanced green fluorescent protein (EGFP), under the control of a 0.8-kb fragment derived from the promoter of the SKALP/elafin gene, which confers high levels of tissue-specific expression at the mRNA level. Induction of the SKALP promoter by tumor necrosis factor-alpha resulted in increased expression levels of the secreted SKALP-EGFP fusion protein as assessed by direct readout of fluorescence and fluorescence polarization in 96-well cell culture plates. The fold stimulation of the reporter gene was comparable to that of the endogenous SKALP gene as assessed by enzyme-linked immunosorbent assay. Although the dynamic range of the screening system is limited, the small standard deviation yields a Z factor of 0.49. This indicates that the assay is suitable as a high-throughput screen, and provides proof of the concept that a secreted EGFP fusion protein under the control of a physiologically relevant endogenous promoter can be used as a fluorescence-based high-throughput screen for differentiation-modifying or antiinflammatory compounds that act via the keratinocyte.

[Some biochemical indices in chicken skin tissue during ontogenesis and under the effect of ionizing radiation]. / Deiaki biokhimichni pokaznyky u tkanynakh shkiry kruei v ontohenezi ta za diï na nykh ionizuiuchoho oprominennia,

The research shows some biochemical indices in ontogenesis in broiler hen skin tissues and also its reaction to inner introduction of different doses of radioactive 137Cs. Namely, the irradiation ionizing irradiation influences actively to the indices of nuclein changes with 1 day and 1 week aged chickens. The quantity of phosphorus of nuclein acids is less with the chickens of the third group (500 Bk pr day) than with the second group (3000 Bk/day). The quantity of soluble protein, activity of the estimated fragments, the quantity of inorganic phosphorus, the concentration of ions of magnesium with chickens of the controlled group is larger than in the studied ones. It proves the influence of radionucleid 137Cs on indices which characterize the protein-nuclein change.