Exogenous glycosaminoglycans (GAG) are able to modulate avian skin differentiation (epithelial keratinization and feather formation

Development ◽  
1984 ◽  
Vol 82 (1) ◽  
pp. 25-40
Author(s):  
E. Becchetti ◽  
G. Stabellini ◽  
A. Caruso ◽  
P. Carinci
Keyword(s):  
Hyaluronic Acid ◽  
Chick Embryo ◽  
Synthetic Medium ◽  
Calf Serum ◽  
Epithelial Differentiation ◽  
Dermal Fibroblasts ◽  
Conditioned Media ◽  
Lung Fibroblasts ◽  
Skin Explants

Several reports have suggested that mesenchymal glycosaminoglycans (GAG) may be involved in the regulatory role of epithelial differentiation. Some researchers have pointed out that exogenousGAG affects extracellular GAG accumulation. We have therefore examined the effect of added GAG on two typical processes of avian skin differentiation: keratinization and feather formation. Glycosaminoglycans, either obtained from fibroblasts cultures (conditioned media) or purified commercially available GAG were administered to 5/6-day chick embryo back skin explants. Control cultures were supported with 199 synthetic medium, chick embryo extract or calf serum. Explants have been examined by histological and histochemical procedures. Skin explants maintained in vitro for 7 days exhibited an epithelial differentiation and a dermal histochemical reactivity which were related to the composition of the culture medium. In conditioned media from dermal fibroblasts, but not from heart or lung fibroblasts, explants always exhibited keratinization. In purified-GAG-containing media, keratinization was observed with condroitinsulphates and not with hyaluronic acid. Keratinization was always related toprevalent accumulation of hyaluronic acid in the underlying mesenchyme whereas feather formation was in relation to deposits of condroitinsulphates in dermis pulp. The above findings demonstrate that exogenous GAG is able to modulate avian skin differentiation and that this regulation is linked to an influence on the mesenchymal GAG pattern.

M2 macrophages have unique transcriptomes but conditioned media does not promote profibrotic responses in lung fibroblasts or alveolar epithelial cells in vitro

2021 ◽  
Author(s):  
Elissa M Hult ◽  
Stephen James Gurczynski ◽  
Bethany B Moore
Keyword(s):  
Pulmonary Fibrosis ◽  
Alveolar Epithelial Cell ◽  
Conditioned Media ◽  
Alveolar Epithelial Cells ◽  
Lung Fibroblasts ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
Alveolar Epithelial ◽  
Fibroblast Migration

Macrophages are critical regulators of pulmonary fibrosis. Their plasticity, proximity, and ability to crosstalk with structural cells of the lung make them a key cell type of interest in the regulation of lung fibrosis. Macrophages can express a variety of phenotypes which have been historically represented through an "M1-like" to "M2-like" delineation. In this classification, M1-like macrophages are proinflammatory and have increased phagocytic capacity compared to alternatively activated M2-like macrophages that are profibrotic and are associated with wound healing. Extensive evidence in the field in both patients and animal models align pulmonary fibrosis with M2 macrophages. In this paper, we performed RNAseq to fully characterize M1 vs. M2-skewed bone marrow-derived macrophages (BMDMs) and investigated the profibrotic abilities of M2 BMDM conditioned media (CM) to promote fibroblast migration, proliferation, alveolar epithelial cell (AEC) apoptosis, and mRNA expression of key fibrotic genes in both fibroblasts and in AECs. Although M2 CM-treated fibroblasts had increased migration and M2 CM-treated fibroblasts and AECs had increased expression of profibrotic proteins over M1 CM-treated cells, all differences can be attributed to M2 polarization reagents IL-4 and IL-13 also present in the CM. Collectively, these data suggest that the profibrotic effects associated with M2 macrophage CM in vitro are attributable to effects of polarization cytokines rather than additional factors secreted in response to those polarizing cytokines.

scholarly journals Comparison of the Effect of Two Hyaluronic Acid Preparations on Fibroblast and Endothelial Cell Functions Related to Angiogenesis

Cells ◽  
2019 ◽  
Vol 8 (12) ◽  
pp. 1479 ◽  
Author(s):  
Valerio Ciccone ◽  
Marco Zazzetta ◽  
Lucia Morbidelli
Keyword(s):  
Hyaluronic Acid ◽  
Inflammatory Infiltrate ◽  
Dermal Fibroblasts ◽  
Low Molecular Weight ◽  
Aesthetic Medicine ◽  
Cell Functions ◽  
Tissue Responses ◽  
Tissue Recovery

Hyaluronic acid (HA) is used in substitutive and aesthetic medicine with various applications. Ultrapure absorbable HA (Bioregen®) and a mix of reticulated and free low molecular weight HA (Regenyal Idea Bioexpander®) (both provided by Regenyal Laboratories Srl, San Benedetto del Tronto (AP), Italy) represent a reliable hydrating device and skin filler, useful for skin blemishes, lines and wrinkles, and lip widening, respectively. The commercial products are known for their safety, but data on the molecular, cellular, and tissue responses are lacking. We aimed to evaluate the bioavailability and the pro-angiogenic features of the products Bioregen® and Bioexpander® in vitro on cultured endothelial cells (ECs) and dermal fibroblasts in vivo when injected into experimental animals. When added to fibroblasts and ECs, Bioexpander® induced cell migration. The two HA preparations were well tolerated, while a transient proangiogenic behavior of Bioexpander®, when implanted subcutaneously in mice, was found. The neovascular response was evident in the first week with higher levels of VEGF and FGF-2 before undergoing regression. In conclusion, our data strengthen the safety of HA synthetic preparations both in vitro and in vivo. Even if a proangiogenic response is documented, it is modest and transient, leading to tissue recovery and absence of an inflammatory infiltrate.

scholarly journals Novel Rotational Combination Regimen of Skin Topicals Improves Facial Photoaging: Efficacy Demonstrated in Double-Blinded Clinical Trials and Laboratory Validation

2021 ◽  
Vol 8 ◽  
Author(s):  
Lisa DiNatale ◽  
Jolanta Idkowiak-Baldys ◽  
Young Zhuang ◽  
Anthony Gonzalez ◽  
Thomas J. Stephens ◽  
...  
Keyword(s):  
Clinical Trials ◽  
Ex Vivo ◽  
Dermal Fibroblasts ◽  
Combination Regimen ◽  
Crow’S Feet ◽  
Skin Explants ◽  
Study Methodology ◽  
Double Blinded

Topical antiaging products are often a first-line intervention to counter visible signs of facial photoaging, aiming for sustained cosmetic improvement. However, prolonged application of a single active topical compound was observed clinically to lead to a plateau effect in improving facial photoaging. In view of this, we set out to reduce this effect systematically using a multi-tiered approach with laboratory evidence and clinical trials. The objective of the study was to evaluate the effects of active topical ingredients applied either alone, in combination, or in a rotational manner on modulation of facial photoaging. The study methodology included in vitro, organotypic, and ex vivo skin explants; in vivo biopsy study; as well as clinical trials. We demonstrate for the first time that a pair of known antiaging ingredients applied rotationally, on human dermal fibroblasts, maximized pro-collagen I production. Indeed, rotational treatment with retinol and phytol/glycolic acid (PGA) resulted in better efficacy than application of each active ingredient alone as shown by explants and in vivo biopsy study, with penetration of active ingredients confirmed by Raman spectroscopy. Furthermore, two split-face, randomized, double-blinded clinical trials were conducted, one for 12 months to compare treated vs. untreated and the other for 6 months followed by a 2-month regression to compare treated vs. commercially marketed products. In both studies, rotational regimen showed superior results to its matching comparison as assessed by clinical grading and image analysis of crow's feet wrinkles. In conclusion, rotational regimen using retinol and PGA is effective in treating facial photoaging signs with long-lasting benefits.

53 FREEZING BULL SEMEN IN A SYNTHETIC MEDIUM

2017 ◽  
Vol 29 (1) ◽  
pp. 134
Author(s):  
L. Gavin-Plagne ◽  
P. Bodranghien ◽  
A. Vachet ◽  
L. Commin ◽  
S. Buff ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Synthetic Medium ◽  
Calf Serum ◽  
Egg Yolk ◽  
Wilcoxon Test ◽  
Computer Assisted ◽  
Sperm Cells ◽  
Bull Semen ◽  
Significant Difference

Animal-derived products are currently used to cryopreserve sperm cells. However, these products represent potential risks of contamination by pathogens. Optidyl® (IMV Technologies, L’Aigle, France), containing egg yolk, is a reference product in Europe used routinely in bovine insemination centers. Commercial media such as soy lecithin or liposome-based media have been used to replace extenders containing products derived from animals. However, their protective effect could be called into question because of their non-synthetic or unstable properties. Despite these innovative extenders on the market, it might be necessary for sanitary reasons to cryopreserve bull semen in a stable and synthetic extender. CRYO3 (Stem Alpha, Saint-Genis l’Argentière, France), a serum-free and protein-free medium used for cryopreserving somatic and stem human cells, is a potential medium to cryopreserve reproductive cells. Recently, CRYO3 improved cryopreservation of in vitro-produced bovine embryos compared with fetal calf serum and BSA-based media. Thus, it could be interesting to test this medium on sperm cells. The objective of this study was to compare 2 in vitro freezing media on bull semen: a commercial egg yolk-based medium and a synthetic medium containing 20% CRYO3. Sperm from 5 bulls were collected in Auriva station (Brindas, France). A sample of each ejaculate was used to assess fresh semen quality. The remaining sperm of each bull was split and diluted in 2 media: Optidyl® and a CRYO3-based medium. Semen was equilibrated at least 4 h at 4°C before being packaged in 0.25-mL French straws, and then frozen into a programmable freezer and plunged into liquid nitrogen. Osmolarity and pH were respectively 1462 mOsm/kg and 6.9 for Optidyl® and 1286 mOsm/kg and 6.8 for CRYO3 medium. Viability (with SYBR-14 and propidium iodide) and high mitochondrial membrane potential (hMMP) (with JC-1) were assessed using flow cytometry. A hypo-osmotic swelling test was performed to assess functional membrane integrity (FMI). The motility parameters were evaluated by computer-assisted sperm analysis. Statistical analysis was performed using Wilcoxon test with the R software. Fresh sperm showed 52% viability, 64% hMMP, 74% FMI, and 62 and 76% progressive and total motility, respectively. For all parameters measured, no significant difference was observed between extenders and between fresh and frozen–thawed sperm (P > 0.05). However, Optidyl® showed clearly better survival than CRYO3 (45% v. 16% for viability, 58% v. 20% for hMMP, 67% v. 25% for FMI, 51% v. 14% for progressive motility and 72% v. 32% for total motility, respectively). These results show that it is possible to freeze bovine semen in synthetic extender though the low survival rate after freezing-thawing. Indeed, it is known that motility and flow cytometry parameters are not necessarily good indicators of fertility. Artificial inseminations will be done to verify the fertility of the sperm cryopreserved in CRYO3. Repetitions with more bulls from different breeds will be performed to complete this preliminary work. This work was supported by grant CRB-ANIM ANR-11-INBS-0003.

Ionic calcium levels in oviduct explant-conditioned media from an Australian marsupial, the brushtail possum (Trichosurus vulpecula) and its relevance to in vitro fertilization

Zygote ◽  
2003 ◽  
Vol 11 (4) ◽  
pp. 285-291 ◽  
Author(s):  
K.S. Sidhu ◽  
K.E. Mate ◽  
F.C. Molinia ◽  
D.K. Berg ◽  
J.C. Rodger
Keyword(s):  
Calf Serum ◽  
Explant Culture ◽  
Conditioned Media ◽  
Brushtail Possum ◽  
Trichosurus Vulpecula ◽  
Large Variability ◽  
Vitro Fertilization ◽  
Ionic Calcium ◽  
Australian Marsupial

Gametes from the brushtail possum (Trichosurus vulpecula), an Australian marsupial, require exposure to oviductal cells and/or their secretions before sperm binding and penetration of the zona pellucida can occur. Sperm-egg fusion, the next critical step in fertilization has not previously been reported in vitro. Here we describe the refinement of an oviduct epithelial cell (OEC) explant culture system using two different media to obtain in vitro sperm-egg fusion in the brushtail possum for the first time. Conditioned media from OEC explant cultures were supplemented with either 1% fetal calf serum (FCS) or 1mg/ml polyvinyl alcohol and used for co-culture of epididymal sperm and superovulated eggs. Under these conditions zona penetration rates varied from 0 to 46% and sperm-egg fusion from 0 to 20%. Analysis of explant conditioned media indicated that qualitative and quantitative differences between batches could account, at least partially, for the large variability in zona penetration rates. Conditioned media that contained approximately 1mM of ionic calcium were most effective for achieving sperm capacitation, zona binding, and penetration and sperm-egg fusion. The reorientation of the sperm head to T-shape, an indicator of capacitation in the brushtail possum, was closely linked with the concentration of calcium present in vitro.

scholarly journals Exosomes of Adipose-derived Stem Cells Conditioned Media Promotes Retinoblastoma and Forkhead-Box M1 Protein Expression

2021 ◽  
Vol 9 (A) ◽  
pp. 422-427
Author(s):  
Sinta Murlistyarini ◽  
Lulus Putri Aninda ◽  
Sri Widyarti ◽  
Agustina Tri Endharti ◽  
Teguh Wahju Sardjono
Keyword(s):  
Protein Expression ◽  
Dermal Fibroblasts ◽  
Conditioned Media ◽  
Control Group ◽  
Forkhead Box ◽  
Forkhead Box M1 ◽  
Post Test ◽  
Foxm1 Protein ◽  
Specific Proteins

BACKGROUND: In the senescence process, the retinoblastoma (Rb) protein binds to E2F in hypophosphorylated conditions, preventing the cell to enter the S-phase in the cell cycle. Human Forkhead Box M1 (FOXM1) protein, key regulator G1/S and G2/M phases, decreases in the senescence process. Many studies have been carried out to reverse this system, one of which used exosomes of adipose-derived stem c ells conditioned media (ADSC-CM). These exosomes contain a variety of specific proteins which have pro-proliferation properties, however, little is known on the role of these exosomes toward the change of phosphorylated Rb and FOXM1. AIM: This study aims to find out the involvement of exosomes of ADSC-CM on these two proteins on senescence human dermal fibroblasts (HDFs). METHODS: In vitro experiment was undergone randomization sample and non-blinded pre-/post-test control group. The primary culture of senescent HDFs was transfected with exosomes of ADSC-CM; then, its effect on migration and senescence reversal was observed through analyzing Sa-β-gal, Rb, and FOXM1 protein expression. RESULTS: The expression of Sa-β-gal was higher in the control group. Our result demonstrated the exosome of ADSC-CM significantly induced the expression of Rb and FOXM1 protein in senescent HDFs (p < 0.05). CONCLUSION: It proved that exosomes of ADSC-CM could shift the senescent fibroblast into metabolically active cells.

Necessary amino acids and vitamins for the growth of human diploid fibroblasts

1979 ◽  
Vol 40 (1) ◽  
pp. 281-291
Author(s):  
J. Litwin
Keyword(s):  
Amino Acids ◽  
Growth Rate ◽  
Folic Acid ◽  
Cell Growth ◽  
Calf Serum ◽  
Life Time ◽  
Lung Fibroblasts ◽  
Human Diploid Fibroblasts ◽  
Support Cell

Only 2 amino acids and one vitamin were found to be essential for the growth of human embryonic diploid lung fibroblasts when 10% undialysed calf serum was used as a medium supplement. These amino acids were either glutamine + cysteine or serine + homocysteine. Replacing cysteine or homocysteine with cystine or homocystine, respectively, reduced growth. The growth rate in the glutamine + cysteine medium was slightly less than that in Eagle's medium, but the in vitro life time was similar. Folic acid was the only vitamin needed to support cell growth in vitro. The addition of other vitamins had no stimulatory effect with the possible exception of nicotinamide. When other amino acids were added to glutamine + cysteine none showed stimulatory effects but tryptophan was either toxic or inhibitory for the 3 human diploid strains examined. Serine was inhibitory for WI-38 but not for MRC5 cells. Subtle nutritional differences appear to exist between fibroblasts of the same type obtained from different embryos.

Organ culture of the anterior pituitary in a synthetic medium. An immunocytochemical study

1983 ◽  
Vol 102 (1) ◽  
pp. 35-41 ◽  
Author(s):  
M. Bégeot ◽  
J. Y. Li ◽  
M. P. Dubois ◽  
P. M. Dubois
Keyword(s):  
Organ Culture ◽  
Anterior Pituitary ◽  
Synthetic Medium ◽  
Calf Serum ◽  
Cell Types ◽  
Foetal Calf Serum ◽  
Pituitary Cells ◽  
The Third ◽  
Different Cell Types

Abstract. Anterior pituitaries removed from human foetuses and rat neonates were maintained in organ culture after using three different media. A comparative immunocytological study of the different cell-types was made in the three media used. In the first medium containing foetal calf serum the different cell-types were present but their frequency in the cultured explants depended upon their representation in the tissue at the beginning of the culture. In the medium without any adjunction, the most characteristic feature was the shrinkage of the cultured tissue. In the third medium in which foetal calf serum was replaced by insulin the evolution of the explants was the same as in the first medium. All the cell-types were represented with a particular development of gonadotrophic cells. In conclusion it seemed that the third synthetic medium can be used in order to study anterior pituitary cells in vitro.

Pharmaceutical evaluation of atorvastatin-loaded nanostructured lipid carriers incorporated into the gelatin/hyaluronic acid/polycaprolactone scaffold for the skin tissue engineering

2020 ◽  
pp. 088532822097076
Author(s):  
Mahsa Ahmadi ◽  
Mehdi Mehdikhani ◽  
Jaleh Varshosaz ◽  
Shadi Farsaei ◽  
Hadis Torabi
Keyword(s):  
Particle Size ◽  
Hyaluronic Acid ◽  
Drug Release ◽  
Zeta Potential ◽  
Mtt Assay ◽  
Dermal Fibroblasts ◽  
Skin Regeneration ◽  
Nanostructured Lipid Carriers ◽  
In Vitro Degradation

In this study, gelatin/hyaluronic acid (HA) scaffolds containing different amounts of atorvastatin-loaded nanostructured lipid carriers (NLCs) coated entirely with polycaprolactone (PCL) film were fabricated for skin regeneration. 12 atorvastatin-loaded NLCs formulations were synthesized, and particle size, zeta potential, drug entrapment efficiency (EE), and drug release of the formulations were determined. The optimum freeze-dried atorvastatin-loaded NLCs were added in 3 different weight percentages to the gelatin and HA membranous scaffolds. Thereafter, the membranes were coated entirely by a thin layer of the PCL. They were characterized, and then mechanical properties, in vitro degradation and in vitro drug release were assessed. Moreover, human dermal fibroblasts (HDF) were cultured on the prepared nanocomposite scaffolds in order to investigate the cytotoxicity by the MTT assay after the first day, third day, and fifth day. Results revealed that the most favorable atorvastatin-loaded NLCs had 99.54 nm average particle size, −24.30 mV zeta potential, 97.98% EE, and 75.24% drug release within 237 hrs. Mechanical tests indicated that all the three scaffolds had approximately a 90 MPa elastic modulus which was more than two-fold of tensile modulus of normal human skin. The in vitro degradation test demonstrated that the membranes were degraded up to 98% after 5 days, and the scaffolds drug release efficiency (DRE) was in a range of 75–79% during those 5 days. The MTT assay results confirmed the cytocompatibility of the scaffolds. The scaffold containing 54.1 wt% NCLs was the optimum sample (S3). Scanning Electron Microscopy (SEM) images of the latter one showed the uniform distribution of the NLCs with an average size of 150 nm, and the images of cultured HDF illustrated the good cell attachment. In conclusion, suitable physicochemical and biological properties of the novel gelatin/HA/PCL nanocomposite scaffold containing 54.1 wt% atorvastatin-loaded NLCs (S3) can be a good candidate for skin regeneration.

scholarly journals Ulva intestinalis Protein Extracts Promote In Vitro Collagen and Hyaluronic Acid Production by Human Dermal Fibroblasts

Molecules ◽  
2020 ◽  
Vol 25 (9) ◽  
pp. 2091 ◽  
Author(s):  
Justine Bodin ◽  
Amandine Adrien ◽  
Pierre-Edouard Bodet ◽  
Delphine Dufour ◽  
Stanislas Baudouin ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Acid Production ◽  
Biochemical Characterization ◽  
Skin Aging ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
Free Process ◽  
Ulva Intestinalis ◽  
Hyaluronic Acid Production

With the increase in life expectancy, reducing the visible signs of skin aging has become a major issue. A reduction in collagen and hyaluronic acid synthesis by fibroblasts is a feature of skin aging. The green seaweed, Ulva intestinalis, is an abundant and rich source of nutrients, especially proteins and peptides. The aim of this study was to assess the potential cosmetic properties of a protein fraction from Ulva intestinalis (PROT-1) containing 51% of proteins and 22% of polysaccharides, and its enzymatic peptide hydrolysates on human dermal fibroblasts. PROT-1 was extracted using a patented acid- and solvent-free process (FR2998894 (B1)). The biochemical characterization and chromatographic analysis showed a main set of proteins (25 kDa). To demonstrate the anti-aging potential of PROT-1, fibroblast proliferation and collagen and hyaluronic acid production were assessed on fibroblast cell lines from donors aged 20 years (CCD-1059Sk) and 46 years (CCD-1090Sk). PROT-1 induced a significant increase in collagen and hyaluronic acid production per cell, and a reduction in cell proliferation without increasing cell mortality. These effects were reversed after protein hydrolysis of PROT-1, showing the central role of proteins in this promising anti-aging property.

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