Attachment and Proliferation of Human Dermal Fibroblasts onto ECM-Immobilized PLGA Films

2005 ◽  
Vol 288-289 ◽  
pp. 291-294
Author(s):  
Hyun Joo Son ◽  
Dong Wook Han ◽  
H.H. Kim ◽  
Hee Joong Kim ◽  
In Seop Lee ◽  
...  
Keyword(s):  
Cell Attachment ◽  
Dermal Fibroblast ◽  
Supporting Cell ◽  
Human Dermal Fibroblast ◽  
Dermal Fibroblasts ◽  
Water Soluble ◽  
Seeding Density ◽  
Tetrazolium Salt ◽  
Cell Seeding ◽  
Plga Film

In this study, human dermal fibroblast behaviors onto non-porous PLGA (75:25) films immobilized with 1, 10 and 100 µg/ml collagen (CN) or fibronectin (FN) were investigated according to different cell-seeding densities (1,000, 10,000 and 100,000 cells/ml). Cell attachment and proliferation were assessed using water soluble tetrazolium salt. The results indicated that 1 µg/ml of FN-immobilized PLGA film demonstrated significantly (p < 0.05) superior cellular attachment to the intact PLGA film after 4 hr of incubation. Moreover, the number of attached cells was shown to be directly proportional to that of initially seeded cells. After 48 hr, the cells showed significantly (p < 0.05) higher proliferation onto 1 or 10 µg/ml of FN-immobilized PLGA films than onto other PLGA films, regardless of the initial cell-seeding density. In terms of CN-immobilization, cell proliferation was appreciably increased but it was relatively lower than FN-immobilization. These results suggested that ECM-immobilization can enhance the cell affinity of hydrophobic scaffolds and be used to potential applications for tissue engineering by supporting cell growth.

scholarly journals Collagen Film Activation with Nanoscale IKVAV-Capped Dendrimers for Selective Neural Cell Response

Nanomaterials ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1157
Author(s):  
Jessica J. Kim ◽  
Daniel V. Bax ◽  
Robert Murphy ◽  
Serena M. Best ◽  
Ruth E. Cameron
Keyword(s):  
Peripheral Nerve ◽  
Cellular Response ◽  
Cell Attachment ◽  
Dermal Fibroblast ◽  
Cellular Responses ◽  
Potential Method ◽  
Human Dermal Fibroblast ◽  
Dermal Fibroblasts ◽  
Fibroblast Proliferation ◽  
Fibroblast Attachment

Biocompatible neural guidance conduits are alternatives to less abundant autologous tissue grafts for small nerve gap injuries. To address larger peripheral nerve injuries, it is necessary to design cell selective biomaterials that attract neuronal and/or glial cells to an injury site while preventing the intrusion of fibroblasts that cause inhibitory scarring. Here, we investigate a potential method for obtaining this selective cellular response by analysing the responses of rat Schwann cells and human dermal fibroblasts to isoleucine-lysine-valine-alanine-valine (IKVAV)-capped dendrimer-activated collagen films. A high quantity of nanoscale IKVAV-capped dendrimers incorporated onto pre-crosslinked collagen films promoted rat Schwann cell attachment and proliferation, and inhibited human dermal fibroblast proliferation. In addition, while pre-crosslinked dendrimer-activated films inhibited fibroblast proliferation, non-crosslinked dendrimer-activated films and films that were crosslinked after dendrimer-activation (post-crosslinked films) did not. The different cellular responses to pre-crosslinked and post-crosslinked films highlight the importance of having fully exposed, non-covalently bound biochemical motifs (pre-crosslinked films) directing certain cellular responses. These results also suggest that high concentrations of nanoscale IKVAV motifs can inhibit fibroblast attachment to biological substrates, such as collagen, which inherently attract fibroblasts. Therefore, this work points toward the potential of IKVAV-capped dendrimer-activated collagen biomaterials in limiting neuropathy caused by fibrotic scarring at peripheral nerve injury sites.

Abstract 189: HuR, RNA Stabilizing Protein, Regulation of Pluripotency and Differentiation in iPSCs

2017 ◽  
Vol 121 (suppl_1) ◽  
Author(s):  
Sahana Suresh Babu ◽  
Johnson Rajasingh ◽  
Wing Tak Wong ◽  
Prasanna Krishnamurthy
Keyword(s):  
Rna Binding ◽  
Rna Binding Proteins ◽  
Dermal Fibroblast ◽  
Critical Role ◽  
Human Dermal Fibroblast ◽  
Dermal Fibroblasts ◽  
General Observation ◽  
Transcript Stability ◽  
Differentiated Cells ◽  
Induced Pluripotent

Background: The Hu family of RNA-binding proteins, HuR (also known as ELAVL1 or human embryonic lethal abnormal vision-like protein), binds to the 3’-untranslated region of mRNAs and regulates transcript stability and translation. Global deletion of HuR is embryonically lethal in mice and plays a critical role in progenitor cell survival and biology. Induced-pluripotent stem cells (iPSC) have distinct transcriptional machinery for the maintenance of pluripotency and achievement of differentiation. However, the exact role of HuR in pluripotency or differentiation of iPSC to cardiomyocytes (iCM) remains unclear. Methods: HuR knockdown in human dermal fibroblast-derived iPSCs was achieved by CRISPR/Cas9 or lentiviral shRNA transduction and subsequently differentiated into cardiomyocytes (iCM). Then, the expression of HuR, pluripotency and cardiomyocyte markers were evaluated on days 0, 1, 3, 6, 8 and 17 following the initiation of differentiation. Results: At basal level, HuR expression was higher in the iPSCs compared to dermal fibroblasts. Upon differentiation of iPSCs into iCM, HuR mRNA expression gradually reduced with significantly lower levels on day 17. As expected, pluripotency markers gradually reduced upon differentiation with significantly lower levels from day 6 onwards. We observed a corresponding increase in ISL1, MESP1 (mesoderm/cardiac progenitor markers) from day 3 through day 8 with a steep fall from day 8 to day 17. This was associated with Myosin light chain-2V and GATA4 expression increases from day 8 through day 17. Interestingly, knockdown of HuR resulted in clumps of colonies with differentiated cells and a corresponding increase in cardiac-troponin positive cells. However, as a general observation, HuR knockdown reduced beating intensity compared to wild type cells. Conclusions: Based on these data, we could speculate that HuR might be necessary for maintenance of pluripotency and loss of which renders cells to differentiate in culture. HuR knockdown yields higher number of c-troponin positive cells but its effect on functional maturity of iCM needs to be further evaluated.

scholarly journals Wound Healing Potential of Spirulina Protein on CCD-986sk Cells

Marine Drugs ◽  
2019 ◽  
Vol 17 (2) ◽  
pp. 130
Author(s):  
Ping Liu ◽  
Jeong-Wook Choi ◽  
Min-Kyeong Lee ◽  
Youn-Hee Choi ◽  
Taek-Jeong Nam
Keyword(s):  
Wound Healing ◽  
Dermal Fibroblast ◽  
Fibroblast Cell ◽  
Underlying Mechanism ◽  
Human Dermal Fibroblast ◽  
Dermal Fibroblasts ◽  
Cyclin Dependent Kinase ◽  
Pi3k Inhibitor Ly294002 ◽  
And Migration ◽  
Proliferation And Migration

Wound healing is a dynamic and complex process. The proliferation and migration of dermal fibroblasts are crucial for wound healing. Recent studies have indicated that the extracts from Spirulina platensis have a positive potential for wound healing. However, its underlying mechanism is not fully understood. Our previous study showed that spirulina crude protein (SPCP) promoted the viability of human dermal fibroblast cell line (CCD-986sk cells). In this study, we further investigated the wound healing effect and corresponding mechanisms of SPCP on CCD-986sk cells. Bromodeoxyuridine (BrdU) assay showed that SPCP promoted the proliferation of CCD-986sk cells. The wound healing assay showed that SPCP promoted the migration of CCD-986sk cells. Furthermore, cell cycle analysis demonstrated that SPCP promoted CCD-986sk cells to enter S and G2/M phases from G0/G1 phase. Western blot results showed that SPCP significantly upregulated the expression of cyclin D1, cyclin E, cyclin-dependent kinase 2 (Cdk2), cyclin-dependent kinase 4 (Cdk4), and cyclin-dependent kinase 6 (Cdk6), as well as inhibited the expression of CDK inhibitors p21 and p27 in CCD-986sk cells. In the meanwhile, SPCP promoted the phosphorylation and activation of phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt). However, the phosphorylation of Akt was significantly blocked by PI3K inhibitor (LY294002), which in turn reduced the SPCP-induced proliferation and migration of CCD-986sk cells. Therefore, the results presenting in this study suggested that SPCP can promote the proliferation and migration of CCD-986sk cells; the PI3K/Akt signaling pathway play a positive and important role in these processes.

scholarly journals Dermal Olfactory Receptor OR51B5 Is Essential for Survival and Collagen Synthesis in Human Dermal Fibroblast (Hs68 Cells)

2021 ◽  
Vol 22 (17) ◽  
pp. 9273
Author(s):  
Bomin Son ◽  
Wesuk Kang ◽  
Soyoon Park ◽  
Dabin Choi ◽  
Taesun Park
Keyword(s):  
Cell Survival ◽  
Collagen Synthesis ◽  
Dermal Fibroblast ◽  
Olfactory Receptors ◽  
Camp Response Element Binding ◽  
Human Dermal Fibroblast ◽  
Skin Aging ◽  
Dermal Fibroblasts ◽  
Nasal Tissue ◽  
Extracellular Matrix Components

Skin dermis comprises extracellular matrix components, mainly collagen fibers. A decrease in collagen synthesis caused by several factors, including ultraviolet (UV) irradiation and stress, eventually causes extrinsic skin aging. Olfactory receptors (ORs) were initially considered to be specifically expressed in nasal tissue, but several ORs have been reported to be present in other tissues, and their biological roles have recently received increasing attention. In this study, we aimed to characterize the role of ORs in cell survival and collagen synthesis in dermal fibroblasts. We confirmed that UVB irradiation and dexamethasone exposure significantly decreased cell survival and collagen synthesis in Hs68 dermal fibroblasts. Moreover, we demonstrated that the mRNA expression of 10 ORs detectable in Hs68 cells was significantly downregulated in aged conditions compared with that in normal conditions. Thereafter, by individual knockdown of the 10 candidate ORs, we identified that only OR51B5 knockdown leads to a reduction of cell survival and collagen synthesis. OR51B5 knockdown decreased cAMP levels and dampened the downstream protein kinase A/cAMP-response element binding protein pathway, downregulating the survival- and collagen synthesis-related genes in the dermal fibroblasts. Therefore, OR51B5 may be an interesting candidate that plays a role in cell survival and collagen synthesis.

scholarly journals Effect of MMP/TIMP Balancing of Cynoglossus semilaevis Shell Extracts on Skin Protection

Fishes ◽  
2021 ◽  
Vol 6 (3) ◽  
pp. 34
Author(s):  
Soo-Cheol Choi ◽  
In-Ah Lee
Keyword(s):  
Skin Diseases ◽  
Dermal Fibroblast ◽  
Functional Materials ◽  
Human Dermal Fibroblast ◽  
Cynoglossus Semilaevis ◽  
Dermal Fibroblasts ◽  
Skin Tissue ◽  
Dependent Manner ◽  
Functional Material ◽  
Concentration Dependent Manner

Cynoglossus semilaevis shell is a by-product of the Cynoglossus semilaevis, a species of fish mainly distributed along the west coast of Korea. As its skin is very tough and difficult to process, it is not useful as food. For this reason, most of it is discarded except for a small amount that is used as feed, which results in environmental pollution. Considering this, there is a need for research on the development of functional materials using Cynoglossus semilaevis shell. This study focused on the mechanism of in vitro expression function of Cynoglossus semilaevis shell extract (CSE) for skin tissue in human dermal fibroblasts that induced or did not induce wrinkles by UV-B irradiation and aims to use it as a functional material for human skin beauty or wrinkle improvement through extraction and purification. According to the ELISA results using human dermal fibroblast cells, CSE reduced MMP-1 and elastase activity by up to 21.89% and 12.04%, respectively, in a concentration-dependent manner, and increased PIP synthesis by up to 62.24% in a concentration-dependent manner. The RT-PCR test results using mRNA showed the MMP-1, 2, and 3 expression levels were suppressed in the CSE-treated group compared to the UVB-induced group and caused a concentration-dependent increase in TIMP-1 in the CSE-treat group. These results suggest that CSE can maintain and improve skin tissue conditions through MMP/TIMP balancing in human dermal fibroblast cell lines and indicate its potential as a functional material for improving skin diseases and suppressing photo-aging.

scholarly journals Pentafluorophenyl Platinum(II) Complexes of PTA and its N-Allyl and N-Benzyl Derivatives: Synthesis, Characterization and Biological Activity

Materials ◽  
2019 ◽  
Vol 12 (23) ◽  
pp. 3907 ◽  
Author(s):  
Sgarbossa ◽  
Śliwińska-Hill ◽  
Guedes da ilva ◽  
Bażanów ◽  
Pawlak ◽  
...  
Keyword(s):  
Dermal Fibroblast ◽  
Human Dermal Fibroblast ◽  
Phosphine Ligands ◽  
Water Soluble ◽  
Breast Adenocarcinoma ◽  
Cytotoxic Activities ◽  
Cervix Carcinoma ◽  
X Ray Diffraction ◽  
Normal Human Dermal Fibroblast ◽  
Human Lung Carcinoma

From the well-known 1,3,5-triaza-phosphaadamantane (PTA, 1a), the novel N-allyl and N-benzyl tetrafuoroborate salts 1-allyl-1-azonia-3,5-diaza-7-phosphaadamantane (APTA(BF4), 1b) and 1-benzyl-1-azonia-3,5-diaza-7-phosphaadamantane (BzPTA(BF4), 1c) were obtained. These phosphines were then allowed to react with (Pt(μ-Cl)(C6F5)(tht))2 (tht = tetrahydrothiophene) affording the water soluble Pt(II) complexes trans-(PtCl(C6F5)(PTA)2) (2a) and its bis-cationic congeners trans-(PtCl(C6F5)(APTA)2)(BF4)2 (2b) and trans-(PtCl(C6F5)(BzPTA)2)(BF4)2 (2c). The compounds were fully characterized by multinuclear NMR, ESI-MS, elemental analysis and (for 2a) also by single crystal X-ray diffraction, which proved the trans configuration of the phosphine ligands. Furthermore, in order to evaluate the cytotoxic activities of all complexes the normal human dermal fibroblast (NHDF) cell culture were used. The antineoplastic activity of the investigated compounds was checked against the human lung carcinoma (A549), epithelioid cervix carcinoma (HeLa) and breast adenocarcinoma (MCF-7) cell cultures. Interactions between the complexes and human serum albumin (HSA) using fluorescence spectroscopy and circular dichroism spectroscopy (CD) were also investigated.

scholarly journals In Vitro Bioeffects of Polyelectrolyte Multilayer Microcapsules Post-Loaded with Water-Soluble Cationic Photosensitizer

Pharmaceutics ◽  
2020 ◽  
Vol 12 (7) ◽  
pp. 610 ◽  
Author(s):  
Alexey V. Ermakov ◽  
Roman A. Verkhovskii ◽  
Irina V. Babushkina ◽  
Daria B. Trushina ◽  
Olga A. Inozemtseva ◽  
...  
Keyword(s):  
Targeted Delivery ◽  
Dermal Fibroblast ◽  
Human Dermal Fibroblast ◽  
Water Soluble ◽  
Zinc Phthalocyanine ◽  
Bacterial Strains ◽  
Normal Human Dermal Fibroblast ◽  
Drug Dosing ◽  
Normal Human

Microencapsulation and targeted delivery of cytotoxic and antibacterial agents of photodynamic therapy (PDT) improve the treatment outcomes for infectious diseases and cancer. In many cases, the loss of activity, poor encapsulation efficiency, and inadequate drug dosing hamper the success of this strategy. Therefore, the development of novel and reliable microencapsulated drug formulations granting high efficacy is of paramount importance. Here we report the in vitro delivery of a water-soluble cationic PDT drug, zinc phthalocyanine choline derivative (Cholosens), by biodegradable microcapsules assembled from dextran sulfate (DS) and poly-l-arginine (PArg). A photosensitizer was loaded in pre-formed [DS/PArg]4 hollow microcapsules with or without exposure to heat. Loading efficacy and drug release were quantitatively studied depending on the capsule concentration to emphasize the interactions between the DS/PArg multilayer network and Cholosens. The loading data were used to determine the dosage for heated and intact capsules to measure their PDT activity in vitro. The capsules were tested using human cervical adenocarcinoma (HeLa) and normal human dermal fibroblast (NHDF) cell lines, and two bacterial strains, Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli. Our results provide compelling evidence that encapsulated forms of Cholosens are efficient as PDT drugs for both eukaryotic cells and bacteria at specified capsule-to-cell ratios.

scholarly journals Electrospun PCL-Surgihoney meshes for skin wound healing applications

2020 ◽  
Vol 318 ◽  
pp. 01029
Author(s):  
Enes Aslan
Keyword(s):  
Wound Dressing ◽  
Cell Attachment ◽  
Positive Impact ◽  
Dermal Fibroblast ◽  
Healing Process ◽  
Human Dermal Fibroblast ◽  
Skin Wound ◽  
Regulatory Function ◽  
Skin Wound Healing ◽  
Skin Healing

Skin is a complex and very important tissue, playing a significant protective and regulatory function. It is also prone to a large number of wounds and defects due to external factors such as temperature, chemical agents, and radiation. Accelerating the skin healing process and minimizing the risk of infection is a relevant research challenge. This paper investigates a novel wound dressing based on polycaprolactone (PCL), a synthetic biocompatible and biodegradable polymer, and honey-Surgihoney® (SH). Solution electrospinning was used to produce the wound dressing meshes. Different polymer solutions were prepared by mixing PCL and SH with acetic acid. Human dermal fibroblast were used to assess the biological characteristics of the electrospun meshes. Results show that the presence of SH1 has a positive impact on cell attachment and proliferation.

scholarly journals Dynamic transcriptome analysis reveals signatures of paradoxical effect of vemurafenib on human dermal fibroblasts

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Eyleen Corrales ◽  
Ella Levit-Zerdoun ◽  
Patrick Metzger ◽  
Silke Kowar ◽  
Manching Ku ◽  
...  
Keyword(s):  
Cancer Progression ◽  
Dermal Fibroblast ◽  
Acquired Resistance ◽  
Braf Inhibitor ◽  
Mek Inhibitor ◽  
Human Dermal Fibroblast ◽  
Dermal Fibroblasts ◽  
Paradoxical Effect ◽  
Combination Therapies ◽  
Stromal Component

Abstract Background Vemurafenib (PLX4032) is one of the most frequently used treatments for late-stage melanoma patients with the BRAFV600E mutation; however, acquired resistance to the drug poses as a major challenge. It remains to be determined whether off-target effects of vemurafenib on normal stroma components could reshape the tumor microenvironment in a way that contributes to cancer progression and drug resistance. Methods By using temporally-resolved RNA- and ATAC-seq, we studied the early molecular changes induced by vemurafenib in human dermal fibroblast (HDF), a main stromal component in melanoma and other tumors with high prevalence of BRAFV600 mutations. Results Transcriptomics analyses revealed a stepwise up-regulation of proliferation signatures, together with a down-regulation of autophagy and proteolytic processes. The gene expression changes in HDF strongly correlated in an inverse way with those in BRAFV600E mutant malignant melanoma (MaMel) cell lines, consistent with the observation of a paradoxical effect of vemurafenib, leading to hyperphosphorylation of MEK1/2 and ERK1/2. The transcriptional changes in HDF were not strongly determined by alterations in chromatin accessibility; rather, an already permissive chromatin landscape seemed to facilitate the early accessibility to MAPK/ERK-regulated transcription factor binding sites. Combinatorial treatment with the MEK inhibitor trametinib did not preclude the paradoxical activation of MAPK/ERK signaling in HDF. When administered together, vemurafenib partially compensated for the reduction of cell viability and proliferation induced by trametinib. These paradoxical changes were restrained by using the third generation BRAF inhibitor PLX8394, a so-called paradox breaker compound. However, the advantageous effects on HDF during combination therapies were also lost. Conclusions Vemurafenib induces paradoxical changes in HDF, enabled by a permissive chromatin landscape. These changes might provide an advantage during combination therapies, by compensating for the toxicity induced in stromal cells by less specific MAPK/ERK inhibitors. Our results highlight the relevance of evaluating the effects of the drugs on non-transformed stromal components, carefully considering the implications of their administration either as mono- or combination therapies.

scholarly journals Bovine vitreous gel can reactivate replicative senescence of human dermal fibroblast

2018 ◽  
Vol 23 (1) ◽  
pp. 48
Author(s):  
Maria Vianny Sansan ◽  
Sunardi Radiono ◽  
Muhamad Eko Irawanto ◽  
Yohanes Widodo Wirohadidjojo
Keyword(s):  
Hyaluronic Acid ◽  
Replicative Senescence ◽  
Dermal Fibroblast ◽  
Human Dermal Fibroblast ◽  
Dermal Fibroblasts ◽  
Influential Factor ◽  
Proliferation Index ◽  
Potential Candidate ◽  
Chronic Ulcers ◽  
Tgf Β1

The most influential factor in the poor healing of chronic ulcers is replicative senescence of fibroblasts that are unresponsive to TGF-β1 stimulation. Cellular replicative senescence can be induced by cultivating normal human dermal fibroblasts (HDFs) in a serum-starved medium. In addition, increasing microenviroment mechanical forces by hyaluronic acid can ameliorate the TGF-β1 signaling of these senescent cells. One of natural resources of hyaluronic acid is bovine vitreous gel. In order to evaluate the effect of bovine-vitreous gel on replicative senescence of fibroblasts, we used various levels of bovine vitreous gel diluted in Dulbecco’s modified Eagle’s medium to stimulate cellular activities of serum-starved HDFs. Those cellular activities were compared to the control media, standardized hyaluronic acid, and to normal HDFs. Our results show that replicative senescence of HDFs treated with 50% bovine vitreous gel exhibited a significantly higher proliferation index, migration rate, and collagen deposition than those cultured in control media, and they displayed an equal level of cellular activity with the HDFs exposed only to standardized hyaluronic acid. We concluded that bovine vitreous gel can be used to stimulate replicative senescence of HDFs and therefore a potential candidate material to stimulate healing of chronic ulcers.

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