Proliferation Of Skin Fibroblasts Research Articles

Overexpression of miR-340-5p Inhibits Skin Fibroblast Proliferation by Targeting Kruppel-like Factor 2.

MicroRNA (miR)-340-5p has been identified to play a key role in several cancers. However, the function of miR-340-5p in skin fibroblasts remains largely unknown. Gain of function experiments were performed by infecting normal skin fibroblast cells with a lentivirus carrying 22-bp miR-340-5p. Cell proliferation was detected by Cell Counting Kit-8 (CCK-8) assay. To uncover the mechanisms, mRNA-seq was used. Differentially expressed mRNAs were further determined by Gene Ontology and KEGG pathway analyses. The protein levels were analysed by Western blotting. A dual-luciferase reporter assay was used to detect the direct binding of miR-340-5p with the 3'UTR of Kruppel-like factor 2 (KLF2). MiR-340-5p lentivirus infection suppressed normal skin fibroblast proliferation. The mRNAseq data revealed that 41 mRNAs were differentially expressed, including 22 upregulated and 19 downregulated transcripts in the miR-340-5p overexpression group compared with those in the control group. Gene Ontology and KEGG pathway analyses revealed that miR-340-5p overexpression correlated with the macromolecule biosynthetic process, cellular macromolecule biosynthetic process, membrane, and MAPK signalling pathway. Bioinformatics analysis and luciferase reporter assays showed that miR-340-5p binds to the 3'UTR of KLF2. Forced expression of miR-340-5p decreased the expression of KLF2 in normal skin fibroblasts. Overexpression of KLF2 restored skin fibroblast proliferation in the miR-340-5p overexpression group. This study demonstrates that miR-340-5p may suppress skin fibroblast proliferation, possibly through targeting KLF2. These findings could help us understand the function of miR-340-5p in skin fibroblasts. miR-340-5p could be a therapeutic target for preventing scarring.

Regenerative effects of spring water-derived bacterial lysates on human skin fibroblast in in vitro culture: preliminary results.

Objective:Previous studies have shown regenerative power of the skin with Comano (Trento, Italy) spring water and resident non-pathogenic microflora. This study investigated the action of bacterial lysates that were isolated from Comano spring water on in vitro culture of human skin fibroblasts.Methods:For this study, we selected the following four bacterial lysates: L1 (closest relative: Rudaea cellulosilytica), L2 (closest relative: Mesorhizobium erdmanii), L3 (closest relative: Herbiconiux ginsengi), and L4 (closest relative: Fictibacillus phosphorivorans). Human fibroblasts were cultured under Dulbecco’s modified Eagle’s medium (DMEM) with bacterial lysates added or DMEM (controls). Cell proliferation was evaluated by spectrophotometric absorbance analysis after the XTT-Microculture Tetrazolium Assay.Results:At 24 hours, cultures with L2, L3, and L4 showed a higher absorbance compared with controls. At 48 hours, cultures with L1, L2, and L3 showed slightly lower absorbance compared with controls, and culture with L4 showed a higher absorbance than in the other experimental conditions. At 72 hours, absorbance was lower in cultures with L1, L2, and L3 than in controls, and absorbance was higher in culture with L4 than in the other experimental conditions.Conclusions:Our study indicates a favorable action of Comano spring water microbiota on proliferation of human skin fibroblasts.

Development of Macroporous Chitosan Scaffolds for Eyelid Tarsus Tissue Engineering.

Reconstruction of large eyelid defects remains challenging due to the lack of suitable eyelid tarsus tissue substitutes. We aimed to evaluate a novel bioengineered chitosan scaffold for use as an eyelid tarsus substitute. Three-dimensional macroporous chitosan hydrogel scaffold were produced via cryogelation with specific biomechanical properties designed to directly match characteristics of native eyelid tarsus tissue. Scaffolds were characterized by confocal microscopy and tensile mechanical testing. To optimise biocompatibility, human eyelid skin fibroblasts were cultured from biopsy-sized samples of fresh eyelid skin. Immunological and gene expression analysis including specific fibroblast-specific markers were used to determine the rate of fibroblast de-differentiation in vitro and characterize cells cultured. Eyelid skin fibroblasts were then cultured over the chitosan scaffolds and the resultant adhesion and growth of cells were characterized using immunocytochemical staining. The chitosan scaffolds were shown to support the attachment and proliferation of NIH 3T3 mouse fibroblasts and human orbital skin fibroblasts in vitro. Our novel bioengineered chitosan scaffold has demonstrated biomechanical compatibility and has the ability to support human eyelid skin fibroblast growth and proliferation. This bioengineered tissue has the potential to be used as a tarsus substitute during eyelid reconstruction, offering the opportunity to pre-seed the patient's own cells and represents a truly personalised approach to tissue engineering.

5-hydroxymethylfurfural-embedded poly (vinyl alcohol)/sodium alginate hybrid hydrogels accelerate wound healing

Treating large acute or chronic wounds remains a challenging task because of a lack of effective methods to accelerate wound healing. Though growth factor-based wound products are found to be effective, they are costly and are potentially associated with increased cancer mortality. Effective, safe, and affordable strategies to tackle large or chronic wounds need to further be designed and developed. Here, we designed a new antioxidant-embedded hydrogel system for speeding up the wound healing process. We prepared multifunctional poly (vinyl alcohol)/sodium alginate (PVA/SA) hydrogels with the incorporation of 5-hydroxymethylfurfural (5-HMF) and silver nanoparticles (Ag-NPs), and investigated their physiochemical and biological properties in vitro and in vivo. PVA, SA, 5-HMF, and Ag-NPs were employed for good mechanical properties, good biocompatibility, anti-inflammation, and anti-bacterial activity, respectively. 5-HMF is commonly found in many food products (e.g. honey, coffee, and black garlic) and is known as an antioxidant. In our in vitro study, 5-HMF was found to effectively facilitate the proliferation and migration of human skin fibroblasts (HSF), and collagen production. Besides, 5-HMF-embedded PVA/SA hybrid hydrogels supported controlled release and good cell compatibility, and more importantly accelerated wound healing in vivo through ameliorated inflammation, enhanced angiogenesis/vascularization, increased collagen production, and promoted re-epithelialization.

Mussel-Inspired Cell/Tissue-Adhesive, Hemostatic Hydrogels for Tissue Engineering Applications.

The combination of multiple physiological (swelling, porosity, mechanical, and biodegradation) and biological (cell/tissue-adhesive, cell proliferation, and hemostatic) properties on a single hydrogel has great potential for skin tissue engineering. Adhesive hydrogels based on polydopamine (PDA) have become the most popular in the biomedical field; however, integrating multiple properties on a single adhesive hydrogel remains a challenge. Here, inspired by the chemistry of mussels, we developed PDA–sodium alginate–polyacrylamide (PDA–SA–PAM)-based hydrogels with multiple physiological and biological properties for skin tissue engineering applications. The hydrogels were prepared by alkali-induced polymerization of DA followed by complexation with SA in PAM networks. The chemical composition of the hydrogels was characterized by X-ray photoelectron spectroscopy. PDA–SA complexed chains were homogeneously dispersed in the PAM network and exhibited good elasticity and excellent mechanical properties, such as a compressive stress of 0.24 MPa at a compression strain of 70% for 0.4PDA–SA–PAM. The adhesive hydrogel also maintained a highly interconnected porous structure (∼94% porosity) along with PDA microfibrils. The hydrogel possesses outstanding swelling and biodegradability properties. Owing to the presence of the PDA–SA complex in the PAM network, the hydrogels show good adhesion to various substrates (plastic, skin, glass, computer screens, and leaves); for example, the adhesive strength of the 0.4PDA–SA–PAM to porcine skin was 24.5 kPa. The adhesive component of the PDA–SA chains in the PAM network significantly improves the cell proliferation, cell attachment, cell spreading, and functional expression of human skin fibroblasts (CCD-986sk) and keratinocytes. Moreover, the PDA chains exhibited good hemostatic properties, resulting in rapid blood coagulation. Considering their excellent cell affinity, and rapid blood coagulation ability, these mussel-inspired hydrogels have substantial potential for skin tissue engineering applications.

Development of halloysite nanotube/carboxylated-cellulose nanocrystal-reinforced and ionically-crosslinked polysaccharide hydrogels.

We report the development of halloysite nanotubes (HNTs)/carboxylated-cellulose nanocrystals (cCNCs) - reinforced and ionically-crosslinked k-carrageenan (k-CG)/xanthan gum (XG) hydrogels. In this study, cCNCs were extracted from microcrystalline cellulose using ammonium persulfate and exhibit 'spindle-like' nanocrystals with approximate diameter of 15-30 nm and length of 30-120 nm. The freeze-dried hydrogels showed highly porous microstructure with good pore-interconnectivity. Further, tunable swelling ratio and in vitro degradation rate of hydrogels under physiological condition (pH 7.4 PBS, 37 °C) were observed. In wet or dry states, the dynamic mechanical analysis of kCXGHN20cCN20 hydrogel showed significantly improved compressive strengths (at 50% strain: 8.1 ± 1.35 kPa or 81.33 ± 1.66 kPa, whereas at 70% strain: 11.84 ± 0.61 kPa or 120.7 ± 1.16 kPa) when reinforced with HNTs (20 wt%)/cCNCs (20 wt%), respectively. The stiffness values are reported at different compressive strains. All hydrogels showed excellent attachment and proliferation of human skin fibroblasts (CCD-986Sk) cells on hydrogels for 7 and 14 days of culture periods. The results showed that these hydrogels may have potential application in soft tissue engineering.

MicroRNA-211-3p has a Role in the Effects of Lipopolysaccharide on Endoplasmic Reticulum Stress in Cultured Human Skin Fibroblasts.

BackgroundLipopolysaccharide (LPS) in bacterial infection of skin wounds delays wound healing. This study aimed to investigate the effects of LPS and endoplasmic reticulum stress in cultured skin fibroblasts and microRNA-211-3p (miR-211-3p) signaling.Material/MethodsHuman skin fibroblasts were cultured in increasing concentrations of LPS at 0 ng/ml, 5 ng/ml, 10 ng/ml, and 20 ng/ml for 0, 12 h, 24 h, 36 h, and 48 h. Cell proliferation was determined using the MTT assay. Protein expression levels of the transcription factors GRP78, CHOP, p-JNK, and the endoplasmic reticulum stress apoptosis proteins, caspase-12 and Bcl-2, were determined by Western blot. The expression of miR-211-3p in human skin fibroblasts was detected by quantitative polymerase chain reaction (qPCR).ResultsCell proliferation of human skin fibroblasts decreased with increasing concentrations of LPS in a dose-dependent and time-dependent way. Protein levels of GRP78, CHOP, p-JNK, caspase-12, and Bcl-2 were increased 8 h and 12 h after LPS treatment compared with 0 h and 4 h after treatment. However, the expression of miR-211-3p was decreased in human skin fibroblasts after treatment with LPS. When miR-211-3p was overexpressed, the endoplasmic reticulum stress/CHOP related proteins, including GRP78, CHOP, p-JNK, caspase-12, and Bcl-2 were unchanged after the addition of LPS. Overexpression of miR-211-3p also reduced inhibitory effects of LPS on the growth of human skin fibroblasts.ConclusionsThis study showed that microRNA-211-3p had a role in the effects of LPS on endoplasmic reticulum stress and CHOP activation in cultured human skin fibroblasts.

Biofabrication of Lysinibacillus sphaericus-reduced graphene oxide in three-dimensional polyacrylamide/carbon nanocomposite hydrogels for skin tissue engineering

The biological synthesis of reduced graphene oxide (rGO) from graphene oxide (GO) is an emerging phenomenon for developing biocompatible nanomaterials for its potential applications in nanomedicine. In this study, we demonstrated a simple, green, and non-toxic method for graphene synthesis using the live biomass of Lysinibacillus sphaericus as the reducing and stabilizing agent under ambient conditions. Ultraviolet-visible spectroscopic analysis confirmed the formation of graphene from GO suspension. X-ray diffraction studies showed the disappearance of the GO peak and the appearance of characteristic graphene broad peak at 2θ = 22.8°. Infrared analysis showed the decrease/disappearance of peaks corresponding to the oxygen-containing functionalities, and appearance of a peak at 1620 cm–1 from unoxidized graphitic domains. Scanning electron microscopic images showed that L. sphaericus-reduced graphene oxide (L-rGO) contains aggregated graphene nanoflakes. Evaluation of the in vitro cytotoxicity of L-rGO nanosheets on human skin fibroblasts using the WST-1 assay did not show any significant effects after 24 h of exposure, which is indicative of biocompatibility. Polyacrylamide hydrogels with L-rGO were synthesized and used as scaffolds to support the growth and proliferation of skin fibroblasts. Cell viability assays and DAPI staining showed proliferation of fibroblasts and exhibited 83% of cell viability even after 28 days. Biofilm formation by Pseudomonas aeruginosa and Staphylococcus aureus was enhanced in nanocomposite hydrogels in the presence of 0.25 mg/mL GO and L-rGO in 48 h. Overall, this study showed that microbially-synthesized L-rGO can be used as a dopant in polymeric scaffolds for tissue engineering and highlighted their role in biofilm formation.

Infrared Skin‐Like Active Stretchable Electronics Based on Organic–Inorganic Composite Structures for Promotion of Cutaneous Wound Healing

AbstractRecent studies on flexible/stretchable medical electronics are almost all focusing on passive monitoring of physiological signs for medical diagnostics, but rare reports have been found on those for active therapeutic purposes. Here, a novel infrared skin‐like active stretchable electronics (ISASE) is introduced with the organic–inorganic composite design for promotion of cutaneous wound healing, which can be conformally mounted anywhere on the human body due to its excellent flexibility and stretchability. Comprehensive experiments, including the proliferation and migration of human skin fibroblasts and rat dermal fibroblasts in vitro, and various tests on wound healing of rats, show favorable effects of the ISASE on promoting the fibroblast migration and proliferation, reducing the inflammatory phase, stimulating the angiogenesis, and shorting the wound healing period. A typical ISASE significantly shortens the cure time for a wounded rat by at least two days compared with the normal treatment with 12 days cure. The mechanical tests confirm the validity of the ISASE when used in extreme large deformations (e.g., stretched by up to 80% strain), which enables the conformability with the human body. The present portable and wearable ISASE demonstrates great potential for cutaneous wound healing, which constitutes an important complement to the current applications in the healthcare field.

Behaviour of cell penetrating peptide TAT-modified liposomes loaded with salvianolic acid B on the migration, proliferation, and survival of human skin fibroblasts

To investigate the efficacy of cell penetrating peptide TAT-modified liposomes loaded with salvianolic acid B (SAB-TAT-LIP) on the proliferation, migration, survival, and TGF-β1 expression of human skin fibroblasts (HSF) and to preliminarily evaluate its effect on the prevention and treatment of hypertrophic scars. SAB-TAT-LIP was prepared using the pH gradient reverse-phase evaporation method. The properties of the liposomes including morphology, percent entrapment efficiency, mean diameter, polydispersity index (PDI), zeta potential, in vitro release, in vitro percutaneous absorption, and serum stability were studied. The proliferation of HSF cells was examined by MTT assays. Cell migration was assessed using the transwell and scratch assays. Cell cycle status was assessed by flow cytometry. Apoptotic induction was assessed through Annexin V-FITC/PI staining. The entrapment efficiency of SAB-TAT-LIP was 86.70 ± 0.85% and the salvianolic acid B (SAB) displayed a unimodal size-distribution with a mean diameter of 183.2 nm ± 4.09, a PDI of 0.190, and a zeta potential of −9.25 mV ± 0.92. The in vitro cumulative release of SAB-TAT-LIP was 62.49% after 24 h. The in vitro 32 h cumulative transdermal rate was 17.21% and the dermal retention was 44.39 μg/cm2±6.87. SAB-TAT-LIP inhibited HSF proliferation in a concentration and time-dependent mode and inhibited the migration and invasion of HSF cells. SAB-TAT-LIP also significantly increased apoptotic induction, the number cells in the G0/G1 phase of the cell cycle, and decreased the levels of TGF-β1 after 48 h of treatment relative to the control group (p < 0.05, p < 0.01). SAB-TAT-LIP offers a promising therapeutic strategy for transdermal delivery during the prevention and treatment of HS.

Enhanced physical, mechanical, and cytocompatibility behavior of polyelectrolyte complex hydrogels by reinforcing halloysite nanotubes and graphene oxide

Polyelectrolyte complex (PEC) hydrogels composed of sodium carboxymethyl cellulose and chitosan reinforced with halloysite nanotubes (HNTs) and graphene oxide (GO) were prepared by polyelectrolyte complexation under acidic environment. Herein, we investigated the effect of HNTs (1.0–10.0 wt%) or HNTs (10.0 wt %)/GO (0.5–1.5 wt%) on the structure-property relationship in microstructural, rheological, mechanical, and cytotoxicity analysis for PEC hydrogels. The results showed highly porous microstructure (varying from 94.3% to 92.8%), stable physical-network and thermal stability of PEC hydrogels and significantly enhanced storage modulus and compressive mechanical properties (in wet state and dry state) of PEC hydrogels reinforced with HNTs/GO. The relative swelling ratio (%) and in vitro degradation of the PEC hydrogels were observed to decrease after the incorporation of HNTs or HNTs/GO content. In addition, PEC hydrogels supported human skin fibroblast cell attachment and proliferation and enhanced effect was observed when HNTs or HNTs/GO were added.

Liposome-encapsulated farnesol accelerated tissue repair in third-degree burns on a rat model

Third-degree or full-thickness burns refer to lesions that extend to the epidermis, dermis, and subcutaneous tissue. The pathophysiology of burn wounds is characterized by tissue inflammation, edema, and hypertrophic scarring. Farnesol is a natural 15-carbon organic compound that possesses many biological effects. We have previously demonstrated that farnesol gel exerts restorative actions on ultraviolet B (UVB)-caused sunburn in vivo. The in vitro results revealed that liposomal farnesol from 0.04mM to 0.8mM significantly enhanced collagen production by murine skin fibroblasts, whereas liposomal farnesol at high (0.8mM) and low concentration (0.04mM) did not show any suppressions on skin fibroblast proliferation. We treated third-degree burns on a rat model with a formulated gel composed of various ratios of 2% hydroxypropyl methylcellulose (HPMC) and 4mM liposomal farnesol for 7 and 14 days. On days 7 and 14 post wounding, histopathological observations revealed that the HPMC:farnesol gel ratios of 1:2 and 2:1 exerted the greatest tissue-repairing effects on the skin after third-degree burns compared with skin untreated or treated with a commercial burn gel and HPMC alone. These findings were consistent with the in vivo quantitative collagen-producing assay, wound healing scoring, and IL-6 Western blot results. These findings demonstrated that the fabricated liposomal farnesol gel is potentially able to promote wound healing after third-degree burns.

Mesenchymal Stem Cell Engineered Nanovesicles for Accelerated Skin Wound Closure

We report development and characterization of cell-engineered nanovesicles made from mesenchymal stem cells (MSCNVs), which have more than 300 times higher productivity than natural extracellular vesicles (EVs). MSCNVs had similar morphological characteristics to MSCEVs but have molecular characteristics that more resemble MSCs than MSCEVs. In vitro MSCNV treatment increased the proliferation and migration of primary skin fibroblasts and showed better effects than treatment using natural MSCEVs. Quantitative real-time PCR analysis showed increased expression of growth factors in MSCNV-treated skin fibroblasts. Intraperitoneal injection of MSCNVs into syngeneic mice induced mild local inflammation, which resulted in recruitment of immune cells to the injection site. In vivo MSCNV treatment of a mouse skin wound accelerated its healing; this acceleration by MSCNVs may occur by promoting blood vessel formation at the wound site. These results indicate the promise of MSCNVs as agents for regenerative medicine.

Effect of cell penetrating peptide TAT-modified liposomes loaded with salvianolic acid B on proliferation,migration and cell cycle of human skin fibroblasts

Hypertrophic scar( HS) is a very common skin fibrosis disorder after human skin injury and wound healing. The objective of this study was to investigate the efficacy of cell penetrating peptide TAT-modified liposomes loaded with salvianolic acid B( SAB-TAT-LIP) on proliferation,migration and cell cycle of human skin fibroblasts( HSF),and preliminarily evaluate its effect on prevention and treatment of HS. HSF were cultured in vitro,and MTT assay was used to detect the inhibitory effect of SAB-TAT-LIP on cell proliferation. Cell migration was assessed by Transwell chamber method and scratch method; and cell cycle change was detected by flow cytometry. In vitro cell studies showed that blank liposome basically had no toxic effect on HSF. Different concentrations of SABTAT-LIP inhibited proliferation on HSF in varying degrees after intervention for different periods in a dose and time dependent manner;meanwhile,SAB-TAT-LIP significantly inhibited the migration and invasion of HSF. At the same time,SAB-TAT-LIP could block the cell cycle at G0/G1 phase after intervention for 48 h,P<0.01 as compared with the blank control group. Conclusively,our experimental data quantitatively demonstrate that SAB-TAT-LIP has significant inhibitory effect on cells proliferation,invasion and migration,with blocking effect on G0/G1 phase. This may offer a promising therapeutic strategy for transdermal delivery in prevention and treatment of HS.

Salvianolic acid B attenuates experimental skin fibrosis of systemic sclerosis

Systemic sclerosis (SSc) is a connective tissue disease characterized mainly by fibrosis of skin and internal organs. Our previous study has shown that salvianolic acid B (SAB), a bioactive component extracted from Salvia miltiorrhiza (SM), was one of the essential ingredients in the traditional Chinese medicine Yiqihuoxue formula, which has been used to treat SSc-related dermal and pulmonary fibrosis. The aim of the present study was to evaluate the effect of SAB on skin fibrosis and explore its underlying anti-fibrotic mechanism. We found that SAB was capable of alleviating skin fibrosis in a bleomycin-induced SSc mouse model, alleviating skin thickness and reducing collagen deposition. in vitro studies indicated that SAB reduced SSc skin fibroblast proliferation and downregulated extracellular matrix gene transcription and collagen protein expression. TGF-β/SMAD and MAPK/ERK pathway activation were also shown to be suppressed in SAB treated fibroblasts. Moreover, RNA-seq revealed that the anti-fibrotic effect of SAB might be related to antioxidant activity, the cell cycle, and the p53 signaling pathway. Taken together, our results suggest that SAB has the ability to alleviate SSc-related skin fibrosis both in vivo and in vitro.

Anti-inflammatory and wound healing properties of polyphenolic extracts from strawberry and blackberry fruits

The polyphenolic profiles by HPLC-TOF-MS of strawberry ‘San Andreas’ and blackberry ‘Black Satin’ crude extracts (CE) were analyzed. Anthocyanin-enriched fractions (AEFs) and proanthocyanidin-enriched fractions (PEFs) were prepared, and all samples were probed for in vitro anti-inflammatory and wound healing effects in a LPS-stimulated RAW 264.7 macrophage model and in a skin fibroblast migration and proliferation assay, respectively. Blackberry samples exhibited higher ROS reduction than strawberry's (up to 50% ROS suppression). Berries CEs exhibited 20% inhibition in Cox-2 gene expression, while AEFs and PEFs were inactive at the same concentration. Strawberry AEF and PEF were more active against IL-1β and IL-6 gene expressions than the similar fractions from blackberry, where PEF was more active than AEF (75% suppression by strawberry PEF). Moreover, berry PEFs were the active polyphenol fraction against iNOS gene expression (50% and 65% gen suppression by strawberry and blackberry PEF, respectively), mirroring results of NO synthesis suppression. The cell migration potential of berry polyphenolics was associated with anthocyanins. AEFs showed fibroblast migration around 50% of that registered for the positive control. Results obtained in this work highlight the anti-inflammatory properties of berry polyphenolics, especially due to proanthocyanidins. Moreover, promising results were obtained about the effects of berry anthocyanins on wound healing.

Effect of Pentacyclic Triterpenoids-Rich Callus Extract of Chaenomeles japonica (Thunb.) Lindl. ex Spach on Viability, Morphology, and Proliferation of Normal Human Skin Fibroblasts.

The effect of the well-characterized callus extract of Chaenomeles japonica on viability, morphology, and proliferation of normal human skin fibroblasts was investigated. The phytochemical analysis was performed using the ultra-high performance liquid chromatography method. The total phenolic, phenolic acid, and flavonoid contents were determined spectrophotometrically. The antioxidant activity was investigated using the DPPH (1,1-Diphenyl-1-picrylhydrazyl Radical Scavenging), FRAP (Ferric Reducing Antioxidant Power), and CUPRAC (CUPric Reducing Antioxidant Capacity) assays. The callus growth index during passages was high as well as the content of pentacyclic triterpenoids. The microscopic observations of the fibroblast viability, morphology and the evaluation of the proliferation ratio (xCELLigence system) proved that the influence of callus extract on the fibroblasts was dose-dependent. The evaluated level of fibroblasts proliferation rate after 72 h of incubation with callus extract at concentration 12.5 µg L−1 was the highest compared to all the analyzed ligands. Moreover, callus extract administrated for 72 h caused a significant increase in the proliferation rate in comparison with the control group (5.7 ± 0.1 vs. 4.4 ± 0.9; p < 0.01). The preliminary studies carried out may suggest that the callus extract rich in triterpenoids may be a potential source of cosmetic ingredients with a beneficial effect on human skin.

New approach for immobilization of 3-aminopropyltrimethoxysilane and TiO2 nanoparticles into cellulose for BJ1 skin cells proliferation

In the present study, tosylcellulose (TC) was used as a key intermediate for the selective coupling with 3-aminopropyltrimethoxysilane (APTMS) affording amino-propylsilane-grafted tosylcellulose (TC-Si). Solid state 13C NMR and FT-IR analyses confirmed the coupling and self-condensation of APTMS along TC. The changes in the surface morphology of the functionalized cellulose were identified by SEM imaging. The thermal stability of TC-Si was significantly improved as compared to MCC and TC. A new organic/inorganic hybrid cellulosic material was fabricated by embedding TiO2 nanoparticles into TC-Si network. The new cellulose polymers were investigated for their ability to promote the proliferation of human skin fibroblast (BJ1). The cell cytotoxicity assay showed that both TC and TC-Si possessed moderate toxicity to BJ1 cells by 17% and 23.8%, respectively at 20 μM. Meanwhile, TC-Si/TiO2 hybrid enhanced the proliferation of BJ1 by 42%. Additionally TC-Si/TiO2 hybrid demonstrated promising antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans.

Influence of Cross-Linking Method and Disinfection/Sterilization Treatment on the Structural, Biophysical, Biochemical, and Biological Properties of Collagen-Based Devices.

Disinfection/sterilization is an essential step during the manufacturing process of any implantable medical device. Cross-linking is also required for biopolymers to control resistance to degradation and enhance mechanical integrity. To date, there is still no single disinfection/sterilization treatment and cross-linking method that can be used universally for collagen-based devices. Herein, we assessed the influence of ethylene oxide, ethanol, gamma irradiation, and gas plasma disinfection/sterilization on the structural, biophysical, biochemical, and biological properties of self-assembled collagen films cross-linked with 4-arms polyethylene glycol succinimidyl glutarate and genipin. Microscopy analysis revealed that gas plasma treatment induced the most profound differences in the non-cross-linked and 4-arms polyethylene glycol succinimidyl glutarate cross-linked collagen films. Gas plasma also significantly increased the swelling ratio of the non-cross-linked and the 4-arms polyethylene glycol succinimidyl glutarate cross-linked films. Non-cross-linked and gas plasma treated 4-arms polyethylene glycol succinimidyl glutarate collagen films exhibited the lowest resistance to collagenase degradation and denaturation temperature. Between the non-cross-linked groups, the gas plasma treatment resulted in the collagen films with the lowest stress at break, strain at break, force at break, and Young's modulus values. Within the 4-arms polyethylene glycol succinimidyl glutarate groups, the ethylene oxide treatment resulted in collagen films with the lowest stress at break, strain at break, force at break, and Young's modulus values. Within the genipin groups, the gas plasma treatment resulted in collagen films with the lowest stress at break, strain at break, force at break, and Young's modulus values. Proliferation, metabolic activity, and viability of human skin fibroblasts were not affected as a function of cross-linking method and disinfection/sterilization treatment. However, proliferation, metabolic activity, and viability of THP1 cells were significantly reduced as a function of the cross-linking method, but they were not affected as a function of the disinfection/sterilization treatment. Overall, our data illustrate that the cross-linking method and the disinfection/sterilization treatment differentially affect the structural, biophysical, biochemical, and biological properties of collagen-based devices, and thus, they should be optimized according to the clinical indication.

Effects of Vitamin B Complex and Vitamin C on Human Skin Cells: Is the Perceived Effect Measurable?

Vitamins are essential for human health. In terms of local application for wound healing, vitamins' positive effect remains unclear. However, because of the regular appearance of nutritional deficiency in chronic wound patients, a favorable impact of locally applied vitamins can be hypothesized. Vitamins B3, B5, B6, B7, B9, B10, B12, and C individually as well as different combinations of B vitamins were investigated regarding their ability to promote proliferation and migration of human skin fibroblasts and keratinocytes. Proliferation assays with and without bacterial challenge, immunocytochemical staining, and scratch assay were used to determine the most effective combination(s). Some vitamin combinations showed a positive impact on proliferation, especially for keratinocytes after 72 hours. In terms of wound closure, the combinations B9 and B12; B3, B5, B6, and B10; and B3, B5, and B7 improved closure rates by 25% to 30%. The improved closure rates are also reflected by immunocytochemically detected upregulation of the migration marker CXCR4 for several combinations. Certain combinations of B vitamins demonstrate a positive influence on human keratinocytes and fibroblasts. Vitamins especially promoted fibroblast migration, and a statistically significant induction of keratinocyte proliferation was observed. Therefore, local vitamin application could benefit the physiologic wound healing process.