Proliferation Of Human Skin Fibroblasts Research Articles

Effects of cerium oxide nanoenzyme-gelatin methacrylate anhydride hydrogel in the repair of infected full-thickness skin defect wounds in mice

Objective: To investigate the effects of cerium oxide nanoenzyme-gelatin methacrylate anhydride (GelMA) hydrogel (hereinafter referred to as composite hydrogel) in the repair of infected full-thickness skin defect wounds in mice. Methods: This study was an experimental study. Cerium oxide nanoenzyme with a particle size of (116±9) nm was prepared by hydrothermal method, and GelMA hydrogel with porous network structure and good gelling performance was also prepared. The 25 μg/mL cerium oxide nanoenzyme which could significantly promote the proliferation of human skin fibroblasts and had high superoxide dismutase activity was screened out. It was added to GelMA hydrogel to prepare composite hydrogel. The percentage of cerium oxide nanoenzyme released from the composite hydrogel was calculated after immersing it in phosphate buffer solution (PBS) for 3 and 7 d. The red blood cell suspension of mice was divided into PBS group, Triton X-100 group, cerium oxide nanoenzyme group, GelMA hydrogel group, and composite hydrogel group, which were treated with corresponding solution. The hemolysis of red blood cells was detected by microplate reader after 1 h of treatment. The bacterial concentrations of methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli were determined after being cultured with PBS, cerium oxide nanoenzyme, GelMA hydrogel, and composite hydrogel for 2 h. The sample size in all above experiments was 3. Twenty-four 8-week-old male BALB/c mice were taken, and a full-thickness skin defect wound was prepared in the symmetrical position on the back and infected with MRSA. The mice were divided into control group without any drug intervention, and cerium oxide nanoenzyme group, GelMA hydrogel group, and composite hydrogel group applied with corresponding solution, with 6 mice in each group. The wound healing was observed on 3, 7, and 14 d after injury, and the remaining wound areas on 3 and 7 d after injury were measured (the sample size was 5). The concentration of MRSA in the wound exudation of mice on 3 d after injury was measured (the sample size was 3), and the blood flow perfusion in the wound of mice on 5 d after injury was observed using a laser speckle flow imaging system (the sample size was 6). On 14 d after injury, the wound tissue of mice was collected for hematoxylin-eosin staining to observe the newly formed epithelium and for Masson staining to observe the collagen situation (the sample size was both 3). Results: After immersion for 3 and 7 d, the release percentages of cerium oxide nanoenzyme in the composite hydrogel were about 39% and 75%, respectively. After 1 h of treatment, compared with that in Triton X-100 group, the hemolysis of red blood cells in PBS group, GelMA hydrogel group, cerium oxide nanoenzyme group, and composite hydrogel group was significantly decreased (P<0.05). Compared with that cultured with PBS, the concentrations of MRSA and Escherichia coli cultured with cerium oxide nanoenzyme, GelMA hydrogel, and composite hydrogel for 2 h were significantly decreased (P<0.05). The wounds of mice in the four groups were gradually healed from 3 to 14 d after injury, and the wounds of mice in composite hydrogel group were all healed on 14 d after injury. On 3 and 7 d after injury, the remaining wound areas of mice in composite hydrogel group were (29±3) and (13±5) mm2, respectively, which were significantly smaller than (56±12) and (46±10) mm2 in control group and (51±7) and (38±8) mm2 in cerium oxide nanoenzyme group (with P values all <0.05), but was similar to (41±5) and (24±9) mm2 in GelMA hydrogel group (with P values both >0.05). On 3 d after injury, the concentration of MRSA on the wound of mice in composite hydrogel group was significantly lower than that in control group, cerium oxide nanoenzyme group, and GelMA hydrogel group, respectively (with P values all <0.05). On 5 d after injury, the volume of blood perfusion in the wound of mice in composite hydrogel group was significantly higher than that in control group, cerium oxide nanoenzyme group, and GelMA hydrogel group, respectively (P<0.05). On 14 d after injury, the wound of mice in composite hydrogel group basically completed epithelization, and the epithelization was significantly better than that in the other three groups. Compared with that in the other three groups, the content of collagen in the wound of mice in composite hydrogel group was significantly increased, and the arrangement was also more orderly. Conclusions: The composite hydrogel has good biocompatibility and antibacterial effect in vivo and in vitro. It can continuously sustained release cerium oxide nanoenzyme, improve wound blood perfusion in the early stage, and promote wound re-epithelialization and collagen synthesis, therefore promoting the healing of infected full-thickness skin defect wounds in mice.

Discovery of indole analogues from Periplaneta americana extract and their activities on cell proliferation and recovery of ulcerative colitis in mice

Background: As an important medicinal insect, Periplaneta americana (PA) has been applied for the treatment of wounds, burns, and ulcers with fewer side effects and a reduced recurrence rate, which provides great potential for developing new drugs based on its active constituents.Materials and methods: The main chromatographic peaks determined by high performance liquid chromatography (HPLC) in the PA concentrated ethanol-extract liquid (PACEL) were separated, purified, and identified by semi-preparative LC, mass spectrum, and 1H NMR spectroscopic analysis. The biological activities of the identified compounds were investigated by methylthiazolyldiphenyl-tetrazolium bromide (MTT) method based on in vitro human skin fibroblasts (HSF) and in vivo experiments based on dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) mouse model. Furthermore, RT-qPCR of six genes related to inflammation or intestinal epithelial cell proliferation was employed to investigate the molecular mechanism of the indole analogues recovering UC in mice.Results: Five indole analogues were purified and identified from PACEL, including tryptophan (Trp), tryptamine (pa01), 1,2,3,4-tetrahydrogen-β-carboline-3-carboxylic acid (pa02), (1S, 3S)-1-methyl-1,2,3,4-tetrahydrogen-β-carboline-3-carboxylic acid (pa03), and (1R, 3S)-1-methyl-1,2,3,4-tetrahydrogen-β-carboline-3-carboxylic acid (pa04), among which the pa02 and pa04 were reported in PA for the first time. In vitro and in vivo experiments showed that PACEL, Trp, and pa02 had promoting HSF proliferation activity and intragastric administration of them could alleviate symptoms of weight loss and colon length shortening in the UC mice. Although recovery activity of the compound pa01 on the colon length was not as obvious as other compounds, it showed anti-inflammatory activity in histological analysis. In addition, The RT-qPCR results indicated that the three indole analogues could alleviate DSS-induced intestinal inflammation in mice by inhibiting pro-inflammatory cytokines (MMP7, IL1α) and down-regulating BMP8B expression.Conclusion: This study reported the isolation, purification, structure identification, and biological activity of the active indole analogues in PACEL. It was found for the first time that the PA extract contained many indole analogues and Trp, which exhibited good proliferation activity on HSF fibroblasts as well as anti-UC activity in mice. These indole analogues probably are important components related to the pharmacological activity in PA.

Adiponectin inhibits TGF-β1-induced skin fibroblast proliferation and phenotype transformation via the p38 MAPK signaling pathway.

The aim of this study was to investigate the effects of adiponectin (APN) on the proliferation and phenotypic transformation of human skin fibroblasts (HSFs) induced by TGF-β1. Primary fibroblast cultures were collected from prepuce surgery, and the cell viability and proliferative activity of HSFs were detected by Cell Counting Kit-8 and EdU assays. In addition, cell migration was detected by Transwell assay. The protein levels of related genes in HSF were detected by Western blotting. The results showed that the proliferation and migration abilities of HSF in the TGF-β1 group were significantly improved, and the relative protein expression levels of PCNA, α-SMA, and Collagen I in the TGF-β1 group were greatly increased. Furthermore, TGF-β1 stimulated the phosphorylation of p38 in HSF, while APN pretreatment significantly inhibited the TGF-β1-induced phosphorylation of p38. Additionally, blocking the p38 MAPK signaling pathway relieved the injury in the HSF induced by TGF-β1 and enhanced the therapeutic effect of APN in the TGF-β1-treated HSF. In conclusion, APN inhibits TGF-β1-induced HSF proliferation and myofibroblast phenotypic transformation by activating the p38 MAPK signaling pathway. APN is expected to become a potential target for preventing and treating skin fibrosis and pathological scars.

Induced skin aging by blue-light irradiation in human skin fibroblasts via TGF-β, JNK and EGFR pathways

BackgroundStudies indicate that blue light (BL) irradiation can damage human skins, but the impact of BL irradiation on skin aging is unknown. ObjectivesThis study aimed to give an insight to phenotypic characteristics and molecular mechanism of blue light-induced skin aging, and thus provide a theoretical basis for the precise protection of photodermatosis. MethodsThe effect of BL on skin photoaging in mice was evaluated by non-invasive measurement equipment and histopathology analysis. The effect of BL irradiation on the proliferation of HFF-1 cells was detected by the Real-Time Cell Analyzer. The expression and protein levels of genes associated with skin aging were examined. ResultsOur studies indicated photoaging caused by BL irradiation, including collagen disorder and increased MMP1. BL irradiation also inhibited cell proliferation and collagen expression in human skin fibroblasts by inhibiting TGF-β signaling pathway, based on in vitro experiments. Importantly, BL irradiation promoted the degradation of collagen by increasing MMP1 activated by the JNK/c-Jun and EGFR pathways. Moreover, ROS levels were significantly increased after BL irradiation in human skin fibroblasts. Yet, the transcriptional change in human skin fibroblasts caused by BL irradiation was unable to be completely restored by ROS scavenger. ConclusionBL irradiation down-regulated expression of type I collagen genes and up-regulated MMP1 expression to inhibit the proliferation of human skin fibroblasts. Multiple key pathways including TGF-β, JNK, and EGFR signaling were involved in BL-induced skin aging. Our results provide theoretical bases for the protection of photoaging caused by BL irradiation.

Hydroxyapatite/hyperbranched polyitaconic acid/chitosan composite scaffold for bone tissue engineering

AbstractIn this study, a promising modified composite scaffold (hydroxyapatite/hyperbranched polyitaconic acid/chitosan) was synthesized for bone tissue engineering. Novel hyperbranched polyitaconic acid was prepared through the polymerization of itaconic acid using reversible addition fragmentation chain transfer using a macro‐RAFT agent. The chemical structure of the prepared hyperbranched polyitaconic acid was characterized by FTIR and 1HNMR and was subsequently embedded into hydroxyapatite/chitosan composite. The obtained modified composite scaffold was evaluated by characterizing its porosity, mechanical properties, bioactivity and cytotoxicity. The results showed that the modified composite scaffold had higher mechanical strength (i.e., 0.56 ± 0.03 MPa) in comparison to chitosan/hydroxyapatite scaffold only (i.e., 0.31 ± 0.01 MPa) and also showed higher bioactivity. In addition, the modified composite scaffold (HAP/HBP‐RAFT‐PI/CS) showed anticancer properties and enhanced human skin fibroblasts proliferation.

Preparation of tanshinone IIA self-soluble microneedles and its inhibition on proliferation of human skin fibroblasts

ObjectiveHypertrophic scars (HS) are a variety of skin tissue fibrosis disease that occurs in human skin, the effective therapeutic method of which is still inaccessible up to now. As a bioactive constituent of a well-known medical plant, Salvia miltiorrhiza (Danshen in Chinese), tanshinone IIA (TSA) is reported to inhibit cell proliferation in HS. Therefore, the aim of this study was to prepare TSA self-soluble microneedles to strengthen its dermal retention and break through the difficulty of significantly thickening epidermal connective tissue and stratum corneum at the HS site. The possible mechanism of action in suppressing HS was studied using human skin fibroblasts (HSF). MethodsTanshinone IIA self-dissolving microneedles (TSA-MN) was prepared using a negative mold casting method. The prescription process of microneedle was optimized by Box-Behnken effect surface method. Different media were selected to investigate the ability of transdermal absorption and in vitro release. Furthermore, according to Cell Counting Kit-8 (CCK8) method as well as the Western blot method, the effect of TSA-MN on the biological characteristics of HSF was investigated. ResultsWith remarkable slow release effect and dermal retention, the release and transdermal properties of TSA-MN in vitro were better than both TSA and ordinary dosage forms. Its effect of HSF confirmed the essential decrease in cell motility during cell proliferation and cell migration in vitro, which plays a significant role in down-regulating the secretion of transforming growth factor-β1 (TGF-β1) in HSF and increasing the expression level of Smad7. ConclusionThe prepared TSA self-soluble microneedles is helpful in solving the problem of hypertrophic scars, with a stable dermal retention effect after process optimization.

Homogeneous preparation of water-soluble products from chitin under alkaline conditions and their cell proliferation in vitro

The purpose of this study was to prepare water-soluble products by homogeneous depolymerization of chitin with H2O2 under alkaline conditions and investigate their potential application in wound healing. For the first time, water-soluble products were successfully prepared using a chitin-NaOH/urea solution; the products were chitosans with molecular weights (Mw) of 3.48–33.5 kDa and degrees of deacetylation (DD) > 0.5. Their Mw, DD and yield were affected by the reaction temperature, reaction time, concentration of H2O2 and chitin DD. The deacetylation and depolymerization of chitin were achieved simultaneously. The depolymerization of chitin was caused by hydrogen abstraction of HO, whereas the deacetylation resulted from the cleavage of amide bonds by HO− and HO2−, although the latter played a more important role. All water-soluble chitosans markedly promoted the proliferation of human skin fibroblast (HSF) cells, but they inhibited the proliferation of human keratinocyte cells. For the proliferation of HSF, a low concentration of chitosans was important. In addition, water-soluble chitosans with an Mw of 3.48–16.4 kDa markedly stimulated the expression of growth factors such as PDGF and TGF-β by macrophages. Water-soluble chitosans could be used as a potential active component in wound dressings.

Highly stretchable porous regenerated silk fibroin film for enhanced wound healing.

Silk fibroin (SF) has received interest in tissue engineering owing to its biocompatibility, biodegradability, and favorable mechanical properties. However, the complex preparation, brittleness, and lack of pores in the structure of the silk fibroin film limit its application. Herein, we show that facile dissolution of SF in aqueous phosphoric acid followed by regeneration in aqueous ammonium sulfate ((NH4)2SO4) could afford highly stretchable films with nano-pores formed in the nonsolvent-induced phase separation process. The named phase separation, which determines the morphology and mechanical properties of the regeneration silk fibroin (RSF) films, is highly dependent on the (NH4)2SO4 concentration as well as the initial concentration of the SF solution. Therefore, the RSF films exhibit a tunable pore size ranging from 230 to 510 nm and excellent stretchability with tensile strain up to 143 ± 16%. Most interestingly, the RSF films were shown to support the proliferation of human skin fibroblasts in vitro as well as speed up full-thickness skin wound healing in a rat model. This work establishes an easy and feasible method to access porous RSF membranes that can be used for wound dressing in clinical settings.

Effects of Chlorhexidine-Iodophor Composite Solution on the Viability and Proliferation of Human Skin Fibroblasts Infected by S. aureus - An in Vitro Experiment.

Background: The chlorhexidine-iodophor (CHX-IP) composite solution is a polymer of chlorhexidine and iodophor, applicable to the control of local microbial load and probably toxic to fibroblasts. However, the effect of CHX-IP on the viability and proliferation of human skin fibroblasts infected by Staphylococcus aureus (S. aureus) remains unknown. Objective: The effects of CHX-IP composite solution on the viability and proliferation of human skin fibroblasts infected by S. aureus were investigated in vitro cell culture in this study. Methods: Optimum multiplicity of infection (MOI) was determined to construct the S. aureus-fibroblast co-culture model. Cell Viability Assay was applied to obtain optical density (OD) value and calculate cell viability. 5-ethynyl-2'- deoxyuridine (EdU) assay was used to investigate the effect of CHX-IP on the proliferation of human skin fibroblasts infected by S. aureus. Results: 10:1 was the optimum MOI for the S. aureus-fibroblast co-culture model. The OD value of human skin fibroblasts infected by S. aureus increased in the blank control group, 0.625 mg/ml, 0.3125 mg/ml, 0.15625 mg/ml, and 0.075625 mg/ml groups after four hours. While that of the negative control group, 5 mg/ml, 2.5 mg/ml, and 1.25 mg/ml groups decreased over time. The two-way ANOVA results indicated that the OD value of human skin fibroblasts infected by S. aureus was significantly different among different CHX-IP concentration groups (F = 34.05, P < .001), and the interaction effect between concentration and time was significant (F = 9.442, P < .001). The results of the EdU cell proliferation assay showed that the blank control group, 0.625 mg/ml CHX-IP group, and 0.075625 mg/ml CHX-IP group had an enhanced fibroblasts cell proliferation, while the fibroblasts cell proliferation of the negative control group and 5 mg/ml CHX-IP group was inhibited. Conclusion: The viability and proliferation of human skin fibroblasts infected by S. aureus were inhibited, while specific concentrations of CHX-IP solution can counteract or even reverse the proliferation inhibition effect.

Zinc alginate hydrogels with embedded RL-QN15 peptide-loaded hollow polydopamine nanoparticles for diabetic wound healing therapy

Chronic wound healing remains a considerable clinical challenge worldwide. We previously identified a pro-regenerative agent, i.e., hollow dopamine nanoparticles (HPDA) loaded with potent pro-healing peptide RL-QN15 (HPDAlR), with significant skin wound healing activity. In the current study, in consideration of clinical application, we successfully prepared a new zinc alginate hydrogel embedded with HPDAlR (HPDAlR&ZA) to treat diabetic wounds. HPDAlR&ZA exhibited no obvious toxicity against keratinocytes, human umbilical vein endothelial cells (HUVECs), human skin fibroblasts (HSFs), and mice. HPDAlR&ZA significantly promoted keratinocyte, HUVEC, and HSF proliferation, as well as HUVEC migration and angiogenesis. HPDAlR&ZA also modulated the expression of cytokines from macrophages. Diabetic mouse full-thickness skin wound and diabetic patient in vitro skin wound models showed that HPDAlR&ZA resisted the inflammatory response by rapidly polarizing M1 macrophages to M2 macrophages, thereby reconstructing blood supply, increasing collagen deposition, and accelerating diabetic wound repair and skin regeneration. In summary, HPDAlR&ZA is a biodegradable hydrogel wound dressing and an efficient treatment strategy for diabetic wound repair, with strong anti-oxidant, anti-inflammatory, and pro-angiogenic features.

LncRNA MALAT1 from human adipose-derived stem cell exosomesaccelerates wound healing via miR-378a/FGF2 axis.

Aim: The effects of MALAT1from human adipose-derived stem cell(ADSC) exosomes in skin wound healingwere investigated. Material & methods: The viability, apoptosisand migration ability of human skin fibroblasts (HSFs) were evaluated by Cell Counting Kit-8 assay, flow cytometryand scratch assay, respectively. A mouse model was established to evaluate the role of exosomal MALAT1 in skin wound healing in vivo. Results: Human ADSC exosomes promoted the proliferation and migration of HSFs and increased MALAT1 expression. MALAT1 silencing in human ADSCs inhibited HSF viability and migration, promoted HSF apoptosisand inhibited angiogenesis by upregulating miR-378a. Overexpression of miR-378a inhibited the migration and proliferation of HSFs by downregulating FGF2expression. ADSC exosomes promoted skin wound healing by mediating MALAT1 in vivo. Conclusion: Exosomal MALAT1 accelerated skin wound healing by regulating the miR-378a/FGF2 axis, suggesting that MALAT1 might be used as a potential target for cutaneous wound treatment.

Investigation of the In Vivo, In Vitro, and In Silico Wound Healing Potential of Pinctada martensii Purified Peptides.

Previous studies found that both oral and topical administration of enzymatic digestion products < 3 K Da ultrafiltration fractions of Pinctada martensii mantle (PMPs) had pro-healing effects. Thus, we further purified them by Sephadex-G25 and screened them by cellular assays to obtain Pinctada martensii purified peptides (PMPPs). In this study, we explored the mechanism of PMPPs on wound healing by in vivo, in vitro, and in silico experiments. LC-MS/MS results showed that PMPPs consisted of 33 peptides with molecular weights ranging from 758.43 to 2014.04 Da, and the characteristic peptide was Leu-Asp. The results of cellular assays showed that PMPPs promoted the proliferation of human skin fibroblasts (HSF) (135%) and human immortalized keratinocyte (HaCaT) cells (125%) very significantly at 12.5 μg/mL. The in vivo results showed that PMPPs could achieve scarless healing by inhibiting the inflammatory response, accelerating the epithelialization process, and regulating collagen I/III ratio. The optimal peptide sequence FAFQAEIAQLMS of PMPPs was screened for key protein receptors in wound healing (EGFR1, FGFR1, and MMP-1) with the help of molecular docking technique, which also showed to be the key pro-healing active peptide sequence. Therefore, it may provide a therapeutic strategy with great potential for wound healing.

A Preliminary Study for the Effect of Coix-seed Reactive Derivatives (CRD) on Human Skin Fibroblast Proliferation and Collagen Production

A Preliminary Study for the Effect of Coix-seed Reactive Derivatives (CRD) on Human Skin Fibroblast Proliferation and Collagen Production

MiR-24-3p obstructs the proliferation and migration of human skin fibroblasts after thermal injury by targeting PPAR-β and positively regulated by NF-κB.

Thermal injury repair is a complex process during which the maintenance of the proliferation and migration of human skin fibroblasts (HSFs) exert a crucial role. MicroRNAs have been proven to exert an essential function in repairing skin burns. This study delves into the regulatory effects of miR-24-3p on the migration and proliferation of HSFs that have sustained a thermal injury, thereby, providing deeper insight into thermal injury repair pathogenesis. The PPAR-β protein expression level progressively increased in a time-dependent manner on the 12th, 24th and 48th hour following the thermal injury of the HSFs. The knockdown of PPAR-β markedly suppressed the proliferation of and migration of HSF. Following thermal injury, the knockdown also promoted the inflammatory cytokine IL-6, TNF-α, PTGS-2 and P65 expression. PPAR-β contrastingly exhibited an opposite trend. A targeted relationship between PPAR-β and miR-24-3p was predicted and verified. miR-24-3p inhibited thermal injured HSF proliferation and migration and facilitated inflammatory cytokine expression through the regulation of PPAR-β. p65 directly targeted the transcriptional precursor of miR-24 and promoted miR-24 expression. A negative correlation between miR-24-3p expression level and PPAR-β expression level in rats' burnt dermal tissues was observed. Our findings reveal that miR-24-3p is conducive to rehabilitating the denatured dermis, which may be beneficial in providing effective therapy of skin burns.

Adipose mesenchymal stem cell-derived exosomes accelerate skin wound healing via the lncRNA H19/miR-19b/SOX9 axis

It has been reported that adipose mesenchymal stem cells (ADSCs) accelerate wound healing. Moreover, exosomes, which serve as paracrine factors, play a vital role in wound healing. However, the mechanism remains unclear. This research aimed to determine the roles of exosomes derived from ADSCs (ADSC-Exos) in wound skin tissue repair. Flow cytometry and electron microscopy were carried out to identify ADSCs and ADSC-Exos, respectively; RT-qPCR was performed to assess the lncRNA H19 (H19), microRNA19b (miR-19b) and SRY-related high-mobility-group box 9 (SOX9) levels; Western blotting was carried out to evaluate collagen and β-catenin expression; CCK-8, scratch and transwell assays were conducted to evaluate human skin fibroblast (HSF) cell proliferation, migration and invasion, respectively; the potential binding sites between H19 and miR-19b, miR-19b and SOX9 were detected by dual-luciferase reporter gene assay and RIP assay; and H&E staining was conducted to observe skin wound tissues. ADSC-Exos accelerated the proliferation, migration and invasion of HSF cells via H19. H19 acts as a molecular sponge towards miR-19b, which targets SOX9. ADSC-Exos inhibited miR-19b expression via H19, resulting in accelerated HSF proliferation, migration and invasion. ADSC-Exos upregulated SOX9 to activate the Wnt/β-catenin pathway, resulting in accelerated HSF cell proliferation, migration and invasion, and ADSC-Exos promoted skin wound healing via H19 in mice.The high expression of H19 in ADSC-Exos may upregulate SOX9 expression via miR-19b to accelerate wound healing of skin tissues. Our study may provide novel perspectives for therapy to accelerate skin wound healing.This study reveals that the long non-coding RNA H19 is highly expressed in exosomes derived from adipose mesenchymal stem cells and accelerates the proliferation, migration and invasion of human skin fibroblasts by upregulation of SOX9 and activation of the Wnt/β-catenin pathway The authors show that H19 affects SOX9 expression via the microRNA miR-19b to promote wound healing in injured skin.

Testing the best mitochondrial source for therapy: Mitochondrial xenotransfer from mice mscs and not from mice muscle tissue induces proliferation of human skin fibroblasts

Testing the best mitochondrial source for therapy: Mitochondrial xenotransfer from mice mscs and not from mice muscle tissue induces proliferation of human skin fibroblasts

Altered CD4+ T cell and cytokine levels in peripheral blood and skin samples from systemic sclerosis patients and IL-35 in CD4+ T cell growth

This study explored the role of IL-35 in CD4+ T lymphocyte and skin fibroblast (HSF) activity and cytokine levels in systemic sclerosis. Blood and skin biopsies were collected from 41 patients and 39 healthy controls to assess CD4+ T lymphocytes and IL-35-related factors. CD4+ T lymphocytes were co-cultured with HSFs, rhIL-35, and IL-35 mAb to evaluate the cell viability, activation of CD4+T lymphocytes, and HSF cells. The proportion of blood Th1/Th2 was lower and Th17/regulatory T cells (Treg) were higher in patients than in controls (p < 0.05). IL-35 and IL-17A levels were higher and IFN-γ, IL-10, and TGF-β levels were lower in patients than in controls. IL-17A, FoxP3, TGF-β1, and COL-1 mRNA and p-STAT1 and p-STAT4 were higher in skin tissues from patients than in those from controls (p < 0.05). IL-6 levels were higher, whereas IL-10 levels were lower in cell culture supernatants. α-SMA and COL-1 proteins and Ki67 positivity were higher in CD4+ T + HSF cells from patients than in those from controls. rhIL-35 treatment inhibited proliferation (p < 0.001), but increased IL-10 and decreased IL-17A, α-SMA, and COL-1 secretion into the conditioned medium of CD4+ T lymphocytes + HSFs from patients compared with those from controls. IL-35 mAb blocked the effects of IL-35 in CD4+ T + HSF cells (p < 0.05). IL-35 plays an inhibitory role in CD4+ T lymphocyte proliferation but induces Treg cell differentiation by STAT1 signalling activation, HSF proliferation, and collagen expression in systemic sclerosis.

Preparation of 6-carboxyl chitin and its effects on cell proliferation in vitro

This study concerns the performance evaluation of 6-carboxyl chitin for its wound healing application. 6-Carboxyl chitins were prepared by the oxidation of chitin at C-6 with NaClO/TEMPO/NaBr after α-chitin was pretreated in NaOH/urea solution. The products with different molecular weights were obtained by changing reaction conditions. They all were completely oxidized at C-6 and N-acetylated at C-2 according to FT-IR and NMR results. 6-Carboxyl chitins could stimulate significantly the proliferation of human skin fibroblasts (HSF) and human keratinocytes (HaCaT), and the bioactivities were concentration and Mws dependent. Within the scope of the study, 10–40 kDa of Mws and 10−100 μg/mL of concentrations were most suitable for the HSF proliferation, but the proliferation of HaCaT increased with decreasing the concentration and Mw. In addition, 6-carboxyl chitins could also induce macrophages and fibroblasts to secrete growth factors. Therefore, 6-carboxyl chitins could be expected to be an active ingredient for wound healing.

2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside-stimulated dental pulp stem cells-derived conditioned medium enhances cell activity and anti-inflammation

Background/purposeDental pulp stem cells (DPSCs) contribute to the regeneration of various tissues and have superior proliferation, immune privilege, and anti-inflammation properties to other mesenchymal stem cells. 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside (THSG) not only enhances the aforementioned properties of DPSCs but also promotes self-renewal and reprogramming-like ability. However, whether THSG enhances the aforementioned properties and abilities through direct or indirect interaction mechanisms remains unclear. To address this knowledge gap, we examined the effects of THSG-stimulated DPSC-derived conditioned medium (THSG-CM) on the activity and anti-inflammation properties of cells. Materials and methodsDPSCs were treated with various concentrations of THSG to produce THSG-CM, which was then collected, analyzed, and lyophilized. A cytokine profiling antibody assay was used to compare protein components between THSG-treated and nontreated CM. Human skin fibroblasts (HSFs) and human gingival fibroblasts (HGFs) were used to investigate the effect of THSG-CM on cell proliferation, anti-inflammation, and wound healing abilities; for this investigation, MTS assay, quantitative real-time PCR analysis, and 2-well silicone inserts wound model were conducted. ResultsWe observed that THSG enhanced the secretion of growth- and immune-associated proteins in THSG-CM and increased the proliferation of HSFs and HGFs. Furthermore, THSG-CM significantly attenuated lipopolysaccharide-stimulated mRNA levels of cytokines in both cells and improved wound healing abilities. ConclusionWe conclude that THSG-CM had more beneficial effects on cell activity and anti-inflammation in the HSFs and HGFs than DPSC-derived CM. DPSC-derived CM can be developed into a cell-free regenerative strategy in the future, and its therapeutic efficacy may be improved by THSG-CM.

Hyperbaric oxygen potentiates diabetic wound healing by promoting fibroblast cell proliferation and endothelial cell angiogenesis

BackgroundDiabetic foot ulcer (DFU), one of the diabetic complications, brings high burden to diabetic patients. Hyperbaric oxygen therapy (HBOT) has been proven to be an effective clinical method for the treatment of DFU. However, the mechanisms still to be elucidated. MethodsDiabetic foot mice model was established, and treated with hyperbaric oxygen. Haematoxylin & eosin (H&E) staining and Masson's trichrome staining were used for the analysis of wound healing. Human skin fibroblast (HSF) and human umbilical vein endothelial cell (HUVECS) were exposed to high glucose and hyperbaric oxygen for studying the mechanism of hyperbaric oxygen promoted wound healing in vitro. Wound healing assay, reactive oxygen species (ROS) assay, cell proliferation assay and tube formation assay were used for the analysis of wound healing. Quantitative-polymerase chain reaction (Q-PCR), Western blotting and enzyme-linked immunosorbent assay (ELISA) were used for the analysis of gene expression. ResultsHBOT facilitated wound healing in DFU mice model, and promoted the expression of HIF-1α, NF-κB, VEGFA, SDF-1, VEGFR2 and CXCR4. Hyperbaric oxygen promoted the proliferation, migration and ROS production, as well as the expression of SDF-1 and VEGFA in HSF. HBOT stimulated the proliferation, migration and tube formation, as well as the expression of CXCR4 and VEGFR2 in HUVECS. ConclusionHyperbaric oxygen potentiates angiogenesis and diabetic wound healing by activating HIF-1α signaling, so as to promote the expression of VEGF/SDF-1 in HSF and the expression of VEGFR/CXCR4 in HUVECS, ultimately to promote the proliferation of HSF and the angiogenesis of HUVECS.