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BACKGROUND: Some scholars have found that dermal papilla spheroid–derived exosomes could promote the development of hair follicles. However, whether adipose-derived stem cell exosomes (ADSC-Exos) have a similar effect on hair growth has not been determined yet. Thus, the purpose of this article was to detect whether ADSC-Exos could promote hair regeneration. METHODS: Adipose-derived stem cells (ADSCs) were isolated from 6-week-old C57BL/6 mice. Then, ADSC-Exos were isolated from the ADSCs. Western blotting was used to detect specific exosome markers. The particle size and distribution of the exosomes were analyzed by NanoSight dynamic light scattering. A total of 12 nude mice were randomly divided into two groups (n = 6 each): the ADSC-Exos group and the control group. For the control group, a mixture of freshly isolated dermal cells (DCs) and epidermal cells (ECs) was grafted. For the ADSC-Exos group, a mixture of DCs, ECs, and 50 μg/ml of ADSC-Exos was grafted. Gross evaluation of the hair regeneration was carried out 2–3 weeks after the transplantation, and the graft site was harvested for histology at the third week. RESULTS: The existence of exosomes derived from ADSCs was evidenced by CD63, ALX1, and CD9 expression. Two or three weeks after the grafting, the number of regenerated hairs in the ADSC-Exos group was higher than that in the control group (p

Background: Idiopathic pulmonary fibrosis (IPF) is a chronic progressive respiratory disease. Arguably, the complex interplay between immune cell subsets, coupled with an incomplete understanding in disease pathophysiology, have contributed to the paucity of successful therapies. In turn, IPF remains a fatal disease at present. Thus, with no effective therapy for either the prevention or treatment of IPF, the need for new therapies is paramount. Mesenchymal stromal/stem cell (MSC) therapy has shown promise in experimental models of IPF. Methods:The aim of this study was to test the therapeutic effect of adipose-derived mesenchymal stem cells in a bleomycin-induced pulmonary fibrosis model. First, MSC cells were obtained from mice, and then flow cytometry and cell differentiation culture methods were uesd to identify them. Then adult C57BL/6 mice were challenged with endotracheal instillation of bleomycin and treated with MSC concurrently for reversal models at day 14. Experimental groups were assessed at day 14, 21, or 28. Finally, lung fibroblasts were challenged with TGF-β1 and treated with MSC supernatant or MSCs to explore the relevant mechanisms of pulmonary fibrosis reversal. Results:We successfully obtained mesenchymal stem cells from mouse adipose tissue and identified their differentiation ability and cell phenotype.We found that the presence of MSC or its supernatant could promote the proliferation and migration of lung fibrotic cells. Meanwhile, MSC supernatant could reduce lung collagen deposition, improve Ashcroft score, and reduce the gene and protein content of lung fibrosis-related substances. Bleomycin-challenged mice presented with severe septal thickening and prominent fibrosis, and this was effectively reversed by MSC treatment.Conclusions: In summary, this study found that MSC therapy can prevent and revert the core features of bleomycin-induced pulmonary fibrosis, and the beneficial effects of MSC may be mediated via its paracrine-related cytokines. It provides further evidence for the importance of the MSC in cell therapy medicine applications.

Background: Platelet-rich plasma (PRP) is a portion of blood plasma enriched with platelets widely investigated for accelerating bone and soft tissue healing. The purpose of this study is to evaluate the effects of PRP on the previous established photoaging model in vitro and to elucidate the potential anti-photoaging mechanisms of PRP. Methods: Murine dermal fibroblasts (MDFs) were isolated and cultured with or without PRP after exposure to repeated UVB irradiation per protocol. The senescent phenotypes were compared among groups, including cell morphology, senescence-associated β-galactosidase (SA-β-gal) expression, cell cycle arrest, production and degradation of extracellular matrix (ECM), ROS generation, and the alteration of major intracellular antioxidants. Results: 1% PRP effectively reduced the amount of flattened cells and significantly decreased the staining intensity as well as the percentage of SA-β-gal-positive cells when compared with UVB group. Furthermore, PRP appeared to prevent cell cycle arrest caused by irradiation by decreasing p53 and p21 expression. The PRP had demonstrated direct effects on ECM, including increase in type I collagen production and decrease in MMPs expression. Additionally, the increase of glutathione, one of the major intracellular antioxidants, induced by PRP provided a possible explanation for its therapeutic effect. Conclusions: Our work confirmed that PRP, a compound of various growth factors, could counteract senescence-like phenotypes at the cellular level, mirroring its rising clinical popularity in anti-aging and regenerative medicine. The promising effects of PRP calls for future study for further exploration of the underlying mechanisms, which will lead to a broadened application of PRP in future clinical use.

This study provides a novel method in injectable tissue engineering which contains porcine extracellular matrix (ECM) powder scaffolds and stromal-vascular fraction (SVF) cells. We fabricated ECM powders from porcine adipose tissue and mixed the SVF cell suspension together with the ECM powders immediately after the isolation. SVF-loaded ECM powders and their controls were respectively injected into the dorsa of mice. After 6 weeks, the interest sites were harvested and analyzed using histology, immunohistochemistry, and quantitative-polymerase chain reaction (qPCR). Histology and immunohistochemistry demonstrated that the presence of the SVF cells loaded in the ECM powders promoted adipogenesis, angiopoiesis and matrix remodeling. This was in parallel with upregulation of host cell recruitment and adipogenesis genes. The results provide cellular and molecular evidence suggesting a novel method in injectable tissue engineering. We believe that SVF-loaded ECM powders could act as efficient injectable biomaterials for tissue engineering and have great potential for meeting clinical challenges in regenerative medicine, particularly in relation to adipose tissue engineering.

GNB2L1 and its O-GlcNAcylation has been reported to play roles in gastric cancer metastasis. However, the roles of GNB2L1 in chemoresistance of gastric cancer has never been determined. In the present study, we found that GNB2L1 was downregulated in chemoresistant patients of gastric cancer, and observed the decrease of GNB2L1 in protein levels instead of mRNA levels in different chemoresistant gastric cancer cell lines. Further we proved that this downregulation of GNB2L1 was resulted from its elevated O-GlcNAcylation catalyzed by OGT in both cell lines and patients. Next, we investigate the function of GNB2L1 and its O-GlcNAcylation on gastric cancer metastasis during chemoresistance, and confirmed Ser124 as the major O-GlcNAcylation site on GNB2L1 that regulated its function on metastasis. Furthermore, our data demonstrated that GNB2L1 modulated EMT via regulating the translation of EMT-related proteins in the process of chemoresistance. In summary, this study indicated that GNB2L1 and its O-GlcNAcylation regulated metastasis via modulating the translation of EMT-related proteins in the chemoresistance of gastric cancer.

Background/Aims: Ultraviolet B (UVB) irradiation alters multiple molecular pathways in the skin, thereby inducing skin photoaging. Murine dermal fibroblasts (MDFs) were subjected to a series of 4 sub-cytotoxic UVB doses (120 mJ/cm2), resulting in changes in cell shape, DNA damage, cell cycle arrest, extracellular matrix variations, reactive oxygen species (ROS) generation, and alterations in major intracellular antioxidant and cellular autophagy levels. Rapamycin (RAPA) is a new macrolide immunosuppressive agent that is primarily used in oncology, cardiology, and transplantation medicine and has been found to extend the lifespan of genetically heterogeneous mice. Several studies have shown that RAPA may have anti-aging effects in cells and organisms. Thus, in this study, we explored the effects and mechanisms of RAPA against the photoaging process using a well-established cellular photoaging model. Methods: We developed a stress-induced premature senescence (SIPS) model through repeated exposure of MDFs to ultraviolet B (UVB) irradiation. The cells were cultured in the absence or presence of RAPA for 48 h. Senescent phenotypes were assessed by examining cell viability, cell morphology, senescence-associated β-galactosidase (SA-β-gal) expression, cell cycle progression, intracellular ROS production, matrix metalloproteinase (MMP) synthesis and degradation, extracellular matrix (ECM) component protein expression, alterations in major intracellular antioxidant levels, and the cellular autophagy level. Results: Compared with the UVB group, pretreatment with RAPA (5 µM) significantly decreased the staining intensity and percentage of SA-β-gal-positive cells and preserved the elongated cell shape. Moreover, cells pretreated with RAPA showed inhibition of the reduction in the type I collagen content by blocking the UVB-induced upregulation of MMP expression. RAPA also decreased photoaging cell cycle arrest and downregulated p53 and p21 expression. RAPA application significantly attenuated irradiation-induced ROS release by modulating intracellular antioxidants and increasing the autophagy level. Conclusions: Our study demonstrated that RAPA elicited oxidative damage in vitro by reducing ROS accumulation in photoaged fibroblasts. The anti-aging effect can be attributed to the maintenance of normal antioxidant and cellular autophagy levels. However, determination of the definitive mechanism requires further study.

The commercial use of epidermal growth factor (EGF) is extensive and has been shown to be effective for skin wound healing in clinical practice. There is evidence to indicate that the topical administration of EGF significantly accelerates re-epithelialization by promoting keratinocyte mitogenesis and migration following acute injury; however, the mechanisms involved remain to be elucidated. Thus, in this study, we focused on Kindlin-1, a four-point-one, ezrin, radixin, moesin (FERM)-domain-containing adaptor protein, and report its contribution to EGF-induced re-epithelialization in skin wound healing. In tissue samples, the expression of Kindlin-1 was induced upon EGF treatment compared to that in the natural healing group. In immortalized human keratinocytes (HaCaT cells), we further proved that Kindlin-1 was necessary for mediating EGF-induced activation signals, including integrin β1 activation, focal adhesion kinase (FAK) phosphorylation and actin re-organization, which finally led to enhanced cell proliferation and migration. These results indicate that Kindlin-1 is essential in EGF-induced re-epithelialization in skin wound healing and provide additional rationale for the clinical application of EGF in the treatment of acute wounds.

Background Histone deacetylase (HDAC) inhibitors are widely used in clinical investigation as novel drug targets. For example, panobinostat and vorinostat have been used to treat patients with melanoma. However, HDAC inhibitors are small-molecule compounds without a specific target, and their mechanism of action is unclear. Therefore, it is necessary to investigate which HDACs are required for the proliferation and metastasis of melanoma cells. Methods We used overexpression and knocking down lentivirus to clarify the influence of HDAC5 and HDAC6 in melanoma development. Also, we introduced stable HDAC5 or HDAC6 knockdown cells into null mice and found that the knockdown cells were unable to form solid tumors. Finally, we tested HDAC5 and HDAC6 expression and sub-location in clinical melanoma tissues and tumor adjacent tissues. Results In this study, and found that HDAC5 and HDAC6 were highly expressed in melanoma cells but exhibited low expression levels in normal skin cells. Furthermore, we knocked down HDAC5 or HDAC6 in A375 cells and demonstrated that both HDAC5 and HDAC6 contributed to the proliferation and metastasis of melanoma cells. Conclusions This study demonstrated both HDAC5 and HDAC6 were required for melanoma cell proliferation and metastasis through different signaling pathways. Electronic supplementary material The online version of this article (doi:10.1186/s12967-015-0753-0) contains supplementary material, which is available to authorized users.

Additional file 4: Figure S4. The effect of HDAC5 and HDAC6 on metastasis didn’t depend on BRAF V600 mutation. a knocking down HDAC5 or HDAC6 in multiple BRAF V600 mutation cells. b Transwell assay to detect metastasis of A375 cells. Cell transwell assays were performed using FalconTM Cell Culture Inserts (BD353097) according to the manufacturer’s instructions. After 24–48 h of incubation, the remaining cells in the upper chamber were removed by cotton swabs. The cells on the lower surface of the membrane were fixed in 4 % paraformaldehyde and stained with 0.5 % crystal violet. Cells in at least 3 random microscopic fields (magnification ×10) were counted and photographed. All experiments were performed in duplicate and repeated 3 times.

Several clinical research studies have demonstrated that chronic cutaneous wounds can be treated with hair follicle grafts. However, the clinical outcomes of hair follicle grafting compared to split-thickness skin grafting have not been examined. This study sought to compare the clinical outcomes of patients with chronic wounds following hair follicle therapy and split-thickness skin graft therapy in a relatively large cohort of patients. Forty patients were enrolled in the study, a retrospective analysis of all patients underwent therapy with hair follicles (cohort A) and split-thickness skin grafts (cohort B) was performed. Safety, healing duration, skin quality (recipient site), scar formation (donor site) and overall postoperative outcome were analyzed. The wound sites were examined using photography at weeks 2, 8, and 12 after surgery. Five non-biased reviewers estimated the above-mentioned clinical outcomes using a five-point Likert scale. The ages and wound areas were similar between cohorts A (n=20) and B (n=20). Total wound closure was observed and adverse events were rare and controllable in both cohorts. The skin and scar quality were rated significantly higher in the hair follicle cohort than the split-thickness skin graft cohort (4.40 vs 3.45, P

We have previously shown alpha-melanocyte stimulating hormone to protect melanocytes and melanoma cells from the proinflammatory actions of tumor necrosis factor-alpha. The aim of the study was to extend this work to look into the influence of tumor necrosis factor-alpha on melanoma cell attachment, invasion, and integrin expression and ask to what extent alpha-melanocyte stimulating hormone might protect cells from tumor necrosis factor-alpha stimulation of increased integrin expression. HBL human melanoma cells were studied under resting and stressed conditions using tumor necrosis factor-alpha as a proinflammatory cytokine. Functional information on the actions of tumor necrosis factor-alpha on melanoma cells was obtained by examining the strength of attachment of melanoma cells to substrates and the ability of melanoma cells to invade through fibronectin. alpha3, alpha4, and beta1 integrin expression was detected by Western immunoblotting and the ability of alpha-melanocyte stimulating hormone to oppose the actions of tumor necrosis factor-alpha was studied on HBL cell attachment, invasion, and integrin subunit expression. Our results show that tumor necrosis factor-alpha increases the number of melanoma cells attaching to collagen (types I and IV) and tissue culture polystyrene, increases ability to invade through fibronectin, and upregulates the expression of alpha3 (28%), alpha4 (90%), and beta1 (65%) integrin subunit expression. In contrast, alpha-melanocyte stimulating hormone reduced cell attachment, invasion, and integrin expression and opposed the stimulatory effects of tumor necrosis factor-alpha. In conclusion this study provides further evidence of alpha-melanocyte stimulating hormone acting to "protect" melanoma cells from proinflammatory cytokine action. Our data support a hypothesis that an inflammatory environment would promote melanoma invasion and that the anti-invasive actions of alpha-melanocyte stimulating hormone are consistent with its working in an anti-inflammatory capacity.