scholarly journals In Vitro Wound Healing Potential of Photobiomodulation Is Possibly Mediated by Its Stimulatory Effect on AKT Expression in Adipose-Derived Stem Cells

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Naresh K. Rajendran ◽  
Nicolette N. Houreld ◽  
Heidi Abrahamse
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Power Density ◽  
Western Blotting ◽  
Akt Signaling ◽  
Enzyme Linked Immunosorbent Assay ◽  
Adipose Derived Stem Cells ◽  
Cell Models ◽  
Trypan Blue Exclusion

Increasing evidence suggests that adipose-derived stem cells (ADSCs) serve as a therapeutic approach for wound healing. The aim of this study was to determine the effect of photobiomodulation (PBM) on antioxidant enzymes in ADSCs. Four ADSC cell models, namely, normal, wounded, diabetic, and diabetic wounded, were irradiated with 660 nm (fluence of 5 J/cm2 and power density of 11.2 mW/cm2) or 830 nm (fluence of 5 J/cm2 and power density of 10.3 mW/cm2). Nonirradiated cells served as controls. Cell morphology and wound migration were determined using light microscopy. Cell viability was determined by the trypan blue exclusion assay. The enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of antioxidants (superoxide dismutase (SOD), catalase (CAT), and heme oxygenase (HMOX1)). AKT activation and FOXO1 levels were determined by immunofluorescence and western blotting. The gaps (wound) in PBM-treated wounded and diabetic wounded cell models closed faster than the controls. PBM treatment significantly increased antioxidant levels in all cell models. This reflects that oxidative stress is reduced on the counterpart of increased antioxidant levels. This might be due to the activation of the AKT signaling pathway as evidenced by the increased AKT signals via western blotting and immunofluorescence. This data suggests that PBM promotes wound healing by increasing antioxidant levels by activating AKT signaling.

Extracellular vesicles derived from adipose-derived stem cells accelerate diabetic wound healing by suppressing the expression of matrix metalloproteinase-9

2021 ◽  
Vol 22 ◽  
Author(s):  
Jiang-wen Wang ◽  
Yuan-zheng Zhu ◽  
Xuan Hu ◽  
Jia-ying Nie ◽  
Zhao-hui Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Mouse Model ◽  
Adipose Derived Stem Cells ◽  
Collagen Deposition ◽  
Diabetic Wound ◽  
Diabetic Wound Healing ◽  
Diabetic Wounds ◽  
Metalloproteinase 9

Background: The healing of diabetic wounds is poor due to a collagen deposition disorder. Matrix metalloproteinase-9 (MMP-9) is closely related to collagen deposition in the process of tissue repair. Many studies have demonstrated that extracellular vesicles derived from adipose-derived stem cells (ADSC-EVs) promote diabetic wound healing by enhancing collagen deposition. Objective: In this study, we explored if ADSC-EVs could downregulate the expression of MMP-9 in diabetic wounds and promote wound healing by improving collagen deposition. The potential effects of ADSC-EVs on MMP-9 and diabetic wound healing were tested both in vitro and in vivo. Methods: We first evaluated the effect of ADSC-EVs on the proliferation and MMP-9 secretion of HaCaT cells treated with advanced glycation end product-bovine serum albumin (AGE-BSA), using CCK-8 western blot and MMP-9 enzyme-linked immunosorbent assay(ELISA). Next, the effect of ADSC-EVs on the healing, re-epithelialisation, collagen deposition, and MMP-9 concentration in diabetic wound fluids was evaluated in an immunodeficient mouse model via MMP-9 ELISA and haematoxylin and eosin, Masson’s trichrome, and immunofluorescence staining for MMP-9. Results: In vitro, ADSC-EVs promoted the proliferation and MMP-9 secretion of HaCaT cells.In vivo, ADSC-EVs accelerated diabetic wound healing by improving re-epithelialisation and collagen deposition and by inhibiting the expression of MMP-9. Conclusion: ADSC-EVs possessed the healing of diabetic wounds in a mouse model by inhibiting downregulating MMP-9 and improving collagen deposition.Thus ,ADSC-EVs are a promising candidate for the treatment of diabetic wounds .

scholarly journals A gelatin/collagen/polycaprolactone scaffold for skin regeneration

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6358 ◽  
Author(s):  
Lin-Gwei Wei ◽  
Hsin-I Chang ◽  
Yiwei Wang ◽  
Shan-hui Hsu ◽  
Lien-Guo Dai ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Dermal Fibroblasts ◽  
Skin Regeneration ◽  
Collagen Content ◽  
Wound Dressings ◽  
Skin Substitute ◽  
Epidermal Keratinocytes ◽  
Adipose Derived Stem Cells

Background A tissue-engineered skin substitute, based on gelatin (“G”), collagen (“C”), and poly(ε-caprolactone) (PCL; “P”), was developed. Method G/C/P biocomposites were fabricated by impregnation of lyophilized gelatin/collagen (GC) mats with PCL solutions, followed by solvent evaporation. Two different GC:PCL ratios (1:8 and 1:20) were used. Results Differential scanning calorimetry revealed that all G/C/P biocomposites had characteristic melting point of PCL at around 60 °C. Scanning electron microscopy showed that all biocomposites had similar fibrous structures. Good cytocompatibility was present in all G/C/P biocomposites when incubated with primary human epidermal keratinocytes (PHEK), human dermal fibroblasts (PHDF) and human adipose-derived stem cells (ASCs) in vitro. All G/C/P biocomposites exhibited similar cell growth and mechanical characteristics in comparison with C/P biocomposites. G/C/P biocomposites with a lower collagen content showed better cell proliferation than those with a higher collagen content in vitro. Due to reasonable mechanical strength and biocompatibility in vitro, G/C/P with a lower content of collagen and a higher content of PCL (GCLPH) was selected for animal wound healing studies. According to our data, a significant promotion in wound healing and skin regeneration could be observed in GCLPH seeded with adipose-derived stem cells by Gomori’s trichrome staining. Conclusion This study may provide an effective and low-cost wound dressings to assist skin regeneration for clinical use.

scholarly journals Paracrine Activity from Adipose-Derived Stem Cells on In Vitro Wound Healing in Human Tympanic Membrane Keratinocytes

2017 ◽  
Vol 26 (6) ◽  
pp. 405-418 ◽  
Author(s):  
Huan Ting Ong ◽  
Sharon L. Redmond ◽  
Robert J. Marano ◽  
Marcus D. Atlas ◽  
Magnus von Unge ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Tympanic Membrane ◽  
Adipose Derived Stem Cells ◽  
Human Tympanic Membrane

scholarly journals Effect of Nanostructured Scaffold on Human Adipose-Derived Stem Cells: Outcome of In Vitro Experiments

Nanomaterials ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1822 ◽  
Author(s):  
Marina Borgese ◽  
Ludovica Barone ◽  
Federica Rossi ◽  
Mario Raspanti ◽  
Roberto Papait ◽  
...  
Keyword(s):  
Stem Cells ◽  
Culture Medium ◽  
Quantitative Polymerase Chain Reaction ◽  
Enzyme Linked Immunosorbent Assay ◽  
Adipose Derived Stem Cells ◽  
In Vitro Experiments ◽  
Fatty Acid Binding ◽  
Beneficial Effects ◽  
Angiogenesis Process

This work is addressed to provide, by in vitro experiments, results on the repercussion that a nanostructured scaffold could have on viability, differentiation and secretion of bioactive factors of human adipose-derived stem cells (hASCs) when used in association to promote angiogenesis, a crucial condition to favour tissue regeneration. To achieve this aim, we evaluated cell viability and morphology by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and microscopy analysis, respectively. We also investigated the expression of some of those genes involved in angiogenesis and differentiation processes utilizing quantitative polymerase chain reaction (qPCR), whereas the amounts of Vascular Endothelial Growth Factor A, Interleukin 6 and Fatty Acid-Binding Protein 4 secreted in the culture medium, were quantified by enzyme-linked immunosorbent assay (ELISA). Results suggested that, in the presence of the scaffold, cell proliferation and the exocytosis of factors involved in the angiogenesis process are reduced; by contrast, the expression of those genes involved in hASC differentiation appeared enhanced. To guarantee cell survival, the construct dimensions are, generally, smaller than clinically required. Furthermore, being the paracrine event the primary mechanism exerting the beneficial effects on injured tissues, the use of conditioned culture medium instead of cells may be convenient.

Effect of HBO therapy on adipose-derived stem cells, fibroblasts and co-cultures: In vitro study of oxidative stress, angiogenic potential and production of pro-inflammatory growth factors in co-cultures1

2020 ◽  
pp. 1-13
Author(s):  
P. Engel ◽  
M. Ranieri ◽  
O. Felthaus ◽  
S. Geis ◽  
F. Haubner ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Stem Cells ◽  
Wound Healing ◽  
Hyperbaric Oxygen ◽  
Hyperbaric Oxygen Therapy ◽  
Oxygen Therapy ◽  
Reactive Oxygen ◽  
Adipose Derived Stem Cells ◽  
Oxygen Species

BACKGROUND: A key moderator of wound healing is oxygen. Wound healing is a dynamic and carefully orchestrated process involving blood cells, cytokines, parenchymal cells (i.e. fibroblasts and mesenchymal stem cells) and extracellular matrix reorganization. Human adipose derived stem cells as well as human fibroblasts produce soluble factors, exhibit diverse effects on inflammation and anti inflammation response and are involved in wound healing processes. Hyperbaric oxygen therapy is an effective adjunct treatment for ischemic disorders such as chronic infection or chronic wounds. In vitro effects of hyperbaric oxygen therapy on human cells were presented in many studies except for those on mono- and co-cultures of human adipose derived stem cells and fibroblasts. OBJECTIVE: The aim of this study was to investigate the effects of hyperbaric oxygen therapy on mono- and co-cultures of human adipose derived stem cells and fibroblasts. METHODS: Mono- and co-cultures from human adipose derived stem cells and fibroblasts were established. These cultures were exposed to hyperbaric oxygen therapy every 24 h for five consecutive days. Measuring experiments were performed on the first, third and fifth day. Therapy effects on the expression of VEGF, IL 6 and reactive oxygen species were investigated. RESULTS: After exposure to hyperbaric oxygen, cell culturess showed a significant increase in the expression of VEGF after 3 and 5 days. All cultures showed significantly reduced formation of reactive oxygen species throughout the experiments. The expression of IL-6 decreased during the experiment in mono-cultures of human adipose derived stem cells and co-cultures. In contrast, mono-cultures of human skin fibroblasts showed an overall significantly increased expression of IL-6. CONCLUSIONS: Hyperbaric oxygen therapy leads to immunmodulatory and proangiogenetic effects in a wound-like enviroment of adipose derived stem cells and fibroblasts.

In vitro Engineering of a Skin Substitute Based on Adipose-Derived Stem Cells

2019 ◽  
Vol 207 (1) ◽  
pp. 46-57 ◽  
Author(s):  
Alessia Paganelli ◽  
Luisa Benassi ◽  
Irena Pastar ◽  
Massimo Pellegrini ◽  
Paola Azzoni ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Stromal Vascular Fraction ◽  
Skin Substitute ◽  
Dermal Substitute ◽  
Adipose Derived Stem Cells ◽  
Dermal Matrix ◽  
Chronic Ulcers ◽  
Main Components

In the field of wound healing, stem cell-based strategies are gaining importance for their regenerative potential. Adipose-derived stem cells (ADSCs) are a particular subset of mesenchymal stem cells present in the stromal-vascular fraction of the adipose tissue, today considered very attractive for their relative abundance and accessibility in the human body. However, ADSCs are still not routinely used in normal clinical practice. Several studies have also reported ADSC transplantation in association with biomaterials in an attempt to enhance the local retention and growth rate of the cells. The aim of our study was to evaluate the ability of ADSCs to build a dermal scaffold to be potentially used as a dermal substitute in the field of wound healing, with optimal biocompatibility and mechanical properties. ADSCs were defined as CD90-, CD73-, and CD105-positive cells. ADSCs turned out to be capable of secreting all the main components of the extracellular matrix (ECM) upon stimulation, thus efficiently producing a collagen and fibronectin-containing dermal matrix. We also checked whether the ADSC-produced dermal scaffold could be seeded with keratinocytes. The scaffolding material directly produced by ADSCs has several advantages when compared to the commercially available ones: it is easily obtained from the patients and it is 100% biocompatible and supports cell-ECM interaction. Moreover, it represents a possible powerful therapeutic tool for patients with chronic ulcers since it appears to be potentially grafted with keratinocytes layers, thus bypassing the classical two-step grafting procedure.

scholarly journals Patterned Polyvinyl Alcohol Hydrogel Dressings with Stem Cells Seeded for Wound Healing

Polymers ◽  
2019 ◽  
Vol 11 (1) ◽  
pp. 171 ◽  
Author(s):  
Tianlin Gao ◽  
Menghui Jiang ◽  
Xiaoqian Liu ◽  
Guoju You ◽  
Wenyu Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Polyvinyl Alcohol ◽  
Adipose Derived Stem Cells ◽  
In Vivo Experiments ◽  
Cell Counts ◽  
Surface Modified ◽  
Testing Instruments

Polyvinyl alcohol (PVA) hydrogel and stem cell therapy have been widely used in wound healing. However, the lack of bioactivity for PVA and security of stem therapy limited their application. In this study, an adipose-derived stem cells (ADSCs)-seeded PVA dressing (ADSCs/PVA) was prepared for wound healing. One side of the PVA dressing was modified with photo-reactive gelatin (Az-Gel) via ultraviolet (UV) irradiation (Az-Gel@PVA), and thus ADSCs could adhere, proliferate on the PVA dressings and keep the other side of the dressings without adhering to the wound. The structure and mechanics of Az-Gel@PVA were determined by scanning electron microscopy (SEM) and material testing instruments. Then, the adhesion and proliferation of ADSCs were observed via cell counts and live-dead staining. Finally, in vitro and in vivo experiments were utilized to confirm the effect of ADSCs/PVA dressing for wound healing. The results showed that Az-Gel was immobilized on the PVA and showed little effect on the mechanical properties of PVA hydrogels. The surface-modified PVA could facilitate ADSCs adhesion and proliferation. Protein released tests indicated that the bioactive factors secreted from ADSCs could penetrated to the wound. Finally, in vitro and in vivo experiments both suggested the ADSCs/PVA could promote the wound healing via secreting bioactive factors from ADSCs. It was speculated that the ADSCs/PVA dressing could not only promote the wound healing, but also provide a new way for the safe application of stem cells, which would be of great potential for skin tissue engineering.

scholarly journals The distribution pattern and growth factor level in platelet-rich fibrin incorporated skin-derived mesenchymal stem cells: An in vitro study

2020 ◽  
Vol 13 (10) ◽  
pp. 2097-2103
Author(s):  
Igo Syaiful Ihsan ◽  
Deya Karsari ◽  
Nora Ertanti ◽  
Aristika Dinaryanti ◽  
Alexander Patera Nugraha ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Growth Factor ◽  
Distribution Pattern ◽  
Enzyme Linked Immunosorbent Assay ◽  
Platelet Rich Fibrin ◽  
Group A ◽  
Group D

Background and Aim: A skin wound in an animal must be cared for to prevent further health issues. Platelet-rich fibrin (PRF) and skin-derived mesenchymal stem cells (SMSCs) have been reported to have potential in increasing the rate of wound healing. This study aimed to analyze the distribution patterns and levels of platelet-derived growth factor (PDGF), insulin-like growth factor (IGF), vascular endothelial growth factor (VEGF), and transforming growth factor-β (TGF-β) in PRF incorporated with SMSCs. Materials and Methods: This study employed a true experiment (in vitro) design with post-test only performed in the control group alone. PRF and SMSCs were extracted from the blood and skin of 16 rabbits. SMSCs were characterized using immunocytochemistry to examine clusters of differentiation for 45, 73, 90, and 105. PRF was incorporated into the SMSCs and then divided into four groups (N=32/n=8): Group A (PRF only), Group B (PRF+SMSCs, incubated for 1 day), Group C (PRF+SMSCs, incubated for 3 days), and Group D (PRF+SMSCs, incubated for 5 days). Scanning electron microscopy was used to examine the distribution pattern of SMSCs between groups. The supernatant serum (Group A) and supernatant medium culture (Group D) were collected for the measurement of PDGF, IGF, VEGF, and TGF-β using an enzyme-linked immunosorbent assay sandwich kit. An unpaired t-test was conducted to analyze the differences between Groups A and D (p<0.01). Results: Group D had the most morphologically visible SMSCs attached to the PRF, with elongated and pseudopodia cells. There was a significant difference between the levels of growth factor in Groups A and D (p=0.0001; p<0.01). Conclusion: SMSCs were able to adhere to and distribute evenly on the surface of PRF after 5 days of incubation. The PRF incorporated SMSCs contained high levels of PDGF, IGF, VEGF, and TGF- β, which may prove to have potential in enhancing wound healing.

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