Evaluation of dermal growth of keratinocytes derived from foreskin in co-culture condition with mesenchymal stem cells on polyurethane/gelatin/amnion scaffold

Staji, Masumeh; Sadeghzadeh, Negar; Benisi, Soheila Zamanlui; Azarani, Mojgan; Golchin, Ali; Soleimani, Masoud; Ardeshirylajimi, Abdolreza; Khojasteh, Arash; Hosseinzadeh, Simzar

doi:10.1080/00914037.2021.2018316

Cited by 7 publications

(19 citation statements)

References 52 publications

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“…However, wound healing with the same closure rate was represented on the fifteenth day in all different groups. Most studies 18,19,40 affirmed that the PU scaffold could adhere uniformly to the wound surface without accumulating wound exudates. At the same time, PU in combination with other polymeric materials such as PU/EEP 39 , PU/ EEP-PCL/Gel 39 , and PU/30WEP 40 www.nature.com/scientificreports/ reported adding other materials to PU-CA polymers, such as curcumin, reduced graphene oxide, and silver which showed a fast healing rate 18 .…”

Section: Discussionmentioning

confidence: 99%

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Overexpression of VEGF in dermal fibroblast cells accelerates the angiogenesis and wound healing function: in vitro and in vivo studies

Shams

Moravvej

Hosseinzadeh

et al. 2022

Sci Rep

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Fibroblasts are the main cells of connective tissue and have pivotal roles in the proliferative and maturation phases of wound healing. These cells can secrete various cytokines, growth factors, and collagen. Vascular endothelial growth factor (VEGF) is a unique factor in the migration process of fibroblast cells through induces wound healing cascade components such as angiogenesis, collagen deposition, and epithelialization. This study aimed to create VEGF165 overexpressing fibroblast cells to evaluate angiogenesis function in wound healing. In vitro, a novel recombinant expression vector, pcDNA3.1(-)-VEGF, was produced and transfected into the fibroblast cells. Following selecting fibroblast cells with hygromycin, recombinant cells were investigated in terms of VEGF expression by quantifying and qualifying methods. Mechanical, physical, and survival properties of polyurethane-cellulose acetate (PU-CA) scaffold were investigated. Finally, in vivo, the angiogenic potential was evaluated in four groups containing control, PU-CA, PU-CA with fibroblast cells, and VEGF-expressing cells on days 0, 2, 5, 12 and 15. Wound biopsies were harvested and the healing process was histopathologically evaluated on different days. qRT-PCR showed VEGF overexpression (sevenfold) in genetically-manipulated cells compared to fibroblast cells. Recombinant VEGF expression was also confirmed by western blotting. Manipulated fibroblast cells represented more angiogenesis than other groups on the second day after surgery, which was also confirmed by the antiCD31 antibody. The percentage of wound closure area on day 5 in genetically-manipulated Hu02 and Hu02 groups showed a significant reduction of wound area compared to other groups. These findings indicate that overexpression of VEGF165 in fibroblast cells results in enhanced angiogenesis and formation of granulated tissue in the early stage of the healing process, which can show its therapeutic potential in patients with impaired wound healing and also provide functional support for gene therapy.

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Section: Discussionmentioning

confidence: 99%

“…Hence, the combination of PU and CA could decrease the limitation of both polymers. The electrospun PU-CA scaffold can be used as external support for cell proliferation and adhesion, which improve tissue regeneration and reorganization 19 .…”

Section: Introductionmentioning

confidence: 99%

Overexpression of VEGF in dermal fibroblast cells accelerates the angiogenesis and wound healing function: in vitro and in vivo studies

Shams

Moravvej

Hosseinzadeh

et al. 2022

Sci Rep

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“…Coculturing cells with biodegradable gransplantable scaffolds that mimic the ECM is a promising approach for delivering cell-based therapeutics to the wound site. 69 Implantable cell/biomaterial constructs for skin regenerating, as an epithelial tissue with two main layers, have been successfully developed using tissue engineering concepts. Current regenerative medicine and tissue engineering methodologies demand combining biomaterials, substances, and cells to construct complex structures.…”

Section: Significance Statementmentioning

confidence: 99%

“…Pasteur's Institute) and cocultured keratinocytes and adipose-derived MSCs as described in our previous study, 69 were used for in vitro investigations (Figure 1B). AMSCs expressed unique CD markers, which were confirmed by flow cytometry (Figure 3C).…”

Section: Cell Culture Processmentioning

confidence: 99%

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Evaluation of in vitro coculture of keratinocytes derived from foreskin and adipose‐derived mesenchymal stem cells (AMSCs) on a multilayer oxygen‐releasing electrospun scaffold based on PU/PCL.Sodium percarbonate (SPC)‐gelatine/PU

Azari

Rahimi

Rajabibazl

et al. 2023

Cell Biochemistry & Function

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Despite significant advancements in tissue engineering and regenerative medicine during the last two decades, the fabrication of proper scaffolds with appropriate cells can still be considered a critical achievement in this field. Hypoxia is a major stumbling block to chronic wound healing, which restrains tissue engineering plans because a lack of oxygen may cause cell death. This study evaluated the cocultured human keratinocytes and human adipose‐derived mesenchymal stem cells (AMSCs) on a multilayer oxygen‐releasing electrospun scaffold based on PU/PCL.Sodium percarbonate (SPC)‐gelatin/PU. The scaffold was characterized using Fourier transform infrared (FTIR) and scanning electron microscopy (SEM) methods. Flow cytometry confirmed mesenchymal stem cells, and then the 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5 diphenyltetrazolium bromide (MTT) assay and DAPI staining were used to assess the in vitro biocompatibility of the scaffold. The experimental results showed that the multilayer electrospun scaffold containing 2.5% SPC could efficiently produce oxygen. Furthermore, according to cell viability results, this structure makes a suitable substrate for the coculture of keratinocytes and AMSCs. Gene expression analysis of various markers such as Involucrin, Cytokeratin 10, and Cytokeratin 14 after 14 days confirmed that keratinocytes and AMSCs coculture on PU/PCL.SPC‐gelatin/PU electrospun scaffold promotes dermal differentiation and epithelial proliferation compared to keratinocytes single‐cell culture. Therefore, our study supports using oxygen‐releasing scaffolds as a potential strategy to hasten skin tissue regeneration. Based on the results, this structure is suggested as a promising candidate for cell‐based skin tissue engineering. Given that the developed oxygen‐generating polymeric electrospun scaffolds could be used as part of a future strategy for skin tissue engineering, the PU/PCL.SPC‐gelatin/PU hybrid electrospun multilayer scaffold in combination with keratinocyte/AMSC coculture is proposed as an effective substrate for skin tissue engineering and regenerative medicine platforms.

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Hydrogel foam dressings with angiogenic and immunomodulatory factors from mesenchymal stem cells

Lan,

Fletcher,

Bender

et al. 2024

J Biomedical Materials Res

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Stem cell therapy and skin substitutes address the stalled healing of chronic wounds in order to promote wound closure; however, the high cost and regulatory hurdles of these treatments limit patient access. A low‐cost method to induce bioactive healing has the potential to substantially improve patient care and prevent wound‐induced limb loss. A previous study reported that bioactive factors derived from apoptotic‐like mesenchymal stem cells (MSCs) demonstrated anti‐inflammatory and proangiogenic effects and improved ischemic muscle regeneration. In this work, these MSC‐derived bioactive factors were loaded into a hydrogel foam to harness immunomodulatory and angiogenic properties from MSC components to facilitate chronic wound healing without the high cost and translational challenges of cell therapies. After incorporation of bioactive factors, the hydrogel foam retained high absorbency, moisture retention, and target water vapor transmission rate. High loading efficiency was confirmed and release studies indicated that over 90% of loaded factors were released within 24 h. Ethylene oxide sterilization and 4‐week storage did not affect the bioactive factor release profile or physical properties of the hydrogel foam dressing. Bioactivity retention of the released factors was also confirmed for as‐sterilized, 4°C‐stored, and −20°C‐stored bioactive hydrogel foams as determined by relevant gene expression levels in treated pro‐inflammatory (M1) macrophages. These results support the use of the bioactive dressings as an off‐the‐shelf product. Overall, this work reports a new method to achieve a first‐line wound dressing with the potential to reduce persistent inflammation and promote angiogenesis in chronic wounds.

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Evaluation of dermal growth of keratinocytes derived from foreskin in co-culture condition with mesenchymal stem cells on polyurethane/gelatin/amnion scaffold

Cited by 7 publications

References 52 publications

Overexpression of VEGF in dermal fibroblast cells accelerates the angiogenesis and wound healing function: in vitro and in vivo studies

Overexpression of VEGF in dermal fibroblast cells accelerates the angiogenesis and wound healing function: in vitro and in vivo studies

Evaluation of in vitro coculture of keratinocytes derived from foreskin and adipose‐derived mesenchymal stem cells (AMSCs) on a multilayer oxygen‐releasing electrospun scaffold based on PU/PCL.Sodium percarbonate (SPC)‐gelatine/PU

Hydrogel foam dressings with angiogenic and immunomodulatory factors from mesenchymal stem cells

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