Platelet lysate as a serum replacement for skin cell culture on biomimetic PCL nanofibers

Sovková, Věra; Vocetková, Karolína; Rampichová, Michala; Míčková, Andrea; Buzgo, Matěj; Lukášová, Věra; Daňková, Jana; Filová, Eva; Nečas, Alois; Amler, Evžen

doi:10.1080/09537104.2017.1316838

Cited by 19 publications

(26 citation statements)

References 37 publications

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“…Furthermore, the samples contained a minimum of spherical defects. The stereology of the scaffolds was in accordance with other findings [14,15]. The scanning electron microscope also showed a fine fibrin network formed on the surface of the plateletfunctionalized samples.…”

Section: Discussionsupporting

confidence: 88%

“…The cell adhesion properties of PVA nanofibers could be improved using other polymers, such as chitosan [20], gelatin [21] or hyaluronic acid [22]. On the other hand, PCL was shown to be effective in fostering the culture of melanocyte, which is in accordance with previously published data [14,18]. The suitability of PCL as a substrate for melanocyte culture was described by Savkovic et al [23].…”

Section: Discussionsupporting

confidence: 81%

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Functionalization of Polymeric Nanofibers Using Platelets for Melanocyte Culture

et al. 2020

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Tissue engineering is an interdisciplinary field that uses a combination of cells, suitable biomaterials and bioactive molecules to engineer the desired tissue and restore lost function. These principles have quickly begun to spread to the therapy of multiple diseases, including depigmentation disorders. The most common depigmentation disorder is vitiligo, a disease with deep psychosocial implications. Thanks to their unique properties, electrospun polymeric nanofibers represent a material suitable for tissue engineering applications. Furthermore, they may be functionalized with platelets, cells that contain a wide spectrum of growth factors and chemokines. The aim of this paper was to evaluate the functionalization of polymeric nanofibers with platelets and their effects in melanocyte culture. The scaffolds were visualized using scanning electron microscopy, the metabolic activity and proliferation of melanocytes was determined using MTS assay and dsDNA quantification, respectively. Furthermore, the melanocytes were stained and visualized using confocal microscopy. The acquired data showed that poly-ε-caprolactone functionalized with platelets promoted the viability and proliferation of melanocytes. According to the results, such a functionalized scaffold combining nanofibers and platelets may be suitable for melanocyte culture.

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Section: Discussionsupporting

confidence: 88%

Section: Discussionsupporting

confidence: 81%

Functionalization of Polymeric Nanofibers Using Platelets for Melanocyte Culture

et al. 2020

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“…In the present study, the cytokine content of HPL was analyzed via a quantitative multiplex immunoassay to identify potentially relevant cytokines contributing to MSC osteogenesis. Although previous studies have measured cytokines in HPL via semi-quantitative assays [ 22 , 36 – 40 ], to our knowledge, only five studies have reported quantitative multiplex-based assessments [ 21 , 39 , 41 – 44 ]. Considerable differences in cytokine concentrations are observed across the different studies (Table 1 ).…”

Section: Discussionmentioning

confidence: 99%

Influence of platelet storage time on human platelet lysates and platelet lysate-expanded mesenchymal stromal cells for bone tissue engineering

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Background Human platelet lysate (HPL) is emerging as the preferred xeno-free supplement for the expansion of mesenchymal stromal cells (MSCs) for bone tissue engineering (BTE) applications. Due to a growing demand, the need for standardization and scaling-up of HPL has been highlighted. However, the optimal storage time of the source material, i.e., outdated platelet concentrates (PCs), remains to be determined. The present study aimed to determine the optimal storage time of PCs in terms of the cytokine content and biological efficacy of HPL. Methods Donor-matched bone marrow (BMSCs) and adipose-derived MSCs (ASCs) expanded in HPL or fetal bovine serum (FBS) were characterized based on in vitro proliferation, immunophenotype, and multi-lineage differentiation. Osteogenic differentiation was assessed at early (gene expression), intermediate [alkaline phosphatase (ALP) activity], and terminal stages (mineralization). Using a multiplex immunoassay, the cytokine contents of HPLs produced from PCs stored for 1–9 months were screened and a preliminary threshold of 4 months was identified. Next, HPLs were produced from PCs stored for controlled durations of 0, 1, 2, 3, and 4 months, and their efficacy was compared in terms of cytokine content and BMSCs’ proliferation and osteogenic differentiation. Results BMSCs and ASCs in both HPL and FBS demonstrated a characteristic immunophenotype and multi-lineage differentiation; osteogenic differentiation of BMSCs and ASCs was significantly enhanced in HPL vs. FBS. Multiplex network analysis of HPL revealed several interacting growth factors, chemokines, and inflammatory cytokines. Notably, stem cell growth factor (SCGF) was detected in high concentrations. A majority of cytokines were elevated in HPLs produced from PCs stored for ≤ 4 months vs. > 4 months. However, no further differences in PC storage times between 0 and 4 months were identified in terms of HPLs’ cytokine content or their effects on the proliferation, ALP activity, and mineralization of BMSCs from multiple donors. Conclusions MSCs expanded in HPL demonstrate enhanced osteogenic differentiation, albeit with considerable donor variation. HPLs produced from outdated PCs stored for up to 4 months efficiently supported the proliferation and osteogenic differentiation of MSCs. These findings may facilitate the standardization and scaling-up of HPL from outdated PCs for BTE applications.

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“…The best results were obtained in the centrifugally spun samples with the highest concentration of platelets (the highest tested concentration of platelet-derived molecules, C10). Nanofibers made of PCL, using electrospinning, were successfully used for the melanocyte culture [ 45 , 57 , 58 ]. Savkovic et al showed PCL nanofibers to be superior to a 2D cell culture substrate, in terms of melanin synthesis [ 57 ].…”

Section: Discussionmentioning

confidence: 99%

A Simple Drug Delivery System for Platelet-Derived Bioactive Molecules, to Improve Melanocyte Stimulation in Vitiligo Treatment

et al. 2020

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Vitiligo is the most common depigmentation disorder of the skin. Currently, its therapy focuses on the halting of the immune response and stimulation of the regenerative processes, leading to the restoration of normal melanocyte function. Platelet-rich plasma (PRP) represents a safe and cheap regenerative therapy option, as it delivers a wide spectrum of native growth factors, cytokines and other bioactive molecules. The aim of this study was to develop a simple delivery system to prolong the effects of the bioactive molecules released from platelets. The surface of electrospun and centrifugally spun poly-ε-caprolactone (PCL) fibrous scaffolds was functionalized with various concentrations of platelets; the influence of the morphology of the scaffolds and the concentration of the released platelet-derived bioactive molecules on melanocytes, was then assessed. An almost two-fold increase in the amount of the released bioactive molecules was detected on the centrifugally spun vs. electrospun scaffolds, and a sustained 14-day release of the bioactive molecules was demonstrated. A strong concentration-dependent response of melanocyte to the bioactive molecules was observed; higher concentrations of bioactive molecules resulted in improved metabolic activity and proliferation of melanocytes. This simple system improves melanocyte viability, offers on-site preparation and is suitable for prolonged topical PRP administration.

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Platelet lysate as a serum replacement for skin cell culture on biomimetic PCL nanofibers

Cited by 19 publications

References 37 publications

Functionalization of Polymeric Nanofibers Using Platelets for Melanocyte Culture

Functionalization of Polymeric Nanofibers Using Platelets for Melanocyte Culture

Influence of platelet storage time on human platelet lysates and platelet lysate-expanded mesenchymal stromal cells for bone tissue engineering

A Simple Drug Delivery System for Platelet-Derived Bioactive Molecules, to Improve Melanocyte Stimulation in Vitiligo Treatment

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