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. 2024 Nov 22;16(11):e74252.
doi: 10.7759/cureus.74252. eCollection 2024 Nov.

Bioactive Effect of Plasma-Rich in Growth Factors (PRGFs) on Cell-Based In Vitro Models of Skin Inflammation in Relation to Inflammatory Skin Disorders

Eduardo Anitua  1   2 , Roberto Tierno  1   2 , Zuriñe Martínez de Lagrán  3 , Mohammad H Alkhraisat  1   2
Affiliations

Bioactive Effect of Plasma-Rich in Growth Factors (PRGFs) on Cell-Based In Vitro Models of Skin Inflammation in Relation to Inflammatory Skin Disorders

Eduardo Anitua et al. Cureus. .

Abstract

Plasma rich in growth factors (PRGFs) has proven potentially beneficial as a bioregenerator in patients with chronic skin disorders due to its anti-inflammatory effect. However, its therapeutic potential may be limited by soluble autoimmune components associated with inflammatory dermatoses in blood plasma. To evaluate the impact of skin health status on cell bioactivity, PRGF was prepared from healthy (H) donors as well as from individuals with atopic dermatitis (AD), psoriasis (PS), or lichen sclerosus (LS). Leukocyte exclusion and heat inactivation (Immunosafe treatment) were evaluated as potential methods to reduce the inflammatory components of the samples under study. The biological effect of PRGF-derived formulations was investigated using cell-based in vitro skin inflammation models, including human dermal fibroblasts (HDFs) and human epidermal keratinocytes (HEKs) exposed to a pro-inflammatory environment. The data confirmed that viability, proliferation, and migration rates were enhanced in inflamed cell cultures supplemented with PRGF formulations compared to those maintained in standard culture media. Nevertheless, significant differences have been identified. About the healthy control, inflamed epidermal keratinocytes supplemented with most PRGF-based formulations obtained from pathological donors (PS/LS) showed lower viability. Heat inactivation significantly promoted cell proliferation in epidermal keratinocytes supplemented with SP (PS/LS) and L-PRP supernatant (LSP) samples (AD), and also cell migration in inflamed HDF (AD/H/LS) and HEK (AD/LS) models supplemented with LSP. Leukocyte exclusion improved cell behavior in terms of migration with the only exception of LSP from individuals with AD added to inflamed HEK cultures. In conclusion, PRGF derived from pathological patients contains autoimmune components that could compromise its effectiveness as a therapy for treating individuals with chronic inflammatory disorders. However, heat inactivation (Immunosafe treatment) or leukocyte exclusion could minimize local adverse effects.

Keywords: bioactivity; heat-inactivation; leukocyte exclusion; plasma rich in growth factors/ prgf; skin inflammation.

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Conflict of interest statement

Human subjects: Consent for treatment and open access publication was obtained or waived by all participants in this study. The Hospital Clinical Research Ethical Committee issued approval BTIIMD-01-IV-23-PRGF. All procedures performed in this study were in accordance with the ethical standards of the Araba University Hospital Clinical Research Ethical Committee (BTIIMD-01-IV-23-PRGF) and with the Helsinki Declaration revised in 2013. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: This work was supported by the funding agency of the Basque Country Government (Spain) under grant Elkartek: KK-2020/00015. Financial relationships: EA is the scientific director of BTI Biotechnology Institute, the company that has developed the Endoret®PRGF® technology. RT and MHA are researchers at BTI Biotechnology Institute. declare(s) employment from BTI Biotechnology Institute. Intellectual property info: Eduardo Anitua Aldecoa has filed for patents to protect Endoret®PRGF® technology. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work.

Figures

Figure 1
Figure 1. Experimental design of the study.
Created with BioRender.com. LS, lichen sclerosus; PRGF, plasma rich in growth factor; LSP, L-PRP supernatant; IS-SP, PRGF Immunosafe supernatant; IS-LSP, L-PRP Immunosafe supernatant; ROS, reactive oxygen species
Figure 2
Figure 2. Barplots representing cell viability of inflamed in vitro models supplemented with plasma-derived formulations obtained from healthy and pathological donors using the following cell lines: (A) HDF and (B) HEK.
Symbols denote statistical significance (P ≤ 0.05): *, differences between pathological donors and healthy donors within a specific formulation; #, differences between raw formulations (SP) and those subjected to the Immunosafe thermal inactivation treatment (IS-SP); and $, differences between PRGF-derived supernatants (SP) and leukocyte-rich supernatants (LSP). PRGF, plasma rich in growth factor
Figure 3
Figure 3. Bar plots representing cell proliferation of inflamed in vitro models supplemented with plasma-derived formulations obtained from healthy and pathological donors using the following cell lines: (A) HDF and (B) HEK.
Symbols denote statistical significance (P ≤ 0.05): *, differences between pathological donors and healthy donors within a specific formulation; #, differences between raw formulations (SP) and those subjected to the Immunosafe thermal inactivation treatment (IS-SP); and $, differences between PRGF-derived supernatants (SP) and leukocyte-rich supernatants (LSP). PRGF, plasma rich in growth factor
Figure 4
Figure 4. Time-lapse analyses of cell migration: (A) high-magnification, phase-contrast (t = 0 hours), and fluorescence images (t = 48 hours) showing HDF and HEK migration in response to cell culture supplementation with multiple PRGF-derived formulations obtained from healthy donors and individuals with different chronic skin disorders; (B) quantification of HDF cell migration assay, and (C) quantification of HEK migration assay.
Scale bars: 25 µm. PRGF, plasma rich in growth factor; HDF, human dermal fibroblast; HEK, human epidermal keratinocyte
See this image and copyright information in PMC

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References

    1. Burden of skin disease and associated socioeconomic status in Europe: an ecologic study from the Global Burden of Disease Study 2017. Chu S, Mehrmal S, Uppal P, Giesey RL, Delost ME, Delost GR. JAAD Int. 2020;1:95–103. - PMC - PubMed
    1. Global epidemiology of atopic dermatitis: a comprehensive systematic analysis and modelling study. Tian J, Zhang D, Yang Y, Huang Y, Wang L, Yao X, Lu Q. Br J Dermatol. 2023;190:55–61. - PubMed
    1. The global, regional, and national burden of psoriasis: results and insights from the Global Burden of Disease 2019 study. Damiani G, Bragazzi NL, Karimkhani Aksut C, et al. Front Med (Lausanne) 2021;8:743180. - PMC - PubMed
    1. Diagnosis and treatment of lichen sclerosus: an update. Fistarol SK, Itin PH. Am J Clin Dermatol. 2013;14:27–47. - PMC - PubMed
    1. Comorbidities of atopic dermatitis-what does the evidence say? Thyssen JP, Halling AS, Schmid-Grendelmeier P, Guttman-Yassky E, Silverberg JI. J Allergy Clin Immunol. 2023;151:1155–1162. - PubMed

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