Anitua Eduardo, Tierno Roberto, Martínez de Lagrán Zuriñe, Alkhraisat Mohammad H
University Institute for Regenerative Medicine and Oral Implantology (UIRMI), Vitoria, Spain; BTI Biotechnology Institute, Vitoria, Spain.
University Institute for Regenerative Medicine and Oral Implantology (UIRMI), Vitoria, Spain; BTI Biotechnology Institute, Vitoria, Spain.
Tissue Cell. 2024 Dec;91:102560. doi: 10.1016/j.tice.2024.102560. Epub 2024 Sep 14.
Plasma rich in growth factors (PRGF) can be used over patients suffering from dermatoses due to its anti-inflammatory effect. However, this population group might present soluble autoimmune components and there is limited information about the effect of chronic skin inflammation on PRGF bioactive properties. With the aim of characterizing PRGF composition, PRGF from healthy (H) donors and patients with atopic dermatitis (AD), psoriasis (PS), or lichen sclerosus (LS) was obtained. In order to reduce the inflammatory component, leukocyte exclusion and heat-inactivation (Immunosafe) were tested. Haematological-serological parameters, platelet functionality, clot microstructure, protein content and bioactivity were determined. Mean values and 95 % confidence intervals (mean[95 % CI]) were computed for key haematological parameters, such as platelet (410×10/mm[371-449]) and leukocyte content (205×10/mm[148-262]), platelet activation (resting: 4.3 %[3.1-5.5] and activated: 97.4 %[96.7-98.0]), the concentration of plasma proteins and morphogens, including immunoglobulins A (210.7 mg/dL[191.8-229.6]), G (933.1 mg/dL[887.2-978.9]), E (783.5 mg/dL[54.4-1512.6]), and M (115.0 mg/dL[97.1-133.0]), Complement Protein (31.6 mg/mL[26.6-36.6]), C-Reactive protein (3.1 mg/L[2.0-4.1]), TGF-β1 (35975.6 pg/mL[34221.3-37729.8]), fibronectin (146410.0 ng/mL[136518.3-156301.7]), PDGF-AB (13308.5 pg/mL[12401.0-14216.0]), CD40L (2389.3 pg/mL[1887.7-2890.8]), IL-4 (0.12 pg/mL[0.07-0.18]), IL-13 (35.4 pg/mL[21.0-49.7]), IL-1β (0.09 pg/mL[0.06-0.11]) and TNF-α (0.31 pg/mL[0.24-0.38]), and also for cell proliferation (332.9ngDNA/mL[317.4-348.3]), viability (135.6 %[132.0-139.2]) and migration (103.8cells/mm[98.3-109.3]). Plasma from AD donors presented increased Immunoglobulin E (IgE) that was significantly reduced after Immunosafe along with the complement system and autoantibodies. Platelet functionality was altered for AD, but no microstructure differences were identified. Pathological groups presented reduced concentration of fibronectin (AD/LS) and Platelet-Derived Growth Factor (PDGF-AB) (P). Immunosafe treatment reduced Cluster of Differentiation 40 Protein (CD40L), interleukin 1β (IL-1β), and Tumor Necrosis Factor α (TNF-α) concentrations. Fibroblasts supplemented with PRGF obtained from pathological patients (PS/AD) showed reduced viability but Immunosafe increased cell proliferation and migration in SP (LS) and L-SP samples (PS/AD). In conclusion, PRGF derived from pathological patients present autoimmune components, but heat-inactivation or leukocyte exclusion could minimize local side effects.
富含生长因子的血浆（PRGF）因其抗炎作用可用于患有皮肤病的患者。然而，这一人群可能存在可溶性自身免疫成分，且关于慢性皮肤炎症对PRGF生物活性特性影响的信息有限。为了表征PRGF的组成，我们获取了来自健康（H）供体以及特应性皮炎（AD）、银屑病（PS）或硬化性苔藓（LS）患者的PRGF。为了减少炎症成分，我们测试了白细胞排除和热灭活（免疫安全）方法。测定了血液学 - 血清学参数、血小板功能、凝块微观结构、蛋白质含量和生物活性。计算了关键血液学参数的平均值和95%置信区间（均值[95%CI]），如血小板（410×10/mm[371 - 449]）和白细胞含量（205×10/mm[148 - 262]）、血小板活化（静息：4.3%[3.1 - 5.5]，活化：97.4%[96.7 - 98.0]）、血浆蛋白和形态发生素的浓度，包括免疫球蛋白A（210.7mg/dL[191.8 - 229.6]）、G（933.1mg/dL[887.2 - 978.9]）、E（783.5mg/dL[54.4 - 1512.6]）和M（115.0mg/dL[97.1 - 133.0]）、补体蛋白（31.6mg/mL[26.6 - 36.6]）、C反应蛋白（3.1mg/L[2.0 - 4.1]）、转化生长因子 - β1（35975.6pg/mL[34221.3 - 37729.8]）、纤连蛋白（146410.0ng/mL[136518.3 - 156301.7]）、血小板衍生生长因子 - AB（13308.5pg/mL[12401.0 - 14216.0]）、CD40L（2389.3pg/mL[1887.7 - 2890.8]）、白细胞介素 - 4（0.12pg/mL[0.07 - 0.18]）、白细胞介素 - 13（35.4pg/mL[21.0 - 49.7]）、白细胞介素 - 1β（0.09pg/mL[0.06 - 0.11]）和肿瘤坏死因子 - α（0.31pg/mL[0.24 - 0.38]），以及细胞增殖（332.9ngDNA/mL[317.4 - 348.3]）、活力（135.6%[132.0 - 139.2]）和迁移（103.8细胞/mm[98.3 - 109.3]）。来自AD供体的血浆中免疫球蛋白E（IgE）增加，经免疫安全处理后与补体系统和自身抗体一起显著降低。AD患者的血小板功能发生改变，但未发现微观结构差异。病理组的纤连蛋白（AD/LS）和血小板衍生生长因子（PDGF - AB）（P）浓度降低。免疫安全处理降低了分化簇40蛋白（CD40L）、白细胞介素1β（IL - 1β）和肿瘤坏死因子α（TNF - α）的浓度。用从病理患者（PS/AD）获得的PRGF补充的成纤维细胞活力降低，但免疫安全处理增加了SP（LS）和L - SP样本（PS/AD）中的细胞增殖和迁移。总之，来自病理患者的PRGF存在自身免疫成分，但热灭活或白细胞排除可将局部副作用降至最低。
