Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan.
Department of Ophthalmology, Taipei Tzu Chi General Hospital, The Buddhist Tzu Chi Medical Foundation, New Taipei City, Taiwan.
PLoS One. 2018 Mar 16;13(3):e0194345. doi: 10.1371/journal.pone.0194345. eCollection 2018.
To evaluate whether lyophilized human platelet lysate (HPL) powder can preserve the growth factor concentrations and epitheliotrophic properties of liquid HPL, and potentially be used as a clinically-friendly treatment option.
Two commercialized liquid HPLs, UltraGRO TM (Helios, Atlanta, GA) and PLTMax (Mill Creek, Rochester, MI), were obtained and converted to lyophilized powder. After redissolution, lyophilized powder HPLs were compared with liquid HPLs, as well as human peripheral serum (HPS) and fetal bovine serum (FBS) in liquid or redissolved lyophilized powder forms. Concentrations of epidermal growth factor (EGF), transforming growth factor-β1 (TGF-β1), platelet-derived growth factor-AB (PDGF-AB) and platelet-derived growth factor-BB (PDGF-BB) were evaluated by enzyme-linked immunosorbent assay (ELISA). Human corneal epithelial cell line was incubated with the blood derivatives and evaluated for cell migration with scratch-induced directional wounding and proliferation with MTS assays. Cell differentiation was examined by transepithelial electrical resistance (TEER). Fluorescein staining and in vivo confocal microscopy were used to evaluate in vivo corneal epithelial wound healing in Sprague-Dawley rats that underwent corneal debridement and topical application of liquid and redissolved powder HPLs.
Liquid form and redissolved lyophilized powder form HPLs had similar concentrations of EGF, TGF-β1, PDGF-AB and PDGF-BB. In vitro experiments on cell migration, proliferation and differentiation and rat models on wound healing demonstrated no significant difference between the liquid and redissolved lyophilized powder forms for HPLs, HPS and FBS. In vivo confocal microscopy revealed similar wound healing process at different layers of cornea after corneal epithelial debridement between liquid form and redissolved lyophilized power form of HPLs.
The redissolved lyophilized powder form of both commercialized HPLs showed similar growth factor concentrations and corneal epitheliotrophic abilities compared to the liquid form. Results suggest that the properties of liquid HPLs can be retained despite lyophilization and that lyophilized HPLs can be a treatment option for corneal epithelial disorders.
评估冻干人血小板裂解物（HPL）粉末是否可以保持液体 HPL 的生长因子浓度和上皮营养特性，并可能作为一种临床友好的治疗选择。
获得两种商业化的液体 HPL，UltraGRO TM（Helios，亚特兰大，GA）和 PLTMax（Mill Creek，罗切斯特，MI），并将其转化为冻干粉末。重溶后，将冻干粉末 HPL 与液体 HPL 以及人外周血清（HPS）和胎牛血清（FBS）在液体或重溶冻干粉末形式下进行比较。通过酶联免疫吸附试验（ELISA）评估表皮生长因子（EGF）、转化生长因子-β1（TGF-β1）、血小板衍生生长因子-AB（PDGF-AB）和血小板衍生生长因子-BB（PDGF-BB）的浓度。用人角膜上皮细胞系孵育血液衍生物，并通过划痕诱导的定向创伤和 MTS 测定评估细胞迁移，通过 transepithelial 电阻（TEER）评估细胞分化。荧光染色和体内共聚焦显微镜用于评估 Sprague-Dawley 大鼠的角膜上皮伤口愈合，大鼠进行角膜清创术并局部应用液体和重溶冻干 HPL。
液体形式和重溶冻干粉末形式的 HPL 具有相似的 EGF、TGF-β1、PDGF-AB 和 PDGF-BB 浓度。细胞迁移、增殖和分化的体外实验以及 HPL、HPS 和 FBS 的大鼠模型实验表明，液体形式和重溶冻干粉末形式的 HPL 之间没有显著差异。体内共聚焦显微镜显示，角膜上皮清创后不同角膜层之间，液体形式和重溶冻干粉末形式的 HPL 具有相似的伤口愈合过程。
与液体形式相比，两种商业化 HPL 的重溶冻干粉末形式均显示出相似的生长因子浓度和角膜上皮营养能力。结果表明，尽管冻干，但液体 HPL 的特性可以保留，并且冻干 HPL 可以作为角膜上皮疾病的治疗选择。
