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The corneal epitheliotrophic abilities of lyophilized powder form human platelet lysates
PLOS ONE ( IF 3.7 ) Pub Date : 2018-03-16 , DOI: 10.1371/journal.pone.0194345
Lily Wei Chen , Chien-Jung Huang , Wen-Hui Tu , Chia-Ju Lu , Yi-Chen Sun , Szu-Yuan Lin , Wei-Li Chen

Purpose

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.

Methods

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.

Results

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.

Conclusions

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.



中文翻译:

冻干粉形成人血小板裂解物的角膜上皮营养能力

目的

为了评估冻干的人血小板裂解物(HPL)粉末是否可以保留液体HPL的生长因子浓度和上皮营养特性,并有可能被用作临床友好的治疗选择。

方法

获得了两种商品化的液体HPL,UltraGRO TM(Helios,佐治亚州亚特兰大)和PLTMax(Mill Creek,罗切斯特,密歇根州),并将其转化为冻干粉。重新溶解后,将冻干的粉末HPL与液体HPL以及液体或重新溶解的冻干粉末形式的人外周血血清(HPS)和胎牛血清(FBS)进行比较。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)的浓度通过酶-连锁免疫吸附测定(ELISA)。将人角膜上皮细胞系与血液衍生物一起孵育,并通过MTS分析评估刮擦诱导的定向伤口和增殖情况下的细胞迁移。通过跨上皮电阻(TEER)检查细胞分化。荧光素染色和体内共聚焦显微镜用于评估经历角膜清创术和局部应用液体和再溶解粉末HPL的Sprague-Dawley大鼠的体内角膜上皮伤口愈合。

结果

液体形式和重新溶解的冻干粉末形式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伤口愈合的大鼠模型表明,液体和重新溶解的冻干粉形式的HPL,HPS和FBS均无显着差异。体内共聚焦显微镜显示液体形式和再溶解的冻干粉末形式之间的角膜上皮清创后,角膜不同层的伤口愈合过程相似。

结论

与液体形式相比,两种商业化的HPL的重新溶解的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的性质仍可保留,冻干的HPL可作为角膜上皮疾病的治疗选择。

更新日期:2018-03-17
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