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人血小板裂解物与其他血液衍生物角膜上皮营养能力的比较。

Comparison of corneal epitheliotrophic capacities among human platelet lysates and other blood derivatives.

作者信息

Huang Chien-Jung, Sun Yi-Chen, Christopher Karen, Pai Amy Shih-I, Lu Chia-Ju, Hu Fung-Rong, Lin Szu-Yuan, Chen Wei-Li

机构信息

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.

出版信息

PLoS One. 2017 Feb 2;12(2):e0171008. doi: 10.1371/journal.pone.0171008. eCollection 2017.

Abstract

PURPOSE

To evaluate the corneal epitheliotropic abilities of two commercialized human platelet lysates (HPLs) and to compare the results with other blood derivatives, including human peripheral serum (HPS) and bovine fetal serum (FBS).

METHODS

In vitro, human corneal epithelial cells were incubated in various concentrations (0%, 3%, 5% and 10%) of blood derivatives. Two commercialized HPLs, including UltraGRO TM (Helios, Atlanta, GA) and PLTMax (Mill Creek, Rochester, MI), were tested and compared with HPS and FBS. Scratch-induced directional wounding assay was performed to evaluate cellular migration. MTS assay was used to evaluate cellular proliferation. Cellular differentiation was examined by scanning electron microscopy, inverted microscopy and transepithelial electrical resistance. Sprague-Dawley rats were used to evaluate the effects of the blood derivatives on corneal epithelial wound healing in vivo. Different blood derivatives were applied topically every 2 hours for 2 days after corneal epithelial debridement. The concentrations of epidermal growth factor (EGF), transforming growth factor -β1 (TGF-β1), fibronectin, platelet-derived growth factor-AB (PDGF-AB), PDGF-BB, and hyaluronic acid in different blood derivatives were evaluated by enzyme-linked immunosorbent assay (ELISA).

RESULTS

In vitro experiments demonstrated statistically comparable epitheliotropic characteristics in cellular proliferation, migration, and differentiation for the two commercialized HPLs compared to FBS and HPS. Cells cultured without any serum were used as control group. The epitheliotropic capacities were statistically higher in the two commercialized HPLs compared to the control group (p<0.05). Among the different concentrations of blood derivatives, the preparations with 3% yielded better outcomes compared to 5% and 10%. In rats, HPLs also caused improved but not statistically significant wound healing compared to HPS. All the blood derivatives had better wound healing ratios than the control group (p<0.05). In the quantification of epitheliotropic factors, UltraGRO and PLTMax had significantly higher levels of EGF, TGF- β1, fibronectin than human peripheral serum (p<0.05).

CONCLUSIONS

Both commercialized HPLs showed comparable corneal epitheliotropic abilities and wound healing rates compared to HPS and FBS in the in vivo and in vitro studies. Our results suggest that HPLs may have the potential to replace HPS in the treatment of corneal epithelial problems.

摘要

目的

评估两种商业化人血小板裂解物(HPLs)的角膜上皮趋化能力,并将结果与其他血液衍生物进行比较,包括人外周血清(HPS)和牛胎血清(FBS)。

方法

在体外,将人角膜上皮细胞置于不同浓度(0%、3%、5%和10%)的血液衍生物中培养。对两种商业化HPLs,即UltraGRO TM(Helios,亚特兰大,佐治亚州)和PLTMax(Mill Creek,罗切斯特,密歇根州)进行测试,并与HPS和FBS进行比较。采用划痕诱导定向损伤试验评估细胞迁移。采用MTS试验评估细胞增殖。通过扫描电子显微镜、倒置显微镜和跨上皮电阻检测细胞分化。使用Sprague-Dawley大鼠评估血液衍生物对体内角膜上皮伤口愈合的影响。角膜上皮清创术后,每2小时局部应用不同的血液衍生物,持续2天。通过酶联免疫吸附测定(ELISA)评估不同血液衍生物中表皮生长因子(EGF)、转化生长因子-β1(TGF-β1)、纤连蛋白、血小板衍生生长因子-AB(PDGF-AB)、PDGF-BB和透明质酸的浓度。

结果

体外实验表明,与FBS和HPS相比,两种商业化HPLs在细胞增殖、迁移和分化方面具有统计学上相当的角膜上皮趋化特性。将未添加任何血清培养的细胞作为对照组。与对照组相比,两种商业化HPLs的角膜上皮趋化能力在统计学上更高(p<0.05)。在不同浓度的血液衍生物中,3%的制剂比5%和10%的制剂效果更好。在大鼠中,与HPS相比,HPLs也能促进伤口愈合,但差异无统计学意义。所有血液衍生物的伤口愈合率均优于对照组(p<0.05)。在角膜上皮趋化因子的定量分析中,UltraGRO和PLTMax的EGF、TGF-β1、纤连蛋白水平显著高于人外周血清(p<0.05)。

结论

在体内和体外研究中,两种商业化HPLs与HPS和FBS相比,均表现出相当的角膜上皮趋化能力和伤口愈合率。我们的结果表明,HPLs在治疗角膜上皮问题方面可能有潜力替代HPS。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5547/5289502/68f9933528a7/pone.0171008.g001.jpg

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