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两种制备富含血小板血浆滴眼液的方法对生长因子释放及去上皮化兔模型的比较

Comparison of Two Preparation Methods for Platelet-Rich Plasma Eye Drops for Release of Growth Factors and De-Epithelization Rabbit Model.

作者信息

Kobayashi Tatsuhiko, Suzuki Takashi, Saito Tomohiko, Itokawa Takashi, Hori Yuichi

机构信息

Department of Ophthalmology, Toho University Graduate School of Medicine, Tokyo, Japan.

Department of Ophthalmology, School of Medicine, Toho University, Tokyo, Japan.

出版信息

J Ophthalmol. 2020 Dec 1;2020:6634744. doi: 10.1155/2020/6634744. eCollection 2020.

Abstract

PURPOSE

To compare two platelet-rich plasma (PRP) preparation methods (double spin (D-PRP) and TriCell PRP (T-PRP)) for eye drops, concentration yields of platelets and other cells, release of growth factors, and efficacy of the de-epithelization rabbit model.

METHODS

PRP was extracted by D-PRP and T-PRP from 30 ml blood from healthy adults. After extraction, platelets and leukocytes were counted. D-PRP and T-PRP were preserved during A: 1 h storage at room temperature, B: 1 h storage at -20°C, C: 24 h storage at 4°C, and D: 24 h storage at -20°C. Platelet-derived growth factor (PDGF) was measured. Freezing/thawing PRP eye drops and control were instilled in the de-epithelization rabbit model four times per day for 5 days. Histology was compared between eyes treated with control, D-PRP, and T-PRP.

RESULTS

14 ml of D-PRP and 4 ml of T-PRP were extracted from 30 ml whole blood samples. D-PRP and T-PRP had 41.36 ± 8.43 × 10 and 67.02 ± 13.55 × 10 platelets and 0.41 ± 0.24 × 10/ml and 10.09 ± 4.29 × 10/ml leucocytes, respectively. In the four storage methods, PDGF concentrations in T-PRP were higher than those in D-PRP eye drops. Freezing/thawing D-PRP and T-PRP increased PDGF concentrations. Histology showed corneal epithelium thickness in T-PRP-treated eyes after healing (38.41 ± 9.10 m) was significantly higher than that in control-treated (27.77 ± 4.76 m) and D-PRP-treated eyes (18.32 ± 3.14 m) ( < 0.05). There was no corneal damage with inflammation in corneal stroma and epithelium of all tested groups. Electron microscopy revealed strong adhesion between cell junctions in T-PRP-treated eyes.

CONCLUSIONS

Freezing/thawing of PRP extracted with the T-PRP kit may result in high platelet and leukocyte concentration and produce high PDGF concentration. PRP eye drops including leucocytes could increase thickness of corneal epithelium without corneal inflammation.

摘要

目的

比较两种用于眼药水的富血小板血浆(PRP)制备方法(双重离心法(D-PRP)和TriCell PRP法(T-PRP)),观察血小板和其他细胞的浓度产量、生长因子的释放以及去上皮化兔模型中的疗效。

方法

从健康成年人的30毫升血液中,用D-PRP和T-PRP提取PRP。提取后,对血小板和白细胞进行计数。D-PRP和T-PRP在以下条件下保存:A:室温保存1小时;B:-20°C保存1小时;C:4°C保存24小时;D:-20°C保存24小时。检测血小板衍生生长因子(PDGF)。将冻融后的PRP眼药水和对照物每天给去上皮化兔模型滴眼4次,共5天。比较用对照物、D-PRP和T-PRP处理的眼睛的组织学情况。

结果

从30毫升全血样本中分别提取出14毫升D-PRP和4毫升T-PRP。D-PRP和T-PRP的血小板计数分别为41.36±8.43×10和67.02±13.55×10,白细胞计数分别为0.41±0.24×10/毫升和10.09±4.29×10/毫升。在四种保存方法中,T-PRP中的PDGF浓度高于D-PRP眼药水。冻融D-PRP和T-PRP可提高PDGF浓度。组织学显示,愈合后T-PRP处理组眼睛的角膜上皮厚度(38.41±9.10微米)显著高于对照处理组(27.77±4.76微米)和D-PRP处理组(18.32±3.14微米)(P<0.05)。所有测试组的角膜基质和上皮均无伴有炎症的角膜损伤。电子显微镜显示T-PRP处理组眼睛的细胞连接处有很强的粘连。

结论

用T-PRP试剂盒提取的PRP冻融后可能会导致高血小板和白细胞浓度,并产生高PDGF浓度。含白细胞的PRP眼药水可增加角膜上皮厚度且无角膜炎症。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/136f/9301758/c870b54c8761/joph2020-6634744.001.jpg

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