Optimization of preparation conditions of platelet-rich plasma in infants and determination of some cytokines
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摘要: 目的 优化幼儿富血小板血浆(platelet-rich plasma，PRP)的制备条件，测定幼儿及成人PRP和贫血小板血浆(platelet-poor plasma, PPP)中部分细胞因子浓度并分析差异，为后续临床研究奠定基础。方法 本研究通过两步法制备PRP，采用不同的离心条件，分别测定制备前全血和制备后PRP中的血小板计数(PLT)、血小板平均容积(MPV)和血小板分布宽度(PDW)等指标，以选择幼儿PRP制备的最优离心条件组合。ELISA法分别测定幼儿PRP及PPP与成人PRP及PPP中表皮细胞生长因子(EGF)、转化生长因子β1(TGF-β1)、胰岛素样生长因子(IGF-1)浓度，并进行统计学分析。结果 可采用全血初次离心600×g(10 min)，上层血浆再次离心2 000×g(10 min)的方法制备幼儿PRP。幼儿PRP组EGF、TGF-β1、IGF-1因子浓度均明显高于成人PRP组，三者比较差异均有统计学意义(P < 0.05)。结论 采用优化离心条件后的两步离心法，可获得稳定、高质量的幼儿PRP。幼儿PRP中EGF、TGF-β1、IGF-1因子浓度均明显高于成人PRP组，为后续相关试验研究提供了方向。Abstract: Objective To optimize the preparation conditions of infant platelet-rich plasma(PRP), measure the concentrations of some cytokines in PRP and platelet-poor plasma(PPP) in infants and adults, and analyze the differences, so as to lay the foundation for subsequent clinical studies.Methods PRP was prepared by a two-step method. Different centrifugation conditions were used to determine platelet count(PLT), mean platelet volume(MPV) and platelet distribution width(PDW) in whole blood PRP before and after preparation, respectively, in order to select the optimal combination of centrifugation conditions for PRP preparation in infants. The concentrations of epidermal cell growth factor(EGF), transforming growth factor β1(TGF-β1) and insulin-like growth factor(IGF-1) in PRP and PPP in infants and PRP and PPP in adults were measured by ELISA, and the concentrations were statistically analyzed.Results Infant PRP could be prepared by centrifugation of whole blood 600×g(10 min) and upper plasma 2 000×g(10 min). The EGF, TGF-β1 and IGF-1 factors in infant PRP were significantly higher than that in PRP group. The difference among the three was statistically significant(P < 0.05).Conclusion Two-step centrifugation method with optimized centrifugation conditions can obtain stable and high quality infant PRP. The concentrations of EGF, TGF-β1 and IGF-1 factors in infant PRP were significantly higher than those in adult PRP group, which provided the direction for subsequent relevant experimental research.
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Key words:
- platelet-rich plasma /
- platelet-poor plasma /
- cytokine
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表 1 第一次制备幼儿PRP离心条件
组别 初次离心 二次离心 离心力/g 离心时间/min 离心力/g 离心时间/min 第一组 500 10 1 500 10 第二组 600 10 2 000 10 第三组 800 10 2 000 10 
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表 2 第二次制备幼儿PRP离心条件
组别 初次离心 二次离心 离心力/g 离心时间/min 离心力/g 离心时间/min 第一组 500 10 1 500 10 第二组 600 10 1 500 10 第三组 600 10 2 000 10
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表 3 第一次制备幼儿PRP的血常规检测结果
检测指标 制备前全血 第一组PRP 第二组PRP 第三组PRP 红细胞/(×1012) 5.16 0.40 0.40 0.40 白细胞/(×109) 9.99 13.54 13.20 8.22 PLT/(×109) 304 1 612 1 620 1 821 MPV/fL 12.8 13.2 13.2 13.2 PDW/% 18.6 20.5 19.0 17.3
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表 4 第二次制备幼儿PRP的血常规检测结果
检测指标 制备前全血 第一组PRP 第二组PRP 第三组PRP 红细胞/(×1012) 5.25 0.22 0.22 0.22 白细胞/(×109) 8.57 1.86 2.34 1.66 PLT/(×109) 361 2 359 2 398 2 557 MPV/fL 11.4 11.8 11.9 11.7 PDW/% 14.1 13.2 13.3 13.2
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表 5 幼儿与成人PRP制备的血常规检测结果
检测指标 幼儿组 成人组 制备前全血 制备后PRP 制备前全血 制备后PRP 红细胞/(×1012) 4.29 0.13 4.52 0.23 白细胞/(×109) 8.28 4.67 6.11 1.34 PLT/(×109) 308 1 560 254 1 551 MPV/fL 10.8 11.4 11.8 12.6 PDW/% 12.5 12.2 14.8 15.7
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[1] Ramaswamy Reddy SH, Reddy R, Babu NC, et al. Stem-cell therapy and platelet-rich plasma in regenerative medicines: A review on pros and cons of the technologies[J]. J Oral Maxillofac Pathol, 2018, 22(3): 367-374. doi: 10.4103/jomfp.JOMFP_93_18
[2] Everts P, Onishi K, Jayaram P, et al. Platelet-rich plasma: new performance understandings and therapeutic considerations in 2020[J]. Int J Mol Sci, 2020, 21(20): 7794. doi: 10.3390/ijms21207794
[3] 张强, 岳宪虎, 黄象艳, 等. 富血小板血浆在髌骨外侧高压症治疗中的应用[J]. 临床输血与检验, 2023, 25(3): 348-353. https://www.cnki.com.cn/Article/CJFDTOTAL-LSXY202303010.htm
[4] Lovisolo F, Carton F, Gino S, et al. Platelet rich plasma-derived microvesicles increased invitro wound healing[J]. Eur Rev Med Pharmacol Sci, 2020, 24(18): 9658-9664.
[5] 杨菁, 周谋, 林放, 等. 负载PRP壳聚糖-明胶复合敷料止血和促愈合效果的初步评价[J]. 临床输血与检验, 2021, 23(5): 612-616. doi: 10.3969/j.issn.1671-2587.2021.05.014
[6] 刘广亚, 许育兵, 单桂秋, 等, 自体富血小板血浆治疗雄激素源性脱发临床疗效的影响因素分析[J]. 中国输血杂志, 2022, 35(12): 1243-1246. https://www.cnki.com.cn/Article/CJFDTOTAL-BLOO202212013.htm
[7] 邝江波, 许喜生, 王永飞, 等, 富血小板血浆联合自体脂肪移植在面部年轻化治疗的Meta分析[J]. 中国美容整形外科杂志, 2023, 34(9): 548-553. doi: 10.3969/j.issn.1673-7040.2023.09.011
[8] Milek T, Nagraba L, Mitek T, et al. Autologous platelet-rich plasma reduces healing time of chronic venous leg ulcers: A prospective ob-servational study[J]. Adv Exp Med Biol, 2019, 1176: 109-117.
[9] 中国输血协会临床输血管理学专业委员会. 自体富血小板血浆制备技术专家共识[J]. 中国输血杂志, 2021, 34(7): 677-683. https://cpfd.cnki.com.cn/Article/CPFDTOTAL-TTYX202311001145.htm
[10] 李少文, 朱展鸿, 单桂秋, 等. 富血小板血浆制备及其质量评价体系初探[J]. 中国输血杂志, 2022, 35(10): 1085-1091. https://www.cnki.com.cn/Article/CJFDTOTAL-BLOO202210024.htm
[11] Tian J, Cheng LH, Cui X, et al. Application of standardized platelet-rich plasma in elderly patients with complex wounds[J]. Wound Repair Regen, 2019, 27(3): 268-276.
[12] Anitua E, Munoz V, Aspe L. In vitro and in vivo effect of platelet-rich plasma-based autologous topical serum on cutaneous wound healing[J]. Skin Pharmacol Physiol, 2022, 35(1): 51-64.
[13] 李文娟, 张树超. 富血小板血浆在慢性难愈性创面治疗中的临床研究进展[J]. 中国输血杂志, 2022, 35(1): 1092-1096. https://www.cnki.com.cn/Article/CJFDTOTAL-BLOO202210025.htm
[14] 吴锦涛, 邹立津, 何霞飞, 等. 血小板浓缩物在创面愈合中的研究进展[J]. 广东医学, 2023, 44(1): 118-123. https://www.cnki.com.cn/Article/CJFDTOTAL-GAYX202301022.htm
[15] 陈勇, 吴培培, 蒋强, 等. 关节腔内注射PRP对膝骨性关节炎化脓性感染疼痛介质与NLRP3/IL-1β通路的影响[J]. 中华医院感染学杂志, 2022, 32(15): 2338-2342. https://www.cnki.com.cn/Article/CJFDTOTAL-ZHYY202215020.htm
[16] 杨亚崇, 帅世超, 朱喜春, 等. 富血小板血浆治疗膝关节骨性关节炎的研究进展[J]. 中国输血杂志2023, 36(1): 90-93. https://www.cnki.com.cn/Article/CJFDTOTAL-BLOO202301023.htm
[17] 宋黎, 陆茂, 唐懿, 等. 点阵CO2激光联合富血小板血浆治疗萎缩性痤疮瘢痕的系统评价[J]. 中国组织工程研究, 2023, 27(35): 5734-5740. https://www.cnki.com.cn/Article/CJFDTOTAL-XDKF202335026.htm
[18] 朱新华, 张二佳, 付梦瑶. 强脉冲光联合富血小板血浆注射治疗玫瑰痤疮疗效观察[J]. 中国美容医学, 2023, 32(10): 102-105. https://www.cnki.com.cn/Article/CJFDTOTAL-MRYX202310049.htm
[19] Dionyssiou D, Demiri E, Foroglou P, et al. The effectiveness of intralesional injection of platelet-rich plasma in accelerating the healing of chronic ulcers: an experimental and clinical study[J]. Int Wound J, 2013, 10(4): 397-406.
[20] Cervelli V, Scioli MG, Gentile P, et al. Platelet-rich plasma greatly potentiates insulin-induced adipogenic differentiation of human adipose-derived stem cells through a serine/threonine kinase Akt-dependent mechanism and promotes clinical fat graft maintenance[J]. Stem Cells Transl Med, 2012, 1(3): 206-220.
[21] 马秉正, 赵而海, 左振柏, 等. 基于Luminex技术对比分析激活后的脐血和成人富血小板血浆研究[J]. 中国输血杂志, 2023, 36(1): 11-15. https://www.cnki.com.cn/Article/CJFDTOTAL-BLOO202301003.htm
[22] 刘建香, 吕孟兴, 屈柯暄, 等. 自体富血小板血浆治愈儿童难治性骨髓炎合并骨折1例[J]. 中国输血杂志, 2023, 36(8): 673-675. https://www.cnki.com.cn/Article/CJFDTOTAL-BLOO202308004.htm
[23] 刘琴, 沈先涛, 胡红兵, 等. 自体富血小板血浆治疗儿童病理性骨折疗效观察[J]. 临床输血与检验, 2023, 25(3): 354-357. https://www.cnki.com.cn/Article/CJFDTOTAL-LSXY202303011.htm
[24] 刘炎, 何颖雨, 刘磊, 等. 富血小板血浆的抑菌作用及其机制的研究进展[J]. 临床血液学杂志, 2023, 36(6): 452-456. https://lcxy.whuhzzs.com/article/doi/10.13201/j.issn.1004-2806.2023.06.016
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