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[自体富血小板血浆治疗早期椎间盘退变的效果实验研究]

[An experimental study on effect of autologous platelet-rich plasma on treatment of early intervertebral disc degeneration].

作者信息

Hu Xinfeng, Wang Chen, Rui Yunfeng

机构信息

Department of Orthopaedics, Affiliated Zhongda Hospital, Southeast University, Nanjing Jiangsu 210009, PR China.

出版信息

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2012 Aug;26(8):977-83.

Abstract

OBJECTIVE

Platelet-rich plasma (PRP) can stimulate intervertebral disc cell proliferation, promote extracellular matrix synthesis, and inhibit annulus fibrosus cell apoptosis. To investigate the effects of autologous PRP on the treatment of the early intervertebral disc degeneration (IDD) so as to provide the experimental basis for its clinical application.

METHODS

Forty-five healthy New Zealand white rabbits (male or female, weighing 2.5-3.0 kg) were randomly divided into the experimental group (n = 15), the control group (n = 15), and the sham group (n = 15). PRP was prepared from the arterial blood of rabbit's ears of the experimental group with Landesberg's method. The platelet concentrations in both whole blood and PRP were detected. The rabbit model of early IDD was established by annulus fibrosus puncture (L4, 5, L5, 6) in both the experimental group and the control group; 100 microL autologous PRP and 100 microL PBS were injected into the degenerative intervertebral discs respectively after 2 weeks of models creation. In sham group, intervertebral discs were separated and exposed without treatment. The general conditions of the rabbits were observed after building models; at 2 weeks after degeneration, 1 and 2 weeks after intervention, 5 rabbits were selected randomly from each group respectively for MRI observation, histological observation by using HE staining and collagen type II immunohistochemical staining. The signal of lumbar MRI was assessed and the contents of collagen type II were detected.

RESULTS

The platelet concentration of PRP was about 4.92 times as much as that of the whole blood. All the animals survived to the end of the experiment. At 2 weeks after degeneration, a lower T2 signal was observed in both the experimental group and the control group; the nucleus pulposus cells decreased and extracellular matrix degenerated; and the expression of collagen type II decreased in both the experimental group and control group. The degenerative grade of lumbar MRI in the experimental group and control group were significantly higher than that in the sham group (P < 0.05), and the content of collagen type II were significantly lower than that in the sham group (P < 0.05). At 1, 2 weeks after intervention, disc degeneration in the experimental group was significantly lower than that in control group (P < 0.05), and significant difference was found between experimental group and sham group (P < 0.05). The nucleus pulposus cells and chondroid matrix in the experimental group were more than those in the control group, showing slight stromal fibrosis; but the expression of collage type II was significantly higher than that in the control group (P < 0.05).

CONCLUSION

The disc injection of autologous PRP may terminate or even reverse the progress of rabbit early IDD, which may be associated with the role of multiple growth factors of PRP in regulating cell function, improving the tissue microenvironment, and promoting tissue regeneration.

摘要

目的

富血小板血浆(PRP)可刺激椎间盘细胞增殖,促进细胞外基质合成,并抑制纤维环细胞凋亡。探讨自体PRP对早期椎间盘退变(IDD)的治疗作用,为其临床应用提供实验依据。

方法

将45只健康新西兰白兔(雌雄不限,体重2.5 - 3.0 kg)随机分为实验组(n = 15)、对照组(n = 15)和假手术组(n = 15)。采用Landesberg法从实验组兔耳动脉血中制备PRP。检测全血和PRP中的血小板浓度。实验组和对照组均通过纤维环穿刺(L4, 5、L5, 6)建立早期IDD兔模型;造模2周后分别向退变的椎间盘内注射100 μL自体PRP和100 μL PBS。假手术组仅分离并暴露椎间盘,不做处理。造模后观察兔的一般情况;退变2周、干预后1周和2周时,每组分别随机选取5只兔进行MRI观察、HE染色及Ⅱ型胶原免疫组化染色进行组织学观察。评估腰椎MRI信号并检测Ⅱ型胶原含量。

结果

PRP中的血小板浓度约为全血的4.92倍。所有动物均存活至实验结束。退变2周时,实验组和对照组T2信号均降低;髓核细胞减少,细胞外基质退变;实验组和对照组Ⅱ型胶原表达均降低。实验组和对照组腰椎MRI退变分级均显著高于假手术组(P < 0.05),Ⅱ型胶原含量均显著低于假手术组(P < 0.05)。干预后1周、2周时,实验组椎间盘退变程度显著低于对照组(P < 0.05),与假手术组相比差异有统计学意义(P < 0.05)。实验组髓核细胞和软骨样基质多于对照组,有轻度基质纤维化;但Ⅱ型胶原表达显著高于对照组(P < 0.05)。

结论

向椎间盘内注射自体PRP可能终止甚至逆转兔早期IDD的进展,这可能与PRP中多种生长因子在调节细胞功能、改善组织微环境及促进组织再生中的作用有关。

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