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软骨碎片程度对自体软骨移植中细胞外基质产生的影响:一项体外研究

The Degree of Chondral Fragmentation Affects Extracellular Matrix Production in Cartilage Autograft Implantation: An In Vitro Study.

作者信息

Bonasia Davide Edoardo, Marmotti Antongiulio, Mattia Silvia, Cosentino Aurelio, Spolaore Simone, Governale Giorgio, Castoldi Filippo, Rossi Roberto

机构信息

Department of Orthopaedics and Traumatology, University of Torino, CTO Hospital, Torino, Italy.

Department of Orthopaedics and Traumatology, University of Torino, Mauriziano "Umberto I" Hospital, Torino, Italy.

出版信息

Arthroscopy. 2015 Dec;31(12):2335-41. doi: 10.1016/j.arthro.2015.06.025. Epub 2015 Aug 28.

Abstract

PURPOSE

To evaluate if the degree of chondral fragmentation affected extracellular matrix (ECM) production in cartilage fragment autograft implantation in vitro.

METHODS

Cartilage was taken from 5 donors undergoing total hip replacement (mean age, 65.6 years; standard deviation [SD], 3). The cartilage was minced to obtain 4 groups with different fragment sizes: (1) "fish scale" (diameter, 8 mm; thickness, 0.3 mm), (2) cubes with 2-mm sides, (3) cubes with 1-mm sides, and (4) cartilage paste (< 0.3 mm). The cultures were maintained in chondrogenic medium for 6 weeks. Biochemically, a proteoglycan (PG):DNA ratio was calculated as the best approximation of ECM production per cell. The ratio between PG released in the culture medium and the PG in the neocartilage (PGrel:PG) was used as a matrix stability index. Histologically, the slides were stained with safranin O fast green and collagen type II immunostaining. The titration of safranin O-positive cells and the Bern score were calculated.

RESULTS

Regarding the PG:DNA ratio, group 4 performed significantly better than groups 1 (P = .001) and 3 (P = .02), whereas group 2 performed better than group 1 (P = .03). No significant difference was found regarding the PGrel:PG ratio and safranin O-positive cells. Regarding the Bern score, group 4 performed significantly better than groups 1 (P = .02), 2 (P = .04), and 3 (P = .03).

CONCLUSIONS

We conclude that human cartilage fragmentation significantly affects ECM production in vitro. Increased fragmentation enhances ECM production.

CLINICAL RELEVANCE

Assuming a similar behavior in vivo, we recommend mincing the cartilage into small pieces when performing the cartilage fragment autograft implantation technique in order to increase ECM production. Further in vitro studies investigating cartilage of younger nonarthritic donors, as well as in vivo studies, are needed.

摘要

目的

评估软骨碎片程度是否会影响体外软骨碎片自体移植中细胞外基质(ECM)的产生。

方法

从5名接受全髋关节置换术的供体(平均年龄65.6岁;标准差[SD]为3)获取软骨。将软骨切碎以获得4组不同碎片大小的样本:(1)“鱼鳞”状(直径8mm;厚度0.3mm),(2)边长为2mm的立方体,(3)边长为1mm的立方体,以及(4)软骨糊(<0.3mm)。将培养物置于软骨形成培养基中培养6周。生化方面,计算蛋白聚糖(PG):DNA比值作为每个细胞ECM产生的最佳近似值。培养基中释放的PG与新软骨中PG的比值(PGrel:PG)用作基质稳定性指标。组织学上,玻片用番红O固绿染色和II型胶原免疫染色。计算番红O阳性细胞的滴定度和伯恩评分。

结果

关于PG:DNA比值,第4组的表现明显优于第1组(P = 0.001)和第3组(P = 0.02),而第2组的表现优于第1组(P = 0.03)。在PGrel:PG比值和番红O阳性细胞方面未发现显著差异。关于伯恩评分,第4组的表现明显优于第1组(P = 0.02)、第2组(P = 0.04)和第3组(P = 0.03)。

结论

我们得出结论,人类软骨碎片化会显著影响体外ECM的产生。碎片化增加会提高ECM的产生。

临床意义

假设在体内有类似表现,我们建议在进行软骨碎片自体移植技术时将软骨切碎成小块,以增加ECM的产生。需要进一步进行针对年轻非关节炎供体软骨的体外研究以及体内研究。

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