Vukasović Andreja, Ivković Alan, Jezek Davor, Cerovecki Ivan, Vnuk Drazen, Kreszinger Mario, Hudetz Damir, Pećina Marko
Zavod za histologiju i embriologiju Medicinskog fakulteta Sveucilista u Zagrebu, Zagreb, Hrvatska.
Acta Med Croatica. 2011;65(3):227-35.
Articular cartilage is an avascular and aneural tissue lacking lymph drainage, hence its inability of spontaneous repair following injury. Thus, it offers an interesting model for scientific research. A number of methods have been suggested to enhance cartilage repair, but none has yet produced significant success. The possible application of the aforementioned methods has brought about the necessity to evaluate their results. The objective of this study was to analyze results of a study of the effects of the use of TGF-beta gene transduced bone marrow clot on articular cartilage defects using ICRS visual histological assessment scale.
The research was conducted on 28 skeletally mature sheep that were randomly assigned to four groups and surgically inflicted femoral chondral defects. The articular surfaces were then treated with TGF-beta1 gene transduced bone marrow clot (TGF group), GFP transduced bone marrow clot (GFP group), untransduced bone marrow clot (BM group) or left untreated (NC group). The analysis was performed by visual examination of cartilage samples and results were obtained using ICRS visual histological assessment scale. The results were subsequently subjected to statistical assessment using Kruskal-Wallis and Mann-Whitney tests.
Kruskal-Wallis test yielded statistically significant difference with respect to cell distribution. Mann-Whitney test showed statistically significant difference between TGF and NC groups (P = 0.002), as well as between BM and NC groups (P = 0.002 with Bonferroni correction).
Twenty-six of the twenty-eight samples were subjected to histologic and subsequent statistical analysis; two were discarded due to faulty histology technique. Our results indicated a level of certainty as to the positive effect of TGF-beta1 gene transduced bone marrow clot in restoration of articular cartilage defects. However, additional research is necessary in the field. One of the significant drawbacks on histologic assessment of cartilage samples were the errors in histologic preparation, for which some samples had to be discarded and significantly impaired the analytical quality of the others. Defects of structures surrounding the articular cartilage, e.g., subchondral bone or connective tissue, might also impair the quality of histologic analysis. Additional analyses, i.e. polarizing microscopy should be performed to determine the degree of integration of the newly formed tissue with the surrounding cartilage. The semiquantitative ICRS scale, although of great practical value, has limitations as to the objectivity of the assessment, taking into account the analytical ability of the evaluator, as well as the accuracy of semiquantitative analysis in comparison to the methods of quantitative analysis.
Overall results of histologic analysis indicated that the application of TGF-beta1 gene transduced bone marrow clot could have measurable clinical effects on articular cartilage repair. The ICRS visual histological assessment scale is a valuable analytical method for cartilage repair evaluation. In this respect, further analyses of the method value would be of great importance.
关节软骨是一种无血管、无神经且缺乏淋巴引流的组织,因此损伤后无法自发修复。故而,它为科学研究提供了一个有趣的模型。人们提出了许多增强软骨修复的方法,但尚无显著成效。上述方法的可能应用使得评估其结果成为必要。本研究的目的是使用国际软骨修复协会(ICRS)视觉组织学评估量表,分析一项关于转化生长因子-β(TGF-β)基因转导骨髓凝块对关节软骨缺损影响的研究结果。
对28只骨骼成熟的绵羊进行研究,将它们随机分为四组,并通过手术造成股骨软骨缺损。然后,用TGF-β1基因转导骨髓凝块(TGF组)、绿色荧光蛋白(GFP)基因转导骨髓凝块(GFP组)、未转导骨髓凝块(BM组)或不进行处理(NC组)对关节表面进行处理。通过对软骨样本进行视觉检查来进行分析,并使用ICRS视觉组织学评估量表获得结果。随后,使用Kruskal-Wallis检验和Mann-Whitney检验对结果进行统计学评估。
Kruskal-Wallis检验在细胞分布方面产生了具有统计学意义的差异。Mann-Whitney检验显示TGF组与NC组之间存在统计学意义的差异(P = 0.002),BM组与NC组之间也存在统计学意义的差异(经Bonferroni校正后P = 0.002)。
28个样本中有26个进行了组织学及后续的统计分析;2个因组织学技术有误而被丢弃。我们的结果表明,对于TGF-β1基因转导骨髓凝块在关节软骨缺损修复中的积极作用有一定程度的确定性。然而,该领域还需要进一步研究。软骨样本组织学评估的一个显著缺点是组织学制备中的误差,为此一些样本不得不被丢弃,这严重影响了其他样本的分析质量。关节软骨周围结构的缺陷,如软骨下骨或结缔组织,也可能损害组织学分析的质量。应进行额外的分析,即偏振显微镜检查,以确定新形成的组织与周围软骨的整合程度。ICRS半定量量表虽然具有很大的实用价值,但考虑到评估者的分析能力以及与定量分析方法相比半定量分析的准确性,其评估的客观性存在局限性。
组织学分析的总体结果表明,应用TGF-β1基因转导骨髓凝块可能对关节软骨修复产生可测量的临床效果。ICRS视觉组织学评估量表是评估软骨修复的一种有价值的分析方法。在这方面,进一步分析该方法的价值将非常重要。