OBJECTIVE:Marrow-stimulation techniques are used by surgeons to repair cartilage lesions although consistent regeneration of hyaline cartilage is rare. We have shown previously that autologous blood can be mixed with a polymer solution containing chitosan in a glycerol phosphate (GP) buffer (chitosan-GP), and that implantation of this polymer/blood composite onto marrow-stimulated chondral defects in rabbit and sheep leads to the synthesis of more chondral repair tissue with greater hyaline character compared to marrow-stimulation alone. In the current study, we examined the modulation of cell recruitment and repair tissue characteristics at early post-surgical time points (from day 1 to 56) in a rabbit model to elucidate potential mechanisms behind this improved repair outcome. DESIGN:Thirty-three skeletally mature New Zealand White rabbits underwent bilateral arthrotomies, with each trochlea receiving a cartilage defect (3.5 mm x 4.5mm) bearing four microdrill holes (0.9 mm diameter, approximately 4 mm deep) into the subchondral bone. One defect per rabbit was treated with a chitosan-GP/blood implant, while the other defect was left as a microdrilled control. Repair tissues were stained by histochemistry, for collagen types I, II, and X by immunohistochemistry and analyzed using quantitative stereological tools. RESULTS:Histological analyses demonstrated that control defects followed a typical healing sequence observed previously in marrow-stimulation animal models while chitosan-GP/blood implants led to three significant modifications in the healing sequence at early stages: (1) increased inflammatory and marrow-derived stromal cell recruitment to the microdrill holes, (2) increased vascularization of the provisional repair tissue in the microdrill holes, and (3) increased intramembranous bone formation and subchondral bone remodeling (BR). CONCLUSIONS:These results suggest that the greater levels of provisional tissue vascularization and BR activity are main factors supporting improved cartilage repair when chitosan-GP/blood implants are applied to marrow-stimulated cartilage lesions.

译文

目的:尽管透明软骨的持续再生很少见,但外科医生仍采用骨髓刺激技术修复软骨损伤。以前我们已经证明,自体血液可以与在磷酸甘油(GP)缓冲液中含有壳聚糖的聚合物溶液(chitosan-GP)混合,并将这种聚合物/血液复合物植入到兔和绵羊的骨髓刺激的软骨缺损中与单独的骨髓刺激相比,可以合成更多的具有更透明的玻璃质特征的软骨修复组织。在当前的研究中,我们在兔模型的手术后早期(从第1天到第56天)检查了细胞募集和修复组织特征的调节,以阐明这种改善的修复结果背后的潜在机制。
设计:对33只骨骼成熟的新西兰白兔进行了双侧关节置换术,每只滑车都接受了软骨缺损(3.5 mm x 4.5mm),并在软骨下骨中带有四个微孔(直径0.9 mm,深约4 mm)。每只兔子的一个缺损用壳聚糖-GP /血液植入物治疗,而另一只缺损留作微钻孔对照。通过组织化学对修复组织进行染色,通过免疫组织化学对I,II和X型胶原进行染色,并使用定量立体工具进行分析。
结果:组织学分析表明,对照缺陷遵循了先前在骨髓刺激动物模型中观察到的典型愈合顺序,而壳聚糖-GP /血液植入物在早期阶段对愈合顺序产生了三个重大改变:(1)炎症和骨髓来源的增加基质细胞募集到微孔中,(2)增加了微孔中临时修复组织的血管形成,(3)膜内骨形成和软骨下骨重塑(BR)增加。
结论:这些结果表明,当将壳聚糖-GP /血液植入物应用于骨髓刺激的软骨病变时,更高水平的临时组织血管生成和BR活性是支持改善软骨修复的主要因素。

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