Redman S N, Dowthwaite G P, Thomson B M, Archer C W
Cardiff School of Biosciences, Cardiff Institute of Tissue Engineering and Repair, Museum Avenue,PO Box 911, CF10 3US, Cardiff, UK.
Osteoarthritis Cartilage. 2004 Feb;12(2):106-16. doi: 10.1016/j.joca.2002.12.001.
To determine the response of immature articular cartilage to both sharp and blunt trauma in terms of cell death, cell proliferation and matrix synthesis.
Blunt wounds were made with a trephine in full depth immature bovine articular cartilage explants which were cut in half through the center of the trephine wound with a sharp scalpel to produce blunt and sharp trauma on the same explant. Explants were maintained in culture for up to 10 days. Prior to fixation at days 2, 5 and 10, medium was supplemented with 10 microCi ml-1 35S-sulphate, [3H]-proline or [3H]-thymidine for 24h to assess matrix synthesis and cell proliferation. Cell death was assessed using a Live/Dead label.
In the case of blunt wounds, a band of cell death was observed adjacent to the lesion edge. Microautoradiography demonstrated little radiolabel incorporation and, therefore, no new matrix synthesis or cell proliferation within this region. In contrast, wounds made with a sharp scalpel showed restricted cell death, with radiolabel incorporation adjacent to the lesion edge at all time points. This demonstrated not only chondrocyte proliferation and new matrix synthesis at the wound margin, but also an up-regulation of matrix synthesis adjacent to the lesion edge.
In terms of clinical relevance, the use of sharp precise instruments during the surgical management of cartilage defects may be necessary to reduce cell death and promote matrix elaboration at the lesion edge in order to facilitate successful integration.
从细胞死亡、细胞增殖和基质合成方面确定未成熟关节软骨对锐性和钝性创伤的反应。
用环钻在未成熟牛关节软骨全层外植体上造成钝性创伤,然后用锋利手术刀通过环钻创伤中心将外植体切成两半,以便在同一外植体上产生钝性和锐性创伤。将外植体培养长达10天。在第2、5和10天固定之前,向培养基中添加10微居里/毫升的35S-硫酸盐、[3H]-脯氨酸或[3H]-胸腺嘧啶核苷24小时,以评估基质合成和细胞增殖。使用活/死标记评估细胞死亡情况。
对于钝性创伤,在损伤边缘附近观察到一条细胞死亡带。显微放射自显影显示几乎没有放射性标记掺入,因此该区域内没有新的基质合成或细胞增殖。相比之下,用锋利手术刀造成的创伤显示细胞死亡受限,在所有时间点损伤边缘附近都有放射性标记掺入。这不仅表明在伤口边缘有软骨细胞增殖和新的基质合成,而且还表明在损伤边缘附近基质合成上调。
就临床相关性而言,在软骨缺损手术治疗过程中使用锋利精密的器械可能是必要的,以减少细胞死亡并促进损伤边缘的基质形成,从而促进成功整合。