Robilotto Anthony T, Clarke Dominic, Baust John M, Van Buskirk Robert G, Gage Andrew A, Baust John G
Institute of Biomedical Technology, State University of New York at Binghamton, Binghamton, NY 13902, USA.
Technol Cancer Res Treat. 2007 Apr;6(2):81-9. doi: 10.1177/153303460700600204.
The study of the effectiveness of cryotherapy as a curative treatment for prostate cancer has often relied on the use of either in vitro cell culture monolayers or animal models. While the data gleaned from these studies have been valuable, each model has inherent limitations. In order to bridge the gap between in vitro studies and clinical applications, we developed a 3-dimensional, tissue engineered human prostate cancer model to simulate and assess the effects of cryotherapy and adjunctive treatments on cell viability and activation of cell death pathways throughout the thermally variable freeze zone. Human prostate cancer cells (PC3) were seeded into collagen based matrices and cryolesions were generated using an Oncura SeedNet Gold cryosurgical device with 17-gauge cryoprobes. Analyses revealed widespread necrosis diminishing towards the edge of the freeze zone, and a time-dependent wave of apoptosis starting as early as 1 hr post-thaw at low temperatures (< -40 degrees C) and moving toward the periphery (-20 degrees C) as recovery times reached 12 and 24 hr. Distal to the -10 degrees C isotherm, minimal cell death was apparent (< 20%) over controls. The adjunctive use of chemotherapeutic agents in conjunction with cryosurgery displayed a similar induction of cell death cascades, but with the zone of cryodestruction extending approximately 10 to 15 degrees C further into the freeze zone periphery. By providing an extracellular environment and a matrix to minimize innate variables, the tissue engineered model yielded a more in vivo-like, tumor-like environment supportive of a deeper understanding of the specific biological responses of cancer cells/tumors to cryotherapeutic intervention.
冷冻疗法作为前列腺癌治疗方法有效性的研究通常依赖于体外细胞培养单层或动物模型。虽然从这些研究中收集的数据很有价值，但每个模型都有其固有的局限性。为了弥合体外研究与临床应用之间的差距，我们开发了一种三维组织工程化人前列腺癌模型，以模拟和评估冷冻疗法及辅助治疗对整个热可变冷冻区内细胞活力和细胞死亡途径激活的影响。将人前列腺癌细胞（PC3）接种到基于胶原蛋白的基质中，并使用带有17号冷冻探头的Oncura SeedNet Gold冷冻手术设备产生冷冻损伤。分析显示，广泛的坏死在冷冻区边缘逐渐减少，并且早在解冻后1小时，在低温（<-40摄氏度）下就开始出现时间依赖性的凋亡波，并随着恢复时间达到12小时和24小时向周边（-20摄氏度）移动。在-10摄氏度等温线远端，与对照组相比，细胞死亡极少（<20%）。化疗药物与冷冻手术联合使用显示出类似的细胞死亡级联诱导，但冷冻破坏区域向冷冻区周边进一步延伸约10至15摄氏度。通过提供细胞外环境和基质以尽量减少固有变量，组织工程模型产生了更类似体内的肿瘤样环境，有助于更深入地了解癌细胞/肿瘤对冷冻治疗干预的特定生物学反应。
