Regenerative, Modular & Developmental Engineering Laboratory (REMODEL), Biomedical Sciences Building, National University of Ireland Galway (NUI Galway), Galway, Ireland. Science Foundation Ireland (SFI) Centre for Research in Medical Devices (CÚRAM), Biomedical Sciences Building, National University of Ireland Galway (NUI Galway), Galway, Ireland.
Biomed Mater. 2017 Oct 25;12(6):065009. doi: 10.1088/1748-605X/aa838d.
Collagen based devices are frequently associated with foreign body response. Although several pre- (e.g. species, state of animal, tissue) and post- (e.g. cross-linking, scaffold architecture) extraction method factors have a profound effect on foreign body response, little is known about which and how during the extraction process factors mediate foreign body response. In this study, we assessed the influence of acetic acid and hydrochloric acid and the utilisation or not of pepsin or salt precipitation during collagen extraction on the yield, purity, free amines, denaturation temperature, resistance to collagenase degradation and macrophage response. Acetic acid/pepsin extracted collagen exhibited the highest yield, purity and free amine content and the lowest denaturation temperature. No differences in resistance to collagenase digestion were detected between the groups. Although all treatments exhibited similar macrophage morphology comprised of round cells (M1 phenotype), elongated cells (M2 phenotype) and cell aggregates (foreign body response), significantly more elongated cells were observed on HC films. Although no differences in metabolic activity were observed between the groups, the DNA concentration was significantly lower for the hydrochloric acid treatments. Further, cytokine analysis revealed that hydrochloric acid treatments induced significantly higher IL-1β and TNF-α release with respect to acetic acid treatments. Salt precipitation did not influence the parameters assessed. Collectively, these data suggest that during the collagen extraction process variables should also be monitored as, evidently, they affect the physicochemical and biological properties of collagen preparations.
胶原基器械常与异物反应有关。尽管有几种预处理（例如物种、动物状态、组织）和后处理（例如交联、支架结构）提取方法因素对异物反应有深远的影响，但对于在提取过程中哪些因素以及如何介导异物反应知之甚少。在这项研究中，我们评估了在胶原提取过程中使用或不使用醋酸和盐酸以及胃蛋白酶或盐沉淀对产量、纯度、游离胺、变性温度、胶原酶降解抗性和巨噬细胞反应的影响。乙酸/胃蛋白酶提取的胶原蛋白表现出最高的产率、纯度和游离胺含量，以及最低的变性温度。各组之间胶原酶消化抗性无差异。尽管所有处理都表现出相似的巨噬细胞形态，包括圆形细胞（M1 表型）、长形细胞（M2 表型）和细胞聚集（异物反应），但在 HC 膜上观察到的长形细胞明显更多。尽管各组之间的代谢活性没有差异，但盐酸处理组的 DNA 浓度明显较低。此外，细胞因子分析表明，与乙酸处理相比，盐酸处理诱导了更高的 IL-1β 和 TNF-α 释放。盐沉淀对评估的参数没有影响。总的来说，这些数据表明，在胶原提取过程中，还应监测变量，因为它们显然会影响胶原制剂的理化和生物学特性。
