Departamento de Innovación en Material Biológico Humano (DIMBIH), Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Ciudad de México, México; Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Ciudad de México, México.
Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Ciudad de México, México.
Forensic Sci Int. 2019 Dec;305:110027. doi: 10.1016/j.forsciint.2019.110027. Epub 2019 Oct 31.
Due the proteins from bone remains are highly resistant to pass of time and environmental conditions, they could tell us about the events that probably happened in the past. In the forensic and physical anthropology context, burnt bone remains are one of the most common pieces of recovered evidence and, generally, they are associated with funerary practices, criminal scenes or massive catastrophic events. In the present study, bone pieces of pigs were calcined at different calcination temperatures, and proteins were searched using biochemical, immunochemical and ultrastructure visualization under these experimentally conditions. For this purpose, it was successfully developed a non-demineralizing protein extraction method from burnt bone remains and the use of specific antibodies permitted the identification of different extracellular matrix and intracellular proteins. While collagen proteins type I and IV were identified and detected under middle and high calcination temperatures (300°C and 600°C); cytoskeletal proteins as actin, tubulin and, the microtubule associated protein Tau, were found under calcination process, even up high calcination temperatures. Under ultrastructural analysis, fibrous materials with a classical disposition of collagens were observed even at high calcination temperatures of the burnt bone remains. The protein identification and characterization in burnt bones as performed in present studies, is clearly demonstrating that using specific strategies for protein characterizations it is possible to found protein biomarkers in burnt bone remains and this strategy could be useful for forensic and anthropological purposes.
由于骨骼中的蛋白质对时间和环境条件的变化具有很强的抵抗力,因此它们可以为我们提供有关过去可能发生的事件的信息。在法医和体质人类学领域,烧焦的骨骼遗骸是最常见的证据之一,通常与丧葬习俗、犯罪现场或大规模灾难性事件有关。在本研究中,我们将猪骨在不同的煅烧温度下进行煅烧,并在这些实验条件下使用生物化学、免疫化学和超微结构可视化技术来寻找蛋白质。为此,我们成功开发了一种从烧焦的骨骼遗骸中提取非脱矿质蛋白质的方法,并且使用特定的抗体可以识别不同的细胞外基质和细胞内蛋白质。在中高温(300°C 和 600°C)下可以鉴定和检测到胶原蛋白 I 型和 IV 型;在煅烧过程中,甚至在高温下,还可以发现肌动蛋白、微管蛋白和微管相关蛋白 Tau 等细胞骨架蛋白。在超微结构分析中,即使在烧焦的骨骼遗骸的高温下,也观察到具有胶原典型排列的纤维材料。本研究中对烧焦骨骼中的蛋白质进行的鉴定和表征,清楚地表明,使用特定的蛋白质特征鉴定策略,可以在烧焦的骨骼遗骸中发现蛋白质生物标志物,并且这种策略可能对法医学和人类学领域具有重要意义。