Department of Oral, Maxillofacial and Plastic Surgery, University Medical Center Rostock, 18057 Rostock, Germany.
Department of Orthopedics, Biomechanics and Implant Technology Research Laboratory, University Medical Center Rostock, 18057 Rostock, Germany.
Int J Mol Sci. 2020 May 28;21(11):3836. doi: 10.3390/ijms21113836.
Chemical and physical processing of allografts is associated with a significant reduction in biomechanics. Therefore, treatment of tissue with high hydrostatic pressure (HHP) offers the possibility to devitalize tissue gently without changing biomechanical properties. To obtain an initial assessment of the effectiveness of HHP treatment, human osteoblasts and chondrocytes were treated with different HHPs (100-150 MPa, 250-300 MPa, 450-500 MPa). Devitalization efficiency was determined by analyzing the metabolic activity via WST-1(water-soluble tetrazolium salt) assay. The type of cell death was detected with an apoptosis/necrosis ELISA (enzyme-linked immune sorbent assay) and flow cytometry. Field emission scanning electron microscopy (FESEM) and transmission electron microscopy (TEM) were carried out to detect the degree of cell destruction. After HHP treatment, the metabolic activities of both cell types decreased, whereas HHP of 250 MPa and higher resulted in metabolic inactivation. Further, the highest HHP range induced mostly necrosis while the lower HHP ranges induced apoptosis and necrosis equally. FESEM and TEM analyses of treated osteoblasts revealed pressure-dependent cell damage. In the present study, it could be proven that a pressure range of 250-300 MPa can be used for cell devitalization. However, in order to treat bone and cartilage tissue gently with HHP, the results of our cell experiments must be verified for tissue samples in future studies.
同种异体移植物的化学和物理处理会导致生物力学显著降低。因此，使用高静水压力（HHP）处理组织为实现温和的组织失活而不改变生物力学特性提供了可能。为了初步评估 HHP 处理的效果，用不同的 HHP（100-150 MPa、250-300 MPa、450-500 MPa）处理人成骨细胞和软骨细胞。通过 WST-1（水溶性四唑盐）测定法分析代谢活性来确定失活效率。用凋亡/坏死 ELISA（酶联免疫吸附试验）和流式细胞术检测细胞死亡类型。进行场发射扫描电子显微镜（FESEM）和透射电子显微镜（TEM）检测以检测细胞破坏程度。HHP 处理后，两种细胞类型的代谢活性均降低，而 250 MPa 及更高 HHP 则导致代谢失活。此外，最高 HHP 范围主要诱导坏死，而较低 HHP 范围则同等地诱导凋亡和坏死。处理后的成骨细胞的 FESEM 和 TEM 分析显示出压力依赖性的细胞损伤。在本研究中，可以证明 250-300 MPa 的压力范围可用于细胞失活。然而，为了温和地用 HHP 处理骨和软骨组织，必须在未来的研究中针对组织样本验证我们的细胞实验结果。
