Moreau M F, Gallois Y, Baslé M F, Chappard D
LHEA: Laboratoire d'Histologie--Embryologie, CHU & Faculté de Médecine, Angers, France.
Biomaterials. 2000 Feb;21(4):369-76. doi: 10.1016/s0142-9612(99)00193-3.
The uncertainties about the transmission of prion proteins from xenogenic grafts prepared from bovine bone has led to the reconsideration of allogenic bone as a grafting material. Allografting is a complementary technique to autografting nowadays when large bone volumes are necessary. Several preparation techniques have been proposed. Fresh-frozen, freeze-dried and gamma irradiation are the most common. However, a large amount of lipids is present in the medullary spaces (near 70% in weight for a human femoral head). They are known to strongly influence the biocompatibility of the bone graft. The exact changes of lipids upon the sterilization and storage processes are poorly known. The aims of the present study were to appreciate the effects of gamma irradiation on medullary lipids and to identify the cytotoxicity of gamma-irradiated bank bone with/without lipid on cultures of osteoblast-like cells. Bone cores from 8 femoral heads retrieved during prosthesis surgery for arthritis were prepared with a drilling trephine. Cores were either sterilized by gamma radiations (25000 gray) or kept frozen until lipid extraction and lipofuschine-like dosage by Folch's method and fluorometric study. Peroxidated lipids appeared 2 to 3-fold higher in the gamma-irradiated cores than in frozen ones. Slices were prepared from bone cores and were transferred on confluent osteoblast-like cell layers (Saos-2). The raw slices (containing lipids) did not induce cell death. On the other hand, cell death was dramatically increased around the gamma-irradiated slices. Defatted slices which had been sterilized by gamma radiations or UV did not induce cell death. Defatting procedures should be added when preparing bone allografts in human bone banks.
牛骨制备的异种移植物中朊病毒蛋白传播的不确定性，已导致人们重新考虑将同种异体骨作为一种移植材料。如今，当需要大量骨组织时，同种异体移植是自体移植的一种补充技术。已经提出了几种制备技术。新鲜冷冻、冻干和伽马辐射是最常见的。然而，骨髓腔中存在大量脂质（人类股骨头中重量近70%）。已知它们会强烈影响骨移植的生物相容性。脂质在灭菌和储存过程中的具体变化尚不清楚。本研究的目的是评估伽马辐射对骨髓脂质的影响，并确定含/不含脂质的伽马辐射骨库骨对成骨样细胞培养的细胞毒性。在关节炎假体手术期间从8个股骨头取出的骨芯用钻孔环锯制备。骨芯要么通过伽马辐射（25000戈瑞）灭菌，要么冷冻保存，直到通过福尔克方法进行脂质提取和脂褐素样定量以及荧光研究。在伽马辐射的骨芯中，过氧化脂质比冷冻骨芯中的高出2至3倍。从骨芯制备切片，并将其转移到汇合的成骨样细胞层（Saos-2）上。未脱脂的切片（含脂质）未诱导细胞死亡。另一方面，伽马辐射切片周围的细胞死亡显著增加。经伽马辐射或紫外线灭菌的脱脂切片未诱导细胞死亡。在人类骨库中制备同种异体骨时应增加脱脂程序。
