Pennock Andrew T, Wagner Ferdinand, Robertson Catherine M, Harwood Frederick L, Bugbee William D, Amiel David
Department of Orthopedic Surgery, University of California, San Diego 92093-0630, USA.
J Knee Surg. 2006 Oct;19(4):265-72. doi: 10.1055/s-0030-1248117.
Osteochondral plugs were harvested from eight fresh human femoral condyles within 96 hours of donor death. The plugs were either stored in a serum-free media containing glucose, salts, and amino acids or 10% fetal bovine serum at 4 degrees C. After 28 days of storage, the osteochondral plugs were analyzed for chondrocyte viability and viable cell density using confocal microscopy, proteoglycan synthesis by (35)SO4 incorporation, and glycosaminoglycan content. Chondrocyte viability and cell density were significantly lower in grafts stored in serum-free media compared to fetal bovine serum, 27% versus 68% (P < .001) and 3250 cells/mm3 versus 8960 cells/mm3, respectively (P < .001). The metabolic activity determined by proteoglycan synthesis was significantly better in the specimens stored in fetal bovine serum (P < .01). No significant difference was detected between the glycosaminoglycan content in any of the specimens. These data suggest that the quality of osteochondral allografts as measured by chondrocyte viability, viable cell density, and proteoglycan synthesis is superior after storage in fetal bovine serum versus serum-free media. These results must be taken cautiously, however, as the clinical ramifications of storage in fetal bovine serum, including potential infectious disease transmission risks and immunogenic factors, have yet to be studied.
在供体死亡后96小时内,从8个新鲜的人类股骨髁采集骨软骨栓。将骨软骨栓保存在含有葡萄糖、盐和氨基酸的无血清培养基中,或保存在4℃的10%胎牛血清中。储存28天后,使用共聚焦显微镜分析骨软骨栓的软骨细胞活力和活细胞密度,通过(35)SO4掺入法分析蛋白聚糖合成,并分析糖胺聚糖含量。与胎牛血清相比,保存在无血清培养基中的移植物中软骨细胞活力和细胞密度显著降低,分别为27%对68%(P <.001)和3250个细胞/mm3对8960个细胞/mm3(P <.001)。通过蛋白聚糖合成测定的代谢活性在保存在胎牛血清中的标本中显著更好(P <.01)。在任何标本的糖胺聚糖含量之间未检测到显著差异。这些数据表明,以软骨细胞活力、活细胞密度和蛋白聚糖合成衡量的同种异体骨软骨移植质量在保存在胎牛血清中比保存在无血清培养基中更优。然而,必须谨慎看待这些结果,因为胎牛血清储存的临床影响,包括潜在的传染病传播风险和免疫原性因素,尚未得到研究。