Liu Yumei, Shi Feng, Bo Lin, Zhi Wei, Weng Jie, Qu Shuxin
Key Lab of Advanced Technologies of Materials, Ministry of Education, School of Materials Science and Engineering, Southwest Jiaotong University, Chengdu 610031, China.
Laboratory of Stem Cell and Tissue Engineering, Regenerative Medicine Research Center, West China Hospital, Sichuan University, Chengdu 610041, China.
Mater Sci Eng C Mater Biol Appl. 2017 Oct 1;79:679-686. doi: 10.1016/j.msec.2017.05.119. Epub 2017 May 17.
The aim of this study was to develop a novel alginate-encapsulated system (Alg beads) to investigate the cell response to critical-sized wear particles of ultra-high molecular weight polyethylene loaded with alendronate sodium (UHMWPE-ALN), one of the most effective drugs to treat bone resorption in clinic. The extrusion method was used to prepare Alg beads encapsulating rat calvarial osteoblasts (RCOs) and critical-sized UHMWPE-ALN wear particles with spherical morphology and uniform size. The morphology, permeability and stability of Alg beads were characterized. The proliferation, ALP activity, cell apoptosis and distribution of live/dead RCOs co-cultured with wear particles in Alg beads were evaluated. RCOs and critical-sized UHMWPE-ALN wear particles distributed evenly and contacted efficiently in Alg beads. Alg beads were both permeable to trypsin and BSA, while the smaller the molecular was, the larger the diffuse was. The proliferation of RCOs in Alg beads increased with time, which indicated that Alg beads provided suitable conditions for cell culture. The long-term stability of Alg beads indicated the possibility for the longer time of co-cultured cells with wear particles. Critical-sized UHMWPE-ALN and UHMWPE wear particles both inhibited the proliferation and differentiation of RCOs, and induced the apoptosis of RCOs encapsulated in Alg beads. However, these effects could be significantly alleviated by the ALN released from the critical-sized UHMWPE-ALN wear particles. The present results suggested that this novel-developed co-culture system was feasible to evaluate the cell response to critical-sized UHMWPE-ALN wear particles for a longer time.
本研究的目的是开发一种新型的海藻酸盐包封系统（Alg珠），以研究细胞对负载阿仑膦酸钠的超高分子量聚乙烯临界尺寸磨损颗粒（UHMWPE-ALN）的反应，阿仑膦酸钠是临床上治疗骨吸收最有效的药物之一。采用挤压法制备包封大鼠颅骨成骨细胞（RCOs）和具有球形形态且尺寸均匀的临界尺寸UHMWPE-ALN磨损颗粒的Alg珠。对Alg珠的形态、渗透性和稳定性进行了表征。评估了与Alg珠中的磨损颗粒共培养的RCOs的增殖、碱性磷酸酶（ALP）活性、细胞凋亡以及活/死RCOs的分布。RCOs和临界尺寸的UHMWPE-ALN磨损颗粒在Alg珠中均匀分布且有效接触。Alg珠对胰蛋白酶和牛血清白蛋白（BSA）均具有渗透性，且分子越小，扩散越大。Alg珠中RCOs的增殖随时间增加，这表明Alg珠为细胞培养提供了合适的条件。Alg珠的长期稳定性表明了共培养细胞与磨损颗粒更长时间接触的可能性。临界尺寸的UHMWPE-ALN和UHMWPE磨损颗粒均抑制RCOs的增殖和分化，并诱导包封在Alg珠中的RCOs凋亡。然而，临界尺寸的UHMWPE-ALN磨损颗粒释放的ALN可显著减轻这些影响。目前的结果表明，这种新开发的共培养系统对于长时间评估细胞对临界尺寸UHMWPE-ALN磨损颗粒的反应是可行的。
