静压力对脂肪干细胞与静电纺丝纳米纤维细胞相容性的影响

Effects of static pressure on the cytocompatibility of adipose-derived stem cells on electrospun nanofibers

背景:静电纺丝聚乳酸/聚己内酯纳米纤维是课题组自行合成的生物可降解材料,前期研究表明该材料生物相容性优越,但缺乏对该材料力学方面的研究。目的:研究静压力对脂肪干细胞与静电纺丝聚乳酸/聚己内酯纳米纤维支架细胞相容性的影响。方法:将脂肪干细胞接种到由聚乳酸和聚己内酯合成的静电纺丝纳米纤维支架上,置于含体积分数为10%胎牛血清的DMEM低糖培养基中培养。利用静压力装置分别对培养板持续加载0,15,30,45 k Pa的静压力,加载时间为4 h。采用MTT法、活死细胞染色检测脂肪干细胞在静电纺丝纳米纤维支架上的增殖、黏附和活性,评价脂肪干细胞与静电纺丝纳米纤维的细胞相容性。结果与结论:(1)在体外培养条件下,经不同静压力持续加载4 h后,MTT法增殖检测得出的各组吸光度值经单因素方差分析差异存在显著性意义。在0-30 k Pa压力范围内,吸光度值随静压力的增加而增加,到45 k Pa时,吸光度值反而下降。各组间多重比较显示差异均有显著性意义;(2)用MTT法进一步检测细胞黏附百分率,各组间差异亦有显著性意义;(3)活死细胞染色结果印证了上述结果,统计分析表明,无论是单因素方差分析还是各组间配对t检验,4组间活细胞百分率的差异均有非常显著性意义;(4)适当大小的静压力可促进脂肪干细胞与静电纺丝纳米纤维的生物相容性,细胞增殖、黏附及活性有所提高,但过高的静压力会抑制细胞的生物学行为,从而影响脂肪干细胞与静电纺丝纳米纤维的相容性。

BACKGROUND： Electrospun polylactic acid/polycaprolactone nanofibers （ENF） are a kind of self-synthesized biodegradable material. Our preliminary studies have indicated that the biomaterial exhibits excellent biocompatibility; however, the research about its mechanics is still little. OBJECTIVE： To explore the effects of static pressure on the cytocompatibility of adipose-derived stem cells on the ENF scaffold. METHODS： Adipose-derived stem cells were seeded onto the ENF scaffold, and then cultured in the low-glucose DMEM supplemented with 10% fetal bovine serum. The mixed constructs were submitted to the static pressure at 0, 15, 30, and 45 kPa for4 hours using a static pressure device, respectively. Subsequently, the proliferation, adhesion and viability of adipose-derived stem cells on the ENF scaffold were detected using MTT assay and living/dead staining to evaluate the cytocompatibility. RESULTS AND CONCLUSION： M＇I-I- assay showed that there were significant differences in absorbance values among groups by one-way analysis of variance after 4 hours of loading with different static pressures in vitro. Under 0-30 kPa static pressure, the absorbance values increased with static pressure, but the absorbance values declined until the pressure reached 45 kPa, and multiple comparisons between groups showed significant difference. The significant differences in the cell attachment percentage by MTT assay could be found among groups. The living/dead staining results supported the above findings. Furthermore, the significant differences in percentage of living cells among groups were shown using either one-way analysis of variance or paired t test. In conclusion, the appropriate static pressure can promote the cytocompatibility, proliferation, adhesion and viability of adipose-derived stem cells on the ENF scaffold. But the excessive pressure is likely to inhibit the cellular biological behaviors, thus affecting cytocompatibility of adipose-derived stem cells with the ENF scaffold.