聚维酮碘作为脱钙骨基质保存剂的实验研究

A study of PVP-I as a long term preservation agent for demineralized bone matrix

目的探讨聚维酮碘(PVP-I)作为脱钙骨基质(demineralized bone matrix,DBM)长期保存剂的可行性。方法选用4种标准菌种进行PVP-I重复5次的灭菌效果验证。分别采用CCK-8法、PNPP法、RT-PCR法检测PVP-I对成骨细胞的增殖、前交叉韧带(anterior cruciate ligament,ACL)活力以及成骨分化标志分子COL1α、ALP、RUNX2、OCN m RNA表达的影响;裸小鼠肌间隙植入实验验证PVP-I长期保存的DBM成骨活性是否受到影响。结果 PVP-I对金黄色葡萄球菌、白色念珠菌、枯草杆菌黑色变种芽孢的杀灭率均为100%,对大肠杆菌的杀灭率为88.7%;与对照组(1±0.09)相比,高浓度PVP-I抑制细胞生长(0.63±0.03)(P<0.05),低浓度PVP-I不促进也不抑制细胞增殖;PVP-I显著增强成骨细胞的ALP活力,上调COL1α、ALP、RUNX2 m RNA的表达量(1.25±0.15、1.38±0.14、1.23±0.09)(P<0.05),对OCN m RNA的表达影响不明显;裸鼠肌间隙植入实验表明,PVP-I长期保存的DBM其成骨活性不受影响。结论 PVP-I有望被开发成DBM的保存剂。

Objective To discuss the feasibility of PVP-I being the preservation of demineralized bone matrix（ DBM）. Methods Four standard strains were used to perform the sterilization experiment in repeated 4 times. PVP-I was added to MC3T3-E1, then cell proliferation and viability were determined by CCK-8 and differentiation status by PNPP and RT-PCR. Muscle implant experiment of nude mice was performed to verify whether the activity of DBM was damaged since it was immersed in PVP-I for a long time. Results The sterilization rates of PVP-I to staphylococcus aureus, monilia albican and bacillus subtilis were all of 100%, while to Escherichia coli 88.7%. CCK-8 assay showed reduced proliferation of the MC3T3-E1 cells when the PVP-I was at a high concentration（ 0.63±0.03）（ P0.05）. When the concentration of PVP-I was down-regulated, a negligible effect was observed on MC3T3-E1 growth. PVP-I significantly enhanced the activity of ALP and up-regulated the expressions of COL1α, ALP and RUNX2（ 1.25±0.15, 1.38±0.14, 1.23±0.09）（ P〈0.05）. However, PVP-I did not affect the expression of OCN. The nude mice experiment suggested that the activity of DBM was not influenced by PVP-I. Conclusions PVP-I may be applied as the preservation of DBM.