可塑形脱矿骨基质/透明质酸骨泥的制备与细胞相容性

背景：应用预制成形的材料修复不规则骨缺损时，常出现材料与骨缺损形状不匹配的问题，制备可任意塑形骨泥对促进不规则骨缺损的愈合具有重要意义。目的：构建可塑形脱钙骨基质/透明质酸骨泥，筛选最佳复合比例并评价其细胞相容性。方法：选取合格供体皮质骨制备脱钙骨基质。分别配置浓度为0.75%，1.5%，3%，4.5%的透明质酸溶液并检测其黏度，以450 mg脱钙骨基质与上述各浓度透明质酸溶液1 mL混合，制备不同比例的脱钙骨基质/透明质酸骨泥。37 ℃条件下将骨泥浸泡在PBS平衡液中，观察其离散时间，筛选最佳复合比例的脱钙骨基质/透明质酸骨泥。应用脱钙骨基质/透明质酸骨泥浸提液培养L-929小鼠成纤维细胞，1，4，7 d采用CCK-8法检测骨泥的细胞毒性。结果与结论：随透明质酸浓度的增加其溶液黏度增大，脱钙骨基质/透明质酸骨泥的离散时间延长，3%浓度的透明质酸与450 mg脱钙骨基质复合制备的骨泥具有良好可塑形能力，离散时间为8 h，为最佳复合比例。脱钙骨基质/透明质酸骨泥浸提液培养L-929小鼠成纤维细胞1，4，7 d的细胞增殖率分别为93.72%，101.65%，97.68%，细胞毒性级别为0或1级。表明脱钙骨基质/透明质酸复合质量比为15∶1制备的骨泥具有良好的可塑形能力及抗离散能力，细胞毒性轻微。

Preparation and cell compatibility of malleable demineralized bone matrix/hyaluronic acid putty

BACKGROUND:It is important to prepare a malleable putty to repair irregular bone defects because the preformed bone substitute material cannot suit to bone defects.OBJECTIVE:To prepare malleable demineralized bone matrix/hyaluronic acid putty and to screen out the optimal composite ratio, as well as to evaluate its cell compatibility.METHODS:Demineralized bone matrix was prepared from the cortical bone of healthy donors. The hyaluronic acid was dissolved at concentrations of 0.75%, 1.5%, 3%, 4.5%, and the viscosity of them was measured. Then 450 mg demineralized bone matrix was compounded with 1 mL hyaluronic acid solution to prepare the demineralized bone matrix/hyaluronic acid putty, and the optimal ratio of demineralized bone matrix/hyaluronic acid was screened by the disintegrated time of the putty at 37 ℃. L-929 mouse fibroblasts were cultured in the leaching liquor of demineralized bone matrix/hyaluronic acid, and the cytotoxicity was tested by cell count kit-8 assay at days 1, 4 and 7.RESULTS AND CONCLUSION:With the increase of hyaluronic acid concentration, the viscosity of hyaluronic acid solution was increased; the demineralized bone matrix/hyaluronic acid putty was strengthened and the disintegrated time was prolonged gradually. When the ratio of demineralized bone matrix/hyaluronic acid was 15/1, demineralized bone matrix/hyaluronic acid was molded easily, and the disintegrated time was 8 hours in 7 ℃ PBS. The results of cytotoxicity showed that cell proliferation rates were 93.72%, 101.65% and 97.68% at days 1, 4 and 7 respectively, while the cytotoxicity was in grade 0 or 1. These findings indicate that demineralized bone matrix/hyaluronic acid putty at a compound ratio of 15:1 can be molded easily and difficult to disintegrate, with low cytotoxicity.