一种新型可塑形骨填充材料的制备及骨修复能力研究

Preparation and bone repair capability of a new plastic bone filler material

目的制备一种黏性载体和成骨基质颗粒均来源于人骨的可塑形骨填充材料,通过动物实验评价其安全性和骨诱导能力。方法取自愿捐赠的人体长骨,采用粉碎、清洗、脱矿等方法制备脱钙骨基质(demineralized bone matrix,DBM)颗粒,再将DBM颗粒经温浴法制备骨基质明胶(bone matrix gelatin,BMG),将BMG及DBM颗粒混合制备成可塑形骨填充材料(实验组);以单纯DBM颗粒作为对照组。取15只6~9周龄健康雄性无胸腺裸小鼠,于两侧臀中肌与臀大肌间制备肌间空隙,均植入实验组材料,术后1、4、6周处死动物,行HE组织学染色评价异位成骨效果。取8只9月龄日本大耳兔,于两侧后肢髁骨处制备直径6 mm骨缺损,左、右侧分别填充实验组和对照组材料,术后12、26周处死动物,行Micro-CT及HE组织学染色评价骨缺损修复效果。结果异位成骨实验HE染色示,术后1周可观察到大量软骨细胞,术后4、6周可观察到明显新生软骨组织。兔髁骨缺损修复实验HE染色示,术后12周,部分材料被吸收,实验组及对照组均可观察到新生软骨;术后26周,大部分材料被吸收,对照组可观察到大量新生骨,实验组可观察到新生骨单元结构。Micro-CT观测示,实验组成骨速率及成骨面积均优于对照组。骨形态计量学参数检测示,两组术后26周各参数均较术后12周显著增加(P<0.05)。术后12周,实验组骨密度、骨体积分数均显著高于对照组(P<0.05),两组骨小梁厚度差异无统计学意义(P>0.05);术后26周,实验组骨密度显著高于对照组(P<0.05),两组骨体积分数、骨小梁厚度差异无统计学意义(P>0.05)。结论新型可塑形骨填充材料具有良好的生物安全性及骨诱导活性,是一种良好的骨填充材料。

Objective To prepare a new plastic bone filler material with adhesive carrier and matrix particles derived from human bone,and evaluate its safety and osteoinductive ability through animal tests.Methods The human long bones donated voluntarily were prepared into decalcified bone matrix(DBM)by crushing,cleaning,and demineralization,and then the DBM was prepared into bone matrix gelatin(BMG)by warm bath method,and the BMG and DBM were mixed to prepare the experimental group’s plastic bone filler material;DBM was used as control group.Fifteen healthy male thymus-free nude mice aged 6-9 weeks were used to prepare intermuscular space between gluteus medius and gluteus maximus muscles,and all of them were implanted with experimental group materials.The animals were sacrificed at 1,4,and 6 weeks after operation,and the ectopic osteogenic effect was evaluated by HE staining.Eight 9-month-old Japanese large-ear rabbits were selected to prepare 6-mm-diameter defects at the condyles of both hind legs,and the left and right sides were filled with the materials of the experimental group and the control group respectively.The animals were sacrificed at 12 and 26 weeks after operation,and the effect of bone defect repair were evaluated by Micro-CT and HE staining.Results In ectopic osteogenesis experiment,HE staining showed that a large number of chondrocytes could be observed at 1 week after operation,and obvious newly formed cartilage tissue could be observed at 4 and 6 weeks after operation.For the rabbit condyle bone filling experiment,HE staining showed that at 12 weeks after operation,part of the materials were absorbed,and new cartilage could be observed in both experimental and control groups;at 26 weeks after operation,the most of the materials were absorbed,and large amount of new bone could be observed in the 2 groups,while new bone unit structure could be observed in the experimental group.Micro-CT observation showed that the bone formation rate and area of the experimental group were better than those of the control group.The measurement of bone morphometric parameters showed that the parameters at 26 weeks after operation in both groups were significantly higher than those at 12 weeks after operation(P<0.05).At 12 weeks after operation,the bone mineral density and bone volume fraction in the experimental group were significantly higher than those in the control group(P<0.05),and there was no significant difference between the two groups in trabecular thickness(P>0.05).At 26 weeks after operation,the bone mineral density of the experimental group was significantly higher than that of the control group(P<0.05).There was no significant difference in bone volume fraction and trabecular thickness between the two groups(P>0.05).Conclusion The new plastic bone filler material is an excellent bone filler material with good biosafety and osteoinductive activity.