中药外敷超微粉体对大鼠骨折后骨痂形成和改建的影响

Effect of traditional Chinese drug ultramicro-powder for external application on the formation and reconstruction of bony callus after fracture in rats

目的:观察中药外敷超微粉体对大鼠骨折后骨痂形成和改建的影响。方法:将60只3月龄SD雌性大鼠制成左侧胫骨骨折模型,并以克氏针固定。造模结束后将大鼠随机分为超微组、散剂组和对照组,每组20只。自术后第1天开始,超微组和散剂组大鼠分别采用中药外敷超微粉体和中药外敷散剂外敷骨折部位,每天2次,共28 d,对照组不进行干预。分别于术后7 d、14 d、21 d和28 d每组各处死5只大鼠,取左侧胫骨分别进行X线、组织学及Micro-CT检查。结果:1X线检查结果。超微组和散剂组术后28 d时骨折处外骨痂密度增加,骨折线模糊,均接近愈合,其中超微组大鼠骨折处外骨痂更致密;对照组术后28 d时仍可看到骨折线。2组织学观察结果。术后7 d骨折处主要为纤维骨痂,3组无明显差别;术后14 d和21 d时,软骨骨痂开始取代纤维性骨痂;术后28 d时软骨骨痂周围不断有原始骨小梁出现,骨折断端表面出现破骨细胞,原始骨小梁不断改建成为成熟骨小梁,其中以超微组骨小梁最多。3Micro-CT检查结果。术后7 d时Micro-CT检查,3组均未见到明显骨痂。术后14 d时3组大鼠骨折部位骨痂体积和骨痂密度组间比较,差异均有统计学意义[(20.2±2.3)mm3,(16.4±1.7)mm3,(14.4±1.3)mm3,F=13.286,P=0.001;(184.6±9.3)mg·m-3,(162.8±5.7)mg·m-3,(148.2±6.8)mg·m-3,F=30.649,P=0.001];超微组的骨痂体积和密度均高于散剂组和对照组(P=0.000,P=0.000;P=0.000,P=0.000),散剂组骨痂体积和密度均高于对照组(P=0.001,P=0.001)。术后21 d时3组大鼠骨折部位骨痂体积和骨痂密度组间比较,差异均有统计学意义[(25.6±3.6)mm3,(21.2±0.8)mm3,(17.6±1.8)mm3,F=41.517,P=0.000;(248.2±16.0)mg·m-3,(222.6±7.8)mg·m-3,(197.8±9.0)mg·m-3,F=23.834,P=0.000];超微组的骨痂体积和密度均高于散剂组和对照组(P=0.000,P=0.000;P=0.000,P=0.000),散剂组骨痂体积和密度均高于对照组(P=0.000,P=0.000)。术后28 d时3组大鼠骨折部位骨痂体积和骨痂密度组间比较,差异均有统计学意义[(30.2±0.8)mm3,(23.4±1.1)mm3,(21.4±1.1)mm3,F=96.727,P=0.000;(292.2±10.9)mg·m-3,(256.8±6.5)mg·m-3,(222.8±4.4)mg·m-3,F=98.826,P=0.000];超微组的骨痂体积和密度均高于散剂组和对照组(P=0.000,P=0.000;P=0.000,P=0.000),散剂组骨痂体积和密度均高于对照组(P=0.000,P=0.000)。结论:中药外敷超微粉体能够促进大鼠骨折后软骨骨痂向骨性骨痂转化,提高骨痂密度,促进骨折愈合,其效果优于中药外敷散剂。

Objective：To observe the effect of traditional Chinese drug（TCD）ultramicro-powder for external application on the forma-tion and reconstruction of bony callus after fracture in rats. Methods：Sixty 3-month-old female SD rats were made into the left tibia fracture models and the fractures were fixed with kirschner wires. Then the rats were randomly divided into ultramicro-powder group,pulvers group and control group,20 cases in each group. The rats in ultramicro-powder group and pulvers group were treated with TCD ultramicro-powder for external application and pulvers for external application at fracture sites respectively on the first day after operation,twice a day for 28 consecutive days,while rats in the control group were not be treated. At 7,14,21 and 28 days after the operation,5 rats were selected from each group and were executed,and their left tibiaes were fetched out for X-ray examination,histology examination and micro-CT examina-tion. Results：At the 28th day after operation,the density of external callus increased at fracture site of ultramicro-powder group and pulvers group,moreover,the fracture lines were fuzzy and the fracture approached clinic healing. The density of external callus of ultramicro-powder group was more dense,while the fracture line can still be seen in control group at the 28th day after operation. At the 7th day after opera-tion,the fiber calluses were found at fracture sites and there was no obvious difference between the 3 groups. At the 14th and 21th day after operation,the fiber calluses began to be replaced by cartilage callus. At the 28th day after operation,original bone trabeculas appeared con-stantly around the cartilage callus,and the osteoclasts were also found on fracture sites. The original bone trabeculas converted to mature bone trabeculas constantly,and the bone trabeculas were the most in ultramicro-powder group. The micro-CT examination showed that no visible bony callus were found in the 3 groups at the 7th day after operation. There were statistical difference in volume and density of bony callus at fracture sites between the 3 groups at the 14th day after operation（20. 2 ＋/﹣ 2. 3,16. 4 ＋/﹣ 1. 7,14. 4 ＋/﹣ 1. 3 mm（3）,F =13. 286,p = 0. 001;184. 6 ＋/﹣ 9. 3,162. 8 ＋/﹣ 5. 7,148. 2 ＋/﹣ 6. 8 mg/ m（3）,F = 30. 649,p = 0. 001）. The volume and density of bony callus were higher in ultramicro-powder group compared with pulvers group and control group（p = 0. 000,p = 0. 000;p = 0. 000,p =0. 000）,and were higher in pulvers group compared with control group（p = 0. 001,p = 0. 001）. There were statistical difference in volume and density of bony callus at fracture sites between the 3 groups at the 21th day after operation（25. 6 ＋/﹣ 3. 6,21. 2 ＋/﹣ 0. 8,17. 6 ＋/﹣ 1. 8 mm（3）,F = 41. 517,p = 0. 000;248. 2 ＋/﹣ 16. 0,222. 6 ＋/﹣ 7. 8,197. 8 ＋/﹣ 9. 0 mg/ m（3）,F = 23. 834,p = 0. 000）. The volume and density of bony callus were higher in ultramicro-powder group compared with pulvers group and control group（p = 0. 000,p = 0. 000;p =0. 000,p = 0. 000）,and were higher in pulvers group compared with control group（p = 0. 000,p = 0. 000）. There were statistical difference in volume and density of bony callus at fracture sites between the 3 groups at the 28th day after operation（30. 2 ＋/﹣ 0. 8,23. 4 ＋/﹣ 1. 1, 21. 4 ＋/﹣ 1. 1 mm（3）,F = 96. 727,p = 0. 000;292. 2 ＋/﹣ 10. 9,256. 8 ＋/﹣ 6. 5,222. 8 ＋/﹣ 4. 4 mg/ m（3）,F = 98. 826,p = 0. 000）. The volume and density of bony callus were higher in ultramicro-powder group compared with pulvers group and control group（p = 0. 000,p =0. 000;p = 0. 000,p = 0. 000）,and were higher in pulvers group compared with control group（p = 0. 000,p = 0. 000）. Conclusion：The TCD ultramicro ﹣ powder for external application can promote the transformation of cartilage callus into osseous callus in rats after fracture and raise the bony callus density. Therefore,it can promote fracture healing. Moreover,its therapeutic effect is better than that of TCD pul-vers for external application.