珍珠层粉复合富血小板纤维蛋白对颅骨缺损模型兔的修复作用

doi:10.13481/j.1671-587X.20210512

摘要/Abstract

摘要： 目的

制备珍珠层粉（NP）和富血小板纤维蛋白（PRF）的复合材料，分析其修复颅骨缺损模型兔效果，阐明其可能的作用机制。

方法

选择12只新西兰兔建立兔颅骨缺损模型，颅骨顶部矢状缝周围3 cm×3 cm区域脱毛处理，全身麻醉后，于矢状缝两侧，造成4个直径 8 mm的全层颅骨缺损孔，并保留完整的硬脑膜。每只新西兰兔4个颅骨缺损孔按照植入物不同分为4组，分别为空白对照组、NP组、PRF组和复合组，于4、8和12周各处死4只新西兰兔，取颅骨缺损标本，HE染色观察各组新西兰兔颅骨缺损部位骨组织病理形态表现，Micro CT扫描检测各组新西兰兔骨矿物质密度（BMD）和新生骨体积/骨总体积百分数（BV/TV），免疫组织化学法检测各组新西兰兔颅骨缺损部位新骨组织中骨形态发生蛋白2（BMP-2）和血管内皮生长因子（VEGF）蛋白表达水平。

结果

4、8和12周观察，与空白对照组、NP组和PRF组比较，复合组新西兰兔大体标本成骨明显丰满。Micro CT扫描，与空白对照组、NP组和PRF组比较，4、8和12周时复合组新西兰兔颅骨缺损部位新骨形成面积更大，BMD和BV/TV明显升高（P<0.05），以第12周最为明显（P<0.05）。HE染色，4周时复合组新西兰兔颅骨缺损部位骨细胞、类骨质和新生血管较空白对照组、NP组和PRF组增多，8周时复合组新西兰兔颅骨缺损部位骨矿化现象及新生骨小梁较空白对照组、NP组和PRF组更加明显，12周时复合组新西兰兔颅骨缺损部位新生骨小梁增粗且成熟程度较空白对照组、NP组和PRF组高。免疫组织化学检测，4、8和12周时复合组新西兰兔颅骨缺损部位新骨组织中BMP-2和VEGF蛋白表达水平较空白对照组、NP组和RFP组均明显升高（P<0.05）。

结论

NP复合PRF制备的复合材料有明显的促进骨组织生长能力，并且可能通过诱导BMP-2和VEGF的增加，促进骨组织的再生。

关键词: 珍珠层粉, 富血小板纤维蛋白, 颅骨缺损, 骨诱导

Abstract: Objective

To prepare the composite material of nacre powder （NP） and platelet-rich fibrin （PRF） and analyze its repair effect in the cranial defect model rabbits， and to clarify its possible mechanism.

Methods

Twelve New Zealand rabbits were selected to set up the rabbit cranial defect models， the 3 cm×3 cm area around the sagittal suture at the top of the cranial was depilated. After general anesthesia， four full-thickness cranial defects with a diameter of 8 mm were created on both sides of the sagittal suture but the intact dura mater was retained. Four cranial defects of each New Zealand rabbit were divided into four groups according to different implants：blank control group（no implant material），NP group （only NP）， PRF group （only PRF），and mixed group （NP+PRF）. Four New Zealand rabbits were sacrificed at 4，8 and 12 weeks respectively， and the cranial defect specimens were collected.Micro CT scan was adopt to detect the bone mineral density（BDM） and the percentages of new bone volome/total bone volume（BV/TV）.The pathomorphology of the bone in the cranial defect areas in various groups were observed with HE staining. The expression levels of bone morphogenetic protein-2 （BMP-2） and vascular endothelial growth factor （VEGF） in the new bones of the New zealand rabbits in various groups were analysed by immunohistochemistry.

Results

At 4， 8 and 12 weeks，compared with blank control group，NP group and PRF group， the bone formation of the gross specimens in mixed group was significantly plump. The results of Micro CT scan showed that compared with blank control group，NP group and PRF group， at 4， 8 and 12 weeks，a larger area of new bone was formed in mixed group， and BMD and BV/TV were significantly increased（P<0.05），especially at 12 weeks （P<0.05）.The results of HE staining showed that the bone cells， osteoid， and new blood vessels of the New Zealand rabbits in mixed group at 4 weeks were higher than those in blank control group，NP group and PRF group； at 8 weeks，bone mineralization and neonatal bone trabecula of the New Zealand rabbits in mixed group were more obvious than those in blank control group，NP group and PRF group； at 12 weeks， the neonatal bone trabecula of the rabbits in mixed group were thicker and the degree of maturity was higher than those in blank control group，NP group and PRF group. The immunohistochemical results showed that the expression levels of BMP-2 and VEGF proteins in the new bone tissue in defect areas of the New Zealand rabbits in mixed group were significantly higher than those in blank control group，NP group and PRF group （P<0.05）.

Conclusion

NP combined with PRF has obvious ability to promote the bone tissue growth and it can promote the bone regeneration by inducing the increase of BMP-2 and VEGF.

Key words: nacre powder, platelet-rich fibrin, cranial defect, osteoinduction

中图分类号:

R782

黄莹,马福娟,MAGED ALI Al-Aroomi,王触灵,谢富强. 珍珠层粉复合富血小板纤维蛋白对颅骨缺损模型兔的修复作用[J]. 吉林大学学报(医学版), 2021, 47(5): 1162-1170.

Ying HUANG,Fujuan MA,ALI Al-Aroomi MAGED,Chuling WANG,Fuqiang XIE. Repair effect of nacre powder combined with platelet-rich fibrin on cranial defect model rabbits[J]. Journal of Jilin University(Medicine Edition), 2021, 47(5): 1162-1170.

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Group	BMD
Group	（week） 4	8	12
Blank control	642.39±4.75	777.03±13.23	867.51±52.02
NP	659.85±34.69	787.34±17.70	929.94±39.54
PRF	635.05±36.71	777.65±11.01	891.43±52.39
Mixed	731.64±23.09^*△#	833.57±17.68^*△#	1 051.65±42.46^*△#

Group	BV/TV
Group	（week）4	8	12
Blank control	4.87±3.88	11.22±2.09	22.25±4.49
NP	9.44±7.66	16.79±4.36	30.09±3.83
PRF	5.33±4.47	14.68±2.55	25.03±4.14
Mixed	24.02±4.58^*△#	29.52±11.96^*△#	43.09±6.99^*△#

Group	BMP-2
Group	（week）4	8	12
Blank control	0.25±0.02	0.26±0.02	0.26±0.01
NP	0.28±0.03	0.31±0.02	0.38±0.03
PRF	0.27±0.02	0.31±0.02	0.33±0.02
Mixed	0.35±0.02^*	0.36±0.02^*	0.40±0.03^*

Group	VEGF
Group	（week）4	8	12
Blank control	0.26±0.01	0.26±0.01	0.26±0.02
NP	0.30±0.01	0.34±0.02	0.38±0.03
PRF	0.30±0.03	0.32±0.01	0.36±0.03
Mixed	0.35±0.02^*	0.38±0.02^*	0.42±0.04^*

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