Calcium hydrogen phosphate dehydrate combined with gelatin and recombinant human bone morphologic protein 2/7 for repair of bone defects in rabbits

doi:10.3969/j.issn.2095-4344.2314

Abstract

Abstract:

BACKGROUND: Studies have shown that the osteogenic ability of 10% strontium-doped calcium hydrogen phosphate dihydrate is higher than that of calcium hydrogen phosphate dihydrate and 5% strontium-doped calcium hydrogen phosphate dihydrate, but also found that the pore structure of 10% strontium-doped calcium hydrogenphosphate dihydrate is not ideal, and the early osteogenic effect is not satisfactory.

OBJECTIVE: To investigate the osteogenic effect of the composite of 10% strontium-doped calcium hydrogenphosphate dehydrate and gelatin and recombinant human bone morphogenetic protein 2/7 (rhBMP2/7).

METHODS: Gelatin-10% strontium-doped calcium hydrogenphosphate dihydrate and gelatin-10% strontium-doped calcium hydrogenphosphate dihydrate material containing 0.04 g/L and 1 g/L rhBMP2/7 were prepared respectively. Forty-five rabbit models of bilateral mandibular defects were prepared and then divided into five groups. In the blank control group, no material was implanted. 10% strontium-doped calcium hydrogen phosphate dihydrate (control group) and gelatin-10% strontium-doped calcium hydrogen phosphate dihydrate (gelatin group), 0.04 g/L rhBMP2/7-gelatin-10% strontium-doped calcium hydrogenphosphate dihydrate (0.04 g/L rhBMP2/7 group), and 1 g/L rhBMP2/7-gelatin-10% strontium-doped calcium hydrogenphosphate dihydrate (1 g/L rhBMP2/7 group) were implanted in the remaining four groups, respectively. Bone defect specimens were taken at 4, 8 and 12 weeks after surgery, and were examined by cone beam CT and immunohistochemistry. This study was approved by the Animal Ethics Committee of North China University of Science and Technology, China.

RESULTS AND CONCLUSION: Cone beam CT examination revealed that at 8 weeks after surgery, bone repair was basically completed and the new bone tissue was almost fused with the surrounding tissue in the 1 g/L rhBMP2/7 group. Most of defect area was repaired, and the edge of new bone was unsmooth in the 0.04 g/L rhBMP2/7 and gelatin groups. Bone defect in the control group partially repaired. At 12 weeks after surgery, bone repair was completed in the gelatin, 0.04 rhBMP2/7 and 1 g/L rhBMP2/7 groups. Immunohistochemistry revealed that at 4 and 8 weeks after surgery, type I collagen expression in the 1 g/L rhBMP2/7 group was significantly higher than that in the other four groups (P < 0.05). Type I collagen expression in the 0.04 g/L rhBMP2/7 and gelatin groups was significantly higher than that in the control group (P < 0.05). At 12 weeks after surgery, there was no significant difference in type I collagen expression between collagen, gelatin, 0.04 rhBMP2/7 and 1 g/L rhBMP2/7 groups (P > 0.05). These results suggest that the addition of gelatin and 1 g/L rhBMP2/7 to 10% strontium-doped calcium hydrogenphosphate dihydrate can promote the repair of bone defects.

Key words: calcium hydrogen phosphate dihydrate, bone defect, gelatin, rhBMP2/7, type I collagen, strontium, osteogenesis, rabbit

CLC Number:

Li Xuewei, Hu Beibei, Zhang Dawei, Quan Lulu, Liang Yongqiang. Calcium hydrogen phosphate dehydrate combined with gelatin and recombinant human bone morphologic protein 2/7 for repair of bone defects in rabbits[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(28): 4533-4539.

Figures/Tables 6

2.2 骨缺损修复大体观察结果术后4周时，缺损区有一层纤维组织覆盖，探针探查空白组有界限清楚的凹陷状骨缺损；对照组材料与骨缺损界限清楚，有少量骨痂形成；明胶组材料与骨缺损界限模糊，有部分骨痂形成；0.04 g/L rhBMP2/7组材料与骨缺损界限模糊，骨痂形成较多；1 g/L rhBMP2/7组骨缺损区减小，表面有纤维骨痂覆盖。术后8周时，空白组较4周时缺损区缩小，边限模糊；对照组缺损区有部分纤维样的瘢痕组织且缺损区缩小；明胶组缺损区减小，表面有纤维骨痂形成；0.04 g/L rhBMP2/7组缺损区大部分由新生硬组织填充；1 g/L rhBMP2/7组凹陷区基本平整，完成修复。术后12周时，空白组缺损区减小，仍有部分凹陷样结构；其余4组缺损均已完成修复。 2.3 骨缺损修复锥形束CT表现将兔下颌骨颊面(缺损处)朝上摆放进行锥形束CT扫描锥形束CT是三维成像，但只有冠状面图像是清晰的，故选择冠状面进行观察，选择测量位置时结合矢状面和横断面，选择缺损区中心的冠状面进行观察，因缺损区中心区域是最后修复的，选取中心位置有助于判断骨修复的效果，避免出现假阳性结果。但有时锥形束CT由于各种原因扫描后的结果不够好，只能在尽量靠近缺损区中心的位置选取断层图片。术后4周时，空白组可见缺损区边界稍清楚的低密度影像；对照组可见比空白组缺损区密度稍高的界限较清晰的影像；明胶组缺损区边界模糊，0.04，1 g/L rhBMP2/7组与明胶组差别不明显，见图3。术后8周时，空白组低密度凹陷区缩小；对照组骨缺损区低密度影像减小，有散在高密度影像；明胶组缺损区有为高密度影像，边缘呈现不规则的低密度影像；0.04 g/L rhBMP2/7组和明胶组差别不明显，缺损区大部分修复，新骨修复边缘不平整；1 g/L rhBMP2/7组基本修复，皮质骨影像不连续，见图4。术后12周时，空白组缺损区仍有部分低密度影像；对照组可见骨皮质不连续影像；其余3组缺损区与正常骨组织影像密度一致，见图5。 "

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