PRP与纳米羟基磷灰石修复兔牙槽骨缺损的实验研究
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摘要
目的:本实验将富血小板血浆(Platelet-rich plasma,PRP)和纳米羟基磷狄石(nanometer-hydroxyapatite,nano-HA)单独或联合用于兔下颌牙槽骨缺损处,观察PRP、nano-HA及PRP/nano-HA复合物对兔牙槽骨组织缺损再生的促进作用;探讨PRP促进缺损牙槽骨组织再生的可能机制,为进一步的动物实验和临床应用提供理论依据。
材料与方法:40只健康家兔,分别在其左右下颌骨体部共建立80个高8mm,宽15mm,深3mm的牙槽骨缺损模型,将缺损区随机分为A、B、C、D四组,每组20个缺损区,A、B、C组分别用PRP、nano-HA、PRP/nano-HA复合物处理,D组设为空白对照组。分别于术后2周、4周、8周、12周处死动物,取材。大体观察动物术后状况:拍摄X片观察缺损区内新骨形成情况;制作常规组织切片,HE染色,进行组织学定性分析,并通过计算机图像分析系统采集切片图像,计算缺损区新骨形成面积占缺损面积的百分比进行统计学分析。
结果:①所有实验动物术后饮食、活动正常,无死亡及术区伤口感染,术后1周伤口一期愈合,伤口缝线自行脱落。②X片示牙槽骨缺损区在术后2周时可见,到术后4周时,缺损区已经有骨组织生长,术后8周时,缺损区基本消失,术后12周时,骨组织恢复良好,达到正常骨组织形态。③组织学分析显示,术后第2周、4周时,成骨活跃,可见大量的成骨细胞及越来越多的新生骨。A、C两实验组新生骨面积(%)与对照组比较有统计学差异(p＜0.05),B组新生骨面积与对照组比较无统计学差异(p＞0.05);术后8周时,成骨现象已很明显,C组与对照组的统计学差异依然存在(p＜0.05),A组与D组的统计学差异消失(p＞0.05),B组与对照组比较依然无统计学差异(p＞0.05);术后12周时,骨组织修复已完成,各实验组与对照组比较均不存在统计学差异(p＞0.05)。实验组之间新生骨面积(%)比较,A组与B组术后第2周时有统计学差异(p＜0.05),第4周、8周时统计学差异消失(p＞0.05);A组与C组术后第2周、4周时无统计学差异(p＞0.05),8周时C组骨形成较明显,与A组有统计学差异(p＜0.05);C组与B组术后第2、4、8周时,有统计学差异(P＜0.05);第12周时,各实验组新生骨面积百分比之间比较均不存在统计学差异(p＞0.05)。
结论:①PRP可以加速骨组织再生,对牙槽骨缺损的愈合有良好的促进作用;②PRP/nano-HA的复合材料具有骨传导和骨诱导能力,可以修复实验性牙槽骨缺损,促进牙槽骨再生;③本实验nano-HA单独用于牙槽骨缺损的填充,未表现出良好的成骨效应,还需要进一步的研究论证;④PRP成骨机理仍需进一步探讨。
Objectives: The aim of the present study was to assess the platelet-rich plasma(Platelet-rich plasma, PRP) and nano-hydroxyapatite (Nanometer-Hydroxyapatite, nano-HA)alone or combined on new bone formation in the mandibular bone defect of family rabbits model, explore the mechanism of PRP to promote bone regeneration, so as to provide a theoretical basis for further animal experiments and clinical applications.
Materials and Methods: 40 healthy family rabbits were were selected and a 15mm×8mm×3mm defect of the mandible was made bilaterally in every rabbit. The bilateral defects were randomly divided into A, B, C, D four groups. A, B, C group respectively with PRP, nano-HA, PRP/nano-HA composite graft, Dgroup as control group. The mandibular healing was evaluated by gross observation, X-ray film and histological dyeing at 2, 4, 8 and 12 weeks postoperatively. HE staining, histological analysis and computer image analysis system for collecting biopsy images to calculate the formation of new bone defect area accounts for the percentage of defect area to carry out a comparative analysis ofstatistics.
Results:①Gorss observation: All experimental animals were normal in diet and activity after surgical operation, and no animals died or wound infection. The wound healed and the suture was off on its own after 1 week.②X-ray evaluation: The alveolar bone defect area ccould be see after 2 weeks, by 4 weeks after operation, the defect area had the growth of bone tissue, but the defect disappeared 4 weeks later, and no significant difference between the groups, by 12 weeks, the bone tissue recovered well.③Histological and histomoprhometric analysis: More mesenchymal cells and osteoblasts, as well as a greater volume of new bone formation were observed in the graft sites that received PRP than the control at 2 and 4 weeks postoperatively. The new bone area(%)in group A and C was significant higher than the control group(p<0.05), but group B was no difference (P>0.05). Osteogenesis phenomenon was already evident after 8 weeks, group C and the control group still existed significant difference (p<0.05), significant difference was disappeared between group A and D (P>0.05) . Group B and control group still had no statistical differences(p>0.05).All the experimental groups and the normal control group did not exist difference at 12 weeks(p<0.05). The new bone area (%)comparison of experimental groups: Group A and group B were significant difference (P<0.05) after two weeks, but the significant difference disappeared (P>0.05)at 4 and 8 weeks. Group A compared with the group C was no significant difference (P>0.05) at 2, 4 weeks,but the difference appeared at 8 weeks(P<0.05). Group C and group B had significant difference (P<0.05)at 2, 4, 8 weeks, the percentage of new bone area of all experimental groups did not exist difference (P>0.05) at 12 weeks.
Conclusions:①PRP could speed up alveolar regeneration bone and promote it healing.②The PRP/nano-HA composite material with the ability of bone conduction and bone induction could repair experimental alveolar bone defect and promote bone regeneration.③Nano-HA alone for the filling of the alveolar bonehad showed no good osteogenic effect, so the further research and feasibility studies were necessary.④The osteogenic mechanism of PRP still needed to be further explored.
材料与方法:40只健康家兔,分别在其左右下颌骨体部共建立80个高8mm,宽15mm,深3mm的牙槽骨缺损模型,将缺损区随机分为A、B、C、D四组,每组20个缺损区,A、B、C组分别用PRP、nano-HA、PRP/nano-HA复合物处理,D组设为空白对照组。分别于术后2周、4周、8周、12周处死动物,取材。大体观察动物术后状况:拍摄X片观察缺损区内新骨形成情况;制作常规组织切片,HE染色,进行组织学定性分析,并通过计算机图像分析系统采集切片图像,计算缺损区新骨形成面积占缺损面积的百分比进行统计学分析。
结果:①所有实验动物术后饮食、活动正常,无死亡及术区伤口感染,术后1周伤口一期愈合,伤口缝线自行脱落。②X片示牙槽骨缺损区在术后2周时可见,到术后4周时,缺损区已经有骨组织生长,术后8周时,缺损区基本消失,术后12周时,骨组织恢复良好,达到正常骨组织形态。③组织学分析显示,术后第2周、4周时,成骨活跃,可见大量的成骨细胞及越来越多的新生骨。A、C两实验组新生骨面积(%)与对照组比较有统计学差异(p＜0.05),B组新生骨面积与对照组比较无统计学差异(p＞0.05);术后8周时,成骨现象已很明显,C组与对照组的统计学差异依然存在(p＜0.05),A组与D组的统计学差异消失(p＞0.05),B组与对照组比较依然无统计学差异(p＞0.05);术后12周时,骨组织修复已完成,各实验组与对照组比较均不存在统计学差异(p＞0.05)。实验组之间新生骨面积(%)比较,A组与B组术后第2周时有统计学差异(p＜0.05),第4周、8周时统计学差异消失(p＞0.05);A组与C组术后第2周、4周时无统计学差异(p＞0.05),8周时C组骨形成较明显,与A组有统计学差异(p＜0.05);C组与B组术后第2、4、8周时,有统计学差异(P＜0.05);第12周时,各实验组新生骨面积百分比之间比较均不存在统计学差异(p＞0.05)。
结论:①PRP可以加速骨组织再生,对牙槽骨缺损的愈合有良好的促进作用;②PRP/nano-HA的复合材料具有骨传导和骨诱导能力,可以修复实验性牙槽骨缺损,促进牙槽骨再生;③本实验nano-HA单独用于牙槽骨缺损的填充,未表现出良好的成骨效应,还需要进一步的研究论证;④PRP成骨机理仍需进一步探讨。
Objectives: The aim of the present study was to assess the platelet-rich plasma(Platelet-rich plasma, PRP) and nano-hydroxyapatite (Nanometer-Hydroxyapatite, nano-HA)alone or combined on new bone formation in the mandibular bone defect of family rabbits model, explore the mechanism of PRP to promote bone regeneration, so as to provide a theoretical basis for further animal experiments and clinical applications.
Materials and Methods: 40 healthy family rabbits were were selected and a 15mm×8mm×3mm defect of the mandible was made bilaterally in every rabbit. The bilateral defects were randomly divided into A, B, C, D four groups. A, B, C group respectively with PRP, nano-HA, PRP/nano-HA composite graft, Dgroup as control group. The mandibular healing was evaluated by gross observation, X-ray film and histological dyeing at 2, 4, 8 and 12 weeks postoperatively. HE staining, histological analysis and computer image analysis system for collecting biopsy images to calculate the formation of new bone defect area accounts for the percentage of defect area to carry out a comparative analysis ofstatistics.
Results:①Gorss observation: All experimental animals were normal in diet and activity after surgical operation, and no animals died or wound infection. The wound healed and the suture was off on its own after 1 week.②X-ray evaluation: The alveolar bone defect area ccould be see after 2 weeks, by 4 weeks after operation, the defect area had the growth of bone tissue, but the defect disappeared 4 weeks later, and no significant difference between the groups, by 12 weeks, the bone tissue recovered well.③Histological and histomoprhometric analysis: More mesenchymal cells and osteoblasts, as well as a greater volume of new bone formation were observed in the graft sites that received PRP than the control at 2 and 4 weeks postoperatively. The new bone area(%)in group A and C was significant higher than the control group(p<0.05), but group B was no difference (P>0.05). Osteogenesis phenomenon was already evident after 8 weeks, group C and the control group still existed significant difference (p<0.05), significant difference was disappeared between group A and D (P>0.05) . Group B and control group still had no statistical differences(p>0.05).All the experimental groups and the normal control group did not exist difference at 12 weeks(p<0.05). The new bone area (%)comparison of experimental groups: Group A and group B were significant difference (P<0.05) after two weeks, but the significant difference disappeared (P>0.05)at 4 and 8 weeks. Group A compared with the group C was no significant difference (P>0.05) at 2, 4 weeks,but the difference appeared at 8 weeks(P<0.05). Group C and group B had significant difference (P<0.05)at 2, 4, 8 weeks, the percentage of new bone area of all experimental groups did not exist difference (P>0.05) at 12 weeks.
Conclusions:①PRP could speed up alveolar regeneration bone and promote it healing.②The PRP/nano-HA composite material with the ability of bone conduction and bone induction could repair experimental alveolar bone defect and promote bone regeneration.③Nano-HA alone for the filling of the alveolar bonehad showed no good osteogenic effect, so the further research and feasibility studies were necessary.④The osteogenic mechanism of PRP still needed to be further explored.
引文
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