口腔医学研究

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (6): 539-543.DOI: 10.13701/j.cnki.kqyxyj.2020.06.009

• 口腔生物学研究 • 上一篇    下一篇

人血小板裂解液对牙髓间充质细胞成骨分化的影响

霍冬梅1, 徐燕1*, 胡韶光1, 汪芹芹1, 王荣海2   

  1. 1.安徽医科大学口腔医学院,安徽医科大学附属口腔医院,安徽省口腔疾病研究中心实验室 安徽 合肥 230032;
    2.安徽安科生物工程(集团)股份有限公司 安徽 合肥 230032;
    3.安徽医科大学人兽共患病安徽省重点实验室 安徽 合肥 230032
  • 收稿日期:2019-10-11 出版日期:2020-07-03 发布日期:2020-07-06
  • 通讯作者: *徐燕,E-mail:173236344@qq.com
  • 作者简介:霍冬梅(1992~ ),女,安徽六安人,硕士在读,研究方向:牙周组织再生。
  • 基金资助:
    2017年度安徽高校自然科学研究项目(编号:KJ2017ZD17)

Effect of Human Platelet Lysate on Osteogenic Differentiation of Dental Pulp Mesenchymal Cells

HUO Dongmei1, XU Yan1*, HU Shaoguang1, WANG Qingqing1, WANG Ronghai2   

  1. 1. Department of Periodontology, Stomatologic Hospital and College, Anhui Medical University, Key Lab of Oral Diseases Research of Anhui Province, Hefei 230032, China;
    2. Anhui Anke Biotechnology (Group) Co. Ltd, Hefei 230032, China;
    3. Key Lab of Zoonoses of Anhui Province, Anhui Medical University, Hefei 230032, China
  • Received:2019-10-11 Online:2020-07-03 Published:2020-07-06

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摘要： 目的: 研究人血小板裂解液(HPL)对人牙髓间充质细胞(hDPSCs)体外及体内成骨能力的影响。方法: 用5%HPL和10%胎牛血清(FBS)对牙髓组织进行培养;茜素红及ALP染色检测细胞体外成骨能力;在10只雄性新西兰大白兔颅骨上造4个8 mm直径的圆形骨缺损,分为四组:A组10%FBS的DPSC接种于明胶海绵(GS)组,B组5%HPL的DPSCs接种于GS组,C组GS组,D组空白组。术后4、8周取标本行Micro-CT三维重建及HE染色观察成骨效果。结果: 茜素红和ALP染色结果显示含5%HPL培养液显著促进hPDSCs成骨分化中的钙结节的形成和ALP的表达(P<0.0001);Micro-CT显示在4、8周B组新骨形成较A组多,C、D均成骨效果不明显;组织学观察发现在4、8周A、B组均有新骨形成,C、D组新骨区域不明显。结论: 5%HPL可以代替10%FBS进行体外培养hDPSCs,并且可以提高hDPSCs的体外及体内成骨能力。

关键词: 血小板, 裂解液, 牙髓组织, 成骨分化

Abstract: Objective: To study the effect of human platelet lysate (HPL) on the in vitro and in vivo osteogenesis of human dental pulp-derived mesenchymal cells (hDPSCs). Methods: The pulp tissues were cultured with 5% HPL and 10% fetal bovine serum (FBS), and the in vitro osteogenic ability of the cells was detected by the alizarin red and ALP staining. Four 8-mm-diameter round bone defects were prepared on the skull of 10 male Japanese white rabbits, and the rabbits were divided into four groups. In group A, the DPSC with 10% FBS was inoculated in the gelatin sponge (GS). In group B, the DPSCs with 5% HPL were inoculated in the GS. Group C was only GS, and group D was blank control. Micro-CT three-dimensional reconstruction and HE staining were used to observe the osteogenic effect 4 and 8 weeks after operation. Results: The results of alizalin red and ALP staining showed that 5%HPL medium significantly promoted the formation of calcium nodules and the expression of ALP in osteogenic differentiation of hPDSCs (P<0.000 1). The imaging results showed that there was a small amount of new bone formation in group A and B at 4 weeks and a large number of new bones at 8 weeks. The osteogenic effect of group C and D was not obvious, and the bone ability of group B was stronger than that of group A. Histologic observation showed that new bone formation was found in group A and B at 4 and 8 weeks, but there was no obvious new bone area in group C and D at 4 and 8 weeks. Conclusion: 5% HPL can be used for the in vitro culture of hDPSCs, and the in vitro and in vivo osteogenic ability of hDPSCs can be improved.

Key words: platelets, lysates, pulp tissue, osteogenic differentiation