中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (30): 4804-4810.doi: 10.3969/j.issn.2095-4344.1419

• 组织工程口腔材料 tissue-engineered oral materials • 上一篇    下一篇

羟基磷灰石结合β-磷酸三钙及海藻酸盐作为牙槽骨修复材料的比较分析

程  扬1，刘  敏1，朱忠焰2，高莎莎3
  

  1. 1西南医科大学附属口腔医院，四川省泸州市  646000；2自贡市第一人民医院口腔科，四川省自贡市  643000；3德阳市口腔医院，四川省德阳市  618000
  • 收稿日期:2019-05-12 出版日期:2019-10-28 发布日期:2019-10-28
  • 通讯作者: 刘敏，主任医师，西南医科大学附属口腔医院，四川省泸州市 646000
  • 作者简介:程扬，女，1985年生，四川省德阳市人，西南医科大学口腔医学院在读硕士，主治医师，主要从事口腔种植修复研究。
  • 基金资助:

    四川省科技厅-泸州市科技局-泸州医学院联合项目(LY-51)，项目负责人：刘敏

Comparative analysis of hydroxyapatite, β-tricalcium phosphate, and carbonated hydroxyapatite alginate dental pulp replacement materials

Cheng Yang1, Liu Min1, Zhu Zhongyan2, Gao Shasha3
  

  1. 1Stomatological Hospital Affiliated to Southwest Medical University, Luzhou 646000, Sichuan Province, China; 2Stomatological Department, Zigong First People's Hospital, Zigong 643000, Sichuan Province, China; 3Deyang Stomatological Hospital, Deyang 618000, Sichuan Province, China
  • Received:2019-05-12 Online:2019-10-28 Published:2019-10-28
  • Contact: Liu Min, Chief physician, Stomatological Hospital Affiliated to Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • About author:Cheng Yang, Master candidate, Attending physician, Stomatological Hospital Affiliated to Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Supported by:

    Sichuan Provincial Department of Science and Technology-Luzhou Bureau of Science and Technology-Luzhou Medical College Joint Project, No. LY-51 (to LM)

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摘要：

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文题释义：
组织工程学：这是一门综合性学科，将细胞生物学和材料科学两者相互融合，在细胞体外或生物体内进行组织构建的一门前沿学科，组成3要素即种子细胞、细胞因子和支架材料，将3者有力的结合并应用于组织缺损再生、器官功能衰竭修复等临床治疗。
牙槽骨修复材料：按其来源主要分为5类，分别是同种异体骨，即相同部位或种类的骨，但来源于其他个体的非自身的人体骨；异种骨，即取自牛，猪等非人体骨，经过脱蛋白等特殊加工工艺处理，去除抗原性，保留无机骨成分的骨替代材料；自体异种骨，即取自人体自身不同部位的骨；人工合成材料，即常见的生物陶瓷和高分子材料；组织工程骨，即将细胞、合成材料、处理过的天然材料和组织、细胞因子及基因治疗广泛地应用于体内的组织再生或体外的组织构建。
 
 
背景：牙槽骨修复材料的种类很多，可以分为天然生物材料，人工合成材料和复合材料，各种类型的材料还可以通过化学生物等方法进行合成，其均有各自的优势和不足。
目的：比较羟基磷灰石/β-磷酸三钙、纳米碳化羟基磷灰石海藻酸盐及单纯羟基磷灰石3种牙槽骨修复材料的细胞毒性及生物相容性。
方法：将羟基磷灰石/β-磷酸三钙浸提液、纳米碳化羟基磷灰石海藻酸盐浸提液、羟基磷灰石浸提液分别与小鼠成骨前细胞、人成骨细胞共培养，XTT实验检测细胞线粒体活性(以单独培养的细胞为对照组)，结晶紫分析实验检测细胞毒性(以DMSO培养的细胞为对照组)。取20只Wistar大鼠(购自北京维通利华实验动物有限公司)，制备上颌右侧中切牙牙槽骨缺损模型，随机分4组干预：对照组不植入任何材料，其余3组分别植入羟基磷灰石/β-磷酸三钙、纳米碳化羟基磷灰石海藻酸盐及单纯羟基磷灰石材料，植入后7，21，42 d，检测血清中RANKL、骨保护素质量浓度。实验已通过西南医科大学动物伦理委员会审批批准，审批号：IACUC20170315-07。
结果与结论：①3组材料浸提液中小鼠成骨前细胞的线粒体活性与对照组比较差异无显著性意义(P > 0.05)，3组材料浸提液中人成骨细胞的线粒体活性与对照组比较差异均无显著性意义(P > 0.05)；3组材料浸提液中，纳米碳化羟基磷灰石海藻酸盐浸提液中两种细胞的线粒体活性最高；②3组材料浸提液对小鼠成骨前细胞与人成骨细胞的毒性均明显低于对照组(P < 0.05)；3组材料浸提液中，纳米碳化羟基磷灰石海藻酸盐浸提液对两种细胞的毒性最低；③纳米碳化羟基磷灰石海藻酸盐材料植入后7 d的RANKL质量浓度低于21 d(P < 0.001)，植入后42 d的骨保护素质量浓度高于植入后7，21 d(P < 0.001)；④结果表明相对于羟基磷灰石/β-磷酸三钙与单纯羟基磷灰石材料，纳米碳化羟基磷灰石海藻酸盐材料具有更好的生物相容性。

关键词: 羟基磷灰石, β-磷酸三钙, 海藻酸盐, 组织工程, 颌骨缺陷, 骨再生, 生物支架材料, 生物相容性

Abstract:

BACKGROUND: There are many kinds of dental pulp replacement materials, which can be divided into natural biological materials, artificial synthetic materials, and composite materials. Various types of materials can also be synthesized by chemical and biological methods, all of which have their own advantages and disadvantages.
OBJECTIVE: To compare cytotoxicity and biological compatibility of three kinds of alveolar bone repair materials including hydroxyapatite/β-tricalcium phosphate, carbonated hydroxyapatite alginate, and pure hydroxyapatite.
METHODS: Mouse pre-osteoblasts and human osteoblasts were cultured together with the leaching liquor of three biomaterials respectively. Cell mitochondrial activity was detected by XTT assay (cells cultured separately were used as the control group), and cytotoxicity was detected by crystal violet assay (cells cultured in DMSO were used as the control group).Twenty Wistar rats purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd were used to prepare the alveolar bone defect model of the right maxillary central incisor. Then the alveolar bone defect models were randomly divided into four groups. In the control group, no material was implanted into the bone defect region. In the other three groups, hydroxyapatite/β-tricalcium phosphate, carbonated hydroxyapatite alginate, and pure hydroxyapatite were implanted into the bone defect region. At 7, 21 and 42 days after implantation, serum concentrations of receptor activator for nuclear factor-κB ligand and osteoprotegerin were measured. This study was approved by Animal Ethics Committee, Southwest Medical University, China (approval No. IACUC20170315-07).
RESULTS AND CONCLUSION: There was no significant difference in the mitochondrial activity of the mouse preosteoblasts and human osteoblasts between three material groups and the control group (P > 0.05). The mitochondrial activity of mouse preosteoblasts and human osteoblasts was higher in the hydroxyapatite/β-tricalcium phosphate and carbonated hydroxyapatite alginate groups than in the pure hydroxyapatite group (P < 0.05). The cytotoxicity of the mouse pre-osteoblasts and human osteoblasts in the three material groups was lower than that in the control group (P < 0.05). Among three material groups, the leaching liquor of carbonated hydroxyapatite alginate had the lowest toxicity to the two kinds of cells. Serum concentration of receptor activator for nuclear factor-κB ligand at 7 days after implantation of carbonated hydroxyapatite alginate was significantly lower than that at 21 days after biomaterial implantation, and serum concentration of osteoprotegerin at 42 days after biomaterial implantation was significantly higher than that at 7 and 21 days (P < 0.001). These results suggest that carbonated hydroxyapatite alginate exhibits better biocompatibility than hydroxyapatite/β-tricalcium phosphate and pure hydroxyapatite.

Key words: hydroxyapatite, β-tricalcium phosphate, carbonated hydroxyapatite alginate, tissue engineering, Jaw defects, bone regeneration, biological scaffold material, biocompatibility

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