掺钴羟基磷灰石的制备及体外成骨诱导性能研究.pdf

DOI：10.13876/J.cnki.ydnse.240004第 43 卷 第 1 期2024 年 3 月延安大学学报（自然科学版）Journal of Yanan University（Natural Science Edition)Vol.43 No.1Mar.2024掺钴羟基磷灰石的制备及体外成骨诱导性能研究张子悦，魏晓博，闫凛，朱苏迪，王成，樊晓霞*（延安大学 医学院，陕西 延安 716000）摘要：为优化羟基磷灰石（HAp）在治疗骨缺损时的诱导成骨能力，采用化学沉淀法制备一系列不同Co/Ca摩尔比的掺钴羟基磷灰石（Co-HAp），表征其形貌和结构，并研究其体外成骨诱导能力。结果表明，合成的Co-HAp具有典型的磷灰石结构。Co/Ca摩尔比为0.1%时，Co-HAp的结晶度最低。当Co/Ca摩尔比为0.1%，浸提液浓度为12.5 g/mL时，Co-HAp促BMSCs增殖能力最佳，具有良好的细胞相容性。该浓度下的Co-HAp可使BMSCs的ALP活性增强，钙结节增多，表现出较好的体外成骨诱导能力。该研究可为骨缺损修复领域探索更丰富的修复材料提供理论依据。关键词：掺钴羟基磷灰石；羟基磷灰石；化学沉淀法；细胞增殖；成骨性能中图分类号：R318.08 文献标识码：A 文章编号：1004-602X（2024）01-0045-06天然骨组织具有较好的自我修复能力，但在临界尺寸（长径比0.05）。培养 2 d 后，与对照组相比，0.05Co-HAp和0.01Co-HAp促BMSCs增殖能力较好。培养3 d后，掺杂不同Co2+比例的Co-HAp和HAp的OD值均高于对照组，其中0.1Co-HAp的OD值最高，说明0.1Co-HAp促BMSCs增殖能力最佳。这证实了Co2+的掺入对 BMSCs的增殖有促进作用，且 0.1Co-HAp的促增殖能力最佳。XRD结果表明0.1Co-HAp的结晶度和结晶尺寸在所有Co-HAp样品中最低，Co2+取代Ca2+最完全，这可能是其促BMSCs增殖能力最佳的一个因素20。图4B是BMSCs在不同浓度的0.1Co-HAp浸提液中培养3 d中的CCK-8测定结果。可知培养2 d和3 d后，25 g/mL的0.1Co-HAp浸提液的促BMSCs促增殖能力均高于对照组；培养3 d后，12.5 g/mL的0.1Co-HAp浸提液的促BMSCs增殖能力最强，因此被选为下一阶段的实验浓度，用于测试和比较HAp和0.1Co-HAp的成骨诱导性能。BMSCs在不同样品浸提液中培养3 d的电子显微镜图如图 5 所示。可知 0.1Co-HAp 与 HAp 样品浸提液中的细胞形态与对照组无明显差异，大多数为梭形和多边形，说明0.1Co-HAp和HAp对BMSCs图3不同Co-HAp样品的SEM图图4BMSCs在不同Co-HAp样品浸提液和不同浓度的0.1Co-HAp浸提液中的CCK-8测定结果48第 1 期张子悦 等：掺钴羟基磷灰石的制备及体外成骨诱导性能研究形态几乎无影响。0.1Co-HAp 浸提液中的细胞生长的密度更大，说明掺Co2+的HAp有较好的促细胞增殖能力。因此，制备的0.1Co-HAp有良好的细胞黏附作用，可促进细胞增殖，并使细胞保持良好形态。2.3成骨诱导性图 6为 BMSCs 在不同培养基中培养 14 d后的茜素红染色结果。由图可知，control组未出现红色钙结节，说明 BMSCs 还未成骨分化。induce 组、induce HAp组和induce Co-HAp组均出现了不同数量的红色结节，钙结节数量多少顺序为induce Co-HApinduce HApinduce，这 说 明 HAp 能 够 诱 导BMSCs成骨分化，而Co2+的掺杂增强了HAp的成骨诱导性能。图 7 为 BMSCs 在不同培养基中的 ALP活性结果，由图 7 可知，ALP 活性按大小排序为induce Co-HAp组induce HAp组induce组control组，即 Co-HAp 的 ALP 活性高于其他各组，此结果同样表明 Co-HAp 比 HAp 具有更强的促 BMSCs 成骨分化能力，证实Co2+的掺杂促进了BMSCs的成骨分化，表现出更优异的成骨诱导性能。茜素红染色实验和ALP活性实验均表明HAp能够促进BMSCs成骨分化，Co2+的掺杂增强了HAp促BMSCs的成骨分化性能，即Co-HAP有望在骨组织缺损修复领域发挥优势。3结论人工生物活性骨修复材料能够促进成骨细胞的增殖和分化，是骨组织缺损修复广泛应用的一类替代材料。本文采用化学沉淀法制备了不同掺钴量的Co-HAp，其具有类似HAp的结构特征，Co/Ca摩尔比为0.1%的Co-HAp的结晶度和结晶尺寸在所有Co-HAp样品中最低。HAp和不同Co2+掺杂比例的Co-HAp样品均能促进BMSCs增殖，而Co2+的掺杂提高了HAp对BMSCs的成骨诱导能力。因此，化学沉淀法合成的Co-HAp有望成为骨替代活性生物材料。参考文献：1 TANG D，TARE R S，YANG L Y，et al.Biofabrication of bone tissue：Approaches，challenges and translation for bone regeneration J.Biomaterials，2016，83：363-382.2 BIEDRZYCKA A，SKWAREK E，HANNA U M.Hydroxyapatite with magnetic core：Synthesis methods，properties，adsorption and medical applications J .Advances in Colloid and Interface Science，2021，291：102401.3 PATHAK D K，PANDEY P M.Evaluation of in vitro corrosion behavior of zinc-hydroxyapatite and zinc-hydroxyapatite-iron 图5BMSCs在不同样品浸提液中的电子显微镜图图6BMSCs在不同培养基中的茜素红染色结果图7BMSCs在不同培养基中的ALP活性(*P0.001，*P0.000 1)49延安大学学报（自然科学版）第 43 卷 as biodegradable composites J.Journal of Biomedical Materials 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716000，China）Abstract：The synthesis process of energetic materials is complex and dangerous.To explore the energetic materials with better safety performance，the crystalline energetic compound Sm4（Hdtztp）2O（OH）4（H2O）7 DMF 2H2O was prepared using 2，5-bis（tetrazolium）terephthalic acid（H4dtztp）as precursor ligand by reaction with samarium nitrate by solvothermic method.It was characterized by elemental analysis，infrared spectroscopy and X-ray powder diffraction，and its thermal decomposition and non-isothermal kinetics were studied.The results showed that the compound belonged to the rhombic space group Pmmn with three-dimensional structure.Meanwhile，the compound had good thermal stability.The thermal ignition point temperature and critical temperature of the compound were 250 and 466.08 respectively，which showed good thermal safety.In addition，the Gibbs free energy of the compound was 154.67 kJ mol-1，indicating that the decomposition process of the compound was non-spontaneous.Key words：Bistetrazole；energetic materials；thermal decomposition；thermal stability；thermal safety（上接第50页）Preparation of cobalt-doped hydroxyapatite and its osteogenic induction properties in vitroZHANG Ziyue，WEI Xiaobo，YAN Lin，ZHU Sudi，WANG Cheng，FAN Xiaoxia*（Medical school of Yan an University，Yan an 716000，China）Abstract：To optimize the bone-inducing ability of hydroxyapatite（HAp）in the treatment of bone defects，a series of cobalt-doped hydroxyapatite（Co-HAp）with different Co/Ca molar ratios were prepared by chemical precipitation method，its morphology and structure were characterized，and its osteogenic induction ability was studied in vitro.The results showed that the synthesized Co-HAp had a typical apatite structure.When the molar ratio of Co/Ca was 0.1%，the crystallinity of Co-HAp was the lowest.When the molar ratio of Co/Ca was 0.1%and the concentration of the extract was 12.5 g/mL，Co-HAp had the best proliferation ability and good cytocompatibility.At this concentration，Co-HAp could enhance the ALP activity of BMSCs and increase the number of calcium nodules，and show better osteogenic induction ability in vitro.This study can provide theoretical basis for exploring more abundant repair materials in the field of bone defect repair.Key words：Cobalt doped hydroxyapatite；Hydroxyapatite；chemical precipitation method；cell proliferation；Osteogenic property56