Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (34): 5537-5543.doi: 10.12307/2021.252
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Liu Rixu, Lü Wenbo, Gao Wenshan
Received:
2020-07-23
Revised:
2020-07-25
Accepted:
2020-08-19
Online:
2021-12-08
Published:
2021-07-28
Contact:
Gao Wenshan, Chief physician, Professor, Master’s supervisor, First Department of Orthopedics, Affiliated Hospital of Hebei University, Baoding 071000, Hebei Province, China
About author:
Liu Rixu, Master candidate, First Department of Orthopedics, Affiliated Hospital of Hebei University, Baoding 071000, Hebei Province, China
CLC Number:
Liu Rixu, Lü Wenbo, Gao Wenshan. Research and clinical application of calcium phosphate cement modification[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(34): 5537-5543.
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2.1 磷酸钙骨水泥研究现状 磷酸钙骨水泥通常是将固相和液相按一定比例混合后先得到一种具备可塑性、注射性的糊状物,然后通过结晶反应最终形成羟基磷灰石或磷酸钙而固化[6]。磷酸钙骨水泥包括自固化型和非自固化型,非自固化型的特点为植入到体内不发生固化反应,双相磷酸钙骨水泥为其典型代表。将自固化型磷酸钙骨水泥移植到体内后可在体液环境下自固化,最终水化形成羟基磷灰石。除可以通过改变固相中的成分或加工工序来改善其表征,如GU等[7]和ROOZBAHANI等[8]分别报告的添加不同比例的磷酸钙来改变骨水泥的性状外,还可以通过在其固相和液相中掺入生物活性物质、金属与非金属离子、具有生物活性的大分子聚合物等来改善其表征,提高机械属性、生物相容性,增加孔隙率,促进骨再生能力。 2.2 磷酸钙骨水泥的改性 2.2.1 掺入生物活性物质 磷酸钙骨水泥与人体骨骼的主要成分相同,新生骨的生成需要体内各种生长因子的共同参与,因此添加生物活性物质会促进新生骨的形成。LEE等[9]将多孔多通道双相磷酸钙颗粒掺入磷酸钙骨水泥中以促进其体内降解速率和骨组织生成,在搭载骨形态发生蛋白2之后,首先用胶原蛋白包被骨水泥,然后将其嵌入其中,与使用普通磷酸钙骨水泥比较,实验组新生骨的生成速度以及降解速度更快。李浩亮等[10]探究搭载骨形态发生蛋白2磷酸钙骨水泥/纤维蛋白胶复合材料的生物学效能,结果表明该复合材料可促进骨折的愈合,提高其机械属性能。肖琦科等[11]将转染骨形态发生蛋白2基因的骨髓间充质干细胞掺入到复合多孔磷酸钙骨水泥中,植入新西兰兔股骨髁缺损4,12周的Van-Gieson染色和免疫荧光染色结果显示,复合多孔的磷酸钙骨水泥注入的缺损模型中有大量新生骨生成。LEE等[12]探究甘露醇成孔剂和藻酸盐在骨修复方面的作用,体外碱性磷酸酶检测表明加入甘露醇和藻酸盐能够促进成骨再生。 血浆中含有丰富的生长因子,这些因子在骨骼愈合中发挥着重要作用[13]。郭瑛[14]将富血小板血浆与骨水泥混合,将其植入体内后可缓解机体的免疫排斥和炎症反应;同时含富血小板血浆的骨水泥具备良好的注射性、黏滞性,不易发生渗漏,可促进骨水泥的降解吸收及诱导新骨形成。BABO等[15]在掺入血小板的磷酸钙骨水泥中加入透明质酸微球,植入骨缺损4,8和12周后通过矿物质量评估其生物性状,结果显示,掺入透明质酸微球虽然会加速骨水泥的降解,但会损害其骨传导性能,并且在术后长达4周的时间里,掺入血小板裂解物的骨水泥表现出良好的骨再生能力。 KLEIN等[16]探究掺入丝素蛋白磷酸钙骨水泥生物复合材料的兔胫骨缺损修复作用,影像学、组织学和扫描电镜检测显示,掺有丝素蛋白的磷酸钙骨水泥具有良好的生物相容性和诱导骨再生的能力。FENG等[17-18]将N-乙酰半胱氨酸掺入丝素蛋白复合骨水泥以提高其抗压强度,结果表明加入N-乙酰半胱氨酸后可提高其抗压强度,改良其机械特性。PALMER等[19]将胶原蛋白掺入磷酸钙骨水泥中,植入动物骨缺损部位未出现炎症或免疫反应,在新生骨裂缝中检测出抗酒石酸酸性磷酸酶,并表现出与聚甲基丙烯酸甲酯相当的机械性能。王松等[20]探讨多孔磷酸钙/骨基质明胶复合骨水泥修复腰椎骨缺损的效果,通过观察术后不同时间的X射线片评估骨融合情况,micro-CT及组织染色观察新生骨形成情况,结果表明复合骨水泥可促进新生骨的生成,增加其抗压强度及孔隙率。 2.2.2 掺入金属和非金属离子 将亲和力高的金属和非金属离子掺入磷酸钙骨水泥中去,以改善其机械性能与其他方面的性能。KUANG等[21]和ZHOU等[22]在制备骨水泥过程中添加锶离子以改善其性状,体内外实验结果显示,掺锶骨水泥具有良好的骨再生能力和碱性磷酸酶活性。REITMAIER等[23]研究含锶磷酸钙骨水泥对重组骨形态发生蛋白2生物活性的影响,结果显示,在含锶骨水泥上骨形态发生蛋白2诱导的成骨活性得到显著提高。LV等[24]将降钙素基因相关肽和壳聚糖锶掺入磷酸钙骨水泥,并对其进行表征,结果显示,含有降钙素基因相关肽和壳聚糖锶的复合骨水泥能改善血管微环境,促进血管生长因子基因、蛋白的表达及血管生成。 GOLDBERG等[25]将镁离子掺入磷酸钙骨水泥中进行实验,结果显示含镁离子骨水泥相较于普通骨水泥具有良好的机械性能。CABKEJOS-AZAMA等[26]的体内外实验显示,含镁骨水泥促进成骨细胞增殖及骨组织再生。XIONG等[27]将掺有锌的磷酸钙骨水泥与小鼠骨髓基质细胞混合,发现含锌骨水泥可以促进细胞增殖及提高碱性磷酸酶活性。WU等[28]将锰离子掺入β-磷酸三钙中探究其成骨能力,发现与含锰离子β-磷酸三钙共培养的骨髓基质细胞相较与普通骨水泥共培养的骨髓基质细胞表现出更好的黏附和铺展行为,并提高了Runx2基因表达水平。QIAN等[29]和WEIWEI等[30]通过将硅酸锌和聚乳酸-乙醇酸共聚物微球掺入磷酸钙骨水泥中以促进新生骨生成、加速其降解及增强其机械性能,结果显示,硅酸锌和聚乳酸-乙醇酸共聚物的添加降低了炎症相关基因(白细胞介素1β、白细胞介素6和肿瘤坏死因子α)的表达,提高了抗炎基因的表达(白细胞介素10);在巨噬细胞条件下,含有硅酸锌和聚乳酸-乙醇酸共聚物复合骨水泥提取物可显著促进小鼠骨髓基质细胞的成骨分化,并抑制破骨细胞生成。LIU等[31]将钡离子与磷酸钙骨水泥混合评估其可注射性、机械强度和生物相容性,结果表明添加钡离子能改善骨水泥的机械强度。 2.2.3 掺入有机物 AMIRIAN等[32]检测含藻酸盐-透明质酸微珠磷酸钙骨水泥与普通磷酸钙骨水泥的形态、化学结构、凝固时间、抗压强度、可注射性、耐冲洗性和体内生物学性能,发现在磷酸钙骨水泥中添加藻酸盐和透明质酸微珠会促进新生骨生成。AHLFELD等[33]将磷酸钙骨水泥和载有血管生成因子的藻酸盐与吉兰糖胶水凝胶糊剂制成双相支架,检测结果显示该双相支架可以促进新生骨生成。WANG等[34]将藻酸盐微囊化的细胞植入骨水泥中后注射于兔肱骨缺损模型中,通过CT评估新合成的骨水泥降解速率和新生骨生成状况,结果显示掺入微囊化细胞的骨水泥复合物极大地促进了新生骨生成。SMITH等[35]在探究磷酸钙骨水泥中掺入葡萄糖微粒和聚乳酸-乙醇酸共聚物对理化性质和孔隙率的影响,结果显示,掺入葡萄糖微粒和聚乳酸-乙醇酸共聚物加快了磷酸钙骨水泥的降解速率。LANAO等[36]探究掺入不同浓度葡糖酸-δ-内酯对磷酸钙骨水泥性状的影响,通过体内植入实验组织形态计量学评估发现,葡糖酸-δ-内酯的溶解加快了骨水泥的降解速率,被新生骨取代,表明适量浓度的葡糖酸-δ-内酯的骨水泥可以加快降解速率和促进新生骨生成。 因壳聚糖良好的相容性及体内良好的降解能力,FANG等[37]开发了一种由磷酸钙骨水泥和壳聚糖组成的生物复合骨水泥,对其进行一系列物理性状检测及体内实验,结果显示该生物复合骨水泥具有更低的固化温度、更长的凝固时间、更高的孔隙率、更低的抗压杨氏模量和更大的抗压强度;细胞增殖测试表明,生物复合骨水泥因其拥有更粗糙的表面有利于细胞的黏附和生长,表现出良好的促进新生骨再生的能力。疏秀林等[38]通过喷雾冷冻干燥法制备了γ-聚谷氨酸/壳聚糖凝胶颗粒,装载骨形态发生蛋白生长因子后与磷酸钙骨水泥的固相混合,再按照比例添加固化液制备骨水泥载体,结果表明,γ-聚谷氨酸/壳聚糖的添加未改变骨水泥的理化性质,各组间初凝时间与终凝时间及抗压强度未见明显差异,但能明显促进骨细胞体外增殖,且具有较好的缓释功能。黄杨等[39]分析壳聚糖纳米短纤维及壳聚糖对磷酸钙骨水泥机械属性和生物相容性的影响,通过红外光谱、X射线衍射、扫描电镜进行骨水泥成分分析和结构观察,利用万能力学试验机检测其生物力学特性,采用免疫荧光染色和MTT法检测成骨细胞在骨水泥材料上的黏附和增殖情况,结果显示壳聚糖纳米短纤维能够促进新生骨生成,并具备良好的细胞相容性。 LODOSO-TORRECILLA等[40]将聚乳酸-乙醇酸共聚物掺入磷酸钙骨水泥,以聚N-乙烯基吡咯烷酮作为成孔剂掺入其中,以增强其机械性能、增加孔隙率,体外降解实验表明,掺入聚N-乙烯基吡咯烷酮虽然会引起初始质量损失,但可以提高体外降解率。AN等[41]将羧甲基纤维素掺入聚乳酸-乙醇酸共聚物与骨水泥复合物中以改善材料性能、增大孔隙率并刺激降解,结果显示含有羧甲基纤维素复合材料能够诱导磷灰石形成磷酸钙达到降解的目的,并伴随新生骨的生成。LI等[42]采用冷冻滴落法制备海岛型聚乳酸-乙醇酸共聚物与骨水泥复合颗粒,在其沉淀物上培养小鼠骨间充质干细胞,细胞数量及碱性磷酸酶活性检测显示,聚乳酸-乙醇酸共聚物与骨水泥复合物具有促进新生骨生成的能力。SCHICKERT 等[43]和PAKNAHAD等[44]合成掺有聚乙烯醇纤维的磷酸钙骨水泥,表征结果表明其机械强度得到增加。LIU等[45]在磷酸钙骨水泥中添加聚多巴胺以提高其抗压强度,发现聚多巴胺的加入缩短了骨水泥凝结时间,并形成了纳米级磷酸钙层,提高了其机械能力。MA等[46]制备聚乙二醇化的癸二酸甘油酯修饰的磷酸钙骨水泥支架,检测其抗压强度和韧性,发现与普通骨水泥相比,该复合骨水泥的抗压强度和韧性分别提高了约5倍和3倍;培育小鼠骨间充质干细胞的体外实验表明,该复合骨水泥可显著促进细胞的附着和增殖,显示出更强的促进骨再生能力。马正宇等[47]采用液相沉淀法制备β-磷酸三钙,两步法合成聚富马酸丙二醇酯高分子,最后制备了可注射掺有聚富马酸丙二醇酯的β-磷酸三钙复合骨水泥,体外实验结果显示骨水泥的抗压强度增加、降解速率加快。 WANG等[48-49]在微波环境中还原掺入了石墨烯/碳纳米管磷酸钙骨水泥的固化过程,发现与普通骨水泥相比,微波处理后复合材料的凝结时间缩短一半,抗压强度得到提高,并具有良好的生物活性。 POOPUT等[50]通过掺入高溶解速率的麦芽糖糊精微带来制造大孔隙率的磷酸钙骨水泥,在生理条件下将掺入麦芽糖糊精微带的骨水泥浸入模拟体液中1 d,检测其凝结时间、机械性能、微结构和降解速率,检测结果显示,材料孔隙率随麦芽糖糊精微带含量的增加而增加并展现出良好的生物相容性。QIAN等[51]探究不同质量比魔芋葡甘露聚糖/瓜尔豆胶混合物对骨水泥抗冲洗能力、抗压强度和细胞相容性的影响,结果表明,当魔芋葡甘露聚糖/瓜尔豆胶质量比为5∶5时能最大限度地加速骨水泥的降解速率,增加其抗压强度;体外培育细胞实验结果显示,魔芋葡甘露聚糖/瓜尔豆胶复合骨水泥具有良好的机械属性、良好的细胞相容性及促进新生骨生成的性能。 2.2.4 掺入干细胞 ZHANG等[52]开发了一种三维培养系统以诱导磷酸钙骨水泥支架的血管形成,在骨水泥支架上对接种的诱导人多能干细胞衍生间充质干细胞进行骨诱导,然后再接种脐带静脉内皮细胞和周细胞,12周后显示新生骨量及血管生成量显著增多,说明三维培养系统能够有效促进血管生成和新生骨生成。WU等[53]针对磷酸钙骨水泥支架强度低下等问题,通过掺入精甘天三肽(Arg-Gly-Asp)来改善支架的生物相容性,发现相较于普通的骨水泥,掺有精甘天肽的壳聚糖纤维骨水泥具备良好的骨再生能力。LIU等[54]将精甘天肽接种到骨水泥支架上,然后接种NELL1基因修饰的诱导多功能间充质干细胞,RT-PCR和Western blot检测结果证实细胞中的NELL1蛋白含量增加,提示骨水泥支架上有大量新生骨生成。 2.2.5 搭载药物 磷酸钙骨水泥在制备过程中无需经受高温,有利于药物在制备过程中不被损坏而均匀分布于材料,被证实是一种良好的药物载体[55-56]。将磷酸钙骨水泥作为载体把药物递送到靶向部位,长时间保持治疗浓度,为治疗疾病开辟了新的治疗途径。 CHEN等[57]将不同浓度的万古霉素掺入骨水泥中,通过抑制环评估万古霉素溶液浓度对金黄色葡萄球菌的抗菌作用,并检测最佳抑菌效果的万古霉素浓度,发现含搭载万古霉素的磷酸钙骨水泥具有抗菌作用,且10%为最佳浓度。 FILIPPO等[58]将通过喷雾凝结获得的庆大霉素加载固体脂质微粒掺入到骨水泥中并对其进行表征,结果表明,将庆大霉素包含在微粒中对革兰阴性细菌和阳性细菌生长表现出强烈的抑制作用。SHI等[59]将载有庆大霉素的骨水泥作为实验组,在低强度脉冲超声条件下观察其与普通骨水泥的疗效的差别,结果显示,超声组中庆大霉素的瞬时浓度和累积释放百分比在每个时间点均高于对照组,表明搭载庆大霉素骨水泥具有良好的抗菌作用。 2.3 临床应用 2.3.1 与骨折治疗相关 GREIMEL等[60]采用结合球囊胫骨成形和生物可吸收磷酸钙骨水泥的新技术治疗胫骨平台凹陷性骨折,评估了术后临床效果和吸收情况,随访结果显示胫骨断端的延伸量增加,骨水泥周围未见骨溶解反应并促进了骨重塑。潘东续等[61]采用骨水泥联合锁定钢板内固定和单纯锁定钢板内固定治疗肱骨近端骨质疏松骨折老年患者,比较两组骨折愈合时间、术后颈干角及肱骨头高度丢失、手术前后肩关节功能,记录两组并发症情况,结果显示骨水泥联合锁定钢板内固定治疗的骨折愈合时间明显缩短,肱骨头高度丢失明显减小,术后随访时患者疼痛、功能、运动限制及解剖复位评分均优于单纯锁定钢板内固定治疗。SCORDINO等[62]对标准胫骨高位截骨骨缺损处注射磷酸钙骨水泥,使用尸体骨标本进行生物力学实验,结果发现显著改善了高位胫骨截骨结构的初始最大抗扭强度和刚度。GIANLUCA等[63]报告10年随访病例发现,对于A型外伤性椎体骨折采用经皮球囊后凸成形和磷酸钙骨水泥注射治疗的临床效果和放射学结果满意,建议对于年轻 A型椎骨骨折患者可采用此方法治疗。 2.3.2 与骨髓炎治疗相关 杨昊飞等[64]分析载万古霉素骨水泥植入治疗慢性骨髓炎的治疗效果,发现观察组(采用载万古霉素骨水泥植入治疗)的治疗有效率明显高于对照组(采用常规治疗),复发率低于对照组。TAKASHI等[65]使用庆大霉素浸渍的骨水泥治疗1例糖尿病跟骨骨髓炎患者,1.5年的随访显示治疗效果良好,表明骨水泥既能作为骨修复材料填充清创过后的骨缺损空间,又能作为载体搭载抗生素让其在局部释放而发挥作用。 2.3.3 与骨肿瘤治疗相关 骨囊肿治疗:杨长远等[66]对42例单纯性骨囊肿患者采取经皮穿刺骨水泥充填术,术后3个月X射线片显示37例囊腔充填完好,5例仅留小囊腔,所有骨缺损部位均有新生骨生成,无并发症,结果表明经皮穿刺骨水泥充填术可作为治疗单纯性骨囊肿的一种方法。TRIPATHI等[67]使用改良的天然骨颗粒(G Graft)和骨水泥来填充37例囊性病变摘除后下颌骨的骨腔,其中30例患者的骨质和数量均在可接受的范围内,而7例患者出现了移植物挤出和术后感染,该结果说明使用改良骨颗粒和骨水泥注射治疗下颌骨囊性病变是替代其他治疗方法的良好选择,但预防术后感染仍是需要关注的问题。NISHIMURA等[68]评估开放手术与内镜手术治疗16例跟骨骨囊肿的优缺点,所有手术(开放式或内镜式)均在刮除骨囊肿内壁后注射骨水泥,结果显示接受内窥镜手术患者恢复运动的时间明显缩短,可尽早康复,并且没有任何不良影响。 骨肿瘤治疗:HIGUCHI等[69]评估了130例良性骨肿瘤手术中填充骨水泥的中长期效果,影像学和临床结果表明骨水泥是良性骨肿瘤手术的良好骨替代物,具有出色的骨传导性和持久稳定性,无需内固定,可明显改善患者的症状。TAKEUCHI等[70]探究彻底刮除酚化和骨水泥填充治疗骨巨细胞瘤26例的效果,采用放射学评估周围骨的骨水泥固结、皮质骨缺损重塑和临床结果,结果显示骨水泥在周围骨骼凝结良好,有11.5%的患者发生局部复发,肺转移发生率为3.8%,皮质骨缺损重塑出现率为84.6%,骨水泥提供了生物界面和持久的稳定性,无需内部固定。RAJEH等[71]研究和评估使用可注射骨水泥填充指、掌骨内生软骨瘤刮除后骨缺损的疗效,平均疼痛评分从术前4.1降低到术后1.6,术后手运动范围已达到对侧手的89.3%,8例患者中有2例骨水泥渗漏,其中1例需要修正。这些结果表明,刮除指、掌骨内软骨瘤后采用可注射骨水泥填充缺损是有效的治疗方法。"
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