不同表面处理的骨整合经皮假体的细胞毒性及对植入后兔背部位感染预防及愈合效果的研究

Cytotoxicity of osteointegrated percutaneous prostheses with different surface treatments and their effects on prevention and healing of posterior infection in rabbits

  • 摘要:
    背景 骨整合假肢可经皮将负荷直接转移到骨骼,相比传统接受腔假肢具有一定优势,但普遍面临经皮部位软组织无法密封导致的感染风险。
    目的 利用机械加工方式制造经皮假体并进行不同表面处理,探讨不同表面处理对机体的影响、感染预防能力及在经皮部位与软组织整合情况。
    方法 利用机加工方式制备实验用经皮假体36枚,随机平均分为A、B、C、D、E和F组,每组各6枚,A ~ E组表面分别采取喷砂+钛铌氮涂层、抛光+钛铌氮涂层、高抛光、喷砂+微弧氧化、酸蚀的处理工艺,F组表面不做处理,通过MTT法分析不同经皮假体浸提液对MC3T3-E1细胞及hFob1.19细胞存活的影响。将18只新西兰大白兔随机分为6组,每组3只,每只实验动物背部对称部位植入同种表面处理的假体2枚,观察实验动物手术部位皮肤生长恢复情况,进行不同时间白细胞、中性粒细胞、血小板、血红蛋白、丙氨酸转氨酶、肌酐、补体成分3及补体成分4的检测,分析不同表面处理经皮假体对兔血液学指标的影响。最后取肝、肾组织及经皮部位素材进行组织学检查。
    结果 MTT方法检测结果显示,各实验组与对照组细胞存活率的差异均无统计学意义(P>0.05)。术后同组不同时间的血小板计数、血红蛋白、丙氨酸转氨酶及肌酐数据差异无统计学意义(P>0.05)。术后第28天,B组白细胞计数低于F组(P=0.019);术后第42天,A组白细胞计数高于F组(P=0.002),B、C、D及E组白细胞计数低于F组(P<0.001);术后第56天,B组补体成分3水平低于F组(P=0.039);术后第28天,B组补体C4水平低于A组(P=0.009)与E组(P=0.006)。肝、肾组织切片中无明显的变性、坏死、炎性变化等情况。经皮部位组织学检查显示抛光+钛铌氮涂层处理的经皮假体与周围软组织贴合紧密。
    结论 不同表面处理的经皮假体是安全的,没有对机体造成明显的损伤。表面未处理的经皮假体抵御感染的能力较弱,表面经过抛光+钛铌氮涂层处理的经皮假体抵御感染的能力较强,而且能够与周围软组织相对紧密的结合,可以作为临床应用的骨整合假肢经皮部位的表面处理工艺。

     

    Abstract:
    Background Osseointegration prostheses can directly transfer the load to the bone through the skin, which has certain advantages compared with traditional prosthetic sockets. However, they generally face the risk of infection caused by the unsealing of the soft tissue in the percutaneous part.
    Objective To fabricate percutaneous prostheses via mechanical machining methods and apply distinct surface treatments to investigate their biological impacts, infection-resistant properties, and integration performance with soft tissues at transcutaneous interfaces.
    Methods Thirty-six experimental percutaneous prostheses were prepared by machining and were randomly divided into groups A, B, C, D, E, and F, with 6 in each group. The surface of groups A and E were treated by sandblasting+ TiNbN coating, polishing+ TiNbN coating, high polishing, sandblasting+micro-arc oxidation, and acid etching, while the surface of group F was not treated. The effects of different percutaneous prosthesis extracts on MC3T3-E1 were analyzed using the MTT method. Eighteen New Zealand white rabbits were randomly divided into 6 groups, with 3 rabbits in each group. Two prostheses with the same surface treatment were implanted in the symmetrical part of the back of each experimental animal. The skin growth and recovery of the surgical site of the experimental animals were observed. At the same time, the white blood cell count, neutrophil count, platelet count, hemoglobin, alanine aminotransferase, creatinine, complement component 3, and complement component 4 were detected at different time points, and the effects of percutaneous prostheses with different surface treatments on hematological indexes of rabbits were analyzed. Finally, the liver, kidney tissue, and percutaneous materials were taken for histological examination.
    Results The results of the MTT assay showed that there was no significant difference in cell survival rate between the experimental groups and the control group (P > 0.05). There was no significant difference in platelet count, hemoglobin, alanine aminotransferase, and creatinine in the same group at different time points after operation (P > 0.05). On the 28th postoperative day, the WBC count in group B was lower than that in group F (P=0.019), and on the 42nd postoperative day, WBC counts in group A was lower than that in group F (P=0.002), groups B, C, D, E (P < 0.001). The level of complement component 3 in group B was lower than that in group F (P=0.039). On the 28th day after operation, the level of complement C4 in group B was lower than that in group A (P=0.009) and group E (P=0.006). There was no obvious degeneration, necrosis, or inflammatory changes in liver and kidney tissue sections. The histological examination of the percutaneous site showed that the percutaneous prosthesis treated with polishing and titanium-niobium-nitrogen coating was closely attached to the surrounding soft tissue.
    Conclusion Percutaneous prostheses with different surface treatments demonstrates biosafety without causing significant tissue damage. Untreated surface prostheses exhibits lower infection resistance, while those subjected to polishing combined with TiNbN coating treatment shows enhanced antimicrobial efficacy and achieves more robust integration with surrounding soft tissues. This optimized surface modification protocol holds clinical potential as a transcutaneous interface processing technique for osseointegrated prostheses.

     

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