Wessel Alex R, Crist Brett D, Stannard James P, Della Rocca Gregory J, Stoker Aaron M, Bozynski Chantelle C, Cook Cristi R, Kuroki Keiichi, Ahner Carin E, Cook James L
Department of Orthopaedic Surgery, University of Missouri, Columbia, USA.
Department of Orthopaedic Surgery, University of Missouri, Columbia, USA; Thompson Laboratory for Regenerative Orthopaedics, University of Missouri, Columbia, USA.
Injury. 2018 Jun;49(6):1046-1051. doi: 10.1016/j.injury.2018.04.010. Epub 2018 Apr 17.
Previous studies indicate that Reamer Irrigator Aspirator (RIA) filtrate contains proteins that have the potential to stimulate bone healing. This study aimed to determine the osteoinductive capabilities of RIA filtrate in a validated in vivo model.
With Institutional Review Board approval, RIA filtrates from 9 patients were collected. The filtrate was processed to remove cells and inorganic particles. A portion of each sample was set aside for protein analysis while the remainder was lyophilized and prepared for implantation. With Animal Care and Use Committee approval, athymic mice (n = 16; 32 hind limbs) were randomly assigned to 1 of 4 groups (n = 8 limbs per group) for percutaneous gastrocnemius muscle injection of demineralized bone matrix (DBM) (10 mg), lyophilized RIA powder (10 mg), RIA liquid (10 mg of lyophilized RIA powder in 100ul phosphate buffered saline (PBS)), or DBM (10 mg) + RIA liquid (10 mg in 100ul PBS). Radiographs were obtained 2, 4, and 8 weeks after injection. At 8 weeks, mice were sacrificed and the entire gastrocnemius muscle from each hind limb was collected and processed for histologic examination. Histological sections and radiographs were assessed for ossification/calcification. Data were compared for statistically significant (p < 0.05) differences among groups and strong (R > 0.7) correlations between outcome measures.
The protein composition of RIA filtrates was consistent among patients and matched previous data. For all groups, radiographic scores were significantly (p < 0.014) higher (more calcification/ossification) at 8 weeks compared to 2 weeks. Radiographic scores for the DBM and DBM + RIA liquid groups were significantly higher than RIA liquid and RIA powder at 4 weeks and 8 weeks (p < 0.019 and p < 0.049, respectively). Histologic scores were significantly (p = 0.004) higher in the DBM + RIA liquid group compared to the RIA liquid group at 8 weeks. Histologic scores showed strong correlations (r > 0.77) to radiographic scores for all groups.
RIA filtrate liquid and powder were osteoinductive in vivo with new bone formation being most abundant using a combination of DBM and RIA filtrate in this validated animal model. RIA filtrate has potential for clinical use in augmenting bone healing treatments.
先前的研究表明，扩髓冲洗吸引器（RIA）滤液中含有可能刺激骨愈合的蛋白质。本研究旨在通过一个经过验证的体内模型来确定RIA滤液的骨诱导能力。
经机构审查委员会批准，收集了9例患者的RIA滤液。对滤液进行处理以去除细胞和无机颗粒。每个样本的一部分留作蛋白质分析，其余部分冻干并准备植入。经动物护理和使用委员会批准，将无胸腺小鼠（n = 16；32只后肢）随机分为4组中的1组（每组n = 8只后肢），用于经皮腓肠肌注射脱矿骨基质（DBM）（10 mg）、冻干RIA粉末（10 mg）、RIA液体（10 mg冻干RIA粉末溶于100 μl磷酸盐缓冲盐水（PBS））或DBM（10 mg）+ RIA液体（10 mg溶于100 μl PBS）。在注射后2、4和8周拍摄X线片。在8周时，处死小鼠，收集每只后肢的整个腓肠肌并进行组织学检查。对组织学切片和X线片进行骨化/钙化评估。比较各组间具有统计学意义（p < 0.05）的差异以及各结局指标之间的强相关性（R > 0.7）。
RIA滤液的蛋白质组成在患者之间是一致的，并且与先前的数据相符。对于所有组，8周时的X线评分显著高于2周时（p < 0.014）（更多的钙化/骨化）。在4周和8周时，DBM组和DBM + RIA液体组的X线评分显著高于RIA液体组和RIA粉末组（分别为p < 0.019和p < 0.049）。在8周时，DBM + RIA液体组的组织学评分显著高于RIA液体组（p = 0.004）。所有组的组织学评分与X线评分均显示出强相关性（r > 0.77）。
在这个经过验证的动物模型中，RIA滤液液体和粉末在体内具有骨诱导性，使用DBM和RIA滤液的组合时新骨形成最为丰富。RIA滤液在增强骨愈合治疗方面具有临床应用潜力。
