Olçar Hacı Ali, Halıcı Mehmet, Kafadar İbrahim Halil, Karaman İbrahim, Lekesizcan Ayça, Gönen Zeynep Burçin
Department of Orthopedics and Traumatology, Yozgat City Hospital, Yozgat, Turkey.
Jt Dis Relat Surg. 2020;31(1):20-7. doi: 10.5606/ehc.2020.71270.
This study aims to evaluate the effects of local adipose stem cell injection on non-union and diabetic non-union of rat femurs.
Forty-eight female Wistar albino rats (weighing mean 200 g and aged 8 weeks) were used in this study. The rats were divided into six groups. Group 1 was chosen as a reference for receptor activator of nuclear factor-kappa (κ) B (RANK), receptor activator of nuclear factor-κ B ligand (RANKL) and osteoprotegerin (OPG) genes and no femur osteotomy was performed in this group. Group 2 underwent femur osteotomy, the osteotomy was fixed with a 1.5 mm K-wire as retrograde from the knee joint, and no gap was left in the osteotomy line. In order to induce non-union, femurs underwent osteotomy fixed with K-wires in groups 3, 4, 5 and 6. In addition, the osteotomy line was measured as 1.8 mm gap with electronic calipers and the gap was fixed with U staple. Before osteotomy, streptozocin was injected intraperitoneally at a dose of 60 mg/kg in 0.1 mol/L citrate buffer solution (Ph 4.4) in groups 4 and 6, in order to induce diabetes mellitus. Left femur anteroposterior and lateral X-rays were taken 10 weeks after the operation and the union in group 2 and non-union in groups 3, 4, 5, and 6 were confirmed. To see if injection of adipose stem cells into the non-union site increases bone union, 2 mL 0.9% sodium chloride (NaCl) in groups 3 and 4 and 2×106 adipose stem cell in groups 5 and 6 were locally injected into the non-union area with fluoroscopy. Femur X-rays were taken eight weeks after the injection and all rats were sacrificed. Femurs of rats were removed for histopathological and gene expression evaluation.
There were significant differences between the groups injected 0.9% NaCI and adipose stem cells in terms of bone healing according to radiological and histopathological evaluations (p<0.05). No statistically significant difference was observed between the groups in terms of gene expression levels.
According to the results of our study, local adipose stem cell injection has positive radiological and histopathological effects in diabetic and non-diabetic femoral non-unions, independently of RANK, RANKL, or OPG gene expression pathways.
本研究旨在评估局部注射脂肪干细胞对大鼠股骨骨不连及糖尿病性骨不连的影响。
本研究选用48只雌性Wistar白化大鼠(平均体重200 g,年龄8周)。将大鼠分为六组。第一组作为核因子κB受体激活剂(RANK)、核因子κB配体受体激活剂(RANKL)和骨保护素(OPG)基因的参照组,该组未进行股骨截骨术。第二组进行股骨截骨术,截骨处用一根1.5 mm克氏针从膝关节逆行固定,截骨线不留间隙。为诱导骨不连,第三、四、五、六组的股骨截骨后用克氏针固定。此外,用电子卡尺将截骨线测量为1.8 mm的间隙,并用U形钉固定。在截骨术前,第四组和第六组腹腔注射链脲佐菌素,剂量为60 mg/kg,溶于0.1 mol/L柠檬酸盐缓冲液(pH 4.4)中,以诱导糖尿病。术后10周拍摄左侧股骨正侧位X线片,确认第二组骨愈合以及第三、四、五、六组骨不连。为观察向骨不连部位注射脂肪干细胞是否能促进骨愈合,第三组和第四组局部注射2 mL 0.9%氯化钠(NaCl),第五组和第六组在透视引导下向骨不连区域局部注射2×10⁶个脂肪干细胞。注射后8周拍摄股骨X线片,然后处死所有大鼠。取出大鼠股骨进行组织病理学和基因表达评估。
根据放射学和组织病理学评估,注射0.9% NaCl组和脂肪干细胞组在骨愈合方面存在显著差异(p<0.05)。各组在基因表达水平方面未观察到统计学显著差异。
根据我们的研究结果,局部注射脂肪干细胞对糖尿病性和非糖尿病性股骨骨不连具有积极的放射学和组织病理学作用,与RANK、RANKL或OPG基因表达途径无关。