重组人成纤维细胞生长因子2联合可溶性肿瘤坏死因子Ⅰ型受体蛋白促进2型糖尿病大鼠骨折修复

背景:有报道1型糖尿病可导致骨再生与修复障碍,局部注射成纤维细胞因子2可明显促进骨折愈合,但其对2型糖尿病的影响尚不十分清楚。目的:观察重组人成纤维细胞生长因子2和可溶性肿瘤坏死因子Ⅰ型受体蛋白联合治疗2型糖尿病引起的骨再生及其修复障碍的作用。设计:随机对照实验。单位:中南大学湘雅三医院骨科。材料:选择雄性Zucker糖尿病大鼠(ZDF/Gmi-fa/fa)20只,10周龄,购自美国查尔斯河实验室。重组人成纤维细胞生长因子2购自美国Orquest公司,可溶性肿瘤坏死因子Ⅰ型受体蛋白购自美国Amgen公司。方法:实验于2006-09/11在中南大学湘雅三医院中心实验室完成。①实验分组:将20只大鼠随机分为对照组和治疗组,每组10只。②实验方法:检测大鼠体质量,血糖、尿糖和尿酮。1周后,接受左胫骨中上段低能截骨,安置外延长固定架。第2天起开始胫骨延长0.2mm/次,2次/d,共14d。期间治疗组大鼠接受截骨处血肿内重组人成纤维细胞生长因子225μg/kg注射1次,可溶性肿瘤坏死因子Ⅰ型受体蛋白8mg/kg皮下注射隔日1次,共14d;对照组仅接受载体和生理盐水注射。③实验评估:测定血液生化指标、胫骨延长间隙中新生骨量及增殖细胞数量。主要观察指标:两组血液生化指标、胫骨延长间隙中新生骨量及增殖细胞数量比较。结果:纳入大鼠20只,全部进入结果分析。①两组血糖、尿糖、尿酮、血清胰岛素和骨钙素比较,差异不明显(P>0.05)。②治疗组大鼠左胫骨延长间隙内新生骨的面积、密度和骨内膜成骨、骨膜成骨均明显高于对照组(P<0.05～0.01)。治疗组大鼠的胫骨牵引间隙中的纤维间区和初始骨基质前沿增殖细胞核抗原阳性细胞及百分比均明显多于对照组(P<0.05～0.01)。结论:2型糖尿病可引起骨再生与修复障碍,重组人成纤维细胞生长因子2和可溶性肿瘤坏死因子Ⅰ型受体蛋白联合治疗可促进成骨细胞增殖,加快牵引成骨速率,有利于2型糖尿病合并骨折的愈合障碍。

Recombinant human fibroblastic growth factor-2 with soluble tumor necrosis factor receptor-1 facilitates fracture repair in rats with type 2 diabetes mellitus

BACKGROUND: It has been reported that type 1 diabetes mellitus can result in impairments of bone regeneration and repair, and local injection of fibroblastic growth factor-2 (FGF-2) can obviously promote fracture healing, but its effect on type 2 diabetes mellitus is still unclear.OBJECTIVE: To observe the effects of recombinant human fibroblastic growth factor-2 (rhFGF-2) combined with soluble tumor necrosis factor receptor-1 (sTNF-R1) on impaired bone regeneration and repair in type 2 diabetes mellitus.DESTGN: A randomized controlled trial.SETTING: Department of Orthopaedics, the Third Xiangya Hospital of Central South University.MATERIALS: Twenty male Zucker diabetic fatty rats (ZDF/Gemi-fa/fa), 10 weeks of age, were purchased from Charles River Laboratory. rhFGF-2 was obtained from Orquest Incorporation; sTNF-R1 protein (PEG-r-metHu-sTNF-R1) was provided by Amgen Incorporation.METHODS: This experiment was finished in the central lab of the Third Xiangya Hospital of Central South University from September to November in 2006. ①Grouping: The 20 rats were randomly assigned into treated group (n =10)and control group (n =10). ② Experimental methods: All rats were examined for body mass, blood glucose, glycosuria and glycosemia. One week later, all the rats underwent the standard DO protocol, including placement of the external fixators and osteotomies to the left tibia. Distraction was initiated in the following morning (one day latency) at 0.2 mm b.I.d. (0.4 mm per day) and continued for 14 days. During surgery, all the rats received an injection of either rhFGF-2(25 mg/kg) for the treated group, or physiological saline (25 mg/kg) for the control group, into the hematoma of the osteotomic gap. The sTNF-R1 (8 mg/kg) or the same. Amount of saline was subcutaneously injected into the treated and control rats respectively every other day for 14 days. Evaluation: The serum biochemical indexes, amount of bone formation and number of proliferative cells in the distraction gaps were determined.MAIN OUTCOME MEASURES: Biochemical indexes, amount of bone formation and number of proliferative cells in the distraction gaps.RESULTS: All the 20 rats were involved in the final analysis of results. ①The blood glucose, glucosuria, ketonuria,serum levels of insulin and osteocalcin were not obviously different between the treated group and control group (P ＞0.05). ② The area and density of mineralization of the distraction gaps, and the endosteal and peristeal new bone formation in the treated group were all obviously higher than those in the control group (P ＜ 0.05-0.01). The number and percentage of the positive cells of proliferating cell nuclear antigen (PCNA) in the distraction gaps were obviously higher in the treated group than in the control group (P ＜ 0.05-0.01).CONCLUSION: The local application of rhFGF-2 combined with sTNF-R1 can enhance bone formation by increasing the proliferation during distraction osteogenesis in ZDF rats. The combination of rhFGF-2 and sTNF-R1 may be an effective treatment for type 2 diabetic patients with fracture healing problem.