佛手苷内酯对磷酸三钙磨损颗粒所致假体周围骨溶解的影响

目的:观察佛手苷内酯对磷酸三钙(TCP)磨损颗粒所致假体周围骨溶解的影响,探讨其可能作用机理。方法:60只雄性ICR小鼠随机分为5组(每组n=12):假手术组(Sham组)、TCP磨损颗粒组(TCP组)、佛手苷内酯5 mg/kg组、佛手苷内酯10 mg/kg组和佛手苷内酯20 mg/kg组。TCP磨损颗粒30 mg包埋于小鼠颅骨顶部建立小鼠假体周围骨溶解模型,Sham组小鼠仅进行原位缝合,不包埋颗粒。Sham组和佛手苷内酯组小鼠分别于术后第2天颅顶局部注射生理盐水和佛手苷内酯,隔日1次。2周后处死动物采血、取颅骨。采用抗酒石酸酸性磷酸酶(TRAP)染色和HE染色观察小鼠颅骨假体周围骨溶解和破骨细胞生成情况;化学比色法检测血清中TRAP和组织蛋白酶K(cathepsin K)活性及乳酸脱氢酶(LDH)含量;ELISA法检测血清中肿瘤坏死因子-alpha(TNF-α)、白介素-1beta(IL-1β)、白介素(IL)-18和核因子-κB受体活化因子配体(RANKL)的水平;western blot检测假体周围骨组织中炎症小体活化相关蛋白Nod样受体蛋白3(NLRP3)和凋亡相关斑点样蛋白(...

Effects of Bergapten on Periprosthetic Osteolysis Caused by TCP Wear Particles in Mice

Objective To observe the effect of bergapten on periprosthetic osteolysis induced by tri⁃calcium phosphate(TCP)wear particles,and to explore its possible mechanism.Methods Sixty male ICR mice were randomly divided into a sham group,a TCP wear particles(TCP)group,a bergapten(5 mg/kg)group,bergapten(10 mg/kg)group and bergapten(20 mg/kg)group,each of 12.A peripros⁃thetic osteolysis model was established by embedding 30 mg of TCP wear particles onto the surface of bilateral parietal bones after removal of the periosteum in all groups except the sham group,which was operated without embedding particles.On the 2nd day postoperatively,normal saline and bergapten were locally injected to the calvarium under the periosteum every other day for the sham and ber⁃gapten groups respectively.Two weeks later,all the mice were sacrificed to obtain the blood and the calvaria.Tartrate resistant acid phosphatase(TRAP)staining and hematoxylin eosin staining were used to observe periprosthetic osteolysis and osteoclastogenesi of the calvaria.The chemical colorimetry was applied to examine the activity of TRAP and cathepsin K and lactate dehydrogenase(LDH)content in serum.Moreover,the enzyme-linked immunosorbent assay(ELISA)was used to measure serum levels of tumor necrosis factor-alpha(TNF-α),interleukin-1beta(IL-1β),interleukin-18(IL-18)and recep⁃tor activator of nuclear factor-kappa B ligand(RANKL),while Western blotting was applied to detect protein expressions of nod-like receptor pyrin domain-containing protein 3(NLRP3),apoptosis-associat⁃ed speck-like protein containing CARD(ASC),cleavage of cysteine aspartate proteinase-1(cleaved cas⁃pase-1)and interleukin(IL)-1βin the periprosthetic bone tissue.Results Compared with the sham group,a significant increase was observed in the average osteolysis area and osteoclast number(P<0.05),the activity of TRAP and cathepsin K,and levels of RANKL,TNF-α,IL-1β,IL-18 and LDH in the serum(P<0.05),as well as the protein expression of NLRP3,ASC,cleaved caspse-1and IL-1βin the calvaria of TCP group(P<0.05).Compared with TCP group,a significant decrease was observed in the average osteolysis area and osteoclast number(P<0.05),the activity of TRAP and ca⁃thepsin K,and levels of RANKL,TNF-α,IL-1β,IL-18 and LDH in the serum(P<0.05),as well as the protein expression of NLRP3,ASC,cleaved caspse-1and IL-1βin the calvaria of bergapten groups(P<0.05).The inhibition of bergapten on the increases in the above indexes caused by TCP wear particles was in a dose-dependent manner.Conclusion Bergapten prevents periprosthetic osteolysis and osteoclastogenesis caused by TCP wear particles through the attenuation of NLRP3 inflammasome activation and pyroptosis.