血友病性关节炎和骨关节炎软骨下骨的组织形态学分析  

Histomorphological analysis of subchondral bone in hemophilic arthritis and osteoarthritis

作　　者：叶厚龙 冯茹 郑刘杰 罗达胜 韩志伟 钟齐刚 戚任飞 荆珏华[1] 姚运峰[1] Ye Houlong;Feng Ru;Zheng Liujie;Han Zhiwei;Zhong Qigang;Qi Rengfei;Jing Juehua;Yao Yunfeng(Department of Orthopaedics,Institute of Orthopaedics,Research Center for Translational Medicine,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601,China;Department of Orthopaedics,The Second People's Hospital of Hefei,Hefei Hospital Affiliated to Anhui Medical University,Hefei 230011,China;Department of Orthopaedics,Puai Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430033,China)

机构地区：[1]安徽医科大学第二附属医院骨科,骨科研究所,合肥230601 [2]合肥市第二人民医院(安徽医科大学附属合肥医院)骨科,合肥230011 [3]华中科技大学同济医学院附属普爱医院骨科,武汉430033

出　　处：《中华骨科杂志》2023年第24期1663-1672,共10页Chinese Journal of Orthopaedics

基　　金：国家自然科学基金面上项目(82071591);安徽省转化医学基金培育项目(2022zhyx-C74);安徽医科大学第二附属医院转化医学研究科研基金(2022ZHYJ09);安徽医科大学第二附属医院临床研究培育计划项目(2021LCZD12)。

摘　　要：目的分析血友病性关节炎(hemophiliac arthritis,HA)与骨关节炎(osteoarthritis,OA)患者软骨下骨组织形态学的差异,探讨HA软骨下骨异常骨重塑的机制。方法选取2021年1月至2023年6月于安徽医科大学第二附属医院因HA行初次全膝关节置换术的15例男性患者,年龄(32.60±7.58)岁(范围22~45岁),均为A型血友病且凝血因子Ⅷ抗体为阴性;选取同期因OA行初次全膝关节置换术的15例男性患者,年龄(75.67±5.09)岁(范围71~87岁)。获取术中截去的胫骨平台骨组织,以MicroCT扫描评估两组胫骨平台标本软骨下骨的形态学参数,HE和蕃红"O"固绿染色评估两组软骨下骨板(subchondral bone plate,SBP)和骨小梁组织结构变化,抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)染色评估软骨下骨破骨细胞分化水平,血管内皮生长因子(vascular endothelial growth factor,VEGF)-A和Osterix免疫组化染色评估软骨下骨血管生成和成骨细胞分化水平。结果MicroCT扫描结果显示:HA组骨体积分数(bonevolume fraction,BV/TV)内侧平台为25.14%±0.70%、外侧平台为22.31%±0.53%;骨小梁连接密度(connectivity density,Conn.D.)内侧平台为(4.20±0.10)1/mm^(3)、外侧平台为(3.27±0.08)1/mm^(3);骨密度(bone mineral density,BMD)内侧平台为(0.288±0.006)g/cm^(3)、外侧平台为(0.285±0.004)g/cm^(3);骨小梁厚度(trabecularthichness,Tb.Th)内侧平台为(0.257±0.008)mm、外侧平台为(0.206±0.008)mm;骨小梁数量(trabecularnumber,Tb.N)内侧平台为(0.984±0.043)1/mm、外侧平台为(0.908±0.026)1/mm;骨小梁间距(trabecularseparation,Tb.Sp)内侧平台为(0.683±0.008)mm、外侧平台为(0.808±0.010)mm;HA组内、外侧平台除Tb.Sp高于OA组,BV/TV、Conn.D.、BMD、Tb.Th和Tb.N均低于OA组,差异均有统计学意义(P<0.05)。HE和蕃红"O"固绿染色结果显示:HA组SBP厚度内侧平台为(177.43±6.42)μm,高于外侧平台的(117.96±5.08)μm,差异有统计学意义(P<0.05)。TRAP染色结果显示:HA组内�Objective To delineate the histomorphological disparities of subchondral bone between hemophilic arthritis(HA)and osteoarthritis(OA)and to explore the mechanisms underpinning aberrant bone remodeling in HA.Methods Fifteen male HA patients,aged 32.60±7.58 years(range 22-45),who underwent total knee arthroplasty at the Second Affiliated Hospital of Anhui Medical University from January 2021 to June 2023,were included.All patients had hemophilia A and tested negative for coagulation factor VIII antibodies.Simultaneously,fifteen male OA patients,aged 75.67±5.09 years(range 71-87),also underwent arthroplasty.Tibial plateau bones were extracted for micro-CT,which assessed morphological parameters.Histological changes in the subchondral bone plate(SBP)and trabecular bone were evaluated with HE and Safranin O-Fast Green staining.TRAP staining determined osteoclast differentiation levels,and VEGF-A and Osterix immunohistochemistry gauged angiogenesis and osteoblast differentiation.Results Micro-CT revealed that HA patients had a BV/TV of 25.14%±0.70%(medial)and 22.31%±0.53%(lateral),Conn.D.of 4.20±0.101/mm^(3)(medial)and 3.27±0.081/mm^(3)(lateral),BMD of 0.288±0.006 g/cm^(3)(medial)and 0.285±0.004 g/cm^(3)(lateral),Tb.Th of 0.257±0.008 mm(medial)and 0.206±0.008 mm(lateral),Tb.N of 0.984±0.0431/mm(medial)and 0.908±0.0261/mm(lateral),and Tb.Sp of 0.683±0.008 mm(medial)and 0.808±0.010 mm(lateral).These parameters were significantly lower than those in the OA group except for Tb.Sp,which was higher(P<0.001).Histological staining indicated that the HA group's SBP thickness was 177.43±6.42μm(medial)and 117.96±5.08μm(lateral)with significant differences observed(P<0.001).TRAP staining showed that TRAP+osteoclasts accounted for 33.4%±3.1%(medial)and 25.1%±2.3%(lateral)in HA subchondral bone,again significantly different(P<0.001).Immunohistochemical staining revealed VEGFA+cells at 34.1%±5.9%(medial)and 25.9%±3.7%(lateral),and Osterix+cells at 14.6%±1.4%(medial)and 5.8%±1.1%(lateral)in HA patients,differing

关 键 词：血友病性关节炎 骨关节炎 软骨下骨 骨重塑 血管生成 

分 类 号：R684.3[医药卫生—骨科学]