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组内侧平台软骨下骨TRAP阳性破骨细胞数为33.4%±3.1%,高于外侧平台的25.1%±2.3%,差异有统计学意义(P<0.05)。免疫组化染色结果显示:HA组内侧平台软骨下骨VEGFA阳性细胞数为34.1%±5.9%,高于外侧平台的25.9%±3.7%;内侧平台软骨下骨Osterix阳性细胞数为14.6%±1.4%,高于外侧平台的5.8%±1.1%,HA组内、外侧平台软骨下骨VEGFA阳性细胞数较OA组多,Osterix阳性细胞数较OA组少,差异均有统计学意义(P<0.05)。结论与OA组相比,HA组软骨下骨的骨破坏程度更高、骨小梁更稀疏,潮线基本消失,破骨细胞分化和血管生成水平更高,成骨细胞分化水平更低,表现出更严重的骨质疏松,但SBP却增厚。结果提示可以通过干预HA软骨下骨异常骨重塑和血管生成达到延缓HA进展和治疗的作用。
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 significantly from the OA group(P<0.001).Conclusion The HA group exhibited more extensive subchondral bone destruction,thinner trabeculae,a nearly absent tidemark,higher osteoclast differentiation,increased angiogenesis,and reduced osteoblast differentiation,indicating severe osteoporosis,despite thicker SBP.These findings suggest that targeting abnormal bone remodeling and angiogenesis in HA could retard its progression and provide therapeutic benefits.
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
姚运峰
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)
出处
《中华骨科杂志》
CAS
CSCD
北大核心
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)。
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
hemophiliacarthritis
osteoarthritis
subchondralbone
bone remodeling
angiogenesis
作者简介
通信作者:姚运峰,Email:cdyaoyunfeng@163.com。
