Clin Osteol 2026; 31(1): 45-48 | DOI: 10.36290/clo.2026.006

An optimized protocol for RNA and microRNA isolation from human mineralized femoral neck bone tissueOriginal contributions

Květa Vrablíková¹, Marcela Slavíčková¹, Helena Párová¹, Petr Hoza^{1, 2}

¹ Ústav klinické biochemie a diagnostiky LF UK a FN Hradec Králové

² Ortopedické oddělení, Pardubická nemocnice, Pardubice

Low-trauma proximal femur fractures are among the most serious complications of osteoporosis. MicroRNAs regulate bone remodeling and cellular senescence, yet extracting high-quality RNA from mineralized human bone remains technically challenging and insufficiently standardized. This study aimed to develop and optimize a reproducible protocol for collecting and processing femoral neck bone biopsies obtained during total hip arthroplasty, enabling isolation of total RNA including the small RNA fraction (< 200 nt) and subsequent targeted expression analysis of specific microRNAs. Forty-six patients were enrolled (21 osteoporotic fractures, 25 osteoarthritis). Samples were immediately stabilized in QIAzol Lysis Reagent (QIAGEN), mechanically homogenized (MagNA Lyser 90 s at 7,000 × g; +30 s if needed), and RNA was isolated using the miRNeasy Mini Kit. RNA was successfully obtained from 44/46 patients (96 %). RNA concentration was higher in the fracture group than in the osteoarthritis group (227.9 ± 158.3 vs. 95.8 ± 81.8 ng/µl; p = 0.0016). The protocol provides a reliable methodological platform for molecular analyses in human mineralized bone tissue.

Keywords: RNA isolation, microRNA, femoral neck biopsy.

Accepted: March 25, 2026; Published: March 30, 2026  Show citation