Cohen David J, Van Duyn Christine M, Deng Jingyao, Lodi Musaddiq K, Gallagher Michelle B, Sugar James T, Rawlinson Jeremy J, Ghosh Preetam, Boyan Barbara D, Schwartz Zvi
Department of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA 23284, USA.
Integrative Life Sciences, Virginia Commonwealth University, Richmond, VA 23284, USA.
Biomimetics (Basel). 2025 Jan 19;10(1):66. doi: 10.3390/biomimetics10010066.
The surface topography and chemistry of titanium-aluminum-vanadium (Ti6Al4V) implants play critical roles in the osteoblast differentiation of human bone marrow stromal cells (MSCs) and the creation of an osteogenic microenvironment. To assess the effects of a microscale/nanoscale (MN) topography, this study compared the effects of MN-modified, anodized, and smooth Ti6Al4V surfaces on MSC response, and for the first time, directly contrasted MN-induced osteoblast differentiation with culture on tissue culture polystyrene (TCPS) in osteogenic medium (OM). Surface characterization revealed distinct differences in microroughness, composition, and topography among the Ti6Al4V substrates. MSCs on MN surfaces exhibited enhanced osteoblastic differentiation, evidenced by increased expression of RUNX2, SP7, BGLAP, BMP2, and BMPR1A (fold increases: 3.2, 1.8, 1.4, 1.3, and 1.2). The MN surface also induced a pro-healing inflammasome with upregulation of anti-inflammatory mediators (170-200% increase) and downregulation of pro-inflammatory factors (40-82% reduction). Integrin expression shifted towards osteoblast-associated integrins on MN surfaces. RNA-seq analysis revealed distinct gene expression profiles between MSCs on MN surfaces and those in OM, with only 199 shared genes out of over 1000 differentially expressed genes. Pathway analysis showed that MN surfaces promoted bone formation, maturation, and remodeling through non-canonical Wnt signaling, while OM stimulated endochondral bone development and mineralization via canonical Wnt3a signaling. These findings highlight the importance of Ti6Al4V surface properties in directing MSC differentiation and indicate that MN-modified surfaces act via signaling pathways that differ from OM culture methods, more accurately mimicking peri-implant osteogenesis in vivo.
钛铝钒（Ti6Al4V）植入物的表面形貌和化学性质在人骨髓基质细胞（MSCs）的成骨细胞分化以及成骨微环境的形成中起着关键作用。为了评估微米级/纳米级（MN）形貌的影响，本研究比较了MN修饰、阳极氧化和光滑Ti6Al4V表面对MSCs反应的影响，并首次将MN诱导的成骨细胞分化与在成骨培养基（OM）中于组织培养聚苯乙烯（TCPS）上培养进行了直接对比。表面表征揭示了Ti6Al4V基底在微观粗糙度、成分和形貌上的明显差异。MN表面上的MSCs表现出增强的成骨细胞分化，这通过RUNX2、SP7、BGLAP、BMP2和BMPR1A表达的增加得以证明（倍数增加：3.2、1.8、1.4、1.3和1.2）。MN表面还诱导了一种促愈合的炎性小体，抗炎介质上调（增加170 - 200%），促炎因子下调（减少40 - 82%）。整合素表达在MN表面向与成骨细胞相关的整合素转变。RNA测序分析揭示了MN表面上的MSCs与OM中的MSCs之间不同的基因表达谱，在超过1000个差异表达基因中只有199个共享基因。通路分析表明，MN表面通过非经典Wnt信号促进骨形成、成熟和重塑，而OM通过经典Wnt3a信号刺激软骨内骨发育和矿化。这些发现突出了Ti6Al4V表面性质在指导MSCs分化中的重要性，并表明MN修饰的表面通过与OM培养方法不同的信号通路起作用，更准确地模拟了体内植入物周围的成骨过程。
