Allogenic demineralized bone matrix has been developed as a reliable alternative to the autologous bone graft. In the present study, we assessed the osteoformation potential of a partially demineralized bone matrix (PDBM) in a paste form obtained without an added carrier. This formulation included the preparation of cancelous bone from femoral heads after decellularision, delipidation, demineralization in HCl and autoclaving at 121 °C. Structural and biochemical characteristics of PDBM were determined using FTIR (Fourier transform infrared spectroscopy), hydroxyproline, DNA content assays, and optical ellipsometry. The osteoformation potential was evaluated in 8-, 6-, and 4-mm-diameter rat-calvarial bone defects by in vivo micro-CT analysis, performed immediately after surgery on days 0, 15, 30, 45, and 60. Moreover, histological and histomorphometric analyses were done on day 60. PDBM was compared to cancelous bone powder (BP) before its partial demineralization. The expression levels of selected inflammation-, angiogenesis-, and bone-related genes were also investigated by RT-PCR, 3, 7, and 14 days after surgery. Compared to the control group, the PDBM group exhibited a significant increase (p < 0.05) in radiopacity in 8-mm- and 6-mm-diameter defects at all time points tested. On day 60, the amount of newly-formed bone was greater (16 and 1.6 folds; p < 0.001; respectively) compared to that in control defects. No bone formation was observed in defects filled with BP regardeless of the size. In 8-mm-diameter defect, PDBM was effective enough to induce the upregulation of genes pertinent to inflammation (i.e., TNFα, IL-6, and IL-8), angiogenesis (i.e., VEGF, VWF), and osteogenesis (ALP, RUNX2, BGLAP, SP7) by day 3 after surgery. This study showed that the tested PDBM deeply influences the early critical events involved in bone regeneration and exhibits efficient osteoformation capacity, making it an attractive graft option for treating defects in periodontal and maxillofacial areas.

同种异体脱矿骨基质已被开发为自体骨移植的可靠替代品。在本研究中，我们评估了在未添加载体的情况下获得的糊状部分脱矿骨基质 (PDBM) 的骨形成潜力。该制剂包括在脱细胞、脱脂、在 HCl 中脱矿质和在 121 °C 高压灭菌后从股骨头制备松质骨。使用 FTIR（傅立叶变换红外光谱）、羟脯氨酸、DNA 含量测定和光学椭偏仪测定 PDBM 的结构和生化特征。在手术后第 0、15、30、45 和 60 天通过体内显微 CT 分析评估 8、6 和 4 毫米直径大鼠颅骨骨缺损的骨形成潜力。此外，在第 60 天进行组织学和组织形态计量学分析。将 PDBM 与其部分脱矿质前的松质骨粉 (BP) 进行比较。在手术后 3、7 和 14 天，还通过 RT-PCR 研究了选定的炎症、血管生成和骨相关基因的表达水平。与对照组相比，PDBM组表现出显着增加（p  < 0.05) 在所有测试时间点的 8 毫米和 6 毫米直径缺陷的射线不透性。在第 60 天，与 对照缺损相比，新形成的骨量更大（分别为 16 倍和 1.6 倍；p < 0.001；）。无论大小如何，在填充有 BP 的缺损中均未观察到骨形成。在 8 毫米直径的缺损中，PDBM 足以有效诱导与炎症相关的基因（即 TNFα、IL-6 和 IL-8）、血管生成（即 VEGF、VWF）和成骨（ALP、 RUNX2、BGLAP、SP7）在手术后第 3 天。这项研究表明，测试的 PDBM 深深影响了骨再生中涉及的早期关键事件，并表现出有效的骨形成能力，使其成为治疗牙周和颌面部缺损的有吸引力的移植选择。