Osteoformation potential of an allogenic partially demineralized bone matrix in critical-size defects in the rat calvarium
Osteoformation potential of an allogenic partially demineralized bone matrix in critical-size defects in the rat calvarium
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DOI：
10.1016/j.msec.2021.112207
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发表时间：
2021-06-11
影响因子：
7.9
通讯作者：
Anagnostou, Fani
中科院分区：
文献类型：
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作者：
Diallo, Ahmad Moustapha;Rota, Solene;Anagnostou, Fani
同种异体脱钙骨基质已被开发为自体骨移植物的可靠替代品。在本研究中，我们评估了部分脱矿骨基质（PDBM）的骨形成潜力，该基质是在没有添加载体的情况下获得的糊状物。该制剂包括在脱细胞、脱脂、HCl脱矿和121 ℃高压灭菌后从股骨头制备松质骨。采用傅里叶变换红外光谱、羟脯氨酸、DNA含量测定和椭圆偏振法对PDBM的结构和生化特性进行了测定。通过体内显微CT分析，在术后第0、15、30、45和60天立即对直径为8、6和4 mm的大鼠颅骨骨缺损进行成骨潜力评价。此外，在第60天进行组织学和组织形态计量学分析。将PDBM与部分脱矿前的松质骨粉（BP）进行比较。选择炎症，血管生成和骨相关基因的表达水平也进行了研究，通过RT-PCR，3，7和14天后手术。与对照组相比，PDBM组在所有测试时间点的8 mm和6 mm直径缺损的不透射线性均显著增加（p < 0.05）。在第60天，与对照缺损相比，新形成的骨的量更大（分别为16和1.6倍; p < 0.001）。BP填充的缺损无骨形成。在8 mm直径的缺损中，PDBM足以有效诱导与炎症相关的基因上调（即，TNF α、IL-6和IL-8）、血管生成（即，VEGF、VWF）和骨生成（ALP、RUNX 2、BG 3、SP 7）。这项研究表明，测试的PDBM深刻影响了骨再生中涉及的早期关键事件，并表现出有效的成骨能力，使其成为治疗牙周和颌面部缺损的有吸引力的移植选择。
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 degrees 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., TNF alpha, 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.