背景:本研究旨在合成藻酸盐表面改性的牙本质粉末,牙髓再生的潜在物质,并评价其对体外培养人牙髓干细胞(hDPSCs)活力和增殖的影响及其体内生物相容性。
方法:在体外阶段,牙本质粉以三个尺寸组(150-250μm,250-500μm,和500-1000μm)在去矿质和去端化后,用于去除牙本质胶原端肽并消除宿主免疫反应。用藻酸盐进行表面改性,然后进行场发射扫描电子显微镜(FE-SEM),能量色散X射线光谱(EDX),以及用研究的hDPSC进行14天的细胞活力和增殖测试。在体内阶段,将牙本质粉末植入大鼠颅骨缺损8周,并进行组织学分析。所有非参数数据都用Kruskal-Wallis检验进行分析,所有定量数据采用SPSS单因素方差分析,P<0.05被认为具有统计学意义。
结果:去矿质和去端化在所有组均成功。所有组的细胞活力最佳且相等(P>0.05)。500-1000μm组细胞增殖明显增高(P<0.05)。组织学评估显示所有组的生物相容性均可接受;250-500和500-1000组的血管生成评分均明显更高,500-1000μm组的炎症反应最小。该组新形成的骨量高于其他组。
结论:藻酸盐对脱矿质和去化牙本质粉进行表面修饰,增强了表面物理性质和细胞增殖,同时显示出良好的组织内生物相容性,降低了宿主免疫反应。这些发现为牙本质-牙髓复合物的再生提供了希望。
BACKGROUND: This study aimed to synthesize dentin powder surface modified with alginate, a potential substance for dental pulp regeneration, and evaluate its effects on the viability and proliferation of human dental pulp stem cells in vitro and its biocompatibility in vivo.
METHODS: In the in vitro phase, dentin powder was synthesized in 3 size groups (150-250 μm, 250-500 μm, and 500-1000 μm) after demineralization and atelopeptidization which is used to remove dentin collagen telopeptides and eliminate host immune response. Surface modification with alginate was performed and followed by field-emission scanning electron microscopy, energy dispersive X-ray spectroscopy, and cell viability and proliferation testing for 14 days with human dental pulp stem cells studied. In the in vivo phase, dentin powders were implanted in rat calvarial defects for 8 weeks, and histologic analysis was conducted. All nonparametric data were analyzed with the Kruskal-Wallis test, and all the quantitative data were analyzed by 1-way analysis of variance using SPSS, and P < .05 was considered statistically significant.
RESULTS: Demineralization and atelopeptidization were successful in all groups. Cell viability was optimal and equal (P > .05) in all groups. The 500- to 1000-μm group exhibited significantly higher cell proliferation (P < .05). Histologic assessment shows acceptable biocompatibility in all groups; the angiogenesis score was significantly greater in both 250-500 and 500-1000, and minimal inflammatory response was noted in the 500- to 1000-μm group, and the amount of newly formed bone in this group was higher than other groups.
CONCLUSIONS: Surface modification of demineralized and atelopeptidized dentin powder with alginate enhanced surface physical properties and cell proliferation while showing great biocompatibility within tissue and reducing the host immune response. These findings hold promise for dentin-pulp complex regeneration.