This study addresses the fabrication of an extracellular matrix material of the acellular sheep periosteum and the systematic evaluation of its biocompatibility to explore its potential application in guided bone regeneration. Sheep periosteum was harvested and decellularized by a combined decellularization protocol. The effectiveness of cell removal was proved and residual α-Gal antigen was also quantitatively detected. Then, mouse MC3T3-E1 cells were seeded onto the acellular periosteum. A scanning electron microscope (SEM) was used to record the whole process of cell adhesion. The CCK-8 assay suggested that the acellular periosteum not only had zero toxic effect on pre-osteoblasts, but played a positive role in cell proliferation. We also tested whether the acellular periosteum possesses favorable osteogenesis induction activity using an alkaline phosphatase (ALP) assay and a quantitative real-time PCR (Col I, Runx2, OCN) assay. An in vivo study of a subcutaneous implantation test using Sprague Dawley (SD) rats was performed to detect the changes in IL-2, IFN-γ and IL-4 in serum and elucidate the host's local response to acellular periosteum through hematoxylin and eosin (HE) and immunohistochemical staining. The results show that acellular sheep periosteum did not elicit a severe immunogenic response via the Th1 pathway, unlike fresh sheep periosteum. In conclusion, acellular sheep periosteum possesses favorable biocompatibility to be employed for guided bone regeneration.

中文翻译：

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脱细胞绵羊骨膜用于引导骨再生的体外和体内生物相容性研究。

这项研究解决了无细胞绵羊骨膜细胞外基质材料的制造及其生物相容性的系统评价，以探索其在引导性骨再生中的潜在应用。收获绵羊骨膜并通过联合脱细胞方案脱细胞。证明了去除细胞的有效性，并且还定量检测了残留的α-Gal抗原。然后，将小鼠MC3T3-E1细胞接种到无细胞骨膜上。使用扫描电子显微镜（SEM）记录细胞粘附的整个过程。CCK-8分析表明，无细胞的骨膜不仅对成骨细胞具有零毒性作用，而且在细胞增殖中起积极作用。我们还使用碱性磷酸酶（ALP）检测和实时定量PCR（Col I，Runx2，OCN）检测来测试无细胞骨膜是否具有良好的成骨诱导活性。使用Sprague Dawley（SD）大鼠进行的皮下植入测试的体内研究旨在检测血清中IL-2，IFN-γ和IL-4的变化，并阐明宿主对苏木精和曙红对脱细胞骨膜的局部反应（ HE）和免疫组化染色。结果表明，与新鲜绵羊骨膜不同，脱细胞绵羊骨膜未通过Th1途径引起严重的免疫原性应答。总之，脱细胞绵羊骨膜具有良好的生物相容性，可用于引导骨再生。使用Sprague Dawley（SD）大鼠进行的皮下植入测试的体内研究旨在检测血清中IL-2，IFN-γ和IL-4的变化，并阐明宿主对苏木精和曙红对脱细胞骨膜的局部反应（ HE）和免疫组化染色。结果表明，与新鲜绵羊骨膜不同，脱细胞绵羊骨膜未通过Th1途径引起严重的免疫原性应答。总之，脱细胞绵羊骨膜具有良好的生物相容性，可用于引导骨再生。使用Sprague Dawley（SD）大鼠进行的皮下植入测试的体内研究旨在检测血清中IL-2，IFN-γ和IL-4的变化，并阐明宿主对苏木精和曙红对脱细胞骨膜的局部反应（ HE）和免疫组化染色。结果表明，与新鲜绵羊骨膜不同，脱细胞绵羊骨膜未通过Th1途径引起严重的免疫原性应答。总之，脱细胞绵羊骨膜具有良好的生物相容性，可用于引导骨再生。结果表明，与新鲜绵羊骨膜不同，脱细胞绵羊骨膜未通过Th1途径引起严重的免疫原性应答。总之，脱细胞绵羊骨膜具有良好的生物相容性，可用于引导骨再生。结果表明，与新鲜绵羊骨膜不同，脱细胞绵羊骨膜未通过Th1途径引起严重的免疫原性应答。总之，脱细胞绵羊骨膜具有良好的生物相容性，可用于引导骨再生。