Department of Oral Biology, University Clinic of Dentistry, Medical University of Vienna, 1090 Vienna, Austria.
Core Facility Proteomics, Medical University of Vienna, 1090 Vienna, Austria.
Int J Mol Sci. 2023 Nov 10;24(22):16181. doi: 10.3390/ijms242216181.
Bone allografts are widely used as osteoconductive support to guide bone regrowth. Bone allografts are more than a scaffold for the immigrating cells as they maintain some bioactivity of the original bone matrix. Yet, it remains unclear how immigrating cells respond to bone allografts. To this end, we have evaluated the response of mesenchymal cells exposed to acid lysates of bone allografts (ALBA). RNAseq revealed that ALBA has a strong impact on the genetic signature of gingival fibroblasts, indicated by the increased expression of IL11, AREG, C11orf96, STC1, and GK-as confirmed by RT-PCR, and for IL11 and STC1 by immunoassays. Considering that transforming growth factor-β (TGF-β) is stored in the bone matrix and may have caused the expression changes, we performed a proteomics analysis, TGF-β immunoassay, and smad2/3 nuclear translocation. ALBA neither showed detectable TGF-β nor was the lysate able to induce smad2/3 translocation. Nevertheless, the TGF-β receptor type I kinase inhibitor SB431542 significantly decreased the expression of IL11, AREG, and C11orf96, suggesting that other agonists than TGF-β are responsible for the robust cell response. The findings suggest that IL11, AREG, and C11orf96 expression in mesenchymal cells can serve as a bioassay reflecting the bioactivity of the bone allografts.
骨同种异体移植物被广泛用作引导骨再生的骨传导性支撑物。骨同种异体移植物不仅仅是细胞迁移的支架，因为它们还保持了原始骨基质的一些生物活性。然而，目前尚不清楚迁移细胞如何对骨同种异体移植物作出反应。为此，我们评估了暴露于骨同种异体移植物酸裂解物（ALBA）的间充质细胞的反应。RNAseq 显示，ALBA 对牙龈成纤维细胞的遗传特征有很强的影响，这表现在 IL11、AREG、C11orf96、STC1 和 GK 的表达增加，这一点通过 RT-PCR 和免疫测定法得到了证实，而 IL11 和 STC1 则通过免疫测定法得到了证实。考虑到转化生长因子-β（TGF-β）储存在骨基质中，并可能导致表达变化，我们进行了蛋白质组学分析、TGF-β免疫测定和 smad2/3 核易位。ALBA 既没有显示出可检测到的 TGF-β，也没有使裂解物能够诱导 smad2/3 易位。尽管如此，TGF-β 受体 I 型激酶抑制剂 SB431542 显著降低了 IL11、AREG 和 C11orf96 的表达，表明除 TGF-β 以外的其他激动剂是导致细胞强烈反应的原因。这些发现表明，间充质细胞中 IL11、AREG 和 C11orf96 的表达可以作为反映骨同种异体移植物生物活性的生物测定。
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