Bou Assaf Rita, Zibara Kazem, Fayyad-Kazan Mohammad, Al-Nemer Fatima, Cordahi Manal, Khairallah Saad, Badran Bassam, Berbéri Antoine
Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Lebanese University, Beirut, Lebanon.
Laboratory of Cancer Biology and Molecular Immunology, Faculty of Sciences-I, Lebanese University, Beirut, Lebanon.
Stem Cells Int. 2019 Sep 30;2019:4185942. doi: 10.1155/2019/4185942. eCollection 2019.
Repairing bone defects, especially in older individuals with limited regenerative capacity, is still a big challenge. The use of biomimetic materials that can enhance the restoration of bone structure represents a promising clinical approach. In this study, we evaluated ectopic bone formation after the transplantation of human maxillary Schneiderian sinus membrane- (hMSSM-) derived cells embedded within various scaffolds in the femur of pigs.
The scaffolds used were collagen, gelatin, and hydroxyapatite/tricalcium phosphate (HA/TCP) where fibrin/thrombin was used as a control. Histological analysis was performed for the new bone formation. Quantitative real-time PCR (qRT-PCR) and immunohistochemistry (IHC) were used to assess mRNA and protein levels of specific osteoblastic markers, respectively.
Histological analysis showed that the three scaffolds we used can support new bone formation with a more pronounced effect observed in the case of the gelatin scaffold. In addition, mRNA levels of the different tested osteoblastic markers Runt-Related Transcription Factor 2 (RUNX-2), osteonectin (ON), osteocalcin (OCN), osteopontin (OPN), alkaline phosphatase (ALP), and type 1 collagen (COL1) were higher, after 2 and 4 weeks, in cell-embedded scaffolds than in control cells seeded within the fibrin/thrombin scaffold. Moreover, there was a very clear and differential expression of RUNX-2, OCN, and vimentin in osteocytes, osteoblasts, hMSSM-derived cells, and bone matrix. Interestingly, the osteogenic markers were more abundant, at both time points, in cell-embedded gelatin scaffold than in other scaffolds (collagen, HA/TCP, fibrin/thrombin).
These results hold promise for the development of successful bone regeneration techniques using different scaffolds embedded with hMSSM-derived cells. This trial is registered with NCT02676921.
修复骨缺损,尤其是对于再生能力有限的老年人而言,仍然是一项巨大的挑战。使用能够促进骨结构恢复的仿生材料是一种很有前景的临床方法。在本研究中,我们评估了嵌入各种支架中的人上颌窦 Schneiderian 膜(hMSSM)来源的细胞移植到猪股骨后异位骨的形成情况。
使用的支架有胶原蛋白、明胶和羟基磷灰石/磷酸三钙(HA/TCP),其中纤维蛋白/凝血酶用作对照。对新骨形成进行组织学分析。分别使用定量实时 PCR(qRT-PCR)和免疫组织化学(IHC)来评估特定成骨标志物的 mRNA 和蛋白质水平。
组织学分析表明,我们使用的三种支架都能支持新骨形成,明胶支架的效果更为显著。此外,在细胞嵌入支架中,不同测试的成骨标志物,即 Runt 相关转录因子 2(RUNX-2)、骨连接蛋白(ON)、骨钙素(OCN)、骨桥蛋白(OPN)、碱性磷酸酶(ALP)和 1 型胶原蛋白(COL1)的 mRNA 水平在 2 周和 4 周后高于接种在纤维蛋白/凝血酶支架中的对照细胞。此外,RUNX-2、OCN 和波形蛋白在骨细胞、成骨细胞、hMSSM 来源的细胞和骨基质中有非常明显的差异表达。有趣的是,在两个时间点,细胞嵌入明胶支架中的成骨标志物比其他支架(胶原蛋白、HA/TCP、纤维蛋白/凝血酶)更丰富。
这些结果为使用嵌入 hMSSM 来源细胞的不同支架开发成功的骨再生技术带来了希望。该试验已在 NCT02676921 注册。