Renaud Matthieu, Bousquet Philippe, Macias Gerard, Rochefort Gael Y, Durand Jean-Olivier, Marsal Lluis F, Cuisinier Frédéric, Cunin Frédérique, Collart-Dutilleul Pierre-Yves
Laboratoire Biosanté et Nanoscience (LBN), Université Montpellier, 34000 Montpellier, France.
Faculty of Dentistry, Université de Tours, 37000 Tours, France.
Bioengineering (Basel). 2023 Jul 19;10(7):852. doi: 10.3390/bioengineering10070852.
To date, bone regeneration techniques use many biomaterials for bone grafting with limited efficiencies. For this purpose, tissue engineering combining biomaterials and stem cells is an important avenue of development to improve bone regeneration. Among potentially usable non-toxic and bioresorbable scaffolds, porous silicon (pSi) is an interesting biomaterial for bone engineering. The possibility of modifying its surface can allow a better cellular adhesion as well as a control of its rate of resorption. Moreover, release of silicic acid upon resorption of its nanostructure has been previously proved to enhance stem cell osteodifferentiation by inducing calcium phosphate formation. In the present study, we used a rat tail model to experiment bone tissue engineering with a critical size defect. Two groups with five rats per group of male Wistar rats were used. In each rat, four vertebrae were used for biomaterial implantation. Randomized bone defects were filled with pSi particles alone or pSi particles carrying dental pulp stem cells (DPSC). Regeneration was evaluated in comparison to empty defect and defects filled with xenogenic bone substitute (Bio-Oss). Fluorescence microscopy and SEM evaluations showed adhesion of DPSCs on pSi particles with cells exhibiting distribution throughout the biomaterial. Histological analyzes revealed the formation of a collagen network when the defects were filled with pSi, unlike the positive control using Bio-Oss. Overall bone formation was objectivated with µCT analysis and showed a higher bone mineral density with pSi particles combining DPSC. Immunohistochemical assays confirmed the increased expression of bone markers (osteocalcin) when pSi particles carried DPSC. Surprisingly, no grafted cells remained in the regenerated area after one month of healing, even though the grafting of DPSC clearly increased bone regeneration for both bone marker expression and overall bone formation objectivated with µCT. In conclusion, our results show that the association of pSi with DPSCs in vivo leads to greater bone formation, compared to a pSi graft without DPSCs. Our results highlight the paracrine role of grafted stem cells by recruitment and stimulation of endogenous cells.
迄今为止,骨再生技术使用多种生物材料进行骨移植,但效率有限。为此,将生物材料与干细胞相结合的组织工程是改善骨再生的重要发展途径。在潜在可用的无毒且可生物吸收的支架中,多孔硅(pSi)是一种用于骨工程的有趣生物材料。对其表面进行修饰的可能性可以实现更好的细胞黏附,并控制其吸收速率。此外,先前已证明其纳米结构吸收时硅酸的释放可通过诱导磷酸钙形成来增强干细胞的成骨分化。在本研究中,我们使用大鼠尾巴模型对具有临界尺寸缺损的骨组织工程进行实验。使用两组雄性Wistar大鼠,每组五只。在每只大鼠中,使用四个椎骨进行生物材料植入。随机的骨缺损单独用pSi颗粒或携带牙髓干细胞(DPSC)的pSi颗粒填充。与空白缺损和填充异种骨替代物(Bio-Oss)的缺损相比,对再生情况进行评估。荧光显微镜和扫描电子显微镜评估显示DPSC黏附在pSi颗粒上,细胞在整个生物材料中呈分布状态。组织学分析显示,当缺损用pSi填充时会形成胶原网络,这与使用Bio-Oss的阳性对照不同。通过微计算机断层扫描(µCT)分析客观评估了整体骨形成情况,结果显示结合DPSC的pSi颗粒具有更高的骨矿物质密度。免疫组织化学分析证实,当pSi颗粒携带DPSC时,骨标志物(骨钙素)的表达增加。令人惊讶的是,愈合一个月后,再生区域中没有残留移植细胞,尽管DPSC的移植明显增加了骨标志物表达和µCT客观评估的整体骨形成方面的骨再生。总之,我们的结果表明,与未添加DPSC的pSi移植相比,体内pSi与DPSC的联合可导致更大的骨形成。我们的结果突出了移植干细胞通过募集和刺激内源性细胞发挥的旁分泌作用。