University Laboratory Animal Resources, The Ohio State University, Columbus, Ohio, United States of America.
Department of Biomedical Engineering, The Ohio State University, Columbus, Ohio, United States of America.
PLoS One. 2022 Dec 30;17(12):e0279519. doi: 10.1371/journal.pone.0279519. eCollection 2022.
Segmental bone defects present complex clinical challenges. Nonunion, malunion, and infection are common sequalae of autogenous bone grafts, allografts, and synthetic bone implants due to poor incorporation with the patient's bone. The current project explores the osteogenic properties of periosteum to facilitate graft incorporation. As tissue area is a natural limitation of autografting, mechanical strain was implemented to expand the periosteum. Freshly harvested, porcine periosteum was strained at 5 and 10% per day for 10 days with non-strained and free-floating samples serving as controls. Total tissue size, viability and histologic examination revealed that strain increased area to a maximum of 1.6-fold in the 10% daily strain. No change in tissue anatomy or viability via MTT or Ki67 staining and quantification was observed among groups. The osteogenic potential of the mechanical expanded periosteum was then examined in vivo. Human cancellous allografts were wrapped with 10% per day strained, fresh, free-floating, or no porcine periosteum and implanted subcutaneously into female, athymic mice. Tissue was collected at 8- and 16-weeks. Gene expression analysis revealed a significant increase in alkaline phosphatase and osteocalcin in the fresh periosteum group at 8-weeks post implantation compared to all other groups. Values among all groups were similar at week 16. Additionally, histological assessment with H&E and Masson-Goldner Trichrome staining showed that all periosteal groups outperformed the non-periosteal allograft, with fresh periosteum demonstrating the highest levels of new tissue mineralization at the periosteum-bone interface. Overall, mechanical expansion of the periosteum can provide increased area for segmental healing via autograft strategies, though further studies are needed to explore culture methodology to optimize osteogenic potential.
节段性骨缺损带来了复杂的临床挑战。由于自体骨移植物、同种异体骨和合成骨植入物与患者骨骼的结合不良，非愈合、畸形愈合和感染是常见的后遗症。本项目探讨了骨膜的成骨特性，以促进移植物的结合。由于组织面积是自体移植的自然限制，因此实施了机械应变以扩展骨膜。新鲜收获的猪骨膜每天以 5%和 10%的应变率拉伸 10 天，未受应变和自由漂浮的样本作为对照。总组织大小、活力和组织学检查显示，应变使 10%日应变量下的面积增加到最大 1.6 倍。各组之间通过 MTT 或 Ki67 染色和定量观察到组织解剖结构或活力没有变化。然后在体内检查机械扩展骨膜的成骨潜力。将人松质同种异体移植物包裹在每天应变 10%的新鲜、自由漂浮或无猪骨膜中，并植入雌性无胸腺小鼠的皮下。在 8 周和 16 周时收集组织。基因表达分析显示，与所有其他组相比，植入后 8 周时新鲜骨膜组碱性磷酸酶和骨钙素的表达显著增加。所有组在第 16 周时的数值相似。此外，用 H&E 和 Masson-Goldner 三色染色进行组织学评估显示，所有骨膜组的表现均优于无骨膜的同种异体移植物，新鲜骨膜在骨膜-骨界面显示出最高水平的新组织矿化。总的来说，骨膜的机械扩张可以通过自体移植策略为节段性愈合提供更大的面积，尽管需要进一步的研究来探索培养方法以优化成骨潜力。
