Nokhbatolfoghahaei Hanieh, Baniameri Sahar, Tabrizi Reza, Yousefi-Koma Amir-Ali, Dehghan Mohammad Mehdi, Derakhshan Samira, Gharehdaghi Niusha, Farzad-Mohajeri Saeed, Behroozibakhsh Marjan, Khojasteh Arash
Dental Research Center, School of Dentistry, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Research Assistance, School of Dentistry, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
In Vitro Cell Dev Biol Anim. 2025 Jan;61(1):67-80. doi: 10.1007/s11626-024-00973-5. Epub 2024 Oct 9.
Vascularization is vital in bone tissue engineering, supporting development, remodeling, and regeneration. Lack of vascularity leads to cell death, necessitating vascularization strategies. Angiogenesis, forming new blood vessels, provides crucial nutrients and oxygen. Pre-vascularized gelatin-coated β-tricalcium phosphate (G/β-TCP) scaffolds show promise in bone regeneration and vascularization. Our study evaluates G/β-TCP scaffolds' osteogenic and angiogenic potential in vitro and a canine model with vascular anastomosis. Channel-shaped G/β-TCP scaffolds were fabricated using foam casting and sintering of a calcium phosphate/silica slurry-coated polyurethane foam, then coated with cross-linked gelatin. Buccal fat pad-derived stem cells (BFPdSCs) were seeded onto scaffolds and assessed over time for adhesion, proliferation, and osteogenic capacity using scanning electron microscopy (SEM), 4,6-diamidino-2-phenylindole (DAPI) staining, Alamar blue, and alkaline phosphatase (ALP) assays. Scaffolds were implanted in a canine model to evaluate osteogenesis and angiogenesis by histology and CT scans at 12 wk. Our studies showed preliminary results for G/β-TCP scaffolds supporting angiogenesis and bone regeneration. In vitro analyses demonstrated excellent proliferation/viability, with BFPdSCs adhering and increasing on the scaffolds. ALP activity and protein levels increased, indicating osteogenic differentiation. Examination of tissue samples revealed granulation tissue with a well-developed vascular network, indicating successful angiogenesis and osteogenesis was further confirmed by a CT scan. In vivo, histology revealed scaffold resorption. However, scaffold placement beneath muscle tissue-restricted bone regeneration. Further optimization is needed for bone regeneration applications.
血管化在骨组织工程中至关重要，它支持骨的发育、重塑和再生。血管缺乏会导致细胞死亡，因此需要血管化策略。血管生成，即形成新的血管，能提供关键的营养物质和氧气。预血管化的明胶包被的β-磷酸三钙（G/β-TCP）支架在骨再生和血管化方面显示出前景。我们的研究评估了G/β-TCP支架在体外以及在具有血管吻合的犬模型中的成骨和血管生成潜力。采用泡沫浇铸法，通过对涂覆有磷酸钙/二氧化硅浆料的聚氨酯泡沫进行烧结来制备通道状G/β-TCP支架，然后用交联明胶进行包被。将颊脂垫来源的干细胞（BFPdSCs）接种到支架上，并随着时间的推移，使用扫描电子显微镜（SEM）、4,6-二脒基-2-苯基吲哚（DAPI）染色、阿拉玛蓝和碱性磷酸酶（ALP）测定法评估细胞的黏附、增殖和成骨能力。将支架植入犬模型中，在12周时通过组织学和CT扫描评估骨生成和血管生成情况。我们的研究显示了G/β-TCP支架在支持血管生成和骨再生方面的初步结果。体外分析表明细胞具有出色的增殖/活力，BFPdSCs在支架上黏附并增殖。ALP活性和蛋白水平升高，表明细胞发生了成骨分化。对组织样本的检查显示有肉芽组织和发育良好的血管网络，表明血管生成成功，CT扫描进一步证实了骨生成。在体内，组织学检查显示支架发生了吸收。然而，将支架放置在肌肉组织下方会限制骨再生。骨再生应用还需要进一步优化。
