Abstract：
OBJECTIVE: The keratinized gingiva plays an important role in maintaining healthy periodontal and peri-implant tissue. Acellular dermal matrix (ADM), as a substitute biomaterial, has a porous structure and good biocompatibility. 3D-bioprinting has the potential for tissue engineering because it enables precise loading of cells layer-by-layer. Herein, we bioprinted ADM scaffold encapsulating gingival fibroblasts (GFs) and evaluated its efficacy in keratinized gingiva augmentation in vivo to assess its potential for clinical periodontal tissue regeneration.
METHODS: GFs were extracted from the gingiva of beagles and transfected with a green fluorescent protein (GFP). The ADM scaffold (ADM cell-free group) was constructed using ADM, gelatin, and sodium alginate mixed at an appropriate ratio via 3D-bioprinting. The ADM cell scaffold (ADM cell group) was established by adding extra GFs in the same manner. Six beagles were divided into blank control, ADM cell-free, and ADM cell groups; and implant surgery was performed. The keratinized gingiva was clinically and histologically evaluated at baseline and after 2 months.
RESULTS: GFs transfected with GFPs expressed green fluorescence and were present in new tissue in the ADM cell group and not observed in the ADM cell-free group. At 2 months after surgery, the keratinized gingival augmentation in the ADM cell group was significantly more than that in the ADM cell-free group. Attached gingival augmentation was also observed more in the ADM cell group than that in the ADM cell-free group. Histological staining showed that the tissue in the ADM cell group displayed a more integrated structure and higher expression of COL I, COL III, and VEGF-A than those in the ADM cell-free group.
CONCLUSIONS: 3D-bioprinted GF-encapsulated ADM scaffolds increased the amount of keratinized gingiva in vivo, suggesting that 3D-bioprinting has great potential for oral soft tissue regeneration.
METHODS: GFs were extracted from the gingiva of beagles and transfected with a green fluorescent protein (GFP). The ADM scaffold (ADM cell-free group) was constructed using ADM, gelatin, and sodium alginate mixed at an appropriate ratio via 3D-bioprinting. The ADM cell scaffold (ADM cell group) was established by adding extra GFs in the same manner. Six beagles were divided into blank control, ADM cell-free, and ADM cell groups; and implant surgery was performed. The keratinized gingiva was clinically and histologically evaluated at baseline and after 2 months.
RESULTS: GFs transfected with GFPs expressed green fluorescence and were present in new tissue in the ADM cell group and not observed in the ADM cell-free group. At 2 months after surgery, the keratinized gingival augmentation in the ADM cell group was significantly more than that in the ADM cell-free group. Attached gingival augmentation was also observed more in the ADM cell group than that in the ADM cell-free group. Histological staining showed that the tissue in the ADM cell group displayed a more integrated structure and higher expression of COL I, COL III, and VEGF-A than those in the ADM cell-free group.
CONCLUSIONS: 3D-bioprinted GF-encapsulated ADM scaffolds increased the amount of keratinized gingiva in vivo, suggesting that 3D-bioprinting has great potential for oral soft tissue regeneration.
摘要：
目的：角化牙龈在维持健康的牙周和种植体周围组织中起着重要作用。脱细胞真皮基质(ADM),作为替代生物材料，具有多孔结构和良好的生物相容性。3D生物打印具有组织工程的潜力，因为它可以逐层精确加载细胞。在这里，我们用生物打印的ADM支架封装牙龈成纤维细胞（GFs），并评估其在角化牙龈增强体内的功效，以评估其临床牙周组织再生的潜力。
方法：从比格犬的牙龈中提取GFs，并用绿色荧光蛋白（GFP）转染。使用ADM构建ADM支架（ADM无细胞组），明胶,和海藻酸钠通过3D生物打印以适当的比例混合。通过以相同方式添加额外的GF来建立ADM细胞支架(ADM细胞组)。六只小猎犬被分为空白对照，ADM无细胞，和ADM细胞组；进行了植入手术。在基线和2个月后对角化牙龈进行临床和组织学评估。
结果：用GFP转染的GFs表达绿色荧光，并存在于ADM细胞组中的新组织中，而在无ADM细胞组中未观察到。手术后2个月,ADM细胞组的角化牙龈增加明显多于无ADM细胞组。与无ADM细胞组相比，在ADM细胞组中也观察到更多的附着牙龈增加。组织学染色显示ADM细胞组的组织呈现更完整的结构和更高的COLⅠ表达,COLIII,和VEGF-A高于无细胞ADM组。
结论：3D生物打印的GF封装的ADM支架增加了体内角化牙龈的量，这表明3D生物打印在口腔软组织再生方面具有巨大的潜力。
方法：从比格犬的牙龈中提取GFs，并用绿色荧光蛋白（GFP）转染。使用ADM构建ADM支架（ADM无细胞组），明胶,和海藻酸钠通过3D生物打印以适当的比例混合。通过以相同方式添加额外的GF来建立ADM细胞支架(ADM细胞组)。六只小猎犬被分为空白对照，ADM无细胞，和ADM细胞组；进行了植入手术。在基线和2个月后对角化牙龈进行临床和组织学评估。
结果：用GFP转染的GFs表达绿色荧光，并存在于ADM细胞组中的新组织中，而在无ADM细胞组中未观察到。手术后2个月,ADM细胞组的角化牙龈增加明显多于无ADM细胞组。与无ADM细胞组相比，在ADM细胞组中也观察到更多的附着牙龈增加。组织学染色显示ADM细胞组的组织呈现更完整的结构和更高的COLⅠ表达,COLIII,和VEGF-A高于无细胞ADM组。
结论：3D生物打印的GF封装的ADM支架增加了体内角化牙龈的量，这表明3D生物打印在口腔软组织再生方面具有巨大的潜力。
