Abstract：
OBJECTIVE: To synthesize casein enzymatic hydrolysate (CEH)-laden gelatin methacryloyl (GelMA) fibrous scaffolds and evaluate the cytocompatibility and anti-inflammatory effects on dental pulp stem cells (DPSCs).
METHODS: GelMA fibrous scaffolds with 10%, 20%, and 30% CEH (w/w) and without CEH (control) were obtained via electrospinning. Chemo-morphological, degradation, and mechanical analyses were conducted to evaluate the morphology and composition of the fibers, mass loss, and mechanical properties, respectively. Adhesion/spreading and viability of DPSCs seeded on the scaffolds were also assessed. The anti-inflammatory potential on DPSCs was tested after the chronic challenge of cells with lipopolysaccharides (LPS), followed by treatment with extracts obtained after immersing the scaffolds in α-MEM. The synthesis of the pro-inflammatory cytokines IL-6, IL-1α, and TNF-α was measured by ELISA. Data were analyzed by ANOVA/post-hoc tests (α = 5%).
RESULTS: CEH-laden electrospun fibers had a larger diameter than pure GelMA (p ≤ 0.036). GelMA scaffolds laden with 20% and 30% CEH had a greater mass loss. Tensile strength was reduced for the 10% CEH fibers (p = 0.0052), whereas no difference was observed for the 20% and 30% fibers (p ≥ 0.6736) compared to the control. Young\'s modulus decreased with CEH (p < 0.0001). Elongation at break increased for the 20% and 30% CEH scaffolds (p ≤ 0.0038). Over time, DPSCs viability increased across all groups, indicating cytocompatibility, with CEH-laden scaffolds exhibiting greater cell viability after seven days (p ≤ 0.0166). Also, 10% CEH-GelMA scaffolds decreased the IL-6, IL-1α, and TNF-α synthesis (p ≤ 0.035).
CONCLUSIONS: CEH-laden GelMA scaffolds facilitated both adhesion and proliferation of DPSCs, and 10% CEH provided anti-inflammatory potential after chronic LPS challenge.
CONCLUSIONS: CEH incorporated in GelMA fibrous scaffolds demonstrated the potential to be used as a cytocompatible and anti-inflammatory biomaterial for vital pulp therapy.
METHODS: GelMA fibrous scaffolds with 10%, 20%, and 30% CEH (w/w) and without CEH (control) were obtained via electrospinning. Chemo-morphological, degradation, and mechanical analyses were conducted to evaluate the morphology and composition of the fibers, mass loss, and mechanical properties, respectively. Adhesion/spreading and viability of DPSCs seeded on the scaffolds were also assessed. The anti-inflammatory potential on DPSCs was tested after the chronic challenge of cells with lipopolysaccharides (LPS), followed by treatment with extracts obtained after immersing the scaffolds in α-MEM. The synthesis of the pro-inflammatory cytokines IL-6, IL-1α, and TNF-α was measured by ELISA. Data were analyzed by ANOVA/post-hoc tests (α = 5%).
RESULTS: CEH-laden electrospun fibers had a larger diameter than pure GelMA (p ≤ 0.036). GelMA scaffolds laden with 20% and 30% CEH had a greater mass loss. Tensile strength was reduced for the 10% CEH fibers (p = 0.0052), whereas no difference was observed for the 20% and 30% fibers (p ≥ 0.6736) compared to the control. Young\'s modulus decreased with CEH (p < 0.0001). Elongation at break increased for the 20% and 30% CEH scaffolds (p ≤ 0.0038). Over time, DPSCs viability increased across all groups, indicating cytocompatibility, with CEH-laden scaffolds exhibiting greater cell viability after seven days (p ≤ 0.0166). Also, 10% CEH-GelMA scaffolds decreased the IL-6, IL-1α, and TNF-α synthesis (p ≤ 0.035).
CONCLUSIONS: CEH-laden GelMA scaffolds facilitated both adhesion and proliferation of DPSCs, and 10% CEH provided anti-inflammatory potential after chronic LPS challenge.
CONCLUSIONS: CEH incorporated in GelMA fibrous scaffolds demonstrated the potential to be used as a cytocompatible and anti-inflammatory biomaterial for vital pulp therapy.
摘要：
目的：合成含酪蛋白酶解产物（CEH）的明胶甲基丙烯酰（GelMA）纤维支架，并评价其对牙髓干细胞（DPSCs）的细胞相容性和抗炎作用。
方法：含有10%的GelMA纤维支架，20%,通过静电纺丝获得30%CEH(w/w)和无CEH(对照)。化学形态学，降解，并进行了力学分析，以评估纤维的形态和组成，质量损失,和机械性能，分别。还评估了接种在支架上的DPSC的粘附/铺展和活力。在用脂多糖（LPS）慢性攻击细胞后，测试了DPSC的抗炎潜力，然后用将支架浸入α-MEM后获得的提取物处理。促炎细胞因子IL-6，IL-1α的合成，用ELISA法测定TNF-α。通过ANOVA/事后检验(α=5%)分析数据。
结果:载有CEH的电纺纤维具有比纯GelMA更大的直径(p≤0.036)。载有20%和30%CEH的GelMA支架具有更大的质量损失。10%CEH纤维的拉伸强度降低(p=0.0052),而与对照相比，20％和30％纤维（p≥0.6736）没有观察到差异。杨氏模量随CEH而降低(p<0.0001)。对于20%和30%CEH支架,断裂伸长率增加(p≤0.0038)。随着时间的推移,DPSC活力在所有组增加，表明细胞相容性，载有CEH的支架在七天后表现出更大的细胞活力(p≤0.0166)。此外，10%CEH-GelMA支架降低IL-6、IL-1α、和TNF-α合成（p≤0.035）。
结论：载有CEH的GelMA支架促进DPSC的粘附和增殖，和10%CEH在慢性LPS攻击后提供抗炎潜力。
结论：在GelMA纤维支架中掺入的CEH证明了作为重要牙髓治疗的细胞相容性和抗炎生物材料的潜力。
方法：含有10%的GelMA纤维支架，20%,通过静电纺丝获得30%CEH(w/w)和无CEH(对照)。化学形态学，降解，并进行了力学分析，以评估纤维的形态和组成，质量损失,和机械性能，分别。还评估了接种在支架上的DPSC的粘附/铺展和活力。在用脂多糖（LPS）慢性攻击细胞后，测试了DPSC的抗炎潜力，然后用将支架浸入α-MEM后获得的提取物处理。促炎细胞因子IL-6，IL-1α的合成，用ELISA法测定TNF-α。通过ANOVA/事后检验(α=5%)分析数据。
结果:载有CEH的电纺纤维具有比纯GelMA更大的直径(p≤0.036)。载有20%和30%CEH的GelMA支架具有更大的质量损失。10%CEH纤维的拉伸强度降低(p=0.0052),而与对照相比，20％和30％纤维（p≥0.6736）没有观察到差异。杨氏模量随CEH而降低(p<0.0001)。对于20%和30%CEH支架,断裂伸长率增加(p≤0.0038)。随着时间的推移,DPSC活力在所有组增加，表明细胞相容性，载有CEH的支架在七天后表现出更大的细胞活力(p≤0.0166)。此外，10%CEH-GelMA支架降低IL-6、IL-1α、和TNF-α合成（p≤0.035）。
结论：载有CEH的GelMA支架促进DPSC的粘附和增殖，和10%CEH在慢性LPS攻击后提供抗炎潜力。
结论：在GelMA纤维支架中掺入的CEH证明了作为重要牙髓治疗的细胞相容性和抗炎生物材料的潜力。
