Sexton Benjamin, Han Yuanyuan, Dal-Fabbro Renan, Xu Jinping, Kaigler Darnell, Bottino Marco C
Department of Biologic and Materials Science, School of Dentistry, University of Michigan, Ann Arbor, MI, United States.
Department of Periodontics and Oral Medicine, School of Dentistry, University of Michigan, Ann Arbor, MI, United States.
Arch Oral Biol. 2024 Sep;165:106027. doi: 10.1016/j.archoralbio.2024.106027. Epub 2024 Jun 8.
This study examined how range concentrations of Fibroblast Growth Factor-2 (FGF-2) influence the differentiation and activity of human-derived periodontal ligament (hPDLSCs) and alveolar bone-derived stem cells (haBMSCs).
hPDLSCs and haBMSCs were cultured with varying concentrations of FGF-2 (0, 1, 2.5, 5, 10, 20 ng/mL) and monitored for osteogenic differentiation through alkaline phosphatase (ALP) activity and quantification of gene expression (qRT-PCR) for osteogenesis markers. Additionally, alizarin red staining and a hydroxyproline colorimetric assay evaluated and quantified osteogenic matrix mineralization and collagen deposition. Statistical analyses were performed using one-way ANOVA or two-way ANOVA for multiple comparisons between groups.
At low FGF-2 concentrations, hPDLSCs differentiated toward an osteogenic lineage, whereas higher concentrations of FGF-2 inhibited osteogenesis and promoted fibroblastic differentiation. The effect of FGF-2 at the lowest concentration tested (1 ng/mL) led to significantly higher ALP activity than osteogenically induced positive controls at early time points and equivalent RUNX2 expression at early and later time points. FGF-2 supplementation of haBMSC cultures was sufficient, at all concentrations, to increase ALP activity at an earlier time point. Mineralization of haBMSC cultures increased significantly within 5-20 ng/mL FGF-2 concentrations under basal growth media conditions (α-minimal essential medium supplemented with 15 % fetal bovine serum and 1 % penicillin/streptomycin).
FGF-2 has a dual capacity in promoting osteogenic and fibroblastic differentiation within hPDLSCs contingent upon the dosage and timing of administration, alongside supporting osteogenic differentiation in haBMSCs. These findings underscore the need for precision growth factors dosing when considering the design of biomaterials for periodontal regeneration.
本研究探讨了不同浓度的成纤维细胞生长因子-2(FGF-2)如何影响人牙周膜干细胞(hPDLSCs)和牙槽骨来源干细胞(haBMSCs)的分化和活性。
将hPDLSCs和haBMSCs与不同浓度的FGF-2(0、1、2.5、5、10、20 ng/mL)一起培养,并通过碱性磷酸酶(ALP)活性以及成骨标志物的基因表达定量(qRT-PCR)监测其成骨分化情况。此外,茜素红染色和羟脯氨酸比色法评估并定量了成骨基质矿化和胶原蛋白沉积。使用单因素方差分析或双因素方差分析进行统计分析,以对各组之间进行多重比较。
在低FGF-2浓度下,hPDLSCs向成骨谱系分化,而较高浓度的FGF-2则抑制成骨并促进成纤维细胞分化。在测试的最低浓度(1 ng/mL)下,FGF-2的作用在早期时间点导致ALP活性显著高于成骨诱导阳性对照,在早期和后期时间点RUNX2表达相当。在所有浓度下,向haBMSC培养物中添加FGF-2足以在更早的时间点增加ALP活性。在基础生长培养基条件下(补充有15%胎牛血清和1%青霉素/链霉素的α-最低必需培养基),在5-20 ng/mL FGF-2浓度范围内,haBMSC培养物的矿化显著增加。
FGF-2具有双重能力,根据给药剂量和时间促进hPDLSCs内的成骨和成纤维细胞分化,同时支持haBMSCs的成骨分化。这些发现强调了在考虑牙周再生生物材料设计时精确使用生长因子剂量的必要性。