Gonçalves Flavia, Letomai Roberta Molisani, Gomes Marjory Muraro, Remédios Aguiar Araújo Maria Dos, Muniz Yasmin Silva, Moreira Maria Stella, Boaro Leticia Cidreira
Faculdade de Odontologia, Universidade Santo Amaro, Av. Prof. Eneas de Siqueira Neto, 340, São Paulo 04829-300, SP, Brazil.
Departamento de Estomatologia, Hospital AC Camargo, São Paulo 01509-010, SP, Brazil.
Bioengineering (Basel). 2025 Jan 30;12(2):130. doi: 10.3390/bioengineering12020130.
The addition of dexamethasone in membranes for guided bone regeneration is promising due to its dual effect: (1) anti-inflammatory action and (2) induction of osteogenesis in host stem cells. Electrospun fiber coating with dexamethasone using the layer-by-layer (LBL) technique offers an interesting alternative for the gradual release of the drug, aiming for enhanced osteodifferentiation activity. This study aimed to develop synthetic poly-L-lactide (PLLA) membranes with dexamethasone incorporated into the fibers or coated on their surface, and to evaluate the drug release rate, as well as the material's ability to promote proliferation, osteoconduction, and osteodifferentiation of human periodontal ligament stem cells (hPDLSCs). PLLA membranes were produced by electrospinning. Dexamethasone was incorporated using three techniques: (A) electrospinning of a co-solution of PLLA with 2.5 /% dexamethasone; (B) deposition of four layers on the PLLA membrane using alternating solutions of chitosan and heparin/dexamethasone; (C) deposition of 10 layers on the PLLA membrane using the same solutions. hPDLSC proliferation was measured via CCK-8 at 1, 7, 14, and 21 days. Cellular differentiation was assessed by alkaline phosphatase activity (7 days) and alizarin red staining (21 days) in clonogenic and osteogenic media (ODM). Data were analyzed using one or two-way ANOVA and Tukey test. Electrospun membranes with dexamethasone and those with 4 layers showed immediate drug release within 24 h, whereas 10 layers exhibited gradual release over 14 days. Cumulative drug release was higher for electrospun membranes at 1 and 7 days, similar to 10 layers at 14 and 21 days. The 4 LBL membrane promoted lower hPDLSC proliferation compared to the 10 LBL and electrospun membranes at 21 days but showed increased extracellular matrix mineralization in osteogenic media. No significant differences in alkaline phosphatase expression were observed between materials. Therefore, the addition of dexamethasone in 10 layers, combined with heparin, enables gradual drug release. However, lower drug release in the first 24 h by four LBL membranes improved the material's osteogenesis properties. None of the materials improved the osteodifferentiation in the clonogenic medium.
在引导骨再生的膜中添加地塞米松很有前景，因为它具有双重作用：（1）抗炎作用和（2）诱导宿主干细胞成骨。使用逐层（LBL）技术制备的载有地塞米松的电纺纤维涂层为药物的逐步释放提供了一种有趣的选择，旨在增强骨分化活性。本研究旨在开发将地塞米松掺入纤维中或涂覆在其表面的合成聚-L-乳酸（PLLA）膜，并评估药物释放速率以及该材料促进人牙周膜干细胞（hPDLSCs）增殖、骨传导和骨分化的能力。PLLA膜通过静电纺丝制备。地塞米松采用三种技术掺入：（A）将PLLA与2.5%地塞米松的共溶液进行静电纺丝；（B）使用壳聚糖和肝素/地塞米松的交替溶液在PLLA膜上沉积四层；（C）使用相同溶液在PLLA膜上沉积十层。在第1、7、14和21天通过CCK-8测量hPDLSC增殖。在克隆形成和成骨培养基（ODM）中，通过碱性磷酸酶活性（7天）和茜素红染色（21天）评估细胞分化。数据使用单因素或双因素方差分析和Tukey检验进行分析。载有地塞米松的电纺膜和四层膜在24小时内显示出药物立即释放，而十层膜在14天内呈现逐步释放。在第1天和第7天，电纺膜的累积药物释放较高，在第14天和第21天与十层膜相似。在第21天，与十层LBL膜和电纺膜相比，四层LBL膜促进的hPDLSC增殖较低，但在成骨培养基中显示出细胞外基质矿化增加。各材料之间未观察到碱性磷酸酶表达的显著差异。因此，十层中添加地塞米松并与肝素结合可实现药物的逐步释放。然而，四层LBL膜在前24小时内较低的药物释放改善了材料的成骨特性。没有一种材料能改善克隆形成培养基中的骨分化。
