A dual-scale micro/nanofibrous PLGA scaffold fabricated by co-electrospinning for regenerative endodontic procedures
DOI:
https://doi.org/10.1590/1678-7765-2026-0524Keywords:
Polymers, Regenerative endodontics, Scaffolds, Stem cellsAbstract
Introduction The unpredictability of blood clots as natural scaffolds in regenerative endodontic procedures (REPs) has driven the search for synthetic alternatives with reproducible properties. This study aimed to develop and characterize a dual-scale micro/nanofibrous poly(lactic-co-glycolic acid) (PLGA) scaffold fabricated by co-electrospinning and evaluate its physicochemical and biological suitability for REPs under lipopolysaccharide (LPS)-induced inflammatory conditions. Methodology A 7.5% (w/v) PLGA solution was co-electrospun using distinct parameters (flow rate and needle-to-collector distance) to generate a dual-scale fiber architecture. Physicochemical characterization included water uptake, in vitro degradation, and morphological analysis by scanning electron microscopy (SEM). Biological properties were evaluated using human apical papilla cells (APCs). Metabolic activity (Alamar Blue), mineralization (Alizarin Red S), and cell adhesion/morphology (SEM) were assessed in the presence or absence of LPS to simulate an inflammatory environment. Results The co-electrospun scaffold showed a hierarchical structure with interconnected micro and nanofibers. Water uptake reached 147.4% within 24 h and increased gradually thereafter. The scaffold showed a biphasic degradation profile—slow initial degradation (21.5% over 28 days) followed by accelerated loss (63.1% at day 45). Hydration induced fiber swelling and pore remodeling. The scaffold supported APC adhesion, spreading, and metabolic activity over 72 h. Under LPS stimulation, APCs maintained metabolic activity and showed robust mineralization potential after 21 days, comparable to the positive control. Conclusion The co-electrospun PLGA scaffold showed physicochemical properties compatible with tissue ingrowth and supported APCs function under inflammatory conditions. By providing a controlled microenvironment, it represents a potential scaffold design for future REPs.
Downloads
References
1- American Association of Endodontists. AAE clinical considerations for a regenerative procedure [Internet]. Chicago (IL): AAE; 2021 [cited 2026 May 30]. Available from: https://www.aae.org/specialty/wp-content/uploads/sites/2/2021/08/ClinicalConsiderationsApprovedByREC062921.pdf
2- Banchs F, Trope M. Revascularization of immature permanent teeth with apical periodontitis: new treatment protocol? J Endod. 2004;30(4):196-200. doi: 10.1097/00004770-200404000-00003
» https://doi.org/10.1097/00004770-200404000-00003
3- Murray PE, Garcia-Godoy F, Hargreaves KM. Regenerative endodontics: a review of current status and a call for action. J Endod. 2007;33(4):377-90. doi: 10.1016/j.joen.2006.09.013
» https://doi.org/10.1016/j.joen.2006.09.013
4- Dohan Ehrenfest DM, Rasmusson L, Albrektsson T. Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF). Trends Biotechnol. 2009;27(3):158-67. doi: 10.1016/j.tibtech.2008.11.009
» https://doi.org/10.1016/j.tibtech.2008.11.009
5- Isobe K, Watanebe T, Kawabata H, Kitamura Y, Okudera T, Okudera H, et al. Mechanical and degradation properties of advanced platelet-rich fibrin (A-PRF), concentrated growth factors (CGF), and platelet-poor plasma-derived fibrin (PPTF). Int J Implant Dent. 2017;3(1):17. doi: 10.1186/s40729-017-0081-7
» https://doi.org/10.1186/s40729-017-0081-7
6- Bottino MC, Pankajakshan D, Nör JE. Advanced scaffolds for dental pulp and periodontal regeneration. Dent Clin North Am. 2017;61(4):689-711. doi: 10.1016/j.cden.2017.06.009
» https://doi.org/10.1016/j.cden.2017.06.009
7- Jazayeri HE, Lee SM, Kuhn L, Fahimipour F, Tahriri M, Tayebi L. Polymeric scaffolds for dental pulp tissue engineering: a review. Dent Mater. 2020;36(2):e47-e58. doi: 10.1016/j.dental.2019.11.005
» https://doi.org/10.1016/j.dental.2019.11.005
8- Bottino MC, Kamocki K, Yassen GH, Platt JA, Vail MM, Ehrlich Y, et al. Bioactive nanofibrous scaffolds for regenerative endodontics. J Dent Res. 2013;92(11):963-9. doi: 10.1177/0022034513505770
» https://doi.org/10.1177/0022034513505770
9- Albuquerque MT, Valera MC, Nakashima M, Nör JE, Bottino MC. Tissue-engineering-based strategies for regenerative endodontics. J Dent Res. 2014;93(12):1222-31. doi: 10.1177/0022034514549809
» https://doi.org/10.1177/0022034514549809
10- Braghirolli DI, Steffens D, Pranke P. Electrospinning for regenerative medicine: a review of the main topics. Drug Discov Today. 2014;19(6):743-53. doi: 10.1016/j.drudis.2014.03.024
» https://doi.org/10.1016/j.drudis.2014.03.024
11- Karczewski A, Feitosa SA, Hamer EI, Pankajakshan D, Gregory RL, Spolnik KJ, et al. Clindamycin-modified triple antibiotic nanofibers: a stain-free antimicrobial intracanal drug delivery system. J Endod. 2018;44(1):155-62. doi: 10.1016/j.joen.2017.08.024
» https://doi.org/10.1016/j.joen.2017.08.024
12- Herrero-Herrero M, Gómez-Tejedor JA, Vallés-Lluch A. Role of electrospinning parameters on poly(lactic-co-glycolic acid) and poly(caprolactone-co-glycolic acid) membranes. Polymers (Basel). 2021;13(5):695. doi: 10.3390/polym13050695
» https://doi.org/10.3390/polym13050695
13- Zhang Q, Yuan C, Liu L, Wen S, Wang X. Effect of 3-dimensional collagen fibrous scaffolds with different pore sizes on pulp regeneration. J Endod. 2022;48(12):1493-501. doi: 10.1016/j.joen.2022.10.007
» https://doi.org/10.1016/j.joen.2022.10.007
14- Bhardwaj N, Kundu SC. Electrospinning: a fascinating fiber fabrication technique. Biotechnol Adv. 2010;28(3):325-47. doi: 10.1016/j.biotechadv.2010.01.004
» https://doi.org/10.1016/j.biotechadv.2010.01.004
15- Kim SJ, Jang DH, Park WH, Min BM. Fabrication and characterization of 3-dimensional PLGA nanofiber/microfiber composite scaffolds. Polymer (Guildf). 2010;51(6):1320-7. doi: 10.1016/j.polymer.2010.01.025
» https://doi.org/10.1016/j.polymer.2010.01.025
16- Braghirolli DI, Zamboni F, Acasigua GA, Pranke P. Association of electrospinning with electrospraying: a strategy to produce 3D scaffolds with incorporated stem cells for use in tissue engineering. Int J Nanomedicine. 2015;10:5159-69. doi: 10.2147/IJN.S84312
17- Meireles AB, Corrêa DK, da Silveira JV, Millás AL, Bittencourt E, Brito-Melo GE, et al. Trends in polymeric electrospun fibers and their use as oral biomaterials. Exp Biol Med (Maywood). 2018;243(8):665-76. doi: 10.1177/1535370218770404
» https://doi.org/10.1177/1535370218770404
18- Huang F, Cheng L, Li J, Ren B. Nanofibrous scaffolds for regenerative endodontics treatment. Front Bioeng Biotechnol. 2022;10:1078453. doi: 10.3389/fbioe.2022.1078453
» https://doi.org/10.3389/fbioe.2022.1078453
19- Bertassoni LE. Progress and challenges in microengineering the dental pulp vascular microenvironment. J Endod. 2020;46(9S):S90-S100. doi: 10.1016/j.joen.2020.06.033
» https://doi.org/10.1016/j.joen.2020.06.033
20- Vishwanat L, Duong R, Takimoto K, Phillips L, Espitia CO, Diogenes A, et al. Effect of bacterial biofilm on the osteogenic differentiation of stem cells of apical papilla. J Endod. 2017;43(6):916-22. doi: 10.1016/j.joen.2017.01.023
» https://doi.org/10.1016/j.joen.2017.01.023
21- Widbiller M, Eidt A, Wölflick M, Lindner SR, Schweikl H, Hiller KA, et al. Interactive effects of LPS and dentine matrix proteins on human dental pulp stem cells. Int Endod J. 2018;51(8):877-88. doi: 10.1111/iej.12897
» https://doi.org/10.1111/iej.12897
22- Fehrmann C, Dörfer CE, Fawzy El-Sayed KM. Toll-like receptor expression profile of human stem/progenitor cells from the apical papilla. J Endod. 2020;46(11):1623-30. doi: 10.1016/j.joen.2020.08.017
» https://doi.org/10.1016/j.joen.2020.08.017
23- Lei S, Liu XM, Liu Y, Bi J, Zhu S, Chen X. Lipopolysaccharide downregulates the osteo-/odontogenic differentiation of stem cells from apical papilla by inducing autophagy. J Endod. 2020;46(4):502-8. doi: 10.1016/j.joen.2020.01.009
» https://doi.org/10.1016/j.joen.2020.01.009
24- Pan T, Song W, Cao X, Wang Y. 3D bioplotting of gelatin/alginate scaffolds for tissue engineering: influence of crosslinking degree and pore architecture on physicochemical properties. J Mater Sci Technol. 2016;32(9):889-900. doi: 10.1016/j.jmst.2016.01.007
25- Li YF, Luo QP, Yang YX, Li AQ, Zhang XC. A novel bi-layered asymmetric membrane incorporating demineralized dentin matrix accelerates tissue healing and bone regeneration in a rat skull defect model. Biomater Sci. 2024;12(16):4226-41. doi: 10.1039/d4bm00350k
» https://doi.org/10.1039/d4bm00350k
26- Sonoyama W, Liu Y, Yamaza T, Tuan RS, Wang S, Shi S, et al. Characterization of the apical papilla and its residing stem cells from human immature permanent teeth: a pilot study. J Endod. 2008;34(2):166-71. doi: 10.1016/j.joen.2007.11.021
» https://doi.org/10.1016/j.joen.2007.11.021
27- Sipert CR, Oliveira AP, Caldeira CL. Cytotoxicity of intracanal dressings on apical papilla cells differ upon activation with E. faecalis LTA. J Appl Oral Sci. 2019;27:e20180291. doi: 10.1590/1678-7757-2018-0291
» https://doi.org/10.1590/1678-7757-2018-0291
28- Zhang J, Zhang Y, Lv H, Yu Q, Zhou Z, Zhu Q, et al. Human stem cells from the apical papilla response to bacterial lipopolysaccharide exposure and anti-inflammatory effects of nuclear factor I C. J Endod. 2013;39(11):1416-22. doi: 10.1016/j.joen.2013.07.018
» https://doi.org/10.1016/j.joen.2013.07.018
29- Pizzatto LN, Meneses CCB, Diniz EA, Dionísio TJ, Santos CF, Sipert CR. Angiotensin II regulates proliferation and function of stem cells of apical papilla. J Endod. 2020;46(6):810-7. doi: 10.1016/j.joen.2020.03.015
» https://doi.org/10.1016/j.joen.2020.03.015
30- Alghofaily M, Torabinejad M, Nosrat A. Regenerative endodontic treatment using periapical blood or circulating blood as scaffold: a volumetric analysis. J Endod. 2022;48(5):625-31. doi: 10.1016/j.joen.2022.01.008
» https://doi.org/10.1016/j.joen.2022.01.008
31- Félix Lanao RP, Jonker AM, Wolke JG, Jansen JA, van Hest JC, Leeuwenburgh SC. Physicochemical properties and applications of poly(lactic-co-glycolic acid) for use in bone regeneration. Tissue Eng Part B Rev. 2013;19(4):380-90. doi: 10.1089/ten.TEB.2012.0443
32- You Y, Min BM, Lee SJ, Lee TS, Park WH. In vitro degradation behavior of electrospun polyglycolide, polylactide, and poly(lactide-co-glycolide). J Appl Polym Sci. 2005;95(2):193-200. doi: 10.1002/app.21116
33- Blackwood KA, McKean R, Canton I, Freeman CO, Franklin KL, Cole D, et al. Development of biodegradable electrospun scaffolds for dermal replacement. Biomaterials. 2008;29(21):3091-104. doi: 10.1016/j.biomaterials.2008.03.037
» https://doi.org/10.1016/j.biomaterials.2008.03.037
34- Chor A, Gonçalves RP, Costa AM, Farina M, Ponche A, Sirelli L, et al. In vitro degradation of electrospun poly(lactic-co-glycolic acid) (PLGA) for oral mucosa regeneration. Polymers (Basel). 2020;12(8):1853. doi: 10.3390/polym12081853
» https://doi.org/10.3390/polym12081853
35- Bakopoulou A, Leyhausen G, Volk J, Tsiftsoglou A, Garefis P, Koidis P, et al. Comparative analysis of in vitro osteo/odontogenic differentiation potential of human dental pulp stem cells (DPSCs) and stem cells from the apical papilla (SCAP). Arch Oral Biol. 2011;56(7):709-21. doi: 10.1016/j.archoralbio.2010.12.008
» https://doi.org/10.1016/j.archoralbio.2010.12.008
36- Liu Q, Gao Y, He J. Stem cells from the apical papilla (SCAPs): past, present, prospects, and challenges. Biomedicines. 2023;11(7):2047. doi: 10.3390/biomedicines11072047
» https://doi.org/10.3390/biomedicines11072047
37- Lovelace TW, Henry MA, Hargreaves KM, Diogenes A. Evaluation of the delivery of mesenchymal stem cells into the root canal space of necrotic immature teeth after clinical regenerative endodontic procedure. J Endod. 2011;37(2):133-8. doi: 10.1016/j.joen.2010.10.009
» https://doi.org/10.1016/j.joen.2010.10.009
38- Bakopoulou A, Kritis A, Andreadis D, Papachristou E, Leyhausen G, Koidis P, et al. Angiogenic potential and secretome of human apical papilla mesenchymal stem cells in various stress microenvironments. Stem Cells Dev. 2015;24(21):2496-512. doi: 10.1089/scd.2015.0197
» https://doi.org/10.1089/scd.2015.0197
39- Chrepa V, Pitcher B, Henry MA, Diogenes A. Survival of the apical papilla and its resident stem cells in a case of advanced pulpal necrosis and apical periodontitis. J Endod. 2017;43(4):561-7. doi: 10.1016/j.joen.2016.09.024
» https://doi.org/10.1016/j.joen.2016.09.024
40- Palma PJ, Ramos JC, Martins JB, Diogenes A, Figueiredo MH, Ferreira P, et al. Histologic evaluation of regenerative endodontic procedures with the use of chitosan scaffolds in immature dog teeth with apical periodontitis. J Endod. 2017;43(8):1279-87. doi: 10.1016/j.joen.2017.03.005
Downloads
Published
Issue
Section
License
Copyright (c) 2026 Stephanie Isabel Diaz Zamalloa; Caroline Carvalho dos Santos, Leticia Martins Santos, Victor Elias Arana-Chavez, Fernando Neves Nogueira, Carla Renata Sipert, Celso Luiz Caldeira

This work is licensed under a Creative Commons Attribution 4.0 International License.
Todo o conteúdo do periódico, exceto onde está identificado, está licenciado sob uma Licença Creative Commons do tipo atribuição CC-BY.