Comprehensive Evaluation of Decellularized Dental Pulp as a New Biomaterial for Regeneration of the Pulp-Dentin Complex
https://doi.org/10.35401/2541-9897-2024-9-4-93-100
Abstract
Objective: To develop a detergent-enzymatic method and evaluate the quality of a decellularized pulp scaffold for regenerative endodontics.
Materials and methods: Biomaterial and mesenchymal stem cells (MSCs) were derived from dental pulp that was obtained following third molar extraction indicated for orthodontic reasons in patients aged 14-18 years. The detergent-enzymatic method enabled to obtain a decellularized scaffold from pulp samples. The proliferative activity and viability of dental pulp-derived MSCs were assessed using trypan blue staining and XTT assay. To assess tissue response, Wistar rats underwent subcutaneous implantation of native and decellularized dental pulp. Explanted samples were stained with hematoxylin-eosin on days 7 and 14.
Results: The detergent-enzymatic treatment of the dental pulp demonstrated the absence of nuclear material, whereas the histoarchitecture of the dental pulp was disturbed. The DNA content in the sample of the decellularized scaffold was 22.79 ± 2.1 ng/mg of tissue; the amount of DNA in the native sample was 78.5 ± 5.4 ng/mg of tissue. According to XTT assay results, no cytotoxicity of the decellularized scaffold against MSCs was found. Biopsy specimens of the rats with implanted decellularized dental pulp were characterized by no signs of inflammation.
Conclusions: The study results will enable to create a biomaterial that can be the base of a tissue-engineered structure of the dental pulp and be used for the regeneration of the pulp-dentin complex.
Keywords
About the Authors
K. I. MelkonianRussian Federation
Karina I. Melkonian - Cand. Sci. (Med.), Associate Professor, Head of the Central Research Laboratory, Kuban State Medical University.
ulitsa M. Sedina 4, Krasnodar, 350063
T. V. Rusinova
Russian Federation
Tatyana V. Rusinova - Cand. Sci. (Bio.), Researcher at the Central Research Laboratory, Kuban State Medical University.
Krasnodar
E. S. Zaporozhskaya-Abramova
Russian Federation
Ekaterina S. Zaporozhskaya-Abramova - Cand. Sci. (Med.), Associate Professor at the Therapeutic Dentistry Department, Kuban State Medical University.
Krasnodar
E. A. Solop
Russian Federation
Elizaveta A. Solop - Laboratory Research Technician, Central Research Laboratory, Kuban State Medical University.
Krasnodar
I. I. Kartashevskiy
Russian Federation
Igor I. Kartashevskiy - 4th Year Student, Faculty of Dentistry, Kuban State Medical University.
Krasnodar
S. A. Knyshova
Russian Federation
Sofiya A. Knyshova - 6th Year Student, Faculty of Dentistry, Kuban State Medical University.
Krasnodar
O. N. Risovannaya
Russian Federation
Olga N. Risovannaya - Dr. Sci. (Med.), Associate Professor, Professor at the Dentistry Department, Kuban State Medical University.
Krasnodar
References
1. Xie Z, Shen Z, Zhan P, et al. Functional dental pulp regeneration: basic research and clinical translation. Int J Mol Sci. 2021;22(16):8991. PMID: 34445703. PMCID: PMC8396610. https://doi.org/10.3390/ijms22168991
2. Hu L, Gao Z, Xu J, et al. Decellularized swine dental pulp as a bioscaffold for pulp regeneration. Biomed Res Int. 2017;2017:9342714. PMID: 29387727. PMCID: PMC5745671. https://doi.org/10.1155/2017/9342714
3. Jakovljevic A, Nikolic N, Jacimovic J, et al. Prevalence of apical periodontitis and conventional nonsurgical root canal treatment in general adult population: an updated systematic review and meta-analysis of cross-sectional studies published between 2012 and 2020. J Endod. 2020;46(10):1371–1386.e8. Published correction appears in J Endod. 2021;47(2):336. PMID: 32673634. https://doi.org/10.1016/j.joen.2020.07.007
4. Galler KM, Eidt A, Schmalz G. Cell-free approaches for dental pulp tissue engineering. J Endod. 2014;40(4 Suppl):S41– S45. PMID: 24698692. https://doi.org/10.1016/j.joen.2014.01.014
5. Duncan HF. Present status and future directions-vital pulp treatment and pulp preservation strategies. Int Endod J. 2022;55 Suppl 3(Suppl 3):497–511. PMID: 35080024. PMCID: PMC9306596. https://doi.org/10.1111/iej.13688
6. Nowwarote N, Petit S, Ferre FC, et al. Extracellular matrix derived from dental pulp stem cells promotes mineralization. Front Bioeng Biotechnol. 2022;9:740712. PMID: 35155398. PMCID: PMC8829122. https://doi.org/10.3389/fbioe.2021.740712
7. Na S, Zhang H, Huang F, et al. Regeneration of dental pulp/ dentine complex with a three-dimensional and scaffold-free stem-cell sheet-derived pellet. J Tissue Eng Regen Med. 2016;10(3):261– 270. PMID: 23365018. https://doi.org/10.1002/term.1686
8. Lopes LB, Neves JA, Botelho J, Machado V, Mendes JJ. Regenerative endodontic procedures: an umbrella review. Int J Environ Res Public Health. 2021;18(2):754. PMID: 33561086. PMCID: PMC7830213. https://doi.org/10.3390/ijerph18020754
9. Hameed MH, Gul M, Ghafoor R, Badar SB. Management of immature necrotic permanent teeth with regenerative endodontic procedures - a review of literature. J Pak Med Assoc. 2019;69(10):1514–1520. PMID: 31622308.
10. Matoug-Elwerfelli M, Duggal MS, Nazzal H, Esteves F, Raïf E. A biocompatible decellularized pulp scaffold for regenerative endodontics. Int Endod J. 2018;51(6):663–673. PMID: 29197101. https://doi.org/10.1111/iej.12882
11. Bakhtiar H, Rajabi S, Pezeshki-Modaress M, et al. Optimizing methods for bovine dental pulp decellularization. J Endod. 2021;47(1):62–68. PMID: 33049226. https://doi.org/10.1016/j.joen.2020.08.027
12. Adanir N, Khurshid Z, Ratnayake J. The regenerative potential of decellularized dental pulp extracellular matrix: a systematic review. Materials (Basel). 2022;15(18):6386. PMID: 36143698. PMCID: PMC9505725. https://doi.org/10.3390/ma15186386
13. Lee DJ, Miguez P, Kwon J, et al. Decellularized pulp matrix as scaffold for mesenchymal stem cell mediated bone regeneration. J. Tissue Eng. 2020;11:2041731420981672. PMID: 33414903. PMCID: PMC7750895. https://doi.org/10.1177/2041731420981672
14. Alqahtani Q, Zaky SH, Patil A, Beniash E, Ray H, Sfeir C. Decellularized swine dental pulp tissue for regenerative root canal therapy. J Dent Res. 2018;97(13):1460–1467. PMID: 30067420. https://doi.org/10.1177/0022034518785124
15. Kim IH, Jeon M, Cheon K, et al. In vivo evaluation of decellularized human tooth scaffold for dental tissue regeneration. Appl Sci (Basel). 2021;11(18):8472. PMID: 36003951. PMCID: PMC9397400. https://doi.org/10.3390/app11188472
16. Bakhtiar H, Pezeshki-Modaress M, Kiaipour Z, et al. Pulp ECM-derived macroporous scaffolds for stimulation of dental-pulp regeneration process. Dent Mater. 2020;36(1):76–87. PMID: 31735424. https://doi.org/10.1016/j.dental.2019.10.011
17. Crapo PM, Gilbert TW, Badylak SF. An overview of tissue and whole organ decellularization processes. Biomaterials. 2011;32(12):3233–3243. PMID: 21296410. https://doi.org/10.1016/j.biomaterials.2011.01.057
18. Nazzal H, Kenny K, Altimimi A, Kang J, Duggal MS. A prospective clinical study of regenerative endodontic treatment of traumatized immature teeth with necrotic pulps using biantibiotic paste. Int Endod J. 2018;51 Suppl 3:e204–e215. PMID: 28653761. https://doi.org/10.1111/iej.12808
19. Soares DG, Bordini EAF, Swanson WB, de Souza Costa CA, Bottino MC. Platform technologies for regenerative endodontics from multifunctional biomaterials to tooth-on-a-chip strategies. Clin Oral Investig. 2021;25(8):4749–4779. PMID: 34181097. PMCID: PMC8546585. https://doi.org/10.1007/s00784-021-04013-4
Review
For citations:
Melkonian K.I., Rusinova T.V., Zaporozhskaya-Abramova E.S., Solop E.A., Kartashevskiy I.I., Knyshova S.A., Risovannaya O.N. Comprehensive Evaluation of Decellularized Dental Pulp as a New Biomaterial for Regeneration of the Pulp-Dentin Complex. Innovative Medicine of Kuban. 2024;(4):93-100. (In Russ.) https://doi.org/10.35401/2541-9897-2024-9-4-93-100