Osteogenic potential of primary stem cells derived from the human dental pulp is enhanced by carboxymethyl cellulose/chitosan scaffold doped with wollastonite particles

Dannie Macrin; Vivek Narayanan; Arikketh Devi

doi:10.7324/JABB.2021.95.1s1

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Abstract

The success of any scaffold assisted regenerative therapy relies on two major factors; a conducive scaffold material and versatile, viable stem cells. In our study, we suggest chitosan (CS)/carboxymethylcellulose (CMC) scaffold functionalized with wollastonite (WS) (CaSiO3 ) particles as a conducive scaffold material which mimics the porous structure of the bone and promotes osteogenesis. We have paired this scaffold material with stem cells derived from dental pulp (DPSCs) which is an ideal cell source with a high percentage of stem cells and have a natural inclination toward forming hard calcified tissues. In our experiments, the cells isolated from human dental pulp produced a cell population with high percentage of cells positive for mesenchymal markers CD73 and CD90. The DPSCs were able to differentiate into osteoblasts when induced with media supplemented with β-glycerophosphate, ascorbic acid, and dexamethasone. Furthermore, physicochemical analysis of CS/CMC/WS scaffold showed that it formed a highly porous structure conducive to cell growth, penetration, and nutrient uptake. Furthermore, CS/CMC/WS scaffold promoted osteogenic differentiation of DPSCs. We propose CS/CMS/WS scaffold paired with DPSCs as an effective system for bone regeneration.

Keyword: Adult stem cells mesenchymal markers bone regeneration osteogenic differentiation

Citation:

Macrin D, Narayanan V, Devi A. Osteogenic potential of primary stem cells derived from the human dental pulp is enhanced by carboxymethyl cellulose/chitosan scaffold doped with wollastonite particles. J Appl Biol Biotech, 2021;9(S1):1-6

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

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1.Sheng G. The developmental basis of mesenchymal stem/stromal cells (MSCs). BMC Dev Biol 2015;15:44. https://doi.org/10.1186/s12861-015-0094-5
2. Lv FJ, Tuan RS, Cheung KM, Leung VY. Concise review: the surface markers and identity of human mesenchymal stem cells. Stem Cells 2014;32:1408-19. https://doi.org/10.1002/stem.1681
3. Stanley HR. The cells of the dental pulp. Oral Surg Oral Med Oral Pathol 1962;15:849-58. https://doi.org/10.1016/0030-4220(62)90337-7
4. Gronthos S, Brahim J, Li W, Fisher LW, Cherman N, Boyde A, et al. Stem cell properties of human dental pulp stem cells. J Dent Res 2002;81:531-5. https://doi.org/10.1177/154405910208100806
5. Shi S, Gronthos S. Perivascular niche of postnatal mesenchymal stem cells in human bone marrow and dental pulp. J Bone Miner Res 2003;18:696-704. https://doi.org/10.1359/jbmr.2003.18.4.696
6. Chang CC, Chang KC, Tsai SJ, Chang HH, Lin CP. Neurogenic differentiation of dental pulp stem cells to neuron-like cells in dopaminergic and motor neuronal inductive media. J Formos Med Assoc 2014;113:956-65. https://doi.org/10.1016/j.jfma.2014.09.003
7. Kawashima N. Characterisation of dental pulp stem cells: a new horizon for tissue regeneration? Arch Oral Biol 2012;57:1439-58. https://doi.org/10.1016/j.archoralbio.2012.08.010
8. Ledesma-Martínez E, Mendoza-Núñez VM, Santiago-Osorio E. Mesenchymal stem cells derived from dental pulp: a review. Stem Cells Int 2016;2016: 4709572. https://doi.org/10.1155/2016/4709572
9. Sivasankar V, Ranganathan K. Growth characteristics and expression of CD73 and CD146 in cells cultured from dental pulp. J Investig Clin Dent 2016;7:278-85. https://doi.org/10.1111/jicd.12155
10. Syed-Picard FN, Du Y, Lathrop KL, Mann MM, Funderburgh ML, Funderburgh JL. Dental pulp stem cells: a new cellular resource for corneal stromal regeneration. Stem Cells Transl Med 2015;4:276-85. https://doi.org/10.5966/sctm.2014-0115
11. Chun SY, Soker S, Jang YJ, Kwon TG, Yoo ES. Differentiation of human dental pulp stem cells into dopaminergic neuron-like cells in vitro. J Korean Med Sci 2016;31:171-7. https://doi.org/10.3346/jkms.2016.31.2.171
12. Bronckaers A, Hilkens P, Fanton Y, Struys T, Gervois P, Politis C, et al. Angiogenic properties of human dental pulp stem cells. PLoS One 2013;8:e71104. https://doi.org/10.1371/journal.pone.0071104
13. Nakashima M, Iohara K, Murakami M. Dental pulp stem cells and regeneration. Endod Topics 2013;28:38-50. https://doi.org/10.1111/etp.12027
14. O'brien FJ. Biomaterials & scaffolds for tissue engineering. Mater Today 2011;14:88-95. https://doi.org/10.1016/S1369-7021(11)70058-X
15. Hollister SJ. Porous scaffold design for tissue engineering. Nat Mater 2005;4:518-24. https://doi.org/10.1038/nmat1421
16. Petrovic V, Zivkovic P, Petrovic D, Stefanovic V. Craniofacial bone tissue engineering. Oral Surg Oral Med Oral Pathol Oral Radiol 2012;114:e1-9. https://doi.org/10.1016/j.oooo.2012.02.030
17. Rezwan K, Chen QZ, Blaker JJ, Boccaccini AR. Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. Biomaterials 2006;27:3413-31. https://doi.org/10.1016/j.biomaterials.2006.01.039
18. Knorr D. Functional properties of chitin and chitosan. J Food Sci 1982;47:593-5. https://doi.org/10.1111/j.1365-2621.1982.tb10131.x
19. Fei Liu X, Lin Guan Y, Zhi Yang D, Li Z, De Yao K. Antibacterial action of chitosan and carboxymethylated chitosan. J Appl Polym Sci 2001;79:1324-35. https://doi.org/10.1002/1097-4628(20010214)79:7<1324::AID-APP210>3.0.CO;2-L
20. Li H, Zhai W, Chang J. Effects of wollastonite on proliferation and differentiation of human bone marrow-derived stromal cells in PHBV/ wollastonite composite scaffolds. J Biomater Appl 2009;24:231-46. https://doi.org/10.1177/0885328208096043
21. Saravanan S, Vimalraj S, Vairamani M, Selvamurugan N. Role of mesoporous wollastonite (calcium silicate) in mesenchymal stem cell proliferation and osteoblast differentiation: a cellular and molecular study. J Biomed Nanotechnol 2015;11:1124-38. https://doi.org/10.1166/jbn.2015.2057
22. Sainitya R, Sriram M, Kalyanaraman V, Dhivya S, Saravanan S, Vairamani M, et al. Scaffolds containing chitosan/carboxymethyl cellulose/mesoporous wollastonite for bone tissue engineering. Int J Biol Macromol 2015;80:481-8. https://doi.org/10.1016/j.ijbiomac.2015.07.016
23. Ramos TL, Sánchez-Abarca LI, Muntión S, Preciado S, Puig N, LópezRuano G, et al. MSC surface markers (CD44, CD73, and CD90) can identify human MSC-derived extracellular vesicles by conventional flow cytometry. Cell Commun Signal 2016;14:1-4. https://doi.org/10.1186/s12964-015-0124-8
24. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006;8:315-7. https://doi.org/10.1080/14653240600855905
25. Murphy CM, Haugh MG, O'Brien FJ. The effect of mean pore size on cell attachment, proliferation and migration in collagenglycosaminoglycan scaffolds for bone tissue engineering. Biomaterials 2010;31:461-6. https://doi.org/10.1016/j.biomaterials.2009.09.063
26. Baier T, Dupeux G, Herbert S, Hardt S, Quéré D. Mesoporous calcium silicate compositions and methods for synthesis of mesoporous calcium silicate for controlled release of bioactive agents, US 8,916,198 B2, 2013.
27. Vladkova TG. Surface engineered polymeric biomaterials with improved biocontact properties. Int J Polym Sci 2010;2010:1-22. https://doi.org/10.1155/2010/296094

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Macrin D [PubMed] [Google Scholar ]

Narayanan V [PubMed] [Google Scholar ]

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1.Sheng G. The developmental basis of mesenchymal stem/stromal cells (MSCs). BMC Dev Biol 2015;15:44. https://doi.org/10.1186/s12861-015-0094-5

2. Lv FJ, Tuan RS, Cheung KM, Leung VY. Concise review: the surface markers and identity of human mesenchymal stem cells. Stem Cells 2014;32:1408-19. https://doi.org/10.1002/stem.1681

3. Stanley HR. The cells of the dental pulp. Oral Surg Oral Med Oral Pathol 1962;15:849-58. https://doi.org/10.1016/0030-4220(62)90337-7

4. Gronthos S, Brahim J, Li W, Fisher LW, Cherman N, Boyde A, et al. Stem cell properties of human dental pulp stem cells. J Dent Res 2002;81:531-5. https://doi.org/10.1177/154405910208100806

5. Shi S, Gronthos S. Perivascular niche of postnatal mesenchymal stem cells in human bone marrow and dental pulp. J Bone Miner Res 2003;18:696-704. https://doi.org/10.1359/jbmr.2003.18.4.696

6. Chang CC, Chang KC, Tsai SJ, Chang HH, Lin CP. Neurogenic differentiation of dental pulp stem cells to neuron-like cells in dopaminergic and motor neuronal inductive media. J Formos Med Assoc 2014;113:956-65. https://doi.org/10.1016/j.jfma.2014.09.003

7. Kawashima N. Characterisation of dental pulp stem cells: a new horizon for tissue regeneration? Arch Oral Biol 2012;57:1439-58. https://doi.org/10.1016/j.archoralbio.2012.08.010

8. Ledesma-Martínez E, Mendoza-Núñez VM, Santiago-Osorio E. Mesenchymal stem cells derived from dental pulp: a review. Stem Cells Int 2016;2016: 4709572. https://doi.org/10.1155/2016/4709572

9. Sivasankar V, Ranganathan K. Growth characteristics and expression of CD73 and CD146 in cells cultured from dental pulp. J Investig Clin Dent 2016;7:278-85. https://doi.org/10.1111/jicd.12155

10. Syed-Picard FN, Du Y, Lathrop KL, Mann MM, Funderburgh ML, Funderburgh JL. Dental pulp stem cells: a new cellular resource for corneal stromal regeneration. Stem Cells Transl Med 2015;4:276-85. https://doi.org/10.5966/sctm.2014-0115

11. Chun SY, Soker S, Jang YJ, Kwon TG, Yoo ES. Differentiation of human dental pulp stem cells into dopaminergic neuron-like cells in vitro. J Korean Med Sci 2016;31:171-7. https://doi.org/10.3346/jkms.2016.31.2.171

12. Bronckaers A, Hilkens P, Fanton Y, Struys T, Gervois P, Politis C, et al. Angiogenic properties of human dental pulp stem cells. PLoS One 2013;8:e71104. https://doi.org/10.1371/journal.pone.0071104

13. Nakashima M, Iohara K, Murakami M. Dental pulp stem cells and regeneration. Endod Topics 2013;28:38-50. https://doi.org/10.1111/etp.12027

14. O'brien FJ. Biomaterials & scaffolds for tissue engineering. Mater Today 2011;14:88-95. https://doi.org/10.1016/S1369-7021(11)70058-X

15. Hollister SJ. Porous scaffold design for tissue engineering. Nat Mater 2005;4:518-24. https://doi.org/10.1038/nmat1421

16. Petrovic V, Zivkovic P, Petrovic D, Stefanovic V. Craniofacial bone tissue engineering. Oral Surg Oral Med Oral Pathol Oral Radiol 2012;114:e1-9. https://doi.org/10.1016/j.oooo.2012.02.030

17. Rezwan K, Chen QZ, Blaker JJ, Boccaccini AR. Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. Biomaterials 2006;27:3413-31. https://doi.org/10.1016/j.biomaterials.2006.01.039

18. Knorr D. Functional properties of chitin and chitosan. J Food Sci 1982;47:593-5. https://doi.org/10.1111/j.1365-2621.1982.tb10131.x

19. Fei Liu X, Lin Guan Y, Zhi Yang D, Li Z, De Yao K. Antibacterial action of chitosan and carboxymethylated chitosan. J Appl Polym Sci 2001;79:1324-35. https://doi.org/10.1002/1097-4628(20010214)79:7<1324::AID-APP210>3.0.CO;2-L

20. Li H, Zhai W, Chang J. Effects of wollastonite on proliferation and differentiation of human bone marrow-derived stromal cells in PHBV/ wollastonite composite scaffolds. J Biomater Appl 2009;24:231-46. https://doi.org/10.1177/0885328208096043

21. Saravanan S, Vimalraj S, Vairamani M, Selvamurugan N. Role of mesoporous wollastonite (calcium silicate) in mesenchymal stem cell proliferation and osteoblast differentiation: a cellular and molecular study. J Biomed Nanotechnol 2015;11:1124-38. https://doi.org/10.1166/jbn.2015.2057

22. Sainitya R, Sriram M, Kalyanaraman V, Dhivya S, Saravanan S, Vairamani M, et al. Scaffolds containing chitosan/carboxymethyl cellulose/mesoporous wollastonite for bone tissue engineering. Int J Biol Macromol 2015;80:481-8. https://doi.org/10.1016/j.ijbiomac.2015.07.016

23. Ramos TL, Sánchez-Abarca LI, Muntión S, Preciado S, Puig N, LópezRuano G, et al. MSC surface markers (CD44, CD73, and CD90) can identify human MSC-derived extracellular vesicles by conventional flow cytometry. Cell Commun Signal 2016;14:1-4. https://doi.org/10.1186/s12964-015-0124-8

24. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006;8:315-7. https://doi.org/10.1080/14653240600855905

25. Murphy CM, Haugh MG, O'Brien FJ. The effect of mean pore size on cell attachment, proliferation and migration in collagenglycosaminoglycan scaffolds for bone tissue engineering. Biomaterials 2010;31:461-6. https://doi.org/10.1016/j.biomaterials.2009.09.063

26. Baier T, Dupeux G, Herbert S, Hardt S, Quéré D. Mesoporous calcium silicate compositions and methods for synthesis of mesoporous calcium silicate for controlled release of bioactive agents, US 8,916,198 B2, 2013.

27. Vladkova TG. Surface engineered polymeric biomaterials with improved biocontact properties. Int J Polym Sci 2010;2010:1-22. https://doi.org/10.1155/2010/296094

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