Effects of Cell Seeding Methods on Chondrogenic Differentiation of Rat Mesenchymal Stem Cells in Polyhydroxybutyrate/Chitosan Scaffolds

Giretová, M.; Medvecký, Ľ.; Petrovová, E.; Čížková, D.; Mudroňová, D.; Danko, J.

doi:10.2478/fv-2019-0002

Abstract

The aim of our study was to examine the effects of passive and active cell seeding techniques on in vitro chondrogenic differentiation of mesenchymal stem cells (MSC) isolated from rat bone marrow and seeded on porous biopolymer scaffolds based on polyhydroxybutyrate/chitosan (PCH) blends. This paper is focused on the distribution of the cells on and in the scaffolds, since it influences the uniformity of the created extracellular matrix (ECM), as well as the homogenity of the distribution of chondrogenic markers in vitro which ultimately affects the quality of the newly created tissue after in vivo implantation. The three types of cell-scaffold constructs were examined by: fluorescence microscopy, SEM, histology and quantitative analysis of the glycosaminoglycans after chondrogenic cultivation. The results demonstrated that the active cells seeded via the centrifugation of the cell suspension onto the scaffold guaranteed an even distribution of cells on the bulk of the scaffold and the uniform secretion of the ECM products by the differentiated cells.

References

1. Barry, F. P., Murphy, J. M., 2004: Review Mesenchymal stem cells: clinical applications and biological characterization. Int. J. Biochem. Cell Biol, 36, 568—584. DOI: 10.1016/j.biocel.2003.11.001.10.1016/j.biocel.2003.11.001
Open DOI Search in Google Scholar Back to article
2. Bornes, T. D., Jomha, N. M., Mulet-Sierra, A., Adesida, A. B., 2016: Optimal seeding densities for in vitro chondrogenesis of two- and three-dimensional-isolated and expanded bone marrow-derived mesenchymal stromal stem cells within a porous collagen scaffold. Tissue Engn. C: Methods, 22, 208— 220. DOI: 10.1089/ten.tec.2015.036558.10.1089/ten.tec.2015.036558
Open DOI Search in Google Scholar Back to article
3. Collins, M. N., Birkinshaw, C., 2013: Hyaluronic acid based scaffolds for tissue engineering—A review. Carb. Pol., 92 (2), 1262—1279. DOI 10.1016/j.carbpol.2012.10.028.10.1016/j.carbpol.2012.10.028
Open DOI Search in Google Scholar Back to article
4. Deng, Y., Lin, X. S., Zheng. Z., Deng. J. G., Chen, J. CH., Ma, H., Chen, G. Q., 2003: Poly (hydroxybutyrate-co-hydroxyhexanoate) promoted production of extracellular matrix of articular cartilage chondrocytes in vitro. Biomaterials, 24, 4273—4281.10.1016/S0142-9612(03)00367-3
Search in Google Scholar Back to article
5. Deng, CH. M., He, L. Z., Zhao, M., Yang, D., Liu, Y. 2007: Biological properties of the chitosan-gelatin sponge wound dressing. Carb. Pol., 69, 583—589. DOI: 10.1016/j.carbpol.2007.01.014.10.1016/j.carbpol.2007.01.014
Open DOI Search in Google Scholar Back to article
6. Dominici, M., Le Blanc, K., Meller, I., Slaper-Cortenbach, I., Marini, F. C., Krause, D. S., et al., 2006: Minimal criteria for defining mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8, 315—317.10.1080/14653240600855905
Search in Google Scholar Back to article
7. Ehlers, E. M., Fuss, M., Rohwedel, J., Russlies, M., Kuehnel, W., Behrens, P., 1999: Development of a biocomposite to fill out articular cartilage lesions. Light, scanning and transmission electron microscopy of sheep chondrocytes cultured on a collagen I/III sponge. Ann. Anat., 181 (6), 513—518. DOI: 10.1016/S0940-9602(99)80055-7.10.1016/S0940-9602(99)80055-7
Open DOI Search in Google Scholar Back to article
8. Fernandez Vallone, V. B., Romaniuk, M. A., Choi, H., Labovsky, V., Otaeugi, J., Chasseing, N. A., 2013: Review: Mesenchymal stem cells and their use in therapy: What has been achieved ? Differentiation, 85 (1—2), 1—10.10.1016/j.diff.2012.08.004
Search in Google Scholar Back to article
9. Godbey, W. T., Hindy, B. S. S., Sherman, M. E., Atala, A., 2004: A novel use of centrifugal force for cell seeding into porous scaffolds. Biomaterials, 25, 2799—2805. DOI: 10.1016/j.biomaterials.2003.09.056.10.1016/j.biomaterials.2003.09.056
Open DOI Search in Google Scholar Back to article
10. Gogolewski, S., 2000: Bioresorbable polymers in trauma and bone surgery. Injury, 31, S‒D28‒32.10.1016/S0020-1383(00)80020-0
Search in Google Scholar Back to article
11. Griffon, D. J., Sedighi, M. R., Schaeffer, D. V., Eurell, J. A., Johnson, A. L., 2006: Chitosan scaffolds: Interconnective pore size and cartilage engineering. Acta Biomaterialia, 2 (3), 313—320.10.1016/j.actbio.2005.12.007
Search in Google Scholar Back to article
12. Harvanova, D., Rosocha, Bakos, D., Švihla, R., Vasko, G., Hornak, S., et al., 2009: Collagen/hyaluronan membrane as a scaffold for chondrocyte cultivation. Biologia, 64 (5), 1032— 1038. DOI: 10.2478/s11756-009-0171-y.10.2478/s11756-009-0171-y
Open DOI Search in Google Scholar Back to article
13. Chen, Y. L., Lee, H. P., Chan, H. Y., Sung, L. Y., Chen, H. CH., Hu, Y. CH., 2007: Composite chondroitin-6-sulfate/dermatan sulfate/chitosan scaffolds for cartilage tissue engineering. Biomaterials, 28 (14), 2294—2305. DOI: 10.1016/j.biomaterials.2007.01.027.10.1016/j.biomaterials.2007.01.027
Search in Google Scholar Back to article
14. Chen, G., Akahane, D., Kawazoe, N., Yamamoto, K., Tateishi, T., 2008: Chondrogenic differentiation of mesenchymal stem cells in a leakproof collagen sponge. Mat. Sci. Engin. C, 28 (1), 195—201. DOI: 10.1016/j.msec.2006.12.009.10.1016/j.msec.2006.12.009
Open DOI Search in Google Scholar Back to article
15. Cho, J. H., Kim, S. H., Park, K. D., Jung, M. CH., Yang, W. I., Han, S. W., et al., 2004: Chondrogenic differentiation of human mesenchymal stem cells using a thermosensitive poly(N-isopropylacrylamide) and water-soluble chitosan copolymer. Biomaterials, 25 (26), 5743—5751. DOI: 10.1016/j.biomaterials.2004.01.051.10.1016/j.biomaterials.2004.01.051
Open DOI Search in Google Scholar Back to article
16. Johnstone, B., Hering, T. M., Caplan, A., Goldberg, V. M., Yoo, J. U., 1998:In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. Exp. Cell Res., 238 (1), 265—272. DOI: 10.1006/excr.1997.3858.10.1006/excr.1997.3858
Open DOI Search in Google Scholar Back to article
17. Kessler, M. W., Grande, D. A, 2008: Review: Tissue engineering and cartilage. Organogenesis, 4 (1), 28—32. https://doi.org/10.4161/org.6116.10.4161/org.6116
Open DOI Search in Google Scholar Back to article
18. Liu, C., Xia, Z., Czernuszka, J. T, 2007: Design and development of three-dimensional scaffolds for tissue engineering. Chem. Eng. Res. Design, 85(A7), 1051—1064.10.1205/cherd06196
Search in Google Scholar Back to article
19. Madihally, S. V., Matthew, H. W. T., 1999: Porous chitosan scaffolds for tissue engineering. Biomaterials, 20 (12), 1133— 1142. DOI: 10.1016/S0142-9612(99)00011-3.10.1016/S0142-9612(99)00011-3
Open DOI Search in Google Scholar Back to article
20. Malafaya, P. B., Silva, G. A., Reis, R. L., 2007: Natural-origin polymers as carriers and scaffolds for biomolecules and cell delivery in tissue engineering applications. Adv. Drug Del. Rev., 59 (4—5), 207—233. DOI:10.1016/j.addr.2007.03.012.10.1016/j.addr.2007.03.012
Open DOI Search in Google Scholar Back to article
21. Medvecky, L., Giretova, M., Stulajterova, R., 2014: Properties and in vitro characterization of polyhydroxybutyratechitosan scaffolds prepared by modified precipitation method. J. Mat. Sci.: Mat. Med., 25 (3), 777—789. DOI:10.1007/s10856-013-5105-0.10.1007/s10856-013-5105-0
Open DOI Search in Google Scholar Back to article
22. Mollon, B., Kandel, R., Chahai, J., Theodoropoulos, J., 2013: The clinical status of cartilage tissue regeneration in humans. Osteoarthritis and Cartilage, 21 (12), 1824—1833. DOI: 10.1016/j.joca.2013.08.024.10.1016/j.joca.2013.08.024
Open DOI Search in Google Scholar Back to article
23. Muzzarelli, R. A. A., 2009: Review: Chitins and chitosans for the repair of wounded skin, nerve, cartilage and bone. Carb. Pol., 76 (2), 167—182. DOI: 10.1016/j.carbpol.2008.11.002.10.1016/j.carbpol.2008.11.002
Open DOI Search in Google Scholar Back to article
24. Nair, S. L., Laurencin, C. T., 2007: Biodegradable polymers as biomaterials. Prog. Polym. Sci., 32, 762—798. http://dx.doiorg/10.1016/j.progpolymsci.2007.05.017.10.1016/j.progpolymsci.2007.05.017
Search in Google Scholar Back to article
25. Nehrer, S., Breinan, H. A., Ramappa, A., Young, G., Shortkroff, S., Louie, L. K., et al., 1997: Matrix collagen type and pore size influence behaviour of seeded canine chondrocytes. Biomaterials, 18 (11), 769—776.10.1016/S0142-9612(97)00001-X
Search in Google Scholar Back to article
26. Pelttari, K., Steck, E., Richter, W., 2008: The use of mesenchymal stem cells for chondrogenesis injury, Int. J. Care Inj., 39S1, S58—S65. DOI:10.1016/j.injury.2008.01.038.10.1016/j.injury.2008.01.038
Open DOI Search in Google Scholar Back to article
27. Puppi, D., Chiellini, F., Piras, A. M., Chiellini, E., 2010: Polymeric materials for bone and cartilage repair. Progress in Polymer Science, 35, 403—444.10.1016/j.progpolymsci.2010.01.006
Search in Google Scholar Back to article
28. Solchaga, L. A., Penick, K. J., Welter, J. F., 2011: Chondrogenic differentiation of bone marrow-derived mesenchymal stem cells: Tips and tricks. Methods Mol. Biol., 698, 253—278. DOI: 10.1007/978-1-60761-999-4_20.10.1007/978-1-60761-999-4_20
Open DOI Search in Google Scholar Back to article
29. Tan, H., Chu, C.R., Payne, K. A., Marra, K. G., 2009: Injectable in situ forming biodegradable chitosan-hyaluronic acid based hydrogels for cartilage tissue engineering. Bio-materials, 30(13), 2499—2506. DOI: 10.1016/j.biomaterials.2008.12.080.10.1016/j.biomaterials.2008.12.080
Open DOI Search in Google Scholar Back to article
30. Tan, L., Ren, Y., Kuijer, R., 2012: A 1-min method for homogenous cell seeding in porous scaffolds. J. Biomat. Applic., 877—898. DOI: 10.1177/0885328210389504.10.1177/0885328210389504
Open DOI Search in Google Scholar Back to article
31. Tang, X., Fan, L., Pei, M., Zeng, L., Ge, Z., 2015: Evolving concepts of chondrogenic differentiation: history, state-ofthe-art and future perspectives. Eur. Cells Mater., 30, 12—27.10.22203/eCM.v030a02
Search in Google Scholar Back to article
32. Vater, C., Kasten, P., Stiehler, M., 2011: Review. Culture media for the differentiation of mesenchymal stromal cells. Acta Biomaterialia, 7 (2), 463—477. DOI: 10.1016/j.actbio.2010.07.037.10.1016/j.actbio.2010.07.037
Open DOI Search in Google Scholar Back to article
33. Wang, Y., Bian, Y. Z., Wu, Q., Chen, G. Q., 2008: Evaluation of three-dimensional scaffolds prepared from poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) for growth of allogeneic chondrocytes for cartilage repair in rabbits. Bio-materials, 29 (19), 2858—2868. DOI: 10.1016/j.biomaterials.2008.03.021.10.1016/j.biomaterials.2008.03.021
Open DOI Search in Google Scholar Back to article
34. Yamane, S., Iwasaki, N., Majima, T., Fukanoshi, T., Masuko, T., Harada, K., et al., 2005: Feasibility of chitosan-based hyaluronic acid hybrid biomaterial for a novel scaffold in cartilage tissue engineering. Biomaterials, 26 (6), 611—619. DOI: 10.1016/j.biomaterials.2004.03.01310.1016/j.biomaterials.2004.03.013
Open DOI Search in Google Scholar Back to article
35. Yang, B., Li, X. Y., Shi, S., Kong, X. Y., Guo, G., Huang, M. J., et al., 2010: Preparation and characterization of a novel chitosan scaffold. Carb. Pol., 80 (3), 860—865.10.1016/j.carbpol.2009.12.044
Search in Google Scholar Back to article

Effects of Cell Seeding Methods on Chondrogenic Differentiation of Rat Mesenchymal Stem Cells in Polyhydroxybutyrate/Chitosan Scaffolds

Abstract

Paradigm

My account