JEAS  Vol.3 No.1 , March 2013
Nanoencapsulation of Human Adipose Mesenchymal Stem Cells: Experimental Factors Role to Successfully Preserve Viability and Functionality of Cells
ABSTRACT

Cell nanoencapsulation is a novel delivery system based on a self-assembly technique mediated by electrostatic interactions called Layer-by-Layer (LbL) deposition, without an increase in volume implant because of the nanometric thickness of its layers. LbL coats the entire surface of individual cells, providing mechanical resistance to cells against manipulation and storage conditions prior to implantation in the patient. In this work, single-cell nanocapsule formation using human adipose-derived mesenchymal stem cells (ADSC) given their potentiality in regenerative medicine was assessed by fluorescence microscopy and Zeta potential assays. Both methodologies were conclusive in showing layer-by-layer nanocapsule formation of every single ADSC. Significant differences in terms of viability and cell functionality preservation were observed depending on the polycation used. Using a combination of fluorescence microscopy and fluorimetric assays, we found that cell survival after nanocapsulation was only efficient when chitosan was added to cells. These results were consistent with other cell types used in this study. Other polycations such as poly(allylamine hydrochloride) (PAH), poly(diallyldimethylammonium chloride) (PDADMAC) and poly-L-lysine (PLL) markedly decreased cell viability (22%, 11% and 15%, respectively). In addition, the use of potassium-enriched saline solutions, such as Hanks and Ringer’s solution, during the nanoencapsulation process on ADSCs was harmful on cell viability compared to standard media (36% vs 79%, respectively). The addition of a mixture of polyanions such as hyaluronic acid and chondroitin sulfate did not affect cell viability (79% and 81%). The combination of chitosan/hyaluronic acid and chondroitin sulfate was also effective in preserving the cell functionality of ADSCs, including the proliferation and differentiation of these cells as assessed by MTT assay and microscopy, respectively. Taken together, these results indicate that ADSCs can be successfully nanoencapsulated using a first layer of chitosan and a second layer of a combination of hyaluronic acid and chondroitin sulfate with a standard potassium concentration in the culture medium.


Cite this paper
D. Hachim, J. Melendez and R. Ebensperger, "Nanoencapsulation of Human Adipose Mesenchymal Stem Cells: Experimental Factors Role to Successfully Preserve Viability and Functionality of Cells," Journal of Encapsulation and Adsorption Sciences, Vol. 3 No. 1, 2013, pp. 1-12. doi: 10.4236/jeas.2013.31001.
References
[1]   H. E. Young, C. Duplaa, R. Katz, T. Thompson, K. C. Hawkins, A. N. Boev, et al., “Adult-Derived Stem Cells and Their Potential for Use in Tissue Repair and Molecular Medicine,” Journal of Cellular and Molecular Medicine, Vol. 9, No. 3, 2005, pp. 753-769. doi:10.1111/j.1582-4934.2005.tb00510.x

[2]   K. Le Blanc, “Mesenchymal Stem Cells (MSCs) in Immunotherapy,” Xenotransplantation, Vol. 14, 2007, pp. 426-427.

[3]   K. Le Blanc and O. Ringden, “Immunomodulation by Mesenchymal Stem Cells and Clinical Experience,” Journal of Internal Medicine, Vol. 262, No. 5, 2007, pp. 509-525. doi:10.1111/j.1365-2796.2007.01844.x

[4]   M. Dominici, K. Le Blanc, I. Mueller, I. Slaper-Cortenbach, F. Marini, D. Krause, et al., “Minimal Criteria for Defining Multipotent Mesenchymal Stromal Cells,” The International Society for Cellular Therapy Position Statement, Cytotherapy, Vol. 8, No. 4, 2006, pp. 315-317. doi:10.1080/14653240600855905

[5]   A. Schaffler and C. Buchler, “Concise Review: Adipose Tissue-Derived Stromal Cells-Basic and Clinical Implications for Novel Cell-Based Therapies,” Stem Cells, Vol. 25, No. 4, 2007, pp. 818-827. doi:10.1634/stemcells.2006-0589

[6]   H. Mizuno, “Adipose-Derived Stem Cells for Tissue Repair and Regeneration: Ten Years of Research and a Literature Review,” Journal of Nippon Medical School, Vol. 76, No. 2, 2009, pp. 56-66. doi:10.1272/jnms.76.56

[7]   A. Diaspro, D. Silvano, S. Krol, O. Cavalleri and A. Gliozzi, “Single Living Cell Encapsulation in Nano-Organized Polyelectrolyte Shells,” Langmuir, Vol. 18, No. 13, 2002, pp. 5047-5050. doi:10.1021/la025646e

[8]   S. Krol, A. Diaspro, R. Magrassi, P. Ballario, B. Grimaldi, P. Filetici, et al., “Nanocapsules: Coating for Living Cells,” IEEE Transactions on Nanobioscience, Vol. 3, No. 1, 2004, pp. 32-38. doi:10.1109/TNB.2004.824279

[9]   S. Krol, S. del Guerra, M. Grupillo, A. Diaspro, A. Gliozzi and P. Marchetti, “Multilayer Nanoencapsulation. New Approach for Immune Protection of Human Pancreatic Islets,” Nano Letters, Vol. 6, No. 9, 2006, pp. 1933-1939. doi:10.1021/nl061049r

[10]   J. T. Wilson and E. L. Chaikof, “Challenges and Emerging Technologies in the Immunoisolation of Cells and Tissues,” Advanced Drug Delivery Reviews, Vol. 60, No. 2, 2008, pp. 124-145. doi:10.1016/j.addr.2007.08.034

[11]   H. Uludag, P. De Vos and P. A. Tresco, “Technology of Mammalian Cell Encapsulation,” Advanced Drug Delivery Reviews, Vol. 42, No. 1-2, 2000, pp. 29-64. doi:10.1016/S0169-409X(00)00053-3

[12]   S. Krol, O. Cavalleri, P. Ramoino, A. Gliozzi and A. Diaspro, “Encapsulated Yeast Cells Inside Paramecium Primaurelia: A Model System for Protection Capability of Polyelectrolyte Shells,” Journal of Microscopy, Vol. 212, No. 3, 2003, pp. 239-243. doi:10.1111/j.1365-2818.2003.01251.x

[13]   S. Krol, M. Nolte, A.Diaspro , D. Mazza, R. Magrassi, A. Gliozzi, et al., “Encapsulated Living Cells on Microstructured Surfaces,” Langmuir, Vol. 21, No. 2, 2005, pp. 705-709. doi:10.1021/la047715q

[14]   A. Diaspro, “Nanocapsules: A European Community Interdisciplinary Network in the Nanobiosciences,” IEEE Transactions on Nanobioscience, Vol. 3, No. 1, 2004, pp. 1-2. doi:10.1109/TNB.2004.824255

[15]   Z. L. Zhi, B. Liu, P. M. Jones and J. C. Pickup, “Polysaccharide Multilayer Nanoencapsulation of Insulin-Producing Beta-Cells Grown as Pseudoislets for Potential Cellular Delivery of Insulin,” Biomacromolecules, Vol. 11, No. 3, 2010, pp. 610-616. doi:10.1021/bm901152k

[16]   N. G. Veerabadran, P. L. Goli, S. S. Stewart-Clark, Y. M. Lvov and D. K. Mills, “Nanoencapsulation of Stem Cells within Polyelectrolyte Multilayer Shells,” Macromolecular Bioscience, Vol. 7, No. 7, 2007, pp. 877-882. doi:10.1002/mabi.200700061

[17]   M. Germain, P. Balaguer, J. C. Nicolas, F. Lopez, J. P. Esteve, G. B. Sukhorukov, et al., “Protection of Mammalian Cell Used in Biosensors by Coating with a Polyelectrolyte Shell,” Biosensors and Bioelectronics, Vol. 21, No. 8, 2006, pp. 1566-1573. doi:10.1016/j.bios.2005.07.011

[18]   P. K. Dutta, M. N. Ravikumar and J. Dutta, “Chitin and Chitosan for Versatile Applications,” Journal of Macromolecular Science, Part C: Polymer Reviews, Vol. 42, No. 3, 2002, pp. 307-354. doi:10.1081/MC-120006451

[19]   P. Angermann, “Glucosamine and Chondroitin Sulfate in the Treatment of Arthritis,” Ugeskrift for Laeger, Vol. 165, No. 5, 2003, pp. 451-454.

[20]   P. Hoffman, K. Meyer and A. Linker, “Transglycosylation during the Mixed Digestion of Hyaluronic Acid and Chondroitin Sulfate by Testicular Hyaluronidase,” Journal of Biological Chemistry, Vol. 219, 1956, pp. 653-663.

[21]   D. K. Singh and A. R. Ray, “Biomedical Applications of Chitin, Chitosan, and Their Derivatives,” Journal of Macromolecular Science, Part C: Polymer Reviews, Vol. 40, No. 1, 2000, pp. 69-83.

[22]   B. A. Bunnell, M. Flaat, C. Gagliardi, B. Patel and C. Ripoll, “Adipose-Derived Stem Cells: Isolation, Expansion and Differentiation,” Methods, Vol. 45, No. 2, 2008, pp. 115-120. doi:10.1016/j.ymeth.2008.03.006

[23]   N. V. Krishnamurthy and B. Gimi, “Encapsulated Cell Grafts to Treat Cellular Deficiencies and Dysfunction,” Critical ReviewsTM in Biomedical Engineering, Vol. 39, No. 6, 2011, pp. 473-491. doi:10.1615/CritRevBiomedEng.v39.i6.10

[24]   A. Goren, N. Dahan, E. Goren, L. Baruch and M. Machluf, “Encapsulated Human Mesenchymal Stem Cells: A Unique Hypoimmunogenic Platform for Long-Term Cellular Therapy,” The FASEB Journal, Vol. 24, No. 1, 2010, pp. 22-31. doi:10.1096/fj.09-131888

[25]   T. Bhaiji, Z. Zhi and J. C. Pickup, “Improving Cellular Function and Immune Protection via Layer-by-Layer Nanocoating of Pancreatic Islet b-Cell Spheroids Cocultured with Mesenchymal Stem Cells,” Journal of Biomedical Materials Research Part A, Vol. 100A, No. 6, 2012, pp. 1628-1636. doi:10.1002/jbm.a.34111

[26]   A. Matsuzawa, M. Matsusaki and M. Akashi, “Effectiveness of Nanometer-Sized Extracellular Matrix Layer-by Layer Assembled Films for a Cell Membrane Coating Protecting Cells from Physical Stress,” Langmuir, 2012 (online). doi:dx.doi.org/10.1021/la303459v

[27]   A. H. Poghosyan, L. H. Arsenyan, H. H. Gharabekyan, J. Koetz and A. A. Shahinyan, “Molecular Dynamics Study of Poly(diallyldimethylammonium chloride) (PDADMAC)/Sodium Dodecyl Sulfate (SDS)/Decanol/Water Systems,” Journal of Physical Chemistry B, Vol. 113, No. 5, 2009, pp. 1303-1310. doi:10.1021/jp806289c

[28]   J. M. Lourenco, P. A. Ribeiro, A. M. B. do Rego, F. M. Braz Fernandes, A. M. Moutinho and M. Raposo, “Counterions in Poly(allylamine hydrochloride) and Poly(styrene sulfonate) Layer-by-Layer Films,” Langmuir, Vol. 20, No. 19, 2004, pp. 8103-8109. doi:10.1021/la049872v

[29]   I. A. Cree, “Cancer Biology,” Cancer Biology, Vol. 731, 2011, pp. 1-11. doi:10.1007/978-1-61779-080-5_1

[30]   M. Chittchang, N. Salamat-Miller, H. H. Alur, D. G. Vander Velde, A. K. Mitra and T. P. Johnston, “POLY(L-lysine) as a Model Drug Macromolecule with which to Investigate Secondary Structure and Microporous Membrane Transport, Part 2: Diffusion Studies,” Journal of Pharmacy and Pharmacology, Vol. 54, No. 11, 2002, pp. 1497-1505. doi:10.1211/002235702108

 
 
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