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Research Papers

Alginate Microspheroid Encapsulation and Delivery of MG-63 Cells Into Polycaprolactone Scaffolds: A New Biofabrication Approach for Tissue Engineering Constructs

[+] Author and Article Information
Lokesh K. Narayanan

Edward P. Fitts Department of Industrial
and Systems Engineering,
North Carolina State University,
400 Daniels Hall,
Raleigh, NC 27616
e-mail: lnaraya@ncsu.edu

Arun Kumar

Edward P. Fitts Department of Industrial
and Systems Engineering,
North Carolina State University,
400 Daniels Hall,
Raleigh, NC 27616
e-mail: akumar9@ncsu.edu

Zhuo (George) Tan

Edward P. Fitts Department of Industrial
and Systems Engineering,
North Carolina State University,
400 Daniels Hall,
Raleigh, NC 27616
e-mail: ztan@ncsu.edu

Susan Bernacki

UNC/NCSU Joint Department
of Biomedical Engineering,
North Carolina State University,
4102C Engineering Building III,
Raleigh, NC 27616
e-mail: shbernac@ncsu.edu

Binil Starly

Edward P. Fitts Department of Industrial
and Systems Engineering;
UNC/NCSU Joint Department of Biomedical
Engineering,
North Carolina State University,
400 Daniels Hall,
Raleigh, NC 27616
e-mail: bstarly@ncsu.edu

Rohan A. Shirwaiker

Edward P. Fitts Department of Industrial
and Systems Engineering;
UNC/NCSU Joint Department of Biomedical
Engineering,
North Carolina State University,
400 Daniels Hall,
Raleigh, NC 27616
e-mail: rashirwaiker@ncsu.edu

1Corresponding author.

Manuscript received June 4, 2015; final manuscript received July 21, 2015; published online September 29, 2015. Assoc. Editor: Ibrahim Ozbolat.

J. Nanotechnol. Eng. Med 6(2), 021003 (Sep 29, 2015) (8 pages) Paper No: NANO-15-1043; doi: 10.1115/1.4031174 History: Received June 04, 2015; Revised July 21, 2015

Scaffolds play an important role in tissue engineering by providing structural framework and a surface for cells to attach, proliferate, and secrete extracellular matrix (ECM). In order to enable efficient tissue formation, delivering sufficient cells into the scaffold three-dimensional (3D) matrix using traditional static and dynamic seeding methods continues to be a critical challenge. In this study, we investigate a new cell delivery approach utilizing deposition of hydrogel-cell encapsulated microspheroids into polycaprolactone (PCL) scaffolds to improve the seeding efficiency. Three-dimensional-bioplotted PCL constructs (0 deg/90 deg lay down, 284 ± 6 μm strand width, and 555 ± 8 μm strand separation) inoculated with MG-63 model bone cells encapsulated within electrostatically generated calcium-alginate microspheroids (Ø 405 ± 13 μm) were evaluated over seven days in static culture. The microspheroids were observed to be uniformly distributed throughout the PCL scaffold cross section. Encapsulated cells remained viable within the constructs over the test interval with the highest proliferation noted at day 4. This study demonstrates the feasibility of the new approach and highlights the role and critical challenges to be addressed to successfully utilize 3D-bioprinting for microencapsulated cell delivery.

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References

Paine, G. F. , Bentley, W. E. , Sun, W. , and Forgacs, G. , 2013, “ Biofabrication,” Encyclopedia of Biophysics, G. C. K. Roberts, ed., Springer, Berlin, pp. 193–194.
Berthiaume, F. , Maguire, T. J. , and Yarmush, M. L. , 2011, “ Tissue Engineering and Regenerative Medicine: History, Progress, and Challenges,” Annu. Rev. Chem. Biomol. Eng., 2, pp. 403–430. [CrossRef] [PubMed]
Shuai, C. , Mao, Z. , and Han, Z. , 2014, “ Fabrication and Characterization of Calcium Silicate Scaffolds for Tissue Engineering,” J. Mech. Med. Biol., 14(4), p. 1450049. [CrossRef]
Chen, C. H. , Shyu, V. B. , and Chen, J. P. , 2014, “ Selective Laser Sintered Poly-Epsilon-Caprolactone Scaffold Hybridized With Collagen Hydrogel for Cartilage Tissue Engineering,” Biofabrication, 6(1), p. 015004. [CrossRef] [PubMed]
Tanodekaew, S. , Channasanon, S. , and Kaewkong, P. , 2013, “ PLA-HA Scaffolds: Preparation and Bioactivity,” Proc. Eng., 59, pp. 144–149. [CrossRef]
Elomaa, L. , Teixeira, S. , and Hakala, R. , 2011, “ Preparation of Poly(Ε-Caprolactone)-Based Tissue Engineering Scaffolds by Stereolithography,” Acta Biomater., 7(11), pp. 3850–3856. [CrossRef] [PubMed]
Jensen, J. , Rolfing, J. H. , and Le, D. Q. , 2014, “ Surface-Modified Functionalized Polycaprolactone Scaffolds for Bone Repair: In Vitro and In Vivo Experiments,” J. Biomed. Mater. Res. A, 102(9), pp. 2993–3003. [CrossRef] [PubMed]
Korpela, J. , Kokkari, A. , and Korhonen, H. , 2013, “ Biodegradable and Bioactive Porous Scaffold Structures Prepared Using Fused Deposition Modeling,” J. Biomed. Mater. Res. B, 101(4), pp. 610–619. [CrossRef]
Ragaert, K. , De Somer, F. , and Van de Velde, S. , 2013, “ Methods for Improved Flexural Mechanical Properties of 3D-Plotted PCL-Based Scaffolds for Heart Valve Tissue Engineering,” Stroj. Vestn., 59(11), pp. 669–676. [CrossRef]
Rücker, M. , Laschke, M. W. , and Junker, D. , 2008, “ Vascularization and Biocompatibility of Scaffolds Consisting of Different Calcium Phosphate Compounds,” J. Biomed. Mater. Res. A, 86(4), pp. 1002–1011. [CrossRef] [PubMed]
Rücker, M. , Laschke, M. W. , and Junker, D. , 2006, “ Angiogenic and Inflammatory Response to Biodegradable Scaffolds in Dorsal Skinfold Chambers of Mice,” Biomaterials, 27(29), pp. 5027–5038. [CrossRef] [PubMed]
Sheshadri, P. , and Shirwaiker, R. A. , 2015, “ Characterization of Material–Process–Structure Interactions in the 3D-Bioplotting of Polycaprolactone,” 3D Print. Addit. Manuf., 2(1), pp. 20–31.
Kim, J. Y. , Yoon, J. J. , and Park, E. K. , 2009, “ Cell Adhesion and Proliferation Evaluation of SFF-Based Biodegradable Scaffolds Fabricated Using a Multi-Head Deposition System,” Biofabrication, 1(1), p. 015002. [CrossRef] [PubMed]
Villalona, G. A. , Udelsman, B. , and Duncan, D. R. , 2010, “ Cell-Seeding Techniques in Vascular Tissue Engineering,” Tissue Eng. Part B, 16(3) pp. 341–350. [CrossRef]
Fedorovich, N. E. , Alblas, J. , and de Wijn, J. R. , 2007, “ Hydrogels as Extracellular Matrices for Skeletal Tissue Engineering: State-of-the-Art and Novel Application in Organ Printing,” Tissue Eng., 13(8), pp. 1905–1925. [CrossRef] [PubMed]
Abbott, A. , 2003, “ Cell Culture: Biology's New Dimension,” Nature, 424(6951), pp. 870–872. [CrossRef] [PubMed]
Bokhari, M. A. , Akay, G. , and Zhang, S. , 2005, “ The Enhancement of Osteoblast Growth and Differentiation In Vitro on a Peptide Hydrogel—polyHIPE Polymer Hybrid Material,” Biomaterials, 26(25), pp. 5198–5208. [CrossRef] [PubMed]
Dixit, V. , Darvasi, R. , and Arthur, M. , 1990, “ Restoration of Liver Function in Gunn Rats Without Immunosuppression Using Transplanted Microencapsulated Hepatocytes,” Hepatology, 12(6), pp. 1342–1349. [CrossRef] [PubMed]
Maguire, T. , Novik, E. , and Schloss, R. , 2006, “ Alginate-PLL Microencapsulation: Effect on the Differentiation of Embryonic Stem Cells Into Hepatocytes,” Biotechnol. Bioeng., 93(3), pp. 581–591. [CrossRef] [PubMed]
Lim, F. , and Sun, A. , 1980, “ Microencapsulated Islets as Bioartificial Endocrine Pancreas,” Science, 210(4472), pp. 908–910. [CrossRef] [PubMed]
De Vos, P. , Faas, M. M. , and Strand, B. , 2006, “ Alginate-Based Microcapsules for Immunoisolation of Pancreatic Islets,” Biomaterials, 27(32), pp. 5603–5617. [CrossRef] [PubMed]
Maysinger, D. , Berezovskaya, O. , and Fedoroff, S. , 1996, “ The Hematopoietic Cytokine Colony Stimulating Factor 1 Is Also a Growth Factor in the CNS: (II) Microencapsulated CSF-1 and LM-10 Cells as Delivery Systems,” Exp. Neurol., 141(1), pp. 47–56. [CrossRef] [PubMed]
Winn, S. R. , Tresco, P. A. , and Zielinski, B. , 1991, “ Behavioral Recovery Following Intrastriatal Implantation of Microencapsulated PC12 Cells,” Exp. Neurol., 113(3), pp. 322–329. [CrossRef] [PubMed]
Cheng, H. W. , Tsui, Y. K. , and Cheung, K. M. , 2009, “ Decellularization of Chondrocyte-Encapsulated Collagen Microspheres: A Three-Dimensional Model to Study the Effects of Acellular Matrix on Stem Cell Fate,” Tissue Eng. Part C, 15(4), pp. 697–706. [CrossRef]
Loty, S. , Sautier, J. , and Loty, C. , 1998, “ Cartilage Formation by Fetal Rat Chondrocytes Cultured in Alginate Beads: A Proposed Model for Investigating Tissue–Biomaterial Interactions,” J. Biomed. Mater. Res., 42(2), pp. 213–222. [CrossRef] [PubMed]
Kaul, G. , Cucchiarini, M. , and Arntzen, D. , 2006, “ Local Stimulation of Articular Cartilage Repair by Transplantation of Encapsulated Chondrocytes Overexpressing Human Fibroblast Growth Factor 2 (FGF-2) In Vivo,” J. Gene Med., 8(1), pp. 100–111. [CrossRef] [PubMed]
Pautke, C. , Schieker, M. , and Tischer, T. , 2004, “ Cell Lines MG-63, Saos-2 and U-2 OS in Comparison to Human Osteoblasts,” Anticancer Res., 24(6), pp. 3743–3748. [PubMed]
Keshaw, H. , Forbes, A. , and Day, R. M. , 2005, “ Release of Angiogenic Growth Factors From Cells Encapsulated in Alginate Beads With Bioactive Glass,” Biomaterials, 26(19), pp. 4171–4179. [CrossRef] [PubMed]
Bian, L. , Zhai, D. Y. , and Tous, E. , 2011, “ Enhanced MSC Chondrogenesis Following Delivery of TGF-Β3 From Alginate Microspheres Within Hyaluronic Acid Hydrogels In Vitro and In Vivo,” Biomaterials, 32(27), pp. 6425–6434. [CrossRef] [PubMed]
Lewis, A. , and Colton, C. K. , 2007, Principles of Tissue Engineering, 3rd ed., Academic Press, Burlington, VT, pp. 405–418.
Croll, T. I. , Gentz, S. , and Mueller, K. , 2005, “ Modeling Oxygen Diffusion and Cell Growth in a Porous, Vascularising Scaffold for Soft Tissue Engineering Applications,” Chem. Eng. Sci., 60(17), pp. 4924–4934. [CrossRef]
Volkmer, E. , Drosse, I. , and Otto, S. , 2008, “ Hypoxia in Static and Dynamic 3D Culture Systems for Tissue Engineering of Bone,” Tissue Eng. Part A, 14(8), pp. 1331–1340. [CrossRef] [PubMed]
Jaasma, M. J. , Plunkett, N. A. , and O'Brien, F. J. , 2008, “ Design and Validation of a Dynamic Flow Perfusion Bioreactor for Use With Compliant Tissue Engineering Scaffolds,” J. Biotechnol., 133(4), pp. 490–496. [CrossRef] [PubMed]

Figures

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Fig. 1

Overview of the experimental methodology used in this study

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Fig. 2

Delivering MG-63 encapsulated alginate microspheroids into PCL scaffolds: (a) scaffold before deposition, (b) location fixture, (c) pipette with microspheroid–media suspension, and (d) scaffold loaded with cell-encapsulated microspheroids

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Fig. 3

Frequency distribution of microspheroid diameters

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Fig. 4

Automated microspheroid image analysis results from matlab. White specks within the spheroid are encapsulated MG-63 cells.

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Fig. 5

Cell proliferation (alamarBlue % reduction) data from MG-63 encapsulated microspheroids-loaded scaffolds over seven days

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Fig. 6

Live/dead fluorescence images of three randomly selected microspheroids retrieved from scaffolds after seven days

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Fig. 7

(a) 50× microscope images (top down) in four quadrants of a microspheroids-deposited scaffold, (b) 100× microscope image of longitudinal cross section (along the 2.5 mm scaffold thickness), and ((c) and (d)) 150× and 200× microscope images, respectively, of a portion of longitudinal scaffold cross section

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