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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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Figures

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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. 1

Overview of the experimental methodology used in this study

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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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