PubMed Health⌕ Search

Biomedical subjects

John J Stankus

Publications and source records attributed to John J Stankus.

4 recordsLinked to original sources

A seeding device for tissue engineered tubular structures.

One of the challenges in the tissue engineering of tubular tissues and organs is the efficient seeding of porous scaffolds with the desired cell type and density in a short period of time, without affecting cell viability. Though different seeding techniques have been investigated, a fast, reproducible, and efficient bulk seeding method with uniform cellular distribution has yet to be reported. In this paper, a novel seeding device utilizing the synergistic effects of vacuum, centrifugal force and flow has been developed and analyzed. The device allows porous tubular scaffolds to be uniformly bulk seeded as well as luminally surface-seeded with cells. Porous tubular polymer scaffolds were bulk and surface-seeded with cell suspensions, and cell viability and seeding efficiency were subsequently assessed. A rigorous quantitative analysis of shear stresses acting on the cells during the seeding process, and of cell location within the scaffolds following seeding was also performed. Our results showed that the scaffolds were uniformly seeded along the longitudinal and circumferential directions within the tube wall without affecting cell viability or exposing them to excessive shear stresses.

Animals↗

Development of composite porous scaffolds based on collagen and biodegradable poly(ester urethane)urea.

Our objective in this work was to develop a flexible, biodegradable scaffold for cell transplantation that would incorporate a synthetic component for strength and flexibility and type I collagen for enzymatic lability and cytocompatibility. A biodegradable poly(ester urethane)urea was synthesized from poly(caprolactone), 1,4-diisocyanatobutane, and putrescine. Using a thermally induced phase separation process, porous scaffolds were created from a mixture containing this polyurethane and 0%, 10%, 20%, or 30% type I collagen. The resulting scaffolds were found to have open, interconnected pores (from 7 to >100 um) and porosities from 58% to 86% depending on the polyurethane/collagen ratio. The scaffolds were also flexible with breaking strains of 82-443% and tensile strengths of 0.97-4.11 MPa depending on preparation conditions. Scaffold degradation was significantly increased when collagenase was introduced into an incubating buffer in a manner that was dependent on the mass fraction of collagen present in the scaffold. Mass losses could be varied from 15% to 59% over 8 weeks. When culturing umbilical artery smooth muscle cells on these scaffolds higher cell numbers were observed over a 4-week culture period in scaffolds containing collagen. In summary, a strong and flexible scaffold system has been developed that can degrade by both hydrolysis and collagenase degradation pathways, as well as support cell growth. This scaffold possesses properties that would make it attractive for future use in soft tissue applications where such mechanical and biological features would be advantageous.

Animals↗

Microintegrating smooth muscle cells into a biodegradable, elastomeric fiber matrix.

Electrospinning permits fabrication of biodegradable elastomers into matrices that can resemble the scale and mechanical behavior of the native extracellular matrix. However, achieving high-cellular density and infiltration with this technique remains challenging and time consuming. We have overcome this limitation by electrospraying vascular smooth muscle cells (SMCs) concurrently with electrospinning a biodegradable, elastomeric poly(ester urethane)urea (PEUU). Trypan blue staining revealed no significant decrease in cell viability from the fabrication process and electrosprayed SMCs spread and proliferated similar to control unprocessed SMCs. The resulting SMC microintegrated PEUU constructs were cultured under static conditions or transmural perfusion. Higher cell numbers resulted with perfusion culture with 131% and 98% more viable cells versus static culture at days 4 and 7 (p<0.05). Fluorescent imaging and hematoxylin and eosin staining further illustrated high cell densities integrated between the elastomeric fibers after perfusion culture. SMC microintegrated PEUU was strong, flexible and anisotropic with tensile strengths ranging from 2.0 to 6.5 MPa and breaking strains from 850 to 1,700% dependent on the material axis. The ability to microintegrate smooth muscle or other cell types into a biodegradable elastomer fiber matrix embodies a novel tissue engineering approach that could be applied to fabricate high cell density elastic tissue mimetics, blood vessels or other cardiovascular tissues.

Absorbable Implants↗

Fabrication of biodegradable elastomeric scaffolds with sub-micron morphologies.

The native extracellular matrix (ECM) of elastic tissues is strong and flexible and supports cell adhesion and enzymatic matrix remodeling. In an attempt to convey these ECM properties to a synthetic scaffold appropriate for soft tissue engineering applications, a biodegradable, elastomeric poly(ester urethane)urea (PEUU) was combined with type I collagen at various ratios (2.5, 5, 10, 20, 50, 60, 70, 80, and 90 wt% collagen) and electrospun to construct elastic matrices. Randomly orientated fibers in the electrospun matrices ranged in diameter from 100-900 nm, dependent on initial polymer concentration. Picrosirius red staining of matrices and CD spectroscopy of released collagen confirmed collagen incorporation and preservation of collagen structure at the higher collagen mass fractions. Matrices were strong and distensible possessing strengths of 2-13 MPa with breaking strains of 160-280% even with low PEUU content. Collagen incorporation significantly enhanced smooth muscle cell adhesion onto electrospun scaffolds. An approach has been demonstrated that mimics elastic extracellular matrices by using a synthetic component to provide mechanical function together with a biomacromolecule, collagen. Such matrices may find application in engineering soft tissue.

Absorbable Implants↗