Biomedical subjects

Yaling Shi

Publications and source records attributed to Yaling Shi.

5 recordsLinked to original sources

PubMed2006

Novel geometries for tissue-engineered tendonous collagen constructs.

A promising approach to addressing the performance limitations of currently available mechanical and bioprosthetic heart valves lies in tissue engineering. Tissue-engineered valves should incorporate the complex microstructure of the native valves to mimic their unique mechanics. This would include a layered topology, mesh networks, and branched collagen fiber bundles. Our approach to heart valve tissue engineering is to develop the functional components of the aortic valve cusps separately in vitro and, once they are mature, integrate them into a composite valve structure. Here we report on our efforts to create more complex collagenous structures, suitable for heart valve tissue engineering. Collagen fiber bundles were fabricated using the principle of directed collagen gel contraction, using neonatal rat aortic smooth muscle cells and acid-soluble type I rat-tail tendon collagen. The collagen gels were cast into rectangular or branched wells with porous end holders that constrained the gels longitudinally but allowed contraction to occur transverse to the long axis. Pairs of such constructs were placed in direct contact with each other and cultured further to determine whether they integrated to form continuous tissue. After 6-8 weeks of culture, highly compacted and aligned collagen fiber bundles formed. Mechanical testing revealed that linear constructs (2 free ends) with an 8:1 aspect ratio were significantly stronger than similar constructs with an aspect ratio of 2:1 (mean +/- SD, 298 +/- 90 kPa vs. 152 +/- 49 kPa; p < .001). Branching reduced mechanical strength considerably. Constructs fabricated with 4 free ends were significantly weaker than constructs with 3 ends (31 +/- 32 kPa vs. 116 +/- 66 kPa; p < .003). Histologic images demonstrated the integration of the crossed collagen bundles, with a bonding strength of 2.1 +/- 1.1 g (0.02 N). We found that the geometry of the molds into which the collagen constructs are cast can greatly affect their mechanical strength: multibranched constructs were the weakest, and long, linear constructs were the strongest. We also found that integration of collagen constructs occurs in vitro and that the fabrication of a composite structure in vitro is probably feasible.

Absorbable Implants↗

PubMed2004

Characterization of statically loaded tissue-engineered mitral valve chordae tendineae.

Chordae tendineae are essential to the proper function of the mitral valve. Native chordae contain a dense collagenous core and an outer elastin sheath. We have been using the principle of directed collagen gel shrinkage to fabricate tissue-engineered mitral valve chordae. Because the microstructure of biologic tissues determines their mechanical behavior, the morphology of collagen and elastin in tissue-engineered chordae should mimic that of native chordae. The objective of this study, therefore, was to examine the morphology of our tissue-engineered constructs in comparison to native chordae. A collagen-cell suspension was cast into silicon rubber wells with microporous anchors at the ends and cultured in an incubator. The anchors allowed shrinkage to occur only transverse to the long axis of the wells, thus creating highly aligned collagen fibril constructs. The collagen constructs were cultured for 8 weeks and characterized mechanically, histologically, and biochemically at different culture time points. Histologic sections showed that in all mature constructs collagen fibers were oriented parallel to the long axis of the constructs. At the edge of the tissue collagen fibers were in general straight, whereas in the middle of the tissue they were wavy. Transmission electron microscopy showed a progressive increase in the density and longitudinal orientation of collagen fibrils with culture time. Light and scanning electron microscopy showed the presence of an elastin sheath around the collagen core. Immunostaining demonstrated that smooth muscle cells differentiate during tissue development and TUNEL assay showed that cells in the interior of the constructs undergo apoptosis. This study has demonstrated that collagen-cell constructs, with material properties and microstructure similar to native mitral valve chordae, can be developed using static culture.

Animals↗

PubMed2003

Loss of chondroitin 6-sulfate and hyaluronan from failed porcine bioprosthetic valves.

Explanted porcine bioprosthetic valves have a thinned spongiosa, partially because of an overall loss of glycosaminoglycans (GAGs). We measured the concentrations of specific GAG classes in explanted bioprosthetic valves (n = 14, implanted 12.0 +/- 4.7 years) compared with glutaraldehyde-fixed porcine controls. After extraction with NaOH, GAGs were analyzed using either a hexuronic acid assay or fluorophore-assisted carbohydrate electrophoresis to quantify the individual GAG classes. The total GAG concentration in explants was 198 +/- 95 pmol/mg wet weight-93% less than freshly fixed controls. Explants also contained altered proportions of the different GAG classes relative to controls. The proportions of hyaluronan and chondroitin/dermatan-6-sulfate were reduced from 39 to 7% and 34 to 18% of total GAGs, respectively. The predominant explant GAG class was chondroitin/dermatan-4-sulfate (proportion elevated from 14 to 70%). This GAG is commonly found in the collagen-associated proteoglycan decorin, which is likely well crosslinked by glutaraldehyde. Chondroitin-6-sulfate is commonly found in the water- and hyaluronan-binding proteoglycan versican, which is likely poorly crosslinked. The loss of versican and its associated water-binding capacity is consistent with the thinned spongiosa. The resultant compromise of hydration, compressive resistance, and viscoelasticity may be responsible for the deterioration of the bioprosthesis in vivo.

Animals↗

PubMed2003

Fabrication of mitral valve chordae by directed collagen gel shrinkage.

The principles of tissue engineering are being used to explore numerous applications in reconstructive surgery. Mitral valve chordae are one such potential area, as mitral valve repair is increasing in popularity and synthetic materials have not been used widely. The use of cells, combined with reconstituted type I collagen, is an attractive option for fabricating materials for the replacement of thin tendonous structures such as mitral valve chordae. We have been using the principle of directed collagen gel shrinkage to fabricate tendinous structures with good mechanical properties. In this study, our objective was to maximize the strength of the collagen constructs by choosing cell type and optimizing cell-seeding density, culture time, and initial collagen concentration. A collagen-cell suspension was cast into silicone rubber wells with microporous anchors at the ends and cultured in an incubator. The anchors allowed shrinkage to occur only transverse to the long axis of the wells, thus creating highly aligned collagenous constructs. Collagen gel contraction increased with higher cell-seeding density. The optimal value was 10(6) cells/mL. The rate of gel contraction decreased with the initial collagen concentration. Fibril density increased with culture time, as the gel contracted. After the system was optimized, the mechanical strength of the constructs increased to 1.1 MPa, a value at least an order of magnitude greater than previously published results with similar systems. This study has demonstrated that collagen-cell constructs, with material properties similar to those of native mitral valve chordae, can be developed using the principle of directed collagen gel shrinkage. These structures may have application in other areas that require small-diameter tendons.

Animals↗

PubMed2002

Towards tissue engineering of a composite aortic valve.

A tissue-engineered valve needs to incorporate the complex microstructure of the native aortic valve if it is to be as durable as existing bioprosthetic valves. Native aortic valve cusps contain large collagen fiber bundles surrounded by tubes of elastin, linked together by elastin sheets and struts. They also contain glycosaminoglycans (GAGs) that bind water and give the valve cusp a gelatinous consistency. Our approach to tissue engineering the aortic valve is to fabricate the cusp from the building blocks described above. We have developed collagen fiber bundles using the principle of directed collagen gel shrinkage, a GAG matrix by crosslinking high molecular weight hyaluronan with divinyl sulfone, and elastin sheets and tubes by culturing neonatal aortic fibroblasts on the appropriate substrates. To make collagen fiber bundles, cells are mixed with solubilized fibrillar collagen and cast into silicon rubber wells fitted with microporous holders to entrap the gel and hold it in place. As the cells interact with the collagen fibrils, they contract and compact the gel. Since the gel is constrained in the longitudinal direction, it can contract only laterally, forming well-aligned, strong (> 1 MPa) collagen fiber bundles. Elastin sheaths from naturally around the collagen fiber bundles and atop the hyaluronan gel when the neonatal aortic fibroblasts are cultured for more than 4 weeks. The individual building blocks of the aortic valve cusp, designed and fabricated according to patterns dictated by the microstructure of the native aortic valve, will be stacked together to make the final composite, tissue-engineered aortic valve cusp.

Absorbable Implants↗