The development and application of an in vitro model of coronary lesion thrombosis

Thesis (Ph. D.)--Harvard-MIT Division of Health Sciences and Technology, 2005.Includes bibliographical references (v. 2, leaves 192-205).Thrombosis is an initiating response to vascular injury. Physiologically, this process aids in the repair and remodeling of the vessel wall. However, if left unchecked, luminal occlusion may rapidly occur. The coronary vascular bed is a life-sustaining environment in which pathological thrombosis can lead to devastating outcomes such as acute coronary syndromes or post-interventional thrombosis. In order to study these coronary thrombotic reactions, it is essential to consider the physical environment in which they occur. We have developed an in vitro method for creating pulsatile flows to mimic the coronary hernodynamic setting on a beat-to-beat basis. Furthermore, he have developed techniques and protocols to parametrically vary both the biological and physical aspects of thrombosis and in doing so, have investigated the effects of real-world temporal and spatial flow perturbations on local site platelet adhesion. Not only do such variations create quantitative differences in local reactions, but qualitative differences as well as various receptors must interact to create stable adhesions in a given hemodynamic environments. These findings have implications on the propensity for certain individuals to form clot under certain conditions, as well as the environment-dependent efficacy of various clinically relevant anti-thrombotic strategies.by Kumaran Kolandaivelu.Ph.D

Solvent-cast direct-writing as a fabrication strategy for radiopaque stents

Bioresorbable stents (BRS) potential in treating coronary heart disease is still to be further developed. Current trends include research with new polymeric materials, the need for thinner struts combined with appropriate mechanical properties, radiopacity and optimized local drug delivery. This work presents a novel solvent-cast direct-write (SC-DW) printing system to manufacture BRS onto a rotating cylinder with poly-l-lactic acid (PLLA) and poly(l-lactic-co- ¿ -caprolactone) (PLCL) inks. Printed stents were characterized in terms of mechanical, thermal and biological properties with human umbilical vein endothelial cells (HUVECs). Expansion assays showed that stents withstood pressures of at least 16 atm and the indirect cytotoxicity test indicated that stents were biocompatible. Polymeric inks were further modified with the addition of 3 radiopaque agents, namely iodine, triiodobenzoic acid (TIBA) and barium sulfate (BaSO) to render stents radiopaque. Subsequent characterization showed a general increase in strut thickness with respect to control PLLA or PLCL stents, which in turn resulted in higher resistance to compression. Microcomputed tomography was used to assess stents’ radiopacity, showing that TIBA and BaSO-containing stents presented high X-ray attenuation values and maintained their radiopacity after 3 months incubation time.Peer ReviewedPostprint (published version

Microbial lysate upregulates host oxytocin

Neuropeptide hormone oxytocin has roles in social bonding, energy metabolism, and wound healing contributing to good physical, mental and social health. It was previously shown that feeding of a human commensal microbe Lactobacillus reuteri (L. reuteri) is sufficient to up-regulate endogenous oxytocin levels and improve wound healing capacity in mice. Here we show that oral L. reuteri-induced skin wound repair benefits extend to human subjects. Further, dietary supplementation with a sterile lysate of this microbe alone is sufficient to boost systemic oxytocin levels and improve wound repair capacity. Oxytocin-producing cells were found to be increased in the caudal paraventricular nucleus [PVN] of the hypothalamus after feeding of a sterile lysed preparation of L. reuteri, coincident with lowered blood levels of stress hormone corticosterone and more rapid epidermal closure, in mouse models. We conclude that microbe viability is not essential for regulating host oxytocin levels. The results suggest that a peptide or metabolite produced by bacteria may modulate host oxytocin secretion for potential public or personalized health goals.Published versio

Stent Thrombogenicity Early in High Risk Interventional Settings is Driven by Stent Design and Deployment, and Protected by Polymer-Drug Coatings

Author Manuscript: 2012 April 5Background—Stent thrombosis is a lethal complication of endovascular intervention. Concern has been raised about the inherent risk associated with specific stent designs and drug-eluting coatings, yet clinical and animal support is equivocal. Methods and Results—We examined whether drug-eluting coatings are inherently thrombogenic and if the response to these materials was determined to a greater degree by stent design and deployment with custom-built stents. Drug/polymer coatings uniformly reduce rather than increase thrombogenicity relative to matched bare metal counterparts (0.65-fold; P=0.011). Thick-strutted (162 μm) stents were 1.5-fold more thrombogenic than otherwise identical thin-strutted (81 μm) devices in ex vivo flow loops (P<0.001), commensurate with 1.6-fold greater thrombus coverage 3 days after implantation in porcine coronary arteries (P=0.004). When bare metal stents were deployed in malapposed or overlapping configurations, thrombogenicity increased compared with apposed, length-matched controls (1.58-fold, P=0.001; and 2.32-fold, P<0.001). The thrombogenicity of polymer-coated stents with thin struts was lowest in all configurations and remained insensitive to incomplete deployment. Computational modeling–based predictions of stent-induced flow derangements correlated with spatial distribution of formed clots. Conclusions—Contrary to popular perception, drug/polymer coatings do not inherently increase acute stent clotting; they reduce thrombosis. However, strut dimensions and positioning relative to the vessel wall are critical factors in modulating stent thrombogenicity. Optimal stent geometries and surfaces, as demonstrated with thin stent struts, help reduce the potential for thrombosis despite complex stent configurations and variability in deployment

The integrity and organization of the human AIPL1 functional domains is critical for its role as a HSP90-dependent co-chaperone for rod PDE6

Biallelic mutations in the photoreceptor-expressed aryl hydrocarbon receptor interacting protein-like 1 (AIPL1) are associated with autosomal recessive Leber congenital amaurosis (LCA), the most severe form of inherited retinopathy in early childhood. AIPL1 functions as a photoreceptor-specific co-chaperone that interacts with the molecular chaperone HSP90 to facilitate the stable assembly of the retinal cyclic GMP (cGMP) phosphodiesterase (PDE6) holoenzyme. In this study, we characterized the functional deficits of AIPL1 variations, some of which induce aberrant pre-mRNA AIPL1 splicing leading to the production of al- ternative AIPL1 isoforms. We investigated the ability of the AIPL1 variants to mediate an interaction with HSP90 and modulate the rod cGMP PDE6 stability and activity. Our data revealed that both the FK506 binding protein (FKBP)-like domain and the tetra- tricopeptide repeat (TPR) domain of AIPL1 are required for interaction with HSP90. We further demonstrate that AIPL1 signifi- cantly modulates the catalytic activity of heterologously expressed rod PDE6. Although the N-terminal FKBP-like domain of AIPL1 binds the farnesylated PDE6a subunit through direct interaction with the farnesyl moiety, mutations compromising the integrity of the C-terminal TPR domain of AIPL1 also failed to modulate PDE6 activity efficiently. These AIPL1 variants moreover failed to promote the HSP90-dependent stabilization of the PDE6a subunit in the cytosol. In summary, we have successfully vali- dated the disease-causing status of the AIPL1 variations in vitro. Our findings provide insight into the mechanism underlying the co-chaperone role of AIPL1 and will be critical for ensuring an early and effective diagnosis of AIPL1 LCA patients

Development of a miniature high frequency ventilator for genetically engineered newborn mice

Thesis (B.S.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 1995.Includes bibliographical references (leaf 49).by Kumaran Kolandaivelu.B.S

The development of an in vitro model of coronary lesion thrombosis

Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2003.Includes bibliographical references (leaves 86-91).by Kumaran Kolandaivelu.S.M

Enhancing physiologic simulations using supervised learning on coarse mesh solutions

Computational modelling of physical and biochemical processes has emerged as a means of evaluating medical devices, offering new insights that explain current performance, inform future designs and even enable personalized use. Yet resource limitations force one to compromise with reduced order computational models and idealized assumptions that yield either qualitative descriptions or approximate, quantitative solutions to problems of interest. Considering endovascular drug delivery as an exemplary scenario, we used a supervised machine learning framework to process data generated from low fidelity coarse meshes and predict high fidelity solutions on refined mesh configurations. We considered two models simulating drug delivery to the arterial wall: (i) two-dimensional drug-coated balloons and (ii) three-dimensional drug-eluting stents. Simulations were performed on computational mesh configurations of increasing density. Supervised learners based on Gaussian process modelling were constructed from combinations of coarse mesh setting solutions of drug concentrations and nearest neighbourhood distance information as inputs, and higher fidelity mesh solutions as outputs. These learners were then used as computationally inexpensive surrogates to extend predictions using low fidelity information to higher levels of mesh refinement. The cross-validated, supervised learner-based predictions improved fidelity as compared with computational simulations performed at coarse level meshes—a result consistent across all outputs and computational models considered. Supervised learning on coarse mesh solutions can augment traditional physics-based modelling of complex physiologic phenomena. By obtaining efficient solutions at a fraction of the computational cost, this framework has the potential to transform how modelling approaches can be applied in the evaluation of medical technologies and their real-time administration in an increasingly personalized fashion.United States. Dept. of Health and Human Services/National Institutes of Health. (U.S.) (P20RR016461)American Heart Association (12FTF12080241)Charles Stark Draper Laboratory (CSDL-29414-005, CSDL-29889-001, CSDL-30716-005 & CSDL-30736-003

Quantification of thrombus formation in malapposed coronary stents deployed in vitro through imaging analysis

Stent thrombosis is a major complication of coronary stent and scaffold intervention. While often unanticipated and lethal, its incidence is low making mechanistic examination difficult through clinical investigation alone. Thus, throughout the technological advancement of these devices, experimental models have been indispensable in furthering our understanding of device safety and efficacy. As we refine model systems to gain deeper insight into adverse events, it is equally important that we continue to refine our measurement methods. We used digital signal processing in an established flow loop model to investigate local flow effects due to geometric stent features and ultimately its relationship to thrombus formation. A new metric of clot distribution on each microCT slice termed normalized clot ratio was defined to quantify this distribution. Three under expanded coronary bare-metal stents were run in a flow loop model to induce clotting. Samples were then scanned in a MicroCT machine and digital signal processing methods applied to analyze geometric stent conformation and spatial clot formation. Results indicated that geometric stent features play a significant role in clotting patterns, specifically at a frequency of 0.6225 Hz corresponding to a geometric distance of 1.606 mm. The magnitude-squared coherence between geometric features and clot distribution was greater than 0.4 in all samples. In stents with poor wall apposition, ranging from 0.27 mm to 0.64 mm maximum malapposition (model of real-world heterogeneity), clots were found to have formed in between stent struts rather than directly adjacent to struts. This early work shows how the combination of tools in the areas of image processing and signal analysis can advance the resolution at which we are able to define thrombotic mechanisms in in vitro models, and ultimately, gain further insight into clinical performance.NIH (Grant R01 GM-49039