Sexual Dimorphism in the Extracellular and Pericellular Matrix of Articular Cartilage.

OBJECTIVE: Women have a higher prevalence and burden of joint injuries and pathologies involving articular cartilage than men. Although knee injuries affecting young women are on the rise, most studies related to sexual dimorphism target postmenopausal women. We hypothesize that sexual dimorphism in cartilage structure and mechanics is present before menopause, which can contribute to sex disparities in cartilage pathologies. DESIGN: Bovine knee was used as a model to study healthy adult cartilage. We compared elastic moduli under compression, abundances of extracellular and pericellular matrix (PCM) proteins using proteomics, and PCM constituency with tissue immunofluorescence. The gene expression of matrix-related genes under basal, anabolic, and catabolic conditions was assessed by quantitative polymerase chain reaction (qPCR). RESULTS: The equilibrium modulus was higher in male cartilage compared with female cartilage. Proteoglycans were not associated with this biomechanical dimorphism. Proteomic and pathway analyses of tissue showed dimorphic enriched pathways in extracellular matrix (ECM)-related proteins in which male cartilage was enriched in matrix interconnectors and crosslinkers that strengthen the ECM network. Moreover, male and female tissue differed in enriched PCM components. Females had more abundance of collagen type VI and decorin, suggesting different PCM mechanics. Furthermore, the activation of regenerative and catabolic function in chondrocytes triggered sex-dependent signatures in gene expression, indicating dimorphic genetic regulation that is dependent on stimulation. CONCLUSIONS: We provide evidence for sexual dimorphism in cartilage before menopause. Some differences are intrinsic to chondrocytes' gene expression defined by their XX versus XY chromosomal constituency.

Bioresorbable Stent to Manage Congenital Heart Defects in Children.

Intravascular stents for pediatric patients that degrade without inhibiting vessel growth remain a clinical challenge. Here, poly(L-lactide) fibers (DH-BDS) at two thicknesses, 250 µm and 300 µm, were assembled into large, pediatric-sized stents (Ø10 - Ø20 mm). Fibers were characterized mechanically and thermally, then stent mechanical properties were compared to metal controls, while mass loss and degradation kinetics modeling estimated total stent degradation time. Thicker fibers displayed lower stiffness (1969 ± 44 vs 2126 ± 37 MPa) and yield stress (117 ± 12 vs 137 ± 5 MPa) than thinner counterparts, but exhibited similar fail strength (478 ± 28 vs 476 ± 16 MPa) at higher strains (47 ± 2 vs 44 ± 2%). Stents all exhibited crystallinity between 51.3 - 54.4% and fiber glass transition temperatures of 88.6 ± 0.5 °C and 84.6 ± 0.5 °C were well above physiological ranges. Radial strength (0.31 ± 0.01 - 0.34 ± 0.02 N/mm) in thinner stents was similar to metal stents (0.24 - 0.41 N/mm) up to Ø14 mm with no foreshortening and thicker coils granted comparable radial strength (0.32 ± 0.02 - 0.34 ± 0.02 N/mm) in stents larger than Ø14 mm. Both 10 mm (1.17 ± 0.02 % and 0.86 ± 0.1 %) and 12 mm (1.1 ± 0.03% and 0.89 ± 0.1%) stents exhibited minimal weight loss over one year. Degradation kinetics models predicted full stent degradation within 2.8 - 4.5 years depending on thickness. DH-BDS exhibiting hoop strength similar to metal stents and demonstrating minimal degradation and strength loss over the first year before completely disappearing within 3 to 4.5 years show promise as a pediatric interventional alternative to current strategies.

Three-dimensional printing of poly(glycerol sebacate fumarate) gadodiamide-poly(ethylene glycol) diacrylate structures and characterization of mechanical properties for soft tissue applications.

Bioresorbable materials have been frequently used to three-dimensional (3D) print biomedical structures. In this study, we developed a technique to 3D print poly(glycerol sebacate fumarate) gadodiamide (Rylar)-poly(ethylene glycol) diacrylate (PEGDA) samples and investigated their mechanical and thermal properties as a function of (PS) and ultraviolet intensity (UVI). The Young's modulus (E), ultimate tensile strength (UTS), failure strain (ɛF ), and glass transition temperature (Tg ) showed strong correlation with PS and UVI. Results showed E to be between 1.31 and 3.12 MPa, UTS between 0.07 and 0.43 MPa, and ɛF between 7 and 20% with brittle failure. The Tg was observed to lie between -54.48 and -49.10 °C without secondary/tertiary transitions. Dominant elastic behavior was observed from the dynamic mechanical testing viscoelastic data. Testing results were used to develop a regression predictive model for E as a function of PS and UVI. The model performance was evaluated experimentally with an average absolute error of 3.62%. The E and stress-strain response of our 3D printed samples show agreement with published data for human tracheal cartilage, and the mechanical properties were comparable to other published soft polymeric scaffolds/patches. The E' moduli were also similar to bovine articular cartilage. We have successfully demonstrated that Rylar, a novel bioresorbable radiopaque polymer, when blended with PEGDA can be 3D printed controllably for soft tissue applications such as airway obstructions. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 664-671, 2019.

Biodegradable Stents for Congenital Heart Disease.

The quest for an ideal biodegradable stent for both adult coronary and pediatric congenital heart disease applications continues. Over the past few years, a lot of progress has been made toward development of a dedicated pediatric biodegradable stent that can be used for congenital heart disease applications. At present, there are no biodegradable stents available for use in congenital heart disease. In this article, the authors review the different biodegradable materials and their limitations and provide an overview of the current biodegradable stents being evaluated for congenital heart disease applications.

Severe Burn-Induced Inflammation and Remodeling of Achilles Tendon in a Rat Model.

Severe burn causes systemic inflammation and hypercatabolism, resulting in damage to multiple organs distant to the burn site, including the musculoskeletal system. Bone mass and muscle loss have been reported. However, tendon that connects bone and muscle has not been studied in comparable detail. Here we aimed to characterize the molecular and functional changes in Achilles tendon triggered by severe burn. Forty male Sprague-Dawley rats received 40% total body surface area scald burn. Achilles tendons were collected up to 14 days postburn. Sham-treated animals served as a control group. We analyzed tendons for changes in expression of IL-6, IL-1ß, TNF, MMP9, MMP13, TGFß1, Collagens I and III, and for morphological and biomechanical changes. Gene expression of IL-6 and IL-1ß as well as MMP9 and MMP13 increased in rat tendon 3 days after burn. Col3a1 increased at day 3 and col1a1 at day 7. At day 14, TGFß1 increased, whereas the protein ratio for collagens I/III decreased, indicating tendon remodeling. Histological analysis with H&E and Picrosirius red staining further revealed a decrease in organized collagen fibers 14 days after burn. Biomechanical analysis showed a decrease in stiffness and ultimate force of tendons in burn rats.We conclude that tendinopathy was observed in Achilles tendon 14 days after severe burn, via the induction of inflammation and remodeling. The present study provides a model of tendinopathy that may be used for the development of therapeutic approaches after burn.

Bacterial sensitivity assessment of multifunctional polymeric coatings for airway stents.

Current interventional technology for pediatric airway obstruction consists of cardiovascular stents and silicon tubes. These devices are composed of permanent materials that have limitations in biocompatibility and mechanical properties that make them controversial for used in pediatrics. Bioresorbable stents offer a temporary intervention that dissolves in the body over time and can serve as a platform for local drug delivery. Here we investigate a novel approach to use an antibiotic, ciprofloxacin, as a polymerization initiator to synthesize poly(ciprofloxacin fumaric acid) (PCFA) and then a second polymer using gadodiamide as an initiator to synthesize poly(gadodiamide ciprofloxacin fumaric acid) (PGCFA). Polymer structure, degradation, thermal properties, and rheological behavior were analyzed. Ciprofloxacin released was determined and polymer degradation extracts were used in bacterial sensitivity assessments with four common airway pathogens. PCFA and PGCFA polymers and drug release properties were compared to our previously published polymer poly(fumaric acid) (PFA). These novel polymers enable new possibilities as coatings for bioresorbable biomedical applications that require antibiotic resistance and imaging capabilities. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2153-2161, 2017.

Paraffin processing of stented arteries using a postfixation dissolution of metallic and polymeric stents.

Studying the morphology of the arterial response to endovascular stent implantation requires embedding the explanted stented artery in rigid materials such as poly(methyl methacrylate) to enable sectioning through both the in situ stent and the arterial wall, thus maintaining the proper anatomic relationships. This is a laborious, time-consuming process. Moreover, the technical quality of stained plastic sections is typically suboptimal and, in some cases, precludes immunohistochemical analysis. Here we describe a novel technique for dissolution of metallic and plastic stents that is compatible with subsequent embedding of "destented" arteries in paraffin, fine sectioning, major staining protocols, and immunohistochemistry.

Bench and initial preclinical results of a novel 8 mm diameter double opposed helical biodegradable stent.

BACKGROUND: Metallic endovascular stents are utilized off-label in congenital heart disease. Biodegradable stents (BDS) offer potential advantages in a growing child. We have previously reported double opposed helical (DH) BDS up to 6 mm diameter (DH-6). The objectives are to investigate the bench characteristics of larger 8 mm diameter BDS (DH-8) manufactured with increasing strut thicknesses and the inflammatory profile in a porcine model. METHODS: DH-8 were manufactured with strut thicknesses 0.10, 0.12, and 0.18 mm and mechanical testing performed. Stents were deployed into the infrarenal descending aorta (DAO) of nine minipigs. At insertion (nonsurvival = 2), 1 week (n = 2), 1 month (n = 2), and 9 months (n = 3) follow-up angiography, intravascular ultrasound and histopathology were performed. RESULTS: There was superior recoil and collapse pressure with increasing strut thickness, with 0.18 mm having 1.0% elastic recoil and collapse pressure 0.75 Atmospheres. There was good wall apposition at insertion with 5 BDS (4 DH-8 and 1 DH-6) but suboptimal in 4 as the minipigs infrarenal DAO were >8 mm (deployed at iliac bifurcation). Structural integrity was maintained in 8 BDS with 1 DH-8 collapsed at 9 months, secondary to strut damage at insertion. No thrombosis was seen. There was mild inflammation and neointimal proliferation at 1 week and 1 month, but a moderate inflammatory response at 9 months. CONCLUSIONS: DH-8 with increased strut thickness had acceptable mechanical properties at the cost of an increased inflammatory response. Miniaturization to improve delivery and further investigation on the long-term inflammatory profile of thicker struts, including through degradation, is needed. © 2016 Wiley Periodicals, Inc.

Mechanical Interaction of an Expanding Coiled Stent with a Plaque-Containing Arterial Wall: A Finite Element Analysis.

Wall injury is observed during stent expansion within atherosclerotic arteries, related in part to stimulation of the inflammatory process. Wall stress and strain induced by stent expansion can be closely examined by finite element analysis (FEA), thus shedding light on procedure-induced sources of inflammation. The purpose of this work was to use FEA to examine the interaction of a coiled polymer stent with a plaque-containing arterial wall during stent expansion. An asymmetric fibrotic plaque-containing arterial wall model was created from intravascular ultrasound (IVUS) images of a diseased artery. A 3D model for a coil stent at unexpanded state was generated in SolidWorks. They were imported into ANSYS for FEA of combined stent expansion and fibrotic plaque-distortion. We simulated the stent expansion in the plaqued lumen by increasing balloon pressure from 0 to 12 atm in 1 atm step. At increasing pressure, we examined how the expanding stent exerts forces on the fibrotic plaque and vascular wall components, and how the latter collectively resist and balance the expansive forces from the stent. Results show the expanding coiled stent creates high stresses within the plaque and the surrounding fibrotic capsule. Lower stresses were observed in adjacent medial and adventitial layers. High principal strains were observed in plaque and fibrotic capsule. The results suggest fibrotic capsule rupture might occur at localized regions. The FEA/IVUS method can be adapted for routine examination of the effects of the expansion of selected furled stents against IVUS-reconstructed diseased vessels, to improve stent deployment practices.

Thermally processed polymeric microparticles for year-long delivery of dexamethasone.

Dexamethasone-releasing poly(lactic-co-glycolic acid) (PLGA) microparticles were formulated using a solvent displacement technique with the addition of distillation aiming to increase drug delivery lifetime. Two PLGA copolymer ratios (50:50 and 75:25) were used to determine the influence of lactic acid and glycolic acid ratio on microparticle characteristics. The addition of distillation significantly slows the release of dexamethasone compared to traditional solvent removal via evaporation while still maintaining a therapeutic dosage. Microparticles formulated with PLGA 50:50 controllably release dexamethasone up to one year and 75:25 release up to two years in-vitro. The ratio of lactic acid to glycolic acid plays a significant role in microparticle stability, drug loading efficiency, and thermal properties. In all, this formulation technique offers new prospects for inflammation suppression in pediatric vascular and airway diseases.

Poly(gadodiamide fumaric acid): A Bioresorbable, Radiopaque, and MRI-Visible Polymer for Biomedical Applications.

Bioresorbable medical devices once implanted into the body are "invisible" to imaging techniques such as X-ray/fluoroscopy and magnetic resonance imagining (MRI). Prior attempts to produce radiopaque polymers have limited success due to their inability to generate homogeneous mixtures of polymer and contrast agent without subsequent alterations in polymer structure. Here we investigate a novel approach in which a MRI contrast medium, gadodiamide, can be used as a polymerization initiator in poly(propylene fumarate) (PPF) synthesis to achieve a radiopaque and MRI-visible polymer poly(gadodiamide fumaric acid) (PGFA). With this method polymer structure, thermal properties, and rheological behavior are conserved with no prior manipulation to monomer units necessary. This unique polymer in combination with poly(lactic-co-glycolic acid) (PLGA) can be formulated into MRI-visible nanoparticles with drug delivery potential. This novel polymer in both liquid and nanoparticle form enables new possibilities in medical device and drug delivery design.

A novel design biodegradable stent for use in congenital heart disease: mid-term results in rabbit descending aorta.

OBJECTIVES: This study evaluates the feasibility of delivery and deployment of low and medium molecular weight (LMW and MMW, respectively) double-opposing helical (DH) poly-l-lactic acid biodegradable stent (BDS) in rabbit descending aorta (DAO). Secondary objectives were to assess patency and inflammation of stented vessels at 9 months and to investigate safety following intentional embolization of stent fragments in DAO. BACKGROUND: A BDS that will relieve aortic obstruction and disappears as the child grows older allowing for preservation of aortic wall elasticity and natural growth of aorta will be ideal to treat Coarctation (CoA). BDS have never been evaluated in the DAO. METHODS: Seven New Zealand white rabbits underwent implantation of DH-LMW (n = 7), DH-MMW (n = 3), and metal stents (n = 7) in DAO. BDS fragments were intentionally embolized into DAO in two rabbits. RESULTS: All stents were deployed via a 6-French sheath. Five BDS covered the origin of major DAO side branches. Angiography and intravascular ultrasound showed good stent apposition to the wall of DAO with minimal luminal loss at 9 months follow-up. All stents had minimal neointimal hyperplasia on histopathology. Adverse events included 1 death, 1 aortic aneurysm, and lower extremity ulceration due to self-mutilation in an embolization rabbit. CONCLUSIONS: Pilot study confirms the feasibility of delivery and deployment of up to 6-millimeter diameter DH BDS in rabbit DAO. Stent integrity with DH design was maintained at 9 months with minimal vessel inflammation. Potential morbidity due to embolized BD fragments cannot be ruled out and needs further evaluation.

Novel bioresorbable stent coating for drug release in congenital heart disease applications.

A novel double opposed helical poly-l-lactic acid (PLLA) bioresorbable stent has been designed for use in pediatrics. The aim was to test the PLLA stent biocompatibility. The PLLA stent was immersed into whole pig's blood in a closed loop circuit then fibrin and platelet association was assessed via enzyme-linked immunosorbent assay. D-Dimer was valued at 0.2 ± 0.002 ng/mL and P-selectin 0.43 ± 00.01 ng/mL indicating limited association of fibrin and platelets on the stent. To improve biocompatibility by targeting inflammatory cells, dexamethasone was incorporated on PLLA fibers with two coating methods. Both coatings were poly(l-lactide-co-glycolide) acid (PLGA) but one was made porous with sucrose while the other remained nonporous. There was no change in mechanical properties of the fiber with either coating of PLGA polymer. The total amount of dexamethasone released was then determined for each coating. The cumulative drug release for the porous fiber was significantly higher (∼100%) over 8 weeks than the nonporous fiber (40%). Surface examination of the fiber with scanning electron microscopy showed more surface microfracturing in coatings that contain pores. The biocompatibility of this novel stent was demonstrated. Mechanical properties of the fiber were not altered by coating with PLGA polymer. Anti-inflammatory drug release was optimized using a porous PLGA polymer.

A novel biodegradable stent applicable for use in congenital heart disease: bench testing and feasibility results in a rabbit model.

OBJECTIVES: A novel double opposed helical (DH) biodegradable stent was designed and fabricated for CHD applications. The primary objective was to evaluate the feasibility of DH stent delivery and deployment in rabbit external iliac arteries (EIA). Secondary objectives were to assess stent patency, thrombosis and inflammation at 1-week and 1-month follow-up. BACKGROUND: Biodegradable stents have largely been designed for adult cardiovascular indications, to avoid long term complications of permanent implants. A growing child with congenital heart disease (CHD) would especially derive substantial benefit from this technology. METHODS: DH stents were manufactured to 3, 4, 5, and 6-mm diameter with poly-l-lactic acid (PLLA) fibers. Bench test analysis was performed. Six DH stents were implanted in rabbit EIA. Vessel patency was assessed at 1-week and 1-month follow-up with repeat angiography, intravascular ultrasound (IVUS). Histopathological evaluation was performed. RESULTS: The elastic recoil and collapse pressure of DH stents were comparable to conventional metal stents. All DH stents were successfully delivered and implanted with good apposition to the vessel wall and no collapse of the proximal, mid or distal ends. All stented vessels remained patent. No acute or early stent thrombosis was noted. Histopathology showed minimal inflammatory response and mild neointimal proliferation at 1 month follow-up. CONCLUSIONS: In vitro results of DH PLLA biodegradable stents are comparable to conventional metal stents. The pilot animal study confirms the delivery and deployment of the DH stents to the desired location. The DH design can be used to fabricate larger diameter stents needed for CHD.

Morphology of interatrial defects created by interventional techniques in a neonatal animal model.

Pathological details are lacking of various techniques used for interatrial defect (IAD) creation in lesions requiring mixing or with left atrial hypertension. Therefore, the morphology of the IAD created by different interventional techniques is described. The atrial septa of ten euthanized piglets (seven at <3 days and three at 2 weeks of age) were surgically exposed. In pigs of both ages, a needle-created communication was dilated with angioplasty, cutting, and cryoplasty balloons. A stent was also implanted in a newborn pig. By way of a patent foramen ovale (PFO) in newborns, angioplasty and septostomy balloons and a stent created IADs. The morphology of the IAD was directly imaged and the size measured. Newborn piglets had noticeably thinner atrial septa, and a PFO was still present. Static balloon dilation created a circular IAD equivalent to the balloon diameter (3 and 6 mm) in both the created defects of pigs of both ages and the neonatal PFO. A 3-mm cutting balloon (CB) produced a 2-mm triangular IAD (corresponding to atherotomes) in pigs of both ages. Premounted stents, by way of a created defect or PFO, led to circular IADs equivalent to balloon diameter. The 3-mm cryoplasty balloon created a large 5 × 4-mm IAD in the newborn pig; however, the IAD measured only 2 × 1-mm at 2 weeks. By way of a neonatal PFO, a septostomy balloon (inflated to 2 cc) created a 3-mm circular IAD. In neonatal piglet hearts, static balloon angioplasty, CBs, and stents created a predictable IAD. Cryoplasty balloons created highly variable defects.

Thymosin ß4 sustained release from poly(lactide-co-glycolide) microspheres: synthesis and implications for treatment of myocardial ischemia.

A sustained release formulation for the therapeutic peptide thymosin ß4 (Tß4) that can be localized to the heart and reduce the concentration and frequency of dose is being explored as a means to improve its delivery in humans. This review contains concepts involved in the delivery of peptides to the heart and the synthesis of polymer microspheres for the sustained release of peptides, including Tß4. Initial results of poly(lactic-co-glycolic acid) microspheres synthesized with specific tolerances for intramyocardial injection that demonstrate the encapsulation and release of Tß4 from double-emulsion microspheres are also presented.

The influence of thermal treatment on the mechanical characteristics of a PLLA coiled stent.

We studied the effects of thermal treatment on the expansive characteristics of a coil-within-coil Poly(L-lactic acid) (PLLA) fiber stent developed at our institution to improve its mechanical performance and reproducibility. Following fabrication, furled stents were thermally treated at 62 degrees C for 25 min. The mechanical characteristics were measured compared with those of untreated stents when both were expanded via sequential balloon catheter pressure loading up to 12 atm. Treated stents reached full diameter at 3 atm and maintained that diameter despite further pressure increases. Using measurements of pressure, diameter, and axial length, we calculated the sequential mechanical work required to unfurl the stent. The mechanical work for complete unfurling of treated stents was significantly less than that required for untreated controls. Little axial dimensional change was observed for treated stents. Treated stents exhibited higher stiffness than controls at all pressure levels and also demonstrated higher resistance to external pressure-induced collapse, as measured in a special apparatus developed in our laboratory. Differential scanning calorimetry measurements indicated higher crystallinity values for fibers used in treated stents compared with controls. SEM examination of striations revealed that treated stents underwent less twist than controls following balloon-induced unfurling. The results indicate that, thermal treatment improves the reorientation and realignment of fiber crystalline structure, and favorably influences on the fiber stress-strain behavior and the expansive mechanical characteristics of the PLLA fiber stents.

Characterizing the expansive deformation of a bioresorbable polymer fiber stent.

Polymeric vascular stents must employ other strategies than malleable deformation, as generally practiced with metal stents, to expand and withstand compressive stresses in situ. The stent expansion strategy must further consider induced flow perturbations and wall stresses that may injure the vessel wall and promote thrombogenesis. Analyzing the stresses furled stents undergo during balloon-assisted expansion is an important first step in achieving a better understanding of stent-wall mechanical interactions, thereby to improve stent function. To this end, we performed finite element (FE) analysis of the balloon-induced unfurling of an internally coiled, bioresorbable polymeric stent employing a 3D FE solid model of a 120 degrees symmetric stent segment and a large deformation finite strain formulation. Uni-axial tensile testing of stent fiber elastic to plastic yielding provided the mechanical property information, and the von Mises criterion was employed to establish the elastic-plastic transition in the FE model. The model was validated with pressure and deformation measurements obtained during stent expansion tests. The internal coils of this inner coil-outer coil design twisted as the stent expanded, leading to plastic yielding at the point of tangency of the inner and outer coils. The remaining stent fiber portions underwent elastic bending. Cross-sections revealed only the outside surface layer of the coiled fiber underwent plastic yielding. The interior elastic fiber was supported by this plastic shell. The analysis suggests that during balloon-induced expansion, local plastic yielding in torsion "sets" the stent fibers, imparting high radial collapse resistance. The results further suggest that the stent exerts non-uniform mechanical forces on the vessel wall during expansion.