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"Hubbell, J.A."
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Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering
New generations of synthetic biomaterials are being developed at a rapid pace for use as three-dimensional extracellular microenvironments to mimic the regulatory characteristics of natural extracellular matrices (ECMs) and ECM-bound growth factors, both for therapeutic applications and basic biological studies. Recent advances include nanofibrillar networks formed by self-assembly of small building blocks, artificial ECM networks from protein polymers or peptide-conjugated synthetic polymers that present bioactive ligands and respond to cell-secreted signals to enable proteolytic remodeling. These materials have already found application in differentiating stem cells into neurons, repairing bone and inducing angiogenesis. Although modern synthetic biomaterials represent oversimplified mimics of natural ECMs lacking the essential natural temporal and spatial complexity, a growing symbiosis of materials engineering and cell biology may ultimately result in synthetic materials that contain the necessary signals to recapitulate developmental processes in tissue- and organ-specific differentiation and morphogenesis.
Journal Article
Molecularly Engineered PEG Hydrogels: A Novel Model System for Proteolytically Mediated Cell Migration
Model systems mimicking the extracellular matrix (ECM) have greatly helped in quantifying cell migration in three dimensions and elucidated the molecular determinants of cellular motility in morphogenesis, regeneration, and disease progression. Here we tested the suitability of proteolytically degradable synthetic poly(ethylene glycol) (PEG)-based hydrogels as an ECM model system for cell migration research and compared this designer matrix with the two well-established ECM mimetics fibrin and collagen. Three-dimensional migration of dermal fibroblasts was quantified by time-lapse microscopy and automated single-cell tracking. A broadband matrix metalloproteinase (MMP) inhibitor and tumor necrosis factor-alpha, a potent MMP-inducer in fibroblasts, were used to alter MMP regulation. We demonstrate a high sensitivity of migration in synthetic networks to both MMP modulators: inhibition led to an almost complete suppression of migration in PEG hydrogels, whereas MMP upregulation increased the fraction of migrating cells significantly. Conversely, migration in collagen and fibrin proved to be less sensitive to the above MMP modulators, as their fibrillar architecture allowed for MMP-independent migration through preexisting pores. The possibility of molecularly recapitulating key functions of the natural extracellular microenvironment and the improved protease sensitivity makes PEG hydrogels an interesting model system that allows correlation between protease activity and cell migration.
Journal Article
Fiber density of collagen grafts impacts rabbit urethral regeneration
There is a need for efficient and “off-the-shelf” grafts in urethral reconstructive surgery. Currently available surgical techniques require harvesting of grafts from autologous sites, with increased risk of surgical complications and added patient discomfort. Therefore, a cost-effective and cell-free graft with adequate regenerative potential has a great chance to be translated into clinical practice. Tubular cell-free collagen grafts were prepared by varying the collagen density and fiber distribution, thereby creating a polarized low fiber density collagen graft (LD-graft). A uniform, high fiber density collagen graft (HD-graft) was engineered as a control. These two grafts were implanted to bridge a 2 cm long iatrogenic urethral defect in a rabbit model. Histology revealed that rabbits implanted with the LD-graft had a better smooth muscle regeneration compared to the HD-graft. The overall functional outcome assessed by contrast voiding cystourethrography showed patency of the urethra in 90% for the LD-graft and in 66.6% for the HD-graft. Functional regeneration of the rabbit implanted with the LD-graft could further be demonstrated by successful mating, resulting in healthy offspring. In conclusion, cell-free low-density polarized collagen grafts show better urethral regeneration than high-density collagen grafts.
Journal Article
Chemisorbed poly(propylene sulphide)-based copolymers resist biomolecular interactions
Alkanethiolates have been widely used as chemisorbates to modify gold surfaces, in spite of their relatively poor oxidative stability. We introduce gold-chemisorbing block copolymers bearing an anchoring block of poly(propylene sulphide) (PPS), selected in the expectation of greater stability. These materials offer a more robust approach to surface modification of gold. As an example, a triblock copolymer with poly(ethylene glycol) (PEG) was selected, with the goal of minimizing biological adsorption and adhesion. The copolymer PEG
17
-
bl
-PPS
25
-
bl
-PEG
9
chemisorbed to form a dense monolayer of 226 ± 26 ng cm
−2
, ∼2.2 nm thick. The copolymeric adlayer was much more stable to oxidation than commonly used alkanethiolates. Its presence greatly reduced protein adsorption (>95%), even after exposure to whole blood serum (>55 mg ml
−1
), as well as cell adhesion over long culture durations (>97%). PPS-containing copolymers are an attractive alternative to alkanethiolates, and PEG-
bl
-PPS-
bl
-PEG presents a powerful example for use in biodiagnostic and bioanalytical devices.
Journal Article
Polymers for tissue engineering
The articles included in this text highlight the important advances in polymer science that impact tissue engineering. The breadth of polymer science is well represented with the relevance of both polymer chemistry and morphology emphasized in terms of cell and tissue response.
eBook
Protein-mediated macrophage adhesion and activation on biomaterials: a model for modulating cell behavior
The elucidation of proteins involved in biomaterial-modulated macrophage behavior is critical for the improvement of material performance and the initial exploration of material design capable of manipulating macrophage function for tissue engineering. In this paper, several in vitro and in vivo techniques are presented to demonstrate means of delineating a part of the complex molecular mechanisms involved in the interaction between biomaterial and macrophage adhesion and phenotypic development. The following conclusions were reached: (1) using radioimmunoassay, complement component C3 was found to be critical in mediating human macrophage adhesion on polyurethanes. (2) The presence of a diphenolic antioxidant additive in polyurethanes increased the propensity for complement upregulation but did not affect adherent macrophage density. (3) The subcutaneous cage-implant system was utilized to delineate interleukin-4 participation in the fusion of adherent macrophages to form foreign body giant cells in vivo in mice. The injection of purified interleukin-4 neutralizing antibody into the implanted cages significantly decreased the giant cell density; conversely, the giant cell density was significantly increased by the injection of recombinant interleukin-4 when compared with the controls. (4) The RGD and PHSRN amino acid sequences of the central cell binding domain and the PRRARV sequence of the C-terminal heparin binding domain of human plasma fibronectin were utilized to study the structure-functional relationship of protein in mediating macrophage behavior. Polyethyleneglycol-based networks grafted with the RGD-containing peptide supported higher adherent human macrophage density than surfaces grafted with other peptides. The formation of foreign body giant cell was highly dependent on the relative orientation between PHSRN and RGD domains located in a single peptide.
Journal Article
Using Orchardgrass and Endophyte-Free Fescue Versus Endophyte-Infected Fescue Overseeded on Bermudagrass for Cow Herds: II. Four-Year Summary of Cow-Calf Performance
A 4-yr trial was initiated in January 2000 to evaluate cow-calf performance on mixed-species pasture systems consisting of (i) endophyte-infected tall fescue (E+; Festuca arundinacea Schreb.) diluted by approximately 50% with common bermudagrass [Cynodon dactylon (L.) Pers.] and other forages; (ii) endophyte-free tall fescue (E-) overseeded into dormant common bermudagrass; and (iii) orchardgrass (OG; Dactylis glomerata L.) established under the same conditions as E-. The E- and OG pastures were grazed with either twice weekly (2W) or twice monthly (2M) rotation schedules, while pastures with E+ were grazed with 2M only. Actual weaning weights tended to be greater (P = 0.096), and age-adjusted 205-d weaning weights and average daily gain from birth to weaning were greater (P
Journal Article
Effects of Forage Management on the Nutritive Value of Stockpiled Bermudagrass
'Common' and 'Tifton 44' bermudagrass [Cynodon dactylon (L.) Pers.] located near Fayetteville and Batesville, AR, respectively, were chosen to evaluate the effects of stockpiling initiation date (August or September), and N fertilization rate (0, 37, 74, or 111 kg N ha-1) on the nutritive value of fall-stockpiled bermudagrass. At the Fayetteville location, there were initiation x harvest date interactions for acid detergent fiber (ADF, P = 0.003), hemicellulose (P = 0.003), cellulose (P = 0.0003), lignin (P = 0.007), and crude protein (CP, P = 0.008) in 2000, and strong interactions (P < or = 0.001) for all response variables in 2001. Generally, neutral detergent fiber (NDF), ADF, cellulose, and lignin increased over harvest dates for both initiation dates in 2000, although increases for lignin were only numerical (P > 0.05) for the September initiation date. Similar responses were observed for the August initiation date in 2001, but fiber components for the September 2001 initiation date declined over time because of contamination by other winter-annual species. For August initiation dates in 2000 and 2001, in vitro organic matter disappearance (IVOMD) declined linearly (P < or = 0.002) over harvest dates, reaching respective minima of 330 and 361 g kg-1. At the Batesville site, an N fertilization effect (P < or = 0.017) was observed for NDF, ADF, and CP in both 2000 and 2001. A similar effect was observed for lignin (P = 0.001) and IVOMD (P < 0.0001) in 2000, and for cellulose (P = 0.0004) during 2001. Fertilization with N at the initiation of stockpiling generally reduced fiber components, and increased CP; however, IVOMD was increased for 2000 only. As observed for the Fayetteville site, most fiber components increased over harvest dates, while IVOMD declined concomitantly. Generally, the nutritive value of stockpiled bermudagrass declines between mid-October and mid-December, and spring-calving beef cows in the Upper South may need to be supplemented with energy to maintain body condition in the late fall or early winter.
Journal Article