Bladder acellular matrix as a substrate for studying in vitro bladder smooth muscle–urothelial cell interactions

Brown, Angela Demke; Brook-Allred, Tamara T; Waddell, Jennifer E; White, John; Werkmeister, Jerome A.; Ramshaw, John A. M.; Bägli, Darius; Woodhouse, Kimberly A.

doi:10.1016/j.biomaterials.2004.02.055

Cited by 68 publications

(53 citation statements)

References 51 publications

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“…Each type of biomaterials has desirable traits which are exclusive of the other. Acellular tissue matrices possess the desired biocompatibility (11)(12)(13), contain biomimetic factors (14)(15)(16)) that promote tissue development and have adhesion domain sequences (e.g., RGD) that may assist in retaining the phenotype and activity of many types of cells (17). These matrices are known to slowly degrade upon implantation and are usually replaced and remodeled by ECM proteins synthesized and secreted by transplanted or ingrowing cells (18)(19)(20)(21)(22)(23)(24)(25).…”

Section: Introductionmentioning

confidence: 99%

Composite scaffolds for the engineering of hollow organs and tissues

Eberli¹,

Filho²,

Atala³

et al. 2009

Methods

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Composite scaffolds for the engineering of hollow organs and tissues AbstractSeveral types of synthetic and naturally derived biomaterials have been used for augmenting hollow organs and tissues. However, each has desirable traits which were exclusive of the other. We fabricated a composite scaffold and tested its potential for the engineering of hollow organs in a bladder tissue model. The composite scaffolds were configured to accommodate a large number of cells on one side and were designed to serve as a barrier on the opposite side. The scaffolds were fabricated by bonding a collagen matrix to PGA polymers with threaded collagen fiber stitches. Urothelial and bladder smooth muscle cells were seeded on the composite scaffolds, and implanted in mice for up to 4 weeks and analyzed. Both cell types readily attached and proliferated on the scaffolds and formed bladder tissue-like structures in vivo. These structures consisted of a luminal urothelial layer, a collagen rich compartment and a peripheral smooth muscle layer. Biomechanical studies demonstrated that the tissues were readily elastic while maintaining their pre-configured structures. This study demonstrates that a composite scaffold can be fabricated with two completely different polymer systems for the engineering of hollow organs. The composite scaffolds are biocompatible, possess adequate physical and structural characteristics for bladder tissue engineering, and are able to form tissues in vivo. This scaffold system may be useful in patients requiring hollow organ replacement. Title: COMPOSITE SCAFFOLDS FOR THE ENGINEERING OF HOLLOW ORGANS AND TISSUES Authors Abstract:Several types of synthetic and naturally derived biomaterials have been used for augmenting hollow organs and tissues. However, each has desirable traits which were exclusive of the other. We fabricated a composite scaffold and tested its potential for the engineering of hollow organs in a bladder tissue model. The composite scaffolds were configured to accommodate a large number of cells on one side and were designed to serve as a barrier on the opposite side. The scaffolds were fabricated by bonding a collagen matrix to PGA polymers with threaded collagen fiber stitches. Urothelial and bladder smooth muscle cells were seeded on the composite scaffolds, and implanted in mice for up to 4 weeks and analyzed. Both cell types readily attached and proliferated on the scaffolds and formed bladder tissue-like structures in vivo. These structures consisted of a luminal urothelial layer, a collagen rich compartment and a peripheral smooth muscle layer. Biomechanical studies demonstrated that the tissues were readily elastic while maintaining their pre-configured structures. This study demonstrates that a composite scaffold can be fabricated with two completely different polymer systems for the engineering of hollow organs. The composite scaffolds are biocompatible, possess adequate physical and structural characteristics for bladder tissue engineering, and are able to form tissues in vivo. Thi...

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Section: Introductionmentioning

confidence: 99%

Composite scaffolds for the engineering of hollow organs and tissues

Eberli¹,

Filho²,

Atala³

et al. 2009

Methods

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“…The urinary bladder matrix is prepared using intralumenal water under pressure to facilitate the separation of the muscle layer from the tunica submucosa. 13 The small intestinal submucosa is processed by physically removing the muscle layer and some portions of the mucosa, followed by chemical treatments to produce an acellular matrix. 8,14,15 More recently, Karim et al 16 used a rotating perfusion bioreactor to decellularize and re-seed porcine heart valves, and Ott et al 17 decellularized whole rat hearts using a modified Langendorff apparatus.…”

Section: Introductionmentioning

confidence: 99%

Preparation of Ex Vivo–Based Biomaterials Using Convective Flow Decellularization

Montoya

McFetridge

2009

Tissue Engineering Part C: Methods

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With advantageous biomechanical properties, materials derived from ex vivo tissues are being actively investigated as scaffolds for tissue engineering applications. However, decellularization treatments are required before implantation to reduce the materials immune impact. The aim of these investigations was to assess a convective flow model as an enhanced methodology to decellularize ex vivo tissue. Isolated human umbilical veins were decellularized using two methods: rotary agitation at 100 rpm on orbital shaker plates, and convective flow run at 5, 50, and 150 mmHg within perfusion bioreactors. Extracted phospholipids and total soluble protein were assessed over time. Histology, SEM, and uniaxial tensile testing analysis were carried out to evaluate variation in the tissues. After 72 h, samples exposed to traditional rotary agitation showed retention of whole cells and cellular components, whereas pressure-based systems showed no visual sign of cells. The convective flow method was significantly more effective at removing phospholipid and total protein than the agitation model. High transmembrane pressure (150 mmHg) resulted in higher phospholipids extraction. However, a more efficient protein extraction occurred at 50 mmHg. Variation in extraction rates was dependent on tissue permeability, which varied as pressure increased. Collectively, these findings show significant improvements in decellularization efficiency that may lead to more immune compliant ex vivo-derived biomaterials.

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“…3,4 Gabouev et al 5 have also shown that cell penetration into SIS takes on the order of weeks. To obtain a construct that may be mechanically stimulated to promote ECM remodeling, cell penetration is necessary.…”

Section: Introductionmentioning

confidence: 99%

Generating Elastin-Rich Small Intestinal Submucosa–Based Smooth Muscle Constructs Utilizing Exogenous Growth Factors and Cyclic Mechanical Stimulation

Heise

Ivanova

Parekh

et al. 2009

Tissue Engineering Part A

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Successful approaches to tissue engineering smooth muscle tissues utilize biodegradable scaffolds seeded with autologous cells. One common problem in using biological scaffolds specifically is the difficulty of inducing cellular penetration and controlling de novo extracellular matrix deposition=remodeling in vitro. Our hypothesis was that small intestinal submucosa (SIS) exposed to specific mechanical stimulation regimes would modulate the synthesis of de novo collagen and elastin by bladder smooth muscle cells (BSMC) within the SIS matrix. We further hypothesized that the cytokines vascular endothelial growth factor (VEGF) and fibroblast growth factor-2 (FGF-2), two key growth factors involved in epithelial mesenchymal signaling, will promote the cellular penetration into SIS necessary for mechanical stimulation. BSMC were seeded at 0.5Â10 6 cells=cm 2 onto the luminal side of SIS specimens. VEGF (10 ng=mL) and FGF-2 (5 ng=mL) were added to each insert in the media every other day for up to 7 days in static culture. Following static culture, specimens were stretched stripbiaxially under 15% peak strain at either 0.5 or 0.1 Hz for an additional 7 days. Following the culture period, specimens were assayed histologically and biochemically for cellular penetration, proliferation, elastin, collagen, and protease activity. Histological analyses demonstrated that in standard culture media, BSMC remained on the surface of the SIS while both FGF-2 and VEGF profoundly promoted ingrowth of the BSMC into the SIS. The penetration of the cells in response to these cytokines was confirmed using a Transwell assay. Following cellular penetration, BSMC produced significant amounts of elastic fibers under cyclic mechanical stretching at 0.1 Hz under 15% stretch, as evidenced by colorimetric assay and histology using a Verhoeff-Van Gieson stain. Protease activity was assessed in the media and found to be statistically increased in static culture following FGF-2 treatment. These findings demonstrate, for the first time, the capability of BSMC to produce histologically apparent elastin fibers in vitro. Moreover, our results suggest that a strategy involving growth factors and controlled mechanical stimulation may be used to engineer functional, elastin-rich tissue replacements using decellularized biologically derived scaffolds.

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Bladder acellular matrix as a substrate for studying in vitro bladder smooth muscle–urothelial cell interactions

Cited by 68 publications

References 51 publications

Composite scaffolds for the engineering of hollow organs and tissues

Composite scaffolds for the engineering of hollow organs and tissues

Preparation of Ex Vivo–Based Biomaterials Using Convective Flow Decellularization

Generating Elastin-Rich Small Intestinal Submucosa–Based Smooth Muscle Constructs Utilizing Exogenous Growth Factors and Cyclic Mechanical Stimulation

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