Mooney DJ, Mazzoni CL, Breuer C, McNamara K, Hern D, Vacanti JP, Langer R. Stabilized polyglycolic acid fibre-based tubes for tissue engineering. Biomaterials. 1996;17 (2) :115-24.Abstract
Polyglycolic acid (PGA) fibre meshes are attractive candidates to transplant cells, but they are incapable of resisting significant compressional forces. To stabilize PGA meshes, atomized solutions of poly(L-lactic acid) (PLLA) and a 50/50 copolymer of poly(D,L-lactic-co-glycolic acid) (PLGA) dissolved in chloroform were sprayed over meshes formed into hollow tubes. The PLLA and PLGA coated the PGA fibres and physically bonded adjacent fibres. The pattern and extent of bonding was controlled by the concentration of polymer in the atomized solution and the total mass of polymer sprayed on the device. The compression resistance of devices increased with the extent of bonding, and PLLA bonded tubes resisted larger compressive forces than PLGA bonded tubes. Tubes bonded with PLLA degraded more slowly than devices bonded with PLGA. Implantation of PLLA bonded tubes into rats revealed that the devices maintained their structure during fibrovascular tissue ingrowth, resulting in the formation of a tubular structure with a central lumen. The potential of these devices to engineer specific tissues was exhibited by the finding that smooth muscle cells and endothelial cells seeded onto devices in vitro formed a tubular tissue with appropriate cell distribution.
Kilmartin DJ, Mooney DJ, Acheson RW, Payne SJ, Maher ER, Eustace P. von Hippel-Lindau disease and familial polyposis coli in the same family. Arch Ophthalmol. 1996;114 (10) :1294.
Mooney DJ, Powell C, Piana J, Rutherford B. Engineering dental pulp-like tissue in vitro. Biotechnol Prog. 1996;12 (6) :865-8.Abstract
Injury or infection of adult dental pulp often necessitates root canal therapy. This terminates dentin formation and subsequent tooth maturation. In addition, the synthetic materials currently utilized to replace lost tooth structure are not capable of completely replacing the function of the lost tissue, and often fail over time. This report describes a technique to engineer new pulp-like tissues utilizing cultured cells and synthetic extracellular matrices. Fibroblasts were obtained from human adult dental pulps and multiplied in culture. These cells were subsequently seeded onto synthetic matrices fabricated from fibers (approximately 15 microns in diameter) of polyglycolic acid (PGA). The pulp-derived fibroblasts adhered to the fibers, proliferated, and formed a new tissue over 60 days in culture with a cellularity similar to that of native pulp. These tissues may find application in the regeneration of oral tissues and may provide novel systems in which to study the biocompatibility of materials and chemicals used in dentistry.
Breuer CK, Shin'oka T, Tanel RE, Zund G, Mooney DJ, Ma PX, Miura T, Colan S, Langer R, Mayer JE, et al. Tissue engineering lamb heart valve leaflets. Biotechnol Bioeng. 1996;50 (5) :562-7.Abstract
Tissue engineered lamb heart valve leaflets (N - 3) were constructed by repeatedly seeding a concentrated suspension of autologous myofibroblasts onto a biodegradable synthetic polymeric scaffold composed of fibers made from polyglycolic acid and polylactic acid. Over a 2-week period the cells attached to the polymer fibers, multiplied, and formed a tissue core in the shape of the matrix. The tissue core was seeded with autologous large-vessel endothelial cells that formed a monolayer which coated the outer surface of the leaflet. The tissue engineered leaflets were surgically implanted in place of the right posterior pulmonary valve leaflet of the donor lamb while on cardiopulmonary bypass. Pulmonary valve function was evaluated by two-dimensional echocardiography with color Doppler which demonstrated valve function without evidence of stenosis and with only trivial regurgitation under normal physiologic conditions. Histologically, the tissue engineered heart valve leaflets resembled native valve leaflet tissue.
Putnam AJ, Mooney DJ. Tissue engineering using synthetic extracellular matrices. Nat Med. 1996;2 (7) :824-6.
Mooney DJ, Park S, Kaufmann PM, Sano K, McNamara K, Vacanti JP, Langer R. Biodegradable sponges for hepatocyte transplantation. J Biomed Mater Res. 1995;29 (8) :959-65.Abstract
Liver cell transplantation may provide a means to replace lost or deficient liver tissue, but devices capable of delivering hepatocytes to a desirable anatomic location and guiding the development of a new tissue from these cells and the host tissue are needed. We have investigated whether sponges fabricated from poly-L-lactic acid (PLA) infiltrated with polyvinyl alcohol (PVA) would meet these requirements. Highly porous sponges (porosity = 90-95%) were fabricated from PLA using a particulate leaching technique. To enable even and efficient cell seeding, the devices were infiltrated with the hydrophilic polymer polyvinyl alcohol (PVA). This reduced their contact angle with water from 79 to 23 degrees, but did not inhibit the ability of hepatocytes to adhere to the polymer. Porous sponges of PLA infiltrated with PVA readily absorbed aqueous solutions into 98% of their pore volume, and could be evenly seeded with high densities (5 x 10(7) cells/mL) of hepatocytes. Hepatocyte-seeded devices were implanted into the mesentery of laboratory rats, and 6 +/- 2 x 10(5) of the hepatocytes engrafted per sponge. Fibrovascular tissue invaded through the devices' pores, leading to a composite tissue consisting of hepatocytes, blood vessels and fibrous tissue, and the polymer sponge.
Mooney DJ, Langer R, Ingber DE. Cytoskeletal filament assembly and the control of cell spreading and function by extracellular matrix. J Cell Sci. 1995;108 ( Pt 6) :2311-20.Abstract
This study was undertaken to analyze how cell binding to extracellular matrix produces changes in cell shape. We focused on the initial process of cell spreading that follows cell attachment to matrix and, thus, cell 'shape' changes are defined here in terms of alterations in projected cell areas, as determined by computerized image analysis. Cell spreading kinetics and changes in microtubule and actin microfilament mass were simultaneously quantitated in hepatocytes plated on different extracellular matrix substrata. The initial rate of cell spreading was highly dependent on the matrix coating density and decreased from 740 microns 2/h to 50 microns 2/h as the coating density was lowered from 1000 to 1 ng/cm2. At approximately 4 to 6 hours after plating, this initial rapid spreading rate slowed and became independent of the matrix density regardless of whether laminin, fibronectin, type I collagen or type IV collagen was used for cell attachment. Analysis of F-actin mass revealed that cell adhesion to extracellular matrix resulted in a 20-fold increase in polymerized actin within 30 minutes after plating, before any significant change in cell shape was observed. This was followed by a phase of actin microfilament disassembly which correlated with the most rapid phase of cell extension and ended at about 6 hours; F-actin mass remained relatively constant during the slow matrix-independent spreading phase. Microtubule mass increased more slowly in spreading cells, peaking at 4 hours, the time at which the transition between rapid and slow spreading rates was observed. However, inhibition of this early rise in microtubule mass using either nocodazole or cycloheximide did not prevent this transition. Use of cytochalasin D revealed that microfilament integrity was absolutely required for hepatocyte spreading whereas interference with microtubule assembly (using nocodazole or taxol) or protein synthesis (using cycloheximide) only partially suppressed cell extension. In contrast, cell spreading could be completely inhibited by combining suboptimal doses of cytochalasin D and nocodazole, suggesting that intact microtubules can stabilize cell form when the microfilament lattice is partially compromised. The physiological relevance of the cytoskeleton and cell shape in hepatocyte physiology was highlighted by the finding that a short exposure (6 hour) of cells to nocodazole resulted in production of smaller cells 42 hours later that exhibited enhanced production of a liver-specific product (albumin). These data demonstrate that spreading and flattening of the entire cell body is not driven directly by net polymerization of either microfilaments or microtubules.(ABSTRACT TRUNCATED AT 400 WORDS)
Mooney DJ, Breuer C, McNamara K, Vacanti JP, Langer R. Fabricating tubular devices from polymers of lactic and glycolic Acid for tissue engineering. Tissue Eng. 1995;1 (2) :107-18.Abstract
Polymers of lactic and glycolic acid are attractive candidates to fabricate devides to transplant cells and engineer new tissues. These polymers are biocompatible, and exhibit a wide range of erosion times and mechanical properties. This manuscript describes the fabrication and characterization, in vitro and in vivo, of hollow, tubular devices from porous films of various polymers of this family. Porous films of these polymers were formed using a particulate leaching technique, and sealed around Teflon cylinders to form hollow tubular devices. The erosion rate of devices was controlled by the specific polymer utilized for fabrication, and ranged from months to years. Devices fabricated from a 50/50 copolymer of D,L-lactic acid and glycolic acid were completely eroded by 2 months, while devices fabricated from a homopolymer of L-lactic acid showed little mass loss after 1 year. Erosion times for devices fabricated from the other polymers [poly-(D,L-lactic acid) and a 85/15 copolymer] were between these two extremes. Devices were capable of resisting significant compressional forces (150 raN) in vitro, and the compression resistance was controlled by the polymer utilized to fabricate the devices. The ability of the devices to maintain their structure after implantation into the mesentery or omentum of laboratory rats was also dependent of the specific polymer utilized to fabricate the device. These results indicate that it is possible to fabricate tubular devices for tissue engineering applications that exhibit a wide range of erosion rates and mechanical properties.
Mooney DJ, Organ G, Vacanti JP, Langer R. Design and fabrication of biodegradable polymer devices to engineer tubular tissues. Cell Transplant. 1994;3 (2) :203-10.Abstract
Engineering new tissues by transplanting cells on polymeric delivery devices is one approach to alleviate the vast shortage of donor tissue. However, it will be necessary to fabricate cell delivery devices that deliver cells to a given location and promote the formation of specific tissue structures from the transplanted cells and the host tissue. This report describes the design and fabrication of a polymeric device for guiding the development of tubular vascularized tissues, which may be useful for engineering a variety of tissues including intestine, blood vessels, tracheas, and ureters. Porous films of poly (D, L-lactic-co-glycolic acid) have been formed and fabricated into tubes capable of resisting compressional forces in vitro and in vivo. These devices promote the ingrowth of fibrovascular tissue following implantation into recipient animals, resulting in a vascularized, tubular tissue. To investigate the utility of these devices as cell delivery devices, enterocytes (intestinal epithelial cells) were seeded onto the devices in vitro. Enterocytes were found to attach to these devices and form an organized epithelial cell layer. These results suggest that these devices may be an appropriate delivery vehicle for transplanting cells and engineering new tubular tissues.
Mooney DJ, Hansen LK, Langer R, Vacanti JP, Ingber DE. Extracellular matrix controls tubulin monomer levels in hepatocytes by regulating protein turnover. Mol Biol Cell. 1994;5 (12) :1281-8.Abstract
Cells have evolved an autoregulatory mechanism to dampen variations in the concentration of tubulin monomer that is available to polymerize into microtubules (MTs), a process that is known as tubulin autoregulation. However, thermodynamic analysis of MT polymerization predicts that the concentration of free tubulin monomer must vary if MTs are to remain stable under different mechanical loads that result from changes in cell adhesion to the extracellular matrix (ECM). To determine how these seemingly contradictory regulatory mechanisms coexist in cells, we measured changes in the masses of tubulin monomer and polymer that resulted from altering cell-ECM contacts. Primary rat hepatocytes were cultured in chemically defined medium on bacteriological petri dishes that were precoated with different densities of laminin (LM). Increasing the LM density from low to high (1-1000 ng/cm2), promoted cell spreading (average projected cell area increased from 1200 to 6000 microns2) and resulted in formation of a greatly extended MT network. Nevertheless, the steady-state mass of tubulin polymer was similar at 48 h, regardless of cell shape or ECM density. In contrast, round hepatocytes on low LM contained a threefold higher mass of tubulin monomer when compared with spread cells on high LM. Furthermore, similar results were obtained whether LM, fibronectin, or type I collagen were used for cell attachment. Tubulin autoregulation appeared to function normally in these cells because tubulin mRNA levels and protein synthetic rates were greatly depressed in round cells that contained the highest level of free tubulin monomer. However, the rate of tubulin protein degradation slowed, causing the tubulin half-life to increase from approximately 24 to 55 h as the LM density was lowered from high to low and cell rounding was promoted. These results indicate that the set-point for the tubulin monomer mass in hepatocytes can be regulated by altering the density of ECM contacts and changing cell shape. This finding is consistent with a mechanism of MT regulation in which the ECM stabilizes MTs by both accepting transfer of mechanical loads and altering tubulin degradation in cells that continue to autoregulate tubulin synthesis.
Hansen LK, Mooney DJ, Vacanti JP, Ingber DE. Integrin binding and cell spreading on extracellular matrix act at different points in the cell cycle to promote hepatocyte growth. Mol Biol Cell. 1994;5 (9) :967-75.Abstract
This study was undertaken to determine the importance of integrin binding and cell shape changes in the control of cell-cycle progression by extracellular matrix (ECM). Primary rat hepatocytes were cultured on ECM-coated dishes in serum-free medium with saturating amounts of growth factors (epidermal growth factor and insulin). Integrin binding and cell spreading were promoted in parallel by plating cells on dishes coated with fibronectin (FN). Integrin binding was separated from cell shape changes by culturing cells on dishes coated with a synthetic arg-gly-asp (RGD)-peptide that acts as an integrin ligand but does not support hepatocyte extension. Expression of early (junB) and late (ras) growth response genes and DNA synthesis were measured to determine whether these substrata induce G0-synchronized hepatocytes to reenter the growth cycle. Cells plated on FN exhibited transient increases in junB and ras gene expression (within 2 and 8 h after plating, respectively) and synchronous entry into S phase. Induction of junB and ras was observed over a similar time course in cells on RGD-coated dishes, however, these round cells did not enter S phase. The possibility that round cells on RGD were blocked in mid to late G1 was confirmed by the finding that when trypsinized and replated onto FN-coated dishes after 30 h of culture, they required a similar time (12-15 h) to reenter S phase as cells that had been spread and allowed to progress through G1 on FN. We have previously shown that hepatocytes remain viable and maintain high levels of liver-specific functions when cultured on these RGD-coated dishes. Thus, these results suggest that ECM acts at two different points in the cell cycle to regulate hepatocyte growth: first, by activating the G0/G1 transition via integrin binding and second, by promoting the G1/S phase transition and switching off the default differentiation program through mechanisms related to cell spreading.
Mooney DJ, Kaufmann PM, Sano K, McNamara KM, Vacanti JP, Langer R. Transplantation of hepatocytes using porous, biodegradable sponges. Transplant Proc. 1994;26 (6) :3425-6.
Mooney DJ. Government policies on a collision course. Health cost containment and the antitrust laws. Med Group Manage J. 1993;40 (5) :46-8, 50, 52.Abstract
The Sherman Act, passed in 1890, was initially enacted to break up the huge "trusts" of that era, writes Donald Mooney, but it has been used more frequently as a weapon in the government's war to slow mounting health care costs. In this era of mergers, acquisitions and joint ventures, groups need to be readily aware of the laws regarding antitrust.
Organ GM, Mooney DJ, Hansen LK, Schloo B, Vacanti JP. Enterocyte transplantation using cell-polymer devices to create intestinal epithelial-lined tubes. Transplant Proc. 1993;25 (1 Pt 2) :998-1001.
Power WJ, Hope-Ross M, Mooney DJ. Preoperative pupil fatigue. J Cataract Refract Surg. 1992;18 (3) :306-9.Abstract
Pupils are frequently dilated on the day before cataract surgery and for retinal detachment surgery so the fundus can be examined. This may, however, interfere with pupil mydriasis on the day of surgery. This study looked at the effect of pupil dilation with tropicamide 1% and with cyclopentolate 1% on pupil mydriasis 24 hours later, using phenylephrine 10% and cyclopentolate 1%, in 40 cataract patients. The pupils dilated with cyclopentolate one day previously demonstrated a mean reduction in subsequent mydriasis of 0.73 mm compared with pupils that had been dilated with tropicamide (P less than .0001). The magnitude of this difference was not related to the patient age (P = .12) or to iris color (P = .21). If it is necessary to dilate pupils on the day before surgery, tropicamide 1% rather than cyclopentolate 1% should be used, as it is less likely to interfere with the pupil mydriasis produced with cyclopentolate 1% and phenylephrine 10% on the day of surgery.
Organ GM, Mooney DJ, Hansen LK, Schloo B, Vacanti JP. Transplantation of enterocytes utilizing polymer-cell constructs to produce a neointestine. Transplant Proc. 1992;24 (6) :3009-11.
Power WJ, Travers SP, Mooney DJ. Welding arc maculopathy and fluphenazine. Br J Ophthalmol. 1991;75 (7) :433-5.Abstract
A 45-year-old male patient presented with a bilateral maculopathy following unprotected exposure of less than two minutes' duration to a manual metal arc welding unit. He had been receiving the drug fluphenazine for the previous 10 years for treatment of depression. We believe that the drug fluphenazine, which had accumulated in his retinal pigment epithelium, may have rendered him particularly susceptible to retinal photic damage.
Doherty PE, Mooney DJ. Deciphering bloody imprints through chemical enhancement. J Forensic Sci. 1990;35 (2) :457-65.Abstract
Obliterated bloody impressions are occasionally submitted to the crime laboratory, and potentially to the document examiner, for decipherment. Nondestructive methods often lead to inconclusive results in these circumstances. With this point in mind, the researchers explored a series of chemical reagents with the intent to enhance bloody imprints to a legible degree. The reagents selected for this comparison include rhodamine dye, luminol, and Coomassie Blue stain.
Power WJ, Coleman K, Curtin DM, Mooney DJ. The pattern ERG in Best's disease. Doc Ophthalmol. 1990;76 (3) :279-84.Abstract
The aim of this study was to assess inner retinal function in patients with Best's disease using the pattern ERG (PERG). Nine patients with Best's disease, who had good visual acuity, were studied. Five of the nine had abnormal PERGs. All five had some reduction in central visual acuity. We believe that the abnormal PERGs in these patients represents photoreceptor cell loss which is occurring at an early stage in Best's disease.
Kinsella FP, Mooney DJ. Anaphylaxis following oral fluorescein angiography. Am J Ophthalmol. 1988;106 (6) :745-6.