Craig Neville

Publications of Craig Neville

• The role of fibroblasts in self-assembled skeletal muscle.

  Authors: Mei Li, Caitlyn E Dickinson, Eric B Finkelstein, Craig M Neville, Cathryn A Sundback

  Tissue engineering. Part A. 06/2011; 17(21-22):2641-50.

  Small facial skeletal muscles often have no autologous donor source to effect surgical reconstruction. Autologously derived muscles could be engineered for replacement tissue, but must beSmall facial skeletal muscles often have no autologous donor source to effect surgical reconstruction. Autologously derived muscles could be engineered for replacement tissue, but must be vascularized and innervated to be functional. As a critical step, engineered muscle must mimic the morphology, protein and gene expression, and function of native muscle. This study utilized a self-assembly process to engineer three-dimensional (3D) muscle from a statically strained muscle cell monolayer. Primary mouse myoblasts (PMMs) and mouse embryonic fibroblasts (MEFs) were separately proliferated and coseeded on a fibrin sheet with anchored sutures. Within 10 days of initiating PMM differentiation, the cell-gel layer contracted, lifted, and rolled into a cylindrical 3D structure around the tendon-like suture anchors; the myotubes longitudinally aligned along the lines of tensile force. The objectives of this study were to characterize these engineered muscles and to elucidate the role of the fibroblasts in the self-assembly process. Fibroblasts maintained myotube viability, mediated fibrin degradation, and assisted in muscle self-assembly. The optimal 1:1 PMM:MEF ratio resulted in tissue morphology remarkably similar to native muscle. Through gene and protein expression assays, the development and maturation of the engineered muscle tissue was demonstrated to recapitulate normal skeletal muscle development.

• Liver-assist device with a microfluidics-based vascular bed in an animal model.

  Authors: Wen-Ming Hsu, Amedeo Carraro, Katherine M Kulig, Mark L Miller, Mohammad Kaazempur-Mofrad, Eli Weinberg, Fateh Entabi, Hassan Albadawi, Michael T Watkins, Jeffrey T Borenstein, Joseph P Vacanti, Craig Neville

  Annals of surgery. 08/2010; 252(2):351-7.

  This study evaluates a novel liver-assist device platform with a microfluidics-modeled vascular network in a femoral arteriovenous shunt in rats. Liver-assist devices in clinical trials that useThis study evaluates a novel liver-assist device platform with a microfluidics-modeled vascular network in a femoral arteriovenous shunt in rats. Liver-assist devices in clinical trials that use pumps to force separated plasma through packed beds of parenchymal cells exhibited significant necrosis with a negative impact on function. Microelectromechanical systems technology was used to design and fabricate a liver-assist device with a vascular network that supports a hepatic parenchymal compartment through a nanoporous membrane. Sixteen devices with rat primary hepatocytes and 12 with human HepG2/C3A cells were tested in athymic rats in a femoral arteriovenous shunt model. Several parenchymal tube configurations were evaluated for pressure profile and cell survival. The blood flow pattern and perfusion status of the devices was examined by laser Doppler scanning. Cell viability and serum protein secretion functions were assessed. Femoral arteriovenous shunt was successfully established in all animals. Blood flow was homogeneous through the vascular bed and replicated native flow patterns. Survival of seeded liver cells was highly dependent on parenchymal chamber pressures. The tube configuration that generated the lowest pressure supported excellent cell survival and function. This device is the first to incorporate a microfluidics network in the systemic circulatory system. The microvascular network supported viability and function of liver cells in a short-term ex vivo model. Parenchymal chamber pressure generated in an arteriovenous shunt model is a critical parameter that affects viability and must be considered in future designs. The microfluidics-based vascular network is a promising platform for generating a large-scale medical device capable of augmenting liver function in a clinical setting.

• Engineered vascularized bone grafts.

  Authors: Olga Tsigkou, Irina Pomerantseva, Joel A Spencer, Patricia A Redondo, Alison R Hart, Elisabeth O'Doherty, Yunfeng Lin, Claudia C Friedrich, Laurence Daheron, Charles P Lin, Cathryn A Sundback, Joseph P Vacanti, Craig Neville

  Proceedings of the National Academy of Sciences of the United States of America. 02/2010; 107(8):3311-6.

  Clinical protocols utilize bone marrow to seed synthetic and decellularized allogeneic bone grafts for enhancement of scaffold remodeling and fusion. Marrow-derived cytokines induce hostClinical protocols utilize bone marrow to seed synthetic and decellularized allogeneic bone grafts for enhancement of scaffold remodeling and fusion. Marrow-derived cytokines induce host neovascularization at the graft surface, but hypoxic conditions cause cell death at the core. Addition of cellular components that generate an extensive primitive plexus-like vascular network that would perfuse the entire scaffold upon anastomosis could potentially yield significantly higher-quality grafts. We used a mouse model to develop a two-stage protocol for generating vascularized bone grafts using mesenchymal stem cells (hMSCs) from human bone marrow and umbilical cord-derived endothelial cells. The endothelial cells formed tube-like structures and subsequently networks throughout the bone scaffold 4-7 days after implantation. hMSCs were essential for stable vasculature both in vitro and in vivo; however, contrary to expectations, vasculature derived from hMSCs briefly cultured in medium designed to maintain a proliferative, nondifferentiated state was more extensive and stable than that with hMSCs with a TGF-beta-induced smooth muscle cell phenotype. Anastomosis occurred by day 11, with most hMSCs associating closely with the network. Although initially immature and highly permeable, at 4 weeks the network was mature. Initiation of scaffold mineralization had also occurred by this period. Some human-derived vessels were still present at 5 months, but the majority of the graft vasculature had been functionally remodeled with host cells. In conclusion, clinically relevant progenitor sources for pericytes and endothelial cells can serve to generate highly functional microvascular networks for tissue engineered bone grafts.

• Bone marrow derived pluripotent cells are pericytes which contribute to vascularization.

  Authors: Xiaoxiao Cai, Yunfeng Lin, Claudia C Friedrich, Craig Neville, Irina Pomerantseva, Cathryn A Sundback, Parul Sharma, Zhiyuan Zhang, Joseph P Vacanti, Peter V Hauschka, Brian E Grottkau

  Stem cell reviews. 12/2009; 5(4):437-45.

  Pericytes are essential to vascularization, but the purification and characterization of pericytes remain unclear. Smooth muscle actin alpha (alpha-SMA) is one marker [corrected] of pericytes. ThePericytes are essential to vascularization, but the purification and characterization of pericytes remain unclear. Smooth muscle actin alpha (alpha-SMA) is one marker [corrected] of pericytes. The aim of this study is to purify the alpha-SMA positive cells from bone marrow and study the characteristics of these cells and the interaction between alpha-SMA positive cells and endothelial cells. The bone marrow stromal cells were harvested from alpha-SMA-GFP transgenic mice, and the alpha-SMA-GFP positive cells were sorted by FACS. The proliferative characteristics and multilineage differentiation ability of the alpha-SMA-GFP positive cells were tested. A 3-D culture model was then applied to test their vascularization by loading alpha-SMA-GFP positive cells and endothelial cells on collagen-fibronectin gel. Results demonstrated that bone marrow stromal cells are mostly alpha-SMA-GFP positive cells which are pluripotent, and these cells expressed alpha-SMA during differentiation. The alpha-SMA-GFP positive cells could stimulate the endothelial cells to form tube-like structures and subsequently robust vascular networks in 3-D culture. In conclusion, the bone marrow derived pluripotent cells include [corrected] pericytes and can contribute to vascularization.

• Erratum to: Bone Marrow Derived Pluripotent Cells are Pericytes which Contribute to Vascularization.

  Authors: Xiaoxiao Cai, Yunfeng Lin, Claudia Friedrich, Craig Neville, Irina Pomerantseva, Cathryn Sundback, Zhiyuan Zhang, Joseph Vacanti, Peter Hauschka, Brian Grottkau

  Stem cell reviews and reports. 11/2009;

• Bone marrow Derived Pluripotent Cells are Pericytes which Contribute to Vascularization.

  Authors: Xiaoxiao Cai, Yunfeng Lin, Claudia Friedrich, Craig Neville, Irina Pomerantseva, Cathryn Sundback, Parul Sharma, Zhiyuan Zhang, Joseph Vacanti, Peter Hauschka, Brian Grottkau

  Stem cell reviews and reports. 10/2009;

  Pericytes are essential to vascularization, but the purification and characterization of pericytes remain unclear. Smooth muscle actin alpha (alpha-SMA) is one maker of pericytes. The aim of thisPericytes are essential to vascularization, but the purification and characterization of pericytes remain unclear. Smooth muscle actin alpha (alpha-SMA) is one maker of pericytes. The aim of this study is to purify the alpha-SMA positive cells from bone marrow and study the characteristics of these cells and the interaction between alpha-SMA positive cells and endothelial cells. The bone marrow stromal cells were harvested from alpha-SMA-GFP transgenic mice, and the alpha-SMA-GFP positive cells were sorted by FACS. The proliferative characteristics and multilineage differentiation ability of the alpha-SMA-GFP positive cells were tested. A 3-D culture model was then applied to test their vascularization by loading alpha-SMA-GFP positive cells and endothelial cells on collagen-fibronectin gel. Results demonstrated that bone marrow stromal cells are mostly alpha-SMA-GFP positive cells which are pluripotent, and these cells expressed alpha-SMA during differentiation. The alpha-SMA-GFP positive cells could stimulate the endothelial cells to form tube-like structures and subsequently robust vascular networks in 3-D culture. In conclusion, the bone marrow derived pluripotent cells are pericytes and can contribute to vascularization.

• Towards regenerating a human thumb in situ.

  Authors: Christian Weinand, Rajiv Gupta, Eli Weinberg, Ijad Madisch, Craig Neville, Jesse B Jupiter, Joseph Vacanti

  Tissue engineering. Part A. 03/2009;

  Regenerative technology promises to alleviate the problem of limited donor supply for bone or organ transplants. Most expensive and time consuming is cell expansion in laboratories. We propose aRegenerative technology promises to alleviate the problem of limited donor supply for bone or organ transplants. Most expensive and time consuming is cell expansion in laboratories. We propose a method of magnetically enriched osteoprogenitor stem cells, dispersed in self-assembling hydrogels and applied onto new ultra-high resolution, jet-based 3D printing of living human bone in a single-step for in-situ bone regeneration. Human mesenchymal stem-cells (hBMSCs) were enriched with CD 117+ osteoprogenitor cells, dispersed in different collagen I, RAD 16I hydrogels mixes and applied onto 3-dimensional printed (3DP) beta-TCP/PLGA scaffolds, printed from ultra-high-resolution volumetric CT (VCT) images of a human thumb. Constructs were directly implanted subcutaneously into nude mice for 6 weeks. In-vivo radiographic VCT scanning and histological evaluations were performed at 1, 2, 4 and 6 weeks, expression of bone-specific genes and biomechanical compression-testing at 6 weeks endpoint. Time dependant accumulation of bone-like extracellular matrix was most evident in CD 117+ hBMSCs using collagen I/RAD 16I hydrogel mix. This was shown histologically by Toluidine blue, von Kossa and alkaline-phosphatase staining, paralleled by increased radiological densities within implants approximating that of human bone, and confirmed by high expression of bone-specific osteonectin and biomechanical stiffness at 6 weeks. Human origin of newly formed tissue was established by expression of human GAPDH using RT-PCR. Statistical analysis confirmed high correlations between biomechanical stiffness, radiological densities and bone-markers. Bone tissue can be successfully regenerated in-situ using a single-step procedure with constructs comprised of RAD 16I /collagen I hydrogel, CD 117+ enriched hBMSCs and porous beta-TCP/PLGA scaffolds.

• In vitro analysis of a hepatic device with intrinsic microvascular-based channels.

  Authors: Amedeo Carraro, Wen-Ming Hsu, Katherine Kulig, Wing Cheung, Mark Miller, Eli Weinberg, Eric Swart, Mohammad Kaazempur-Mofrad, Jeffrey Borenstein, Joseph Vacanti, Craig Neville

  Biomedical microdevices. 08/2008;

  A novel microfluidics-based bilayer device with a discrete parenchymal chamber modeled upon hepatic organ architecture is described. The microfluidics network was designed using computational modelsA novel microfluidics-based bilayer device with a discrete parenchymal chamber modeled upon hepatic organ architecture is described. The microfluidics network was designed using computational models to provide appropriate flow behavior based on physiological data from human microvasculature. Patterned silicon wafer molds were used to generate films with the vascular-based microfluidics network design and parenchymal chamber by soft lithography. The assembled device harbors hepatocytes behind a nanoporous membrane that permits transport of metabolites and small proteins while protecting them from the effects of shear stress. The device can sustain both human hepatoma cells and primary rat hepatocytes by continuous in vitro perfusion of medium, allowing proliferation and maintaining hepatic functions such as serum protein synthesis and metabolism. The design and fabrication processes are scalable, enabling the device concept to serve as both a platform technology for drug discovery and toxicity, and for the continuing development of an improved liver-assist device.