Bryan Hilbert | Charles Sturt University (original) (raw)

Papers by Bryan Hilbert

Research paper thumbnail of Direct Conversion of Equine Adipose-Derived Stem Cells into Induced Neuronal Cells Is Enhanced in Three-Dimensional Culture

Cellular reprogramming, Jan 18, 2015

The ability to culture neurons from horses may allow further investigation into equine neurologic... more The ability to culture neurons from horses may allow further investigation into equine neurological disorders. In this study, we demonstrate the generation of induced neuronal cells from equine adipose-derived stem cells (EADSCs) using a combination of lentiviral vector expression of the neuronal transcription factors Brn2, Ascl1, Myt1l (BAM) and NeuroD1 and a defined chemical induction medium, with βIII-tubulin-positive induced neuronal cells displaying a distinct neuronal morphology of rounded and compact cell bodies, extensive neurite outgrowth, and branching of processes. Furthermore, we investigated the effects of dimensionality on neuronal transdifferentiation, comparing conventional two-dimensional (2D) monolayer culture against three-dimensional (3D) culture on a porous polystyrene scaffold. Neuronal transdifferentiation was enhanced in 3D culture, with evenly distributed cells located on the surface and throughout the scaffold. Transdifferentiation efficiency was increased ...

Research paper thumbnail of Prognosis in equine colic patients using multivariate analysis

Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire

Research paper thumbnail of In Vitro Mechanical Testing of Braided Polyurethane Elastic Fiber and Braided Polyester for Equine Laryngoplasty

Veterinary Surgery, 2014

In vitro comparison of the mechanical properties of braided polyurethane elastomer (Lycra®) and b... more In vitro comparison of the mechanical properties of braided polyurethane elastomer (Lycra®) and braided polyester (Ethibond™) (1) when inserted into the muscular process of the arytenoid cartilage and (2) as suture loops. Experimental. Equine cadaver larynges (n = 15). The muscular processes (n = 30) of the arytenoid cartilages were dissected from each larynx and embedded in a resin base. Lycra® and Ethibond™ prostheses were randomly allocated to the left or right muscular process and each underwent cyclic fatigue (25-50 N) followed by load-to-failure testing. Isolated suture loops of Lycra® (n = 25) and Ethibond™ (n = 25) also underwent the same cyclic fatigue followed by load-to-failure testing (n = 20) or a creep testing protocol (25 N for 10 min; n = 5). Lycra® prostheses pulled through the cartilage in a significantly greater proportion of cyclic tests (P = .015) and at lower mean (±SD) loads, (95.9 ± 23.4 N) during load-to-failure testing than Ethibond™ prostheses (155.2 ± 24.4 N; P = .0041). Lycra® had a significantly greater displacement with and without a cartilage interface when compared to Ethibond™ (P < .001, P < .002). The Lycra® isolated suture loops failed at significantly greater loads (233.0 ± 38.7 N) during load-to-failure testing than Ethibond™ loops (201.6 ± 47.4 N; P = .042). Lycra® prostheses embedded in laryngeal cartilage pulled through the cartilage at lower loads than Ethibond™ prostheses. Lycra® suture loops were stronger than Ethibond™ suture loops. Lycra® had greater displacement than Ethibond™ in all tests as suture loops or when embedded in cartilage.

Research paper thumbnail of Jejuno- or ileocecal anastomosis performed in seven horses exhibiting colic

The Canadian veterinary journal. La revue vétérinaire canadienne, 1975

Research paper thumbnail of Efficient transduction of equine adipose-derived mesenchymal stem cells by VSV-G pseudotyped lentiviral vectors

Research in veterinary science, Jan 6, 2014

Equine adipose-derived mesenchymal stem cells (EADMSC) provide a unique cell-based approach for t... more Equine adipose-derived mesenchymal stem cells (EADMSC) provide a unique cell-based approach for treatment of a variety of equine musculoskeletal injuries, via regeneration of diseased or damaged tissue, or the secretion of immunomodulatory molecules. These capabilities can be further enhanced by genetic modification using lentiviral vectors, which provide a safe and efficient method of gene delivery. We investigated the suitability of lentiviral vector technology for gene delivery into EADMSC, using GFP expressing lentiviral vectors pseudotyped with the G glycoprotein from the vesicular stomatitis virus (V-GFP) or, for the first time, the baculovirus gp64 envelope protein (G-GFP). In this study, we produced similarly high titre V-GFP and G-GFP lentiviral vectors. Flow cytometric analysis showed efficient transduction using V-GFP; however G-GFP exhibited a poor ability to transduce EADMSC. Transduction resulted in sustained GFP expression over four passages, with minimal effects on c...

Research paper thumbnail of Association of age, sex and breed factors in acute equine colic: A retrospective study of 320 cases admitted to a veterinary teaching hospital in the U.S.A

Preventive Veterinary Medicine, 1989

Research paper thumbnail of A paper-based scaffold for enhanced osteogenic differentiation of equine adipose-derived stem cells

Biotechnology Letters, 2015

We investigated the applicability of single layer paper-based scaffolds for the three-dimensional... more We investigated the applicability of single layer paper-based scaffolds for the three-dimensional (3D) growth and osteogenic differentiation of equine adipose-derived stem cells (EADSC), with comparison against conventional two-dimensional (2D) culture on polystyrene tissue culture vessels. Viable culture of EADSC was achieved using paper-based scaffolds, with EADSC grown and differentiated in 3D culture retaining high cell viability (>94 %), similarly to EADSC in 2D culture. Osteogenic differentiation of EADSC was significantly enhanced in 3D culture, with Alizarin Red S staining and quantification demonstrating increased mineralisation (p < 0.0001), and an associated increase in expression of the osteogenic-specific markers alkaline phosphatase (p < 0.0001), osteopontin (p < 0.0001), and runx2 (p < 0.01). Furthermore, scanning electron microscopy revealed a spherical morphology of EADSC in 3D culture, compared to a flat morphology of EADSC in 2D culture. Single layer paper-based scaffolds provide an enhanced environment for the in vitro 3D growth and osteogenic differentiation of EADSC, with high cell viability, and a spherical morphology.

Research paper thumbnail of Direct Conversion of Equine Adipose-Derived Stem Cells into Induced Neuronal Cells Is Enhanced in Three-Dimensional Culture

Cellular reprogramming, Jan 18, 2015

The ability to culture neurons from horses may allow further investigation into equine neurologic... more The ability to culture neurons from horses may allow further investigation into equine neurological disorders. In this study, we demonstrate the generation of induced neuronal cells from equine adipose-derived stem cells (EADSCs) using a combination of lentiviral vector expression of the neuronal transcription factors Brn2, Ascl1, Myt1l (BAM) and NeuroD1 and a defined chemical induction medium, with βIII-tubulin-positive induced neuronal cells displaying a distinct neuronal morphology of rounded and compact cell bodies, extensive neurite outgrowth, and branching of processes. Furthermore, we investigated the effects of dimensionality on neuronal transdifferentiation, comparing conventional two-dimensional (2D) monolayer culture against three-dimensional (3D) culture on a porous polystyrene scaffold. Neuronal transdifferentiation was enhanced in 3D culture, with evenly distributed cells located on the surface and throughout the scaffold. Transdifferentiation efficiency was increased ...

Research paper thumbnail of Prognosis in equine colic patients using multivariate analysis

Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire

Research paper thumbnail of In Vitro Mechanical Testing of Braided Polyurethane Elastic Fiber and Braided Polyester for Equine Laryngoplasty

Veterinary Surgery, 2014

In vitro comparison of the mechanical properties of braided polyurethane elastomer (Lycra®) and b... more In vitro comparison of the mechanical properties of braided polyurethane elastomer (Lycra®) and braided polyester (Ethibond™) (1) when inserted into the muscular process of the arytenoid cartilage and (2) as suture loops. Experimental. Equine cadaver larynges (n = 15). The muscular processes (n = 30) of the arytenoid cartilages were dissected from each larynx and embedded in a resin base. Lycra® and Ethibond™ prostheses were randomly allocated to the left or right muscular process and each underwent cyclic fatigue (25-50 N) followed by load-to-failure testing. Isolated suture loops of Lycra® (n = 25) and Ethibond™ (n = 25) also underwent the same cyclic fatigue followed by load-to-failure testing (n = 20) or a creep testing protocol (25 N for 10 min; n = 5). Lycra® prostheses pulled through the cartilage in a significantly greater proportion of cyclic tests (P = .015) and at lower mean (±SD) loads, (95.9 ± 23.4 N) during load-to-failure testing than Ethibond™ prostheses (155.2 ± 24.4 N; P = .0041). Lycra® had a significantly greater displacement with and without a cartilage interface when compared to Ethibond™ (P < .001, P < .002). The Lycra® isolated suture loops failed at significantly greater loads (233.0 ± 38.7 N) during load-to-failure testing than Ethibond™ loops (201.6 ± 47.4 N; P = .042). Lycra® prostheses embedded in laryngeal cartilage pulled through the cartilage at lower loads than Ethibond™ prostheses. Lycra® suture loops were stronger than Ethibond™ suture loops. Lycra® had greater displacement than Ethibond™ in all tests as suture loops or when embedded in cartilage.

Research paper thumbnail of Jejuno- or ileocecal anastomosis performed in seven horses exhibiting colic

The Canadian veterinary journal. La revue vétérinaire canadienne, 1975

Research paper thumbnail of Efficient transduction of equine adipose-derived mesenchymal stem cells by VSV-G pseudotyped lentiviral vectors

Research in veterinary science, Jan 6, 2014

Equine adipose-derived mesenchymal stem cells (EADMSC) provide a unique cell-based approach for t... more Equine adipose-derived mesenchymal stem cells (EADMSC) provide a unique cell-based approach for treatment of a variety of equine musculoskeletal injuries, via regeneration of diseased or damaged tissue, or the secretion of immunomodulatory molecules. These capabilities can be further enhanced by genetic modification using lentiviral vectors, which provide a safe and efficient method of gene delivery. We investigated the suitability of lentiviral vector technology for gene delivery into EADMSC, using GFP expressing lentiviral vectors pseudotyped with the G glycoprotein from the vesicular stomatitis virus (V-GFP) or, for the first time, the baculovirus gp64 envelope protein (G-GFP). In this study, we produced similarly high titre V-GFP and G-GFP lentiviral vectors. Flow cytometric analysis showed efficient transduction using V-GFP; however G-GFP exhibited a poor ability to transduce EADMSC. Transduction resulted in sustained GFP expression over four passages, with minimal effects on c...

Research paper thumbnail of Association of age, sex and breed factors in acute equine colic: A retrospective study of 320 cases admitted to a veterinary teaching hospital in the U.S.A

Preventive Veterinary Medicine, 1989

Research paper thumbnail of A paper-based scaffold for enhanced osteogenic differentiation of equine adipose-derived stem cells

Biotechnology Letters, 2015

We investigated the applicability of single layer paper-based scaffolds for the three-dimensional... more We investigated the applicability of single layer paper-based scaffolds for the three-dimensional (3D) growth and osteogenic differentiation of equine adipose-derived stem cells (EADSC), with comparison against conventional two-dimensional (2D) culture on polystyrene tissue culture vessels. Viable culture of EADSC was achieved using paper-based scaffolds, with EADSC grown and differentiated in 3D culture retaining high cell viability (>94 %), similarly to EADSC in 2D culture. Osteogenic differentiation of EADSC was significantly enhanced in 3D culture, with Alizarin Red S staining and quantification demonstrating increased mineralisation (p < 0.0001), and an associated increase in expression of the osteogenic-specific markers alkaline phosphatase (p < 0.0001), osteopontin (p < 0.0001), and runx2 (p < 0.01). Furthermore, scanning electron microscopy revealed a spherical morphology of EADSC in 3D culture, compared to a flat morphology of EADSC in 2D culture. Single layer paper-based scaffolds provide an enhanced environment for the in vitro 3D growth and osteogenic differentiation of EADSC, with high cell viability, and a spherical morphology.