The peripheral nervous system has a limited innate capacity for self-repair following injury, and... more The peripheral nervous system has a limited innate capacity for self-repair following injury, and surgical intervention is often required. For injuries greater than a few millimeters autografting is standard practice although it is associated with donor site morbidity and is limited in its availability. Because of this, nerve guidance conduits (NGCs) can be viewed as an advantageous alternative, but currently have limited efficacy for short and large injury gaps in comparison to autograft. Current commercially available NGC designs rely on existing regulatory approved materials and traditional production methods, limiting improvement of their design. The aim of this study was to establish a novel method for NGC manufacture using a custom built laser-based microstereolithography (mSL) setup that incorporated a 405nmlaser source to produce 3D constructs with ~50 mm resolution from a photocurable poly(ethylene glycol) resin. These were evaluated by SEM, in vitro neuronal, Schwann and dorsal root ganglion culture and in vivo using a thy-1-YFP-H mouse common fibular nerve injurymodel. NGCs with dimensions of 1mminternal diameter5mmlength with awall thickness of 250 mm were fabricated and capable of supporting re-innervation across a 3 mm injury gap after 21 days, with results close to that of an autograft control. The study provides a technology platform for the rapid microfabrication of biocompatible materials, a novel method for in vivo evaluation, and a benchmark for future development in more advanced NGC designs, biodegradable and larger device sizes, and longer-term implantation studies.
Current nerve tissue engineering applications are adopting xenogeneic nerve tissue as potential n... more Current nerve tissue engineering applications are adopting xenogeneic nerve tissue as potential nerve grafts to help aid nerve regeneration. However, there is little literature that describes the exact location, anatomy and physiology of these nerves to highlight their potential as a donor graft. The aim of this study was to identify and characterise the structural and extracellular matrix (ECM) components of porcine peripheral nerves in the hind leg. Methods included the dissection of porcine nerves, localisation, characterisation and quantification of the ECM components and identification of nerve cells. Results showed a noticeable variance between porcine and rat nerve (a commonly studied species) in terms of fascicle number. The study also revealed that when porcine peripheral nerves branch, a decrease in fascicle number and size was evident. Porcine ECM and nerve fascicles were found to be predominately comprised of collagen together with glycosaminoglycans, laminin and fibrone...
In this study, we report the production of amine functionalized nanodiamond. The amine functional... more In this study, we report the production of amine functionalized nanodiamond. The amine functionalized nanodiamond forms a conformal monolayer on a negatively charged surface produced via plasma polymerization of acrylic acid. Nanodiamond terminated surfaces were studied as substrates for neuronal cell culture. NG108-15 neuroblastoma-glioma hybrid cells were successfully cultured upon amine functionalized nanodiamond coated surfaces for between 1 and 7 d. Additionally, primary dorsal root ganglion (DRG) neurons and Schwann cells isolated from Wistar rats were also successfully cultured over a period of 21 d illustrating the potential of the coating for applications in the treatment of peripheral nerve injury.
Uncoated and poly(styrene sulphonate) (PSS)-coated gold nanorods were taken up by NG108-15 neuron... more Uncoated and poly(styrene sulphonate) (PSS)-coated gold nanorods were taken up by NG108-15 neuronal cells. Exposure to 780 nm laser light at the plasmon resonance wavelength of the gold nanorods was found to induce intracellular Ca(2+) transients. The higher Ca(2+) peaks were observed at lower laser doses, with the highest levels obtained at a radiant exposure of 0.33 J/cm(2) . In contrast, the cells without nanoparticles showed a consistently small response, independent of the laser dose. These initial results open up new opportunities for peripheral nerve regeneration treatments and for more efficient optical stimulation techniques.
Epithelial renewal in skin is achieved by the constant turnover and differentiation of keratinocy... more Epithelial renewal in skin is achieved by the constant turnover and differentiation of keratinocytes. Three popular hypotheses have been proposed to explain basal keratinocyte regeneration and epidermal homeostasis: 1) asymmetric division (stem-transit amplifying cell); 2) populational asymmetry (progenitor cell with stochastic fate); and 3) populational asymmetry with stem cells. In this study, we investigated lineage dynamics using these hypotheses with a 3D agent-based model of the epidermis. The model simulated the growth and maintenance of the epidermis over three years. The offspring of each proliferative cell was traced. While all lineages were preserved in asymmetric division, the vast majority were lost when assuming populational asymmetry. The third hypothesis provided the most reliable mechanism for self-renewal by preserving genetic heterogeneity in quiescent stem cells, and also inherent mechanisms for skin ageing and the accumulation of genetic mutation.
Advances in experimental medicine and biology, 2010
The melanocortin family are a series of very potent neuropeptides that derive from a parent propo... more The melanocortin family are a series of very potent neuropeptides that derive from a parent propopiomelanocortin molecule (POMC). They are expressed predominantly in the brain, specifically the pituitary gland and also in the central nervous system. Interestingly, recent research also suggests the existence of regulatory functions outside of the brain, in a wide range of peripheral tissues. Several important melanocortin peptides with differing functions are created by the tissue-specific proteolytic cleavage of POMC, generating peptides including ACTH and α-MSH. For many years the major recognised function of α-MSH was an ability to stimulate melanocyte cells of the skin to pigment. However, a number of parallel functions unrelated to melanogenesis have been described in the literature for several years. A more complete understanding of this work arose after the discovery and cloning of the melanocortin receptors in 1992, which lead to the recognition of many wider roles of the mel...
We reported recently that the inflammatory cytokine tumour necrosis factor alpha (TNF-alpha) can ... more We reported recently that the inflammatory cytokine tumour necrosis factor alpha (TNF-alpha) can upregulate integrin expression, cell attachment and invasion of cells through fibronectin in a human melanoma cell line (HBL). Furthermore, the actions of TNF-alpha were suppressed by the addition of an anti-inflammatory peptide alpha-melanocyte-stimulating hormone (alpha-MSH). In the current study, we extend this work investigating to what extent TNF-alpha might stimulate melanoma invasion by promoting cell migration and whether alpha-MSH is also inhibitory. Two human melanoma cell lines were examined in vitro (HBL and C8161) using a scratch migration assay. Analysis using either time-lapse video microscopy or imaging software analysis of migrating 'fronts' of cells revealed that C8161 cells migrated more rapidly than HBL cells. However, when cells were stimulated with TNF-alpha both cell types responded with a significant increase in migration distance over a 16-26 h incubation...
Alpha-melanocyte stimulating hormone (alpha-MSH) was originally documented as a peptide that indu... more Alpha-melanocyte stimulating hormone (alpha-MSH) was originally documented as a peptide that induced pigmentation in skin. However, more recent work suggests that it acts as a potent anti-inflammatory molecule in several tissues including nerve. Alpha-MSH works by directly inhibiting cytokines that cause inflammation, in particular tumour necrosis factor-alpha, interleukin-1beta, interleukin-6 and gamma-interferon. A common mechanism of inhibition is via the NF-kappaB transcription factor. We investigated the ability of alpha-MSH to inhibit the activation of NF-kappaB in cultured rat primary olfactory ensheathing cells stimulated with tumour necrosis factor-alpha or gamma-interferon. Both cytokines activated NF-kappaB rapidly (after 60 min incubation), observed as a translocation from cytoplasm to nucleus. alpha-MSH inhibited this activation (and hence nuclear translocation) by approximately 50% for both cytokines. The anti-inflammatory properties of this peptide in neural cells may...
Alpha-melanocyte stimulating hormone (alpha-MSH) is known to have pleiotrophic functions includin... more Alpha-melanocyte stimulating hormone (alpha-MSH) is known to have pleiotrophic functions including pigmentary, anti-inflammatory, antipyretic and immunoregulatory roles in the mammalian body. It is also reported to influence melanoma invasion with levels of alpha-, beta- and gamma-MSH correlated clinically with malignant melanoma development, but other studies suggest alpha-MSH acts to retard invasion. In the present study, we investigated the action of alpha-MSH on three human melanoma cell lines (HBL, A375-SM and C8161) differing in metastatic potential. alpha-melanocyte-simulating hormone reduced invasion through fibronectin and also through a human reconstructed skin composite model for the HBL line, and inhibited proinflammatory cytokine-stimulated activation of the NF-kappaB transcription factor. However, A375-SM and C8161 cells did not respond to alpha-MSH. Immunofluorescent microscopy and Western blotting identified melanocortin-1 receptor (MC-1R) expression for all three li...
The Journal of neuroscience : the official journal of the Society for Neuroscience, 2000
While the transporters that accumulate classical neurotransmitters in synaptic vesicles have been... more While the transporters that accumulate classical neurotransmitters in synaptic vesicles have been identified, little is known about how their expression regulates synaptic transmission. We have used adenoviral-mediated transfection to increase expression of the brain vesicular monoamine transporter VMAT2 and presynaptic amperometric recordings to characterize the effects on quantal release. In presynaptic axonal varicosities of ventral midbrain neurons in postnatal culture, VMAT2 overexpression in small synaptic vesicles increased both quantal size and frequency, consistent with the recruitment of synaptic vesicles that do not normally release dopamine. This was confirmed using noncatecholaminergic AtT-20 cells, in which VMAT2 expression induced the quantal release of dopamine. The ability to increase quantal size in vesicles that were already competent for dopamine release was shown in PC12 cells, in which VMAT2 expression increased the quantal size but not the number of release ev...
The Journal of neuroscience : the official journal of the Society for Neuroscience, 1998
The modulation of motor behavior by protein kinase C (PKC) signaling pathways in nigrostriatal ne... more The modulation of motor behavior by protein kinase C (PKC) signaling pathways in nigrostriatal neurons was examined by using a genetic intervention approach. Herpes simplex virus type 1 (HSV-1) vectors that encode a catalytic domain of rat PKCbetaII (PkcDelta) were developed. PkcDelta exhibited a constitutively active protein kinase activity with a substrate specificity similar to that of rat brain PKC. As demonstrated in cultured sympathetic neurons, PkcDelta caused a long-lasting, activation-dependent increase in neurotransmitter release. In the rat brain, microinjection of HSV-1 vectors that contain the tyrosine hydroxylase promoter targeted expression to dopaminergic nigrostriatal neurons. Expression of pkcDelta in a small percentage of nigrostriatal neurons (approximately 0.1-2%) was sufficient to produce a long-term (>/=1 month) change in apomorphine-induced rotational behavior. Nigrostriatal neurons were the only catecholaminergic neurons that contained PkcDelta, and the a...
... 6301Effects of Catecholaminergic Drugs upon MemoryStorage Processes in Mice1,2JOHN W. HAYCOCK... more ... 6301Effects of Catecholaminergic Drugs upon MemoryStorage Processes in Mice1,2JOHN W. HAYCOCK,a RODERICK VAN BUSKIRK,4AND JAMES ... Numerous studies have previously noted similar effects of DDC uponbrain catecholamine levels (Collins, 1965; Carlsson et al ...
INTRODUCTION: The use of primary Schwann cells in conjunction with nerve guidance conduits has be... more INTRODUCTION: The use of primary Schwann cells in conjunction with nerve guidance conduits has been shown to improve regeneration and remyelination of injured peripheral nerve [1]. However, clinical application of primary Schwann cells for nerve repair is limited due to the requirement of nerve biopsies, and slow growth of Schwann cells in vitro. Adipose-derived stem cells (ASCs) may be an optional cell source for derivation towards Schwann-like cells. In this study adipose-derived stem cells from three anatomically different sites were explored for differentiation towards a Schwann phenotype which included protein expression, morphology and function. METHODS: Stem cells were isolated from: i) subcutaneous; ii) perinephric adipose and iii) epididymis adipose tissue of adult male Wistar rats according to the method of Bjorntorp et al [2]. Plastic-adherent multipotent cells were then cultured according to an experimental differentiation protocol using β-mercaptoethanol, retinoic and g...
The peripheral nervous system has a limited innate capacity for self-repair following injury, and... more The peripheral nervous system has a limited innate capacity for self-repair following injury, and surgical intervention is often required. For injuries greater than a few millimeters autografting is standard practice although it is associated with donor site morbidity and is limited in its availability. Because of this, nerve guidance conduits (NGCs) can be viewed as an advantageous alternative, but currently have limited efficacy for short and large injury gaps in comparison to autograft. Current commercially available NGC designs rely on existing regulatory approved materials and traditional production methods, limiting improvement of their design. The aim of this study was to establish a novel method for NGC manufacture using a custom built laser-based microstereolithography (mSL) setup that incorporated a 405nmlaser source to produce 3D constructs with ~50 mm resolution from a photocurable poly(ethylene glycol) resin. These were evaluated by SEM, in vitro neuronal, Schwann and dorsal root ganglion culture and in vivo using a thy-1-YFP-H mouse common fibular nerve injurymodel. NGCs with dimensions of 1mminternal diameter5mmlength with awall thickness of 250 mm were fabricated and capable of supporting re-innervation across a 3 mm injury gap after 21 days, with results close to that of an autograft control. The study provides a technology platform for the rapid microfabrication of biocompatible materials, a novel method for in vivo evaluation, and a benchmark for future development in more advanced NGC designs, biodegradable and larger device sizes, and longer-term implantation studies.
Current nerve tissue engineering applications are adopting xenogeneic nerve tissue as potential n... more Current nerve tissue engineering applications are adopting xenogeneic nerve tissue as potential nerve grafts to help aid nerve regeneration. However, there is little literature that describes the exact location, anatomy and physiology of these nerves to highlight their potential as a donor graft. The aim of this study was to identify and characterise the structural and extracellular matrix (ECM) components of porcine peripheral nerves in the hind leg. Methods included the dissection of porcine nerves, localisation, characterisation and quantification of the ECM components and identification of nerve cells. Results showed a noticeable variance between porcine and rat nerve (a commonly studied species) in terms of fascicle number. The study also revealed that when porcine peripheral nerves branch, a decrease in fascicle number and size was evident. Porcine ECM and nerve fascicles were found to be predominately comprised of collagen together with glycosaminoglycans, laminin and fibrone...
In this study, we report the production of amine functionalized nanodiamond. The amine functional... more In this study, we report the production of amine functionalized nanodiamond. The amine functionalized nanodiamond forms a conformal monolayer on a negatively charged surface produced via plasma polymerization of acrylic acid. Nanodiamond terminated surfaces were studied as substrates for neuronal cell culture. NG108-15 neuroblastoma-glioma hybrid cells were successfully cultured upon amine functionalized nanodiamond coated surfaces for between 1 and 7 d. Additionally, primary dorsal root ganglion (DRG) neurons and Schwann cells isolated from Wistar rats were also successfully cultured over a period of 21 d illustrating the potential of the coating for applications in the treatment of peripheral nerve injury.
Uncoated and poly(styrene sulphonate) (PSS)-coated gold nanorods were taken up by NG108-15 neuron... more Uncoated and poly(styrene sulphonate) (PSS)-coated gold nanorods were taken up by NG108-15 neuronal cells. Exposure to 780 nm laser light at the plasmon resonance wavelength of the gold nanorods was found to induce intracellular Ca(2+) transients. The higher Ca(2+) peaks were observed at lower laser doses, with the highest levels obtained at a radiant exposure of 0.33 J/cm(2) . In contrast, the cells without nanoparticles showed a consistently small response, independent of the laser dose. These initial results open up new opportunities for peripheral nerve regeneration treatments and for more efficient optical stimulation techniques.
Epithelial renewal in skin is achieved by the constant turnover and differentiation of keratinocy... more Epithelial renewal in skin is achieved by the constant turnover and differentiation of keratinocytes. Three popular hypotheses have been proposed to explain basal keratinocyte regeneration and epidermal homeostasis: 1) asymmetric division (stem-transit amplifying cell); 2) populational asymmetry (progenitor cell with stochastic fate); and 3) populational asymmetry with stem cells. In this study, we investigated lineage dynamics using these hypotheses with a 3D agent-based model of the epidermis. The model simulated the growth and maintenance of the epidermis over three years. The offspring of each proliferative cell was traced. While all lineages were preserved in asymmetric division, the vast majority were lost when assuming populational asymmetry. The third hypothesis provided the most reliable mechanism for self-renewal by preserving genetic heterogeneity in quiescent stem cells, and also inherent mechanisms for skin ageing and the accumulation of genetic mutation.
Advances in experimental medicine and biology, 2010
The melanocortin family are a series of very potent neuropeptides that derive from a parent propo... more The melanocortin family are a series of very potent neuropeptides that derive from a parent propopiomelanocortin molecule (POMC). They are expressed predominantly in the brain, specifically the pituitary gland and also in the central nervous system. Interestingly, recent research also suggests the existence of regulatory functions outside of the brain, in a wide range of peripheral tissues. Several important melanocortin peptides with differing functions are created by the tissue-specific proteolytic cleavage of POMC, generating peptides including ACTH and α-MSH. For many years the major recognised function of α-MSH was an ability to stimulate melanocyte cells of the skin to pigment. However, a number of parallel functions unrelated to melanogenesis have been described in the literature for several years. A more complete understanding of this work arose after the discovery and cloning of the melanocortin receptors in 1992, which lead to the recognition of many wider roles of the mel...
We reported recently that the inflammatory cytokine tumour necrosis factor alpha (TNF-alpha) can ... more We reported recently that the inflammatory cytokine tumour necrosis factor alpha (TNF-alpha) can upregulate integrin expression, cell attachment and invasion of cells through fibronectin in a human melanoma cell line (HBL). Furthermore, the actions of TNF-alpha were suppressed by the addition of an anti-inflammatory peptide alpha-melanocyte-stimulating hormone (alpha-MSH). In the current study, we extend this work investigating to what extent TNF-alpha might stimulate melanoma invasion by promoting cell migration and whether alpha-MSH is also inhibitory. Two human melanoma cell lines were examined in vitro (HBL and C8161) using a scratch migration assay. Analysis using either time-lapse video microscopy or imaging software analysis of migrating 'fronts' of cells revealed that C8161 cells migrated more rapidly than HBL cells. However, when cells were stimulated with TNF-alpha both cell types responded with a significant increase in migration distance over a 16-26 h incubation...
Alpha-melanocyte stimulating hormone (alpha-MSH) was originally documented as a peptide that indu... more Alpha-melanocyte stimulating hormone (alpha-MSH) was originally documented as a peptide that induced pigmentation in skin. However, more recent work suggests that it acts as a potent anti-inflammatory molecule in several tissues including nerve. Alpha-MSH works by directly inhibiting cytokines that cause inflammation, in particular tumour necrosis factor-alpha, interleukin-1beta, interleukin-6 and gamma-interferon. A common mechanism of inhibition is via the NF-kappaB transcription factor. We investigated the ability of alpha-MSH to inhibit the activation of NF-kappaB in cultured rat primary olfactory ensheathing cells stimulated with tumour necrosis factor-alpha or gamma-interferon. Both cytokines activated NF-kappaB rapidly (after 60 min incubation), observed as a translocation from cytoplasm to nucleus. alpha-MSH inhibited this activation (and hence nuclear translocation) by approximately 50% for both cytokines. The anti-inflammatory properties of this peptide in neural cells may...
Alpha-melanocyte stimulating hormone (alpha-MSH) is known to have pleiotrophic functions includin... more Alpha-melanocyte stimulating hormone (alpha-MSH) is known to have pleiotrophic functions including pigmentary, anti-inflammatory, antipyretic and immunoregulatory roles in the mammalian body. It is also reported to influence melanoma invasion with levels of alpha-, beta- and gamma-MSH correlated clinically with malignant melanoma development, but other studies suggest alpha-MSH acts to retard invasion. In the present study, we investigated the action of alpha-MSH on three human melanoma cell lines (HBL, A375-SM and C8161) differing in metastatic potential. alpha-melanocyte-simulating hormone reduced invasion through fibronectin and also through a human reconstructed skin composite model for the HBL line, and inhibited proinflammatory cytokine-stimulated activation of the NF-kappaB transcription factor. However, A375-SM and C8161 cells did not respond to alpha-MSH. Immunofluorescent microscopy and Western blotting identified melanocortin-1 receptor (MC-1R) expression for all three li...
The Journal of neuroscience : the official journal of the Society for Neuroscience, 2000
While the transporters that accumulate classical neurotransmitters in synaptic vesicles have been... more While the transporters that accumulate classical neurotransmitters in synaptic vesicles have been identified, little is known about how their expression regulates synaptic transmission. We have used adenoviral-mediated transfection to increase expression of the brain vesicular monoamine transporter VMAT2 and presynaptic amperometric recordings to characterize the effects on quantal release. In presynaptic axonal varicosities of ventral midbrain neurons in postnatal culture, VMAT2 overexpression in small synaptic vesicles increased both quantal size and frequency, consistent with the recruitment of synaptic vesicles that do not normally release dopamine. This was confirmed using noncatecholaminergic AtT-20 cells, in which VMAT2 expression induced the quantal release of dopamine. The ability to increase quantal size in vesicles that were already competent for dopamine release was shown in PC12 cells, in which VMAT2 expression increased the quantal size but not the number of release ev...
The Journal of neuroscience : the official journal of the Society for Neuroscience, 1998
The modulation of motor behavior by protein kinase C (PKC) signaling pathways in nigrostriatal ne... more The modulation of motor behavior by protein kinase C (PKC) signaling pathways in nigrostriatal neurons was examined by using a genetic intervention approach. Herpes simplex virus type 1 (HSV-1) vectors that encode a catalytic domain of rat PKCbetaII (PkcDelta) were developed. PkcDelta exhibited a constitutively active protein kinase activity with a substrate specificity similar to that of rat brain PKC. As demonstrated in cultured sympathetic neurons, PkcDelta caused a long-lasting, activation-dependent increase in neurotransmitter release. In the rat brain, microinjection of HSV-1 vectors that contain the tyrosine hydroxylase promoter targeted expression to dopaminergic nigrostriatal neurons. Expression of pkcDelta in a small percentage of nigrostriatal neurons (approximately 0.1-2%) was sufficient to produce a long-term (>/=1 month) change in apomorphine-induced rotational behavior. Nigrostriatal neurons were the only catecholaminergic neurons that contained PkcDelta, and the a...
... 6301Effects of Catecholaminergic Drugs upon MemoryStorage Processes in Mice1,2JOHN W. HAYCOCK... more ... 6301Effects of Catecholaminergic Drugs upon MemoryStorage Processes in Mice1,2JOHN W. HAYCOCK,a RODERICK VAN BUSKIRK,4AND JAMES ... Numerous studies have previously noted similar effects of DDC uponbrain catecholamine levels (Collins, 1965; Carlsson et al ...
INTRODUCTION: The use of primary Schwann cells in conjunction with nerve guidance conduits has be... more INTRODUCTION: The use of primary Schwann cells in conjunction with nerve guidance conduits has been shown to improve regeneration and remyelination of injured peripheral nerve [1]. However, clinical application of primary Schwann cells for nerve repair is limited due to the requirement of nerve biopsies, and slow growth of Schwann cells in vitro. Adipose-derived stem cells (ASCs) may be an optional cell source for derivation towards Schwann-like cells. In this study adipose-derived stem cells from three anatomically different sites were explored for differentiation towards a Schwann phenotype which included protein expression, morphology and function. METHODS: Stem cells were isolated from: i) subcutaneous; ii) perinephric adipose and iii) epididymis adipose tissue of adult male Wistar rats according to the method of Bjorntorp et al [2]. Plastic-adherent multipotent cells were then cultured according to an experimental differentiation protocol using β-mercaptoethanol, retinoic and g...
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intervention is often required. For injuries greater than a few millimeters autografting is standard practice
although it is associated with donor site morbidity and is limited in its availability. Because of this, nerve
guidance conduits (NGCs) can be viewed as an advantageous alternative, but currently have limited efficacy
for short and large injury gaps in comparison to autograft. Current commercially available NGC designs rely
on existing regulatory approved materials and traditional production methods, limiting improvement of
their design. The aim of this study was to establish a novel method for NGC manufacture using a custom
built laser-based microstereolithography (mSL) setup that incorporated a 405nmlaser source to produce 3D
constructs with ~50 mm resolution from a photocurable poly(ethylene glycol) resin. These were evaluated
by SEM, in vitro neuronal, Schwann and dorsal root ganglion culture and in vivo using a thy-1-YFP-H mouse
common fibular nerve injurymodel. NGCs with dimensions of 1mminternal diameter5mmlength with
awall thickness of 250 mm were fabricated and capable of supporting re-innervation across a 3 mm injury
gap after 21 days, with results close to that of an autograft control. The study provides a technology platform
for the rapid microfabrication of biocompatible materials, a novel method for in vivo evaluation, and a
benchmark for future development in more advanced NGC designs, biodegradable and larger device sizes,
and longer-term implantation studies.
intervention is often required. For injuries greater than a few millimeters autografting is standard practice
although it is associated with donor site morbidity and is limited in its availability. Because of this, nerve
guidance conduits (NGCs) can be viewed as an advantageous alternative, but currently have limited efficacy
for short and large injury gaps in comparison to autograft. Current commercially available NGC designs rely
on existing regulatory approved materials and traditional production methods, limiting improvement of
their design. The aim of this study was to establish a novel method for NGC manufacture using a custom
built laser-based microstereolithography (mSL) setup that incorporated a 405nmlaser source to produce 3D
constructs with ~50 mm resolution from a photocurable poly(ethylene glycol) resin. These were evaluated
by SEM, in vitro neuronal, Schwann and dorsal root ganglion culture and in vivo using a thy-1-YFP-H mouse
common fibular nerve injurymodel. NGCs with dimensions of 1mminternal diameter5mmlength with
awall thickness of 250 mm were fabricated and capable of supporting re-innervation across a 3 mm injury
gap after 21 days, with results close to that of an autograft control. The study provides a technology platform
for the rapid microfabrication of biocompatible materials, a novel method for in vivo evaluation, and a
benchmark for future development in more advanced NGC designs, biodegradable and larger device sizes,
and longer-term implantation studies.