Functional recovery in penetrating neurological injury is hampered by a lack of clinical regenerative therapies.Biomaterial therapies show promise as medical materials for neural repair through immunomodulation,struct...Functional recovery in penetrating neurological injury is hampered by a lack of clinical regenerative therapies.Biomaterial therapies show promise as medical materials for neural repair through immunomodulation,structural support,and delivery of therapeutic biomolecules.However,a lack of facile and pathology-mimetic models for therapeutic testing is a bottleneck in neural tissue engineering research.We have deployed a two-dimensional,high-density multicellular cortical brain sheet to develop a facile model of injury(macrotransection/scratch wound)in vitro.The model encompasses the major neural cell types involved in pathological responses post-injury.Critically,we observed hallmark pathological responses in injury foci including cell scarring,immune cell infiltration,precursor cell migration,and shortrange axonal sprouting.Delivering test magnetic particles to evaluate the potential of the model for biomaterial screening shows a high uptake of introduced magnetic particles by injury-activated immune cells,mimicking in vivo findings.Finally,we proved it is feasible to create reproducible traumatic injuries in the brain sheet(in multielectrode array devices in situ)characterized by focal loss of electrical spiking in injury sites,offering the potential for longer term,electrophysiology plus histology assays.To our knowledge,this is the first in vitro simulation of transecting injury in a two-dimensional multicellular cortical brain cell sheet,that allows for combined histological and electrophysiological readouts of damage/repair.The patho-mimicry and adaptability of this simplified model of brain injury could benefit the testing of biomaterial therapeutics in regenerative neurology,with the option for functional electrophysiological readouts.展开更多
The use of live animal models for testing new therapies for brain and spinal cord repair is a controversial area. Live animal models have associated ethical issues and scientific concerns regarding the predictability ...The use of live animal models for testing new therapies for brain and spinal cord repair is a controversial area. Live animal models have associated ethical issues and scientific concerns regarding the predictability of human responses. Alternative models that replicate the 3 D architecture of the central nervous system have prompted the development of organotypic neural injury models. However, the lack of reliable means to access normal human neural tissue has driven reliance on pathological or post-mortem tissue which limits their biological utility. We have established a protocol to use donor cerebellar tonsillar tissue surgically resected from patients with Chiari malformation(cerebellar herniation towards the foramen magnum, with ectopic rather than diseased tissue) to develop an in vitro organotypic model of traumatic brain injury. Viable tissue was maintained for approximately 2 weeks with all the major neural cell types detected. Traumatic injuries could be introduced into the slices with some cardinal features of post-injury pathology evident. Biomaterial placement was also feasible within the in vitro lesions. Accordingly, this ‘proof-of-concept’ study demonstrates that the model offers potential as an alternative to the use of animal tissue for preclinical testing in neural tissue engineering. To our knowledge, this is the first demonstration that donor tissue from patients with Chiari malformation can be used to develop a benchtop model of traumatic brain injury. However, significant challenges in relation to the clinical availability of tissue were encountered, and we discuss logistical issues that must be considered for model scale-up.展开更多
基金supported by awards from the EPSRC Centre for Doctoral Training in Regenerative Medicine(EP/L014904/1,to JW)an NHS bursary(to RHB)an EPSRC Healthcare Technologies award(EP/T013885/1,to DMC)。
文摘Functional recovery in penetrating neurological injury is hampered by a lack of clinical regenerative therapies.Biomaterial therapies show promise as medical materials for neural repair through immunomodulation,structural support,and delivery of therapeutic biomolecules.However,a lack of facile and pathology-mimetic models for therapeutic testing is a bottleneck in neural tissue engineering research.We have deployed a two-dimensional,high-density multicellular cortical brain sheet to develop a facile model of injury(macrotransection/scratch wound)in vitro.The model encompasses the major neural cell types involved in pathological responses post-injury.Critically,we observed hallmark pathological responses in injury foci including cell scarring,immune cell infiltration,precursor cell migration,and shortrange axonal sprouting.Delivering test magnetic particles to evaluate the potential of the model for biomaterial screening shows a high uptake of introduced magnetic particles by injury-activated immune cells,mimicking in vivo findings.Finally,we proved it is feasible to create reproducible traumatic injuries in the brain sheet(in multielectrode array devices in situ)characterized by focal loss of electrical spiking in injury sites,offering the potential for longer term,electrophysiology plus histology assays.To our knowledge,this is the first in vitro simulation of transecting injury in a two-dimensional multicellular cortical brain cell sheet,that allows for combined histological and electrophysiological readouts of damage/repair.The patho-mimicry and adaptability of this simplified model of brain injury could benefit the testing of biomaterial therapeutics in regenerative neurology,with the option for functional electrophysiological readouts.
基金funded by a grant from the North Staffordshire Medical Institute,UK (to DMC and NT)。
文摘The use of live animal models for testing new therapies for brain and spinal cord repair is a controversial area. Live animal models have associated ethical issues and scientific concerns regarding the predictability of human responses. Alternative models that replicate the 3 D architecture of the central nervous system have prompted the development of organotypic neural injury models. However, the lack of reliable means to access normal human neural tissue has driven reliance on pathological or post-mortem tissue which limits their biological utility. We have established a protocol to use donor cerebellar tonsillar tissue surgically resected from patients with Chiari malformation(cerebellar herniation towards the foramen magnum, with ectopic rather than diseased tissue) to develop an in vitro organotypic model of traumatic brain injury. Viable tissue was maintained for approximately 2 weeks with all the major neural cell types detected. Traumatic injuries could be introduced into the slices with some cardinal features of post-injury pathology evident. Biomaterial placement was also feasible within the in vitro lesions. Accordingly, this ‘proof-of-concept’ study demonstrates that the model offers potential as an alternative to the use of animal tissue for preclinical testing in neural tissue engineering. To our knowledge, this is the first demonstration that donor tissue from patients with Chiari malformation can be used to develop a benchtop model of traumatic brain injury. However, significant challenges in relation to the clinical availability of tissue were encountered, and we discuss logistical issues that must be considered for model scale-up.
