Biomaterials and tissue engineering in traumatic brain injury:novel perspectives on promoting neural regeneration

Traumatic brain injury is a serious medical condition that can be attributed to falls, motor vehicle accidents, sports injuries and acts of violence, causing a series of neural injuries and neuropsychiatric symptoms. However, limited accessibility to the injury sites, complicated histological and anatomical structure, intricate cellular and extracellular milieu, lack of regenerative capacity in the native cells, vast variety of damage routes, and the insufficient time available for treatment have restricted the widespread application of several therapeutic methods in cases of central nervous system injury. Tissue engineering and regenerative medicine have emerged as innovative approaches in the field of nerve regeneration. By combining biomaterials, stem cells, and growth factors, these approaches have provided a platform for developing effective treatments for neural injuries, which can offer the potential to restore neural function, improve patient outcomes, and reduce the need for drugs and invasive surgical procedures. Biomaterials have shown advantages in promoting neural development, inhibiting glial scar formation, and providing a suitable biomimetic neural microenvironment, which makes their application promising in the field of neural regeneration. For instance, bioactive scaffolds loaded with stem cells can provide a biocompatible and biodegradable milieu. Furthermore, stem cells-derived exosomes combine the advantages of stem cells, avoid the risk of immune rejection, cooperate with biomaterials to enhance their biological functions, and exert stable functions, thereby inducing angiogenesis and neural regeneration in patients with traumatic brain injury and promoting the recovery of brain function. Unfortunately, biomaterials have shown positive effects in the laboratory, but when similar materials are used in clinical studies of human central nervous system regeneration, their efficacy is unsatisfactory. Here, we review the characteristics and properties of various bioactive materials, followed by the introduction of applications based on biochemistry and cell molecules, and discuss the emerging role of biomaterials in promoting neural regeneration. Further, we summarize the adaptive biomaterials infused with exosomes produced from stem cells and stem cells themselves for the treatment of traumatic brain injury. Finally, we present the main limitations of biomaterials for the treatment of traumatic brain injury and offer insights into their future potential.

A benchtop brain injury model using resected donor tissue from patients with Chiari malformation

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.

Etiology, Prevalence, and Management of Oral and Maxillofacial Soft Tissue Injuries in Children at the Komfo Anokye Teaching Hospital, Kumasi-Ghana

Background: Maxillofacial injury may vary from simple soft tissue lacerations to complex fractures of the orofacial region. Soft tissue injuries, whether isolated or in combination with other injuries, form part of the frequent traumatic craniofacial injuries seen at the emergency department. The force of impact and the injury type is directly related to the severity of the injury sustained. This study aimed to analyze the etiological factors, prevalence, and management of oral and maxillofacial soft tissue injuries at the Komfo Anokye Teaching Hospital. Methods: This was a prospective study that involved children presenting with oral and maxillofacial injuries at the Accident and Emergency Department and the Oral and Maxillofacial Surgery unit of the Komfo Anokye Teaching Hospital in Kumasi between the period of April to October 2020 (6 months). Patient selection was by convenience sampling targeting all children with injuries who met the inclusion criteria. Inclusion criteria were children below the age of 18 years whose parents or caregivers consent to participation. Children with maxillofacial injuries as a result of burns were excluded from the study. Results: During the study period a total of 134 children were reviewed with oral and maxillofacial injuries at KATH. Of these, 107 (78.9%) were recorded cases of orofacial soft tissue injuries. There were 63 (58.9%) males and 44 (41.1%) females and the male-to-female ratio was 1.5:1. The age range of patients studied was 8 months – 17 years, with mean age ± SD being 9.5 ± 5.3 years. Road Traffic Crash (50.5%) was the most common etiology of which Motor cycle crash constituted 24.3% and Pedestrian knockdown was 19.6%. Falls (42.1%) were the next most common etiology. The lips (19.8%) and the forehead (18.5%) were the most frequently injured sites on the face whiles the tongue (3.3%) had the most injuries intraorally. Laceration (45.7%) was the most frequent injury reviewed, followed by abrasions (35.8%). Most of the soft tissue injuries underwent primary closure (56.3%). A complication rate of 21.2% was recorded in this study and hypertrophic scarring (11.3%) was the most observed.

Therapeutic Effect and Mechanism of Jipei Dilong Ointment on Acute Soft Tissue Injury in Rats

[Objectives]This study was conducted to observe the therapeutic effect of Jipei Dilong Ointment on rats with acute soft tissue injury caused by heavy objects and to explore its action mechanism.[Methods]Thirty six rats were randomly divided into six groups(control group,model group,high-dose Jipei Dilong Ointment group(JP-H),medium-dose Jipei Dilong Ointment group(JP-M),low-dose Jipei Dilong Ointment group(JP-L)and diclofenac group).Except for the Control group,other groups were subjected to modeling of acute soft tissue injury by the weight impact method.All administration was performed once a day for nine consecutive days.The local appearance score and activity disorder score were determined after soft tissue injury in rats.HE staining was used to detect the pathological changes of injured soft tissues in rats.RT-PCR was used to detect the relative mRNA expressions of Bax,Bcl-2,MMP-9 and TIMP-1 in injured soft tissues of rats.Western Blot was used to detect the protein expressions of MMP-9,TIMP-1,TLR4,MyD88 and NF-κB p65 in injured soft tissues of rats.Results were statistically analyzed.[Results]Compared with the model group,Jipei Dilong Ointment could significantly improve the appearance symptoms such as swelling and ecchymosis in the injured area and the movement function of the affected limb(P<0.05).It could also improve the infiltration of inflammatory cells and widening of the intermuscular space caused by injury.Among them,the JP-H group and the diclofenac group had more significant curative effects.After 9 d of administration,each administration group could significantly up-regulate the ratio of Bcl-2/Bax mRNA expression level(P<0.05 or P<0.01),and the ratio of MMP-9/TIMP-1 mRNA expression level showed a downward trend(P>0.05).The expression level of NF-κB p65 protein in each administration group was significantly decreased(P<0.01).The protein expression levels of TLR4 and MyD88 and the ratio of MMP-9/TIMP-1 protein expression level in each administration group decreased to varying degrees.Among them,the JP-H group and diclofenac group significantly decreased(P<0.05).[Conclusions]Jipei Dilong Ointment has the functions of relieving pain,swelling and inflammation.It could improve the local appearance,functional activity and tissue morphology of affected limbs in rats,and has a therapeutic effect on acute soft tissue injury in rats.Its mechanism of action might be related to the inhibition of TLR4/MyD88/NF-κB p65 signaling pathway and the regulation of Bcl-2/Bax and MMP-9/TIMP-1 balance.

Inhibiting endogenous tissue plasminogen activator enhanced neuronal apoptosis and axonal injury after traumatic brain injury

Tissue plasminogen activator is usually used for the treatment of acute ischemic stroke,but the role of endogenous tissue plasminogen activator in traumatic brain injury has been rarely reported.A rat model of traumatic brain injury was established by weight-drop method.The tissue plasminogen activator inhibitor neuroserpin(5μL,0.25 mg/mL)was injected into the lateral ventricle.Neurological function was assessed by neurological severity score.Neuronal and axonal injuries were assessed by hematoxylin-eosin staining and Bielschowsky silver staining.Protein level of endogenous tissue plasminogen activator was analyzed by western blot assay.Apoptotic marker cleaved caspase-3,neuronal marker neurofilament light chain,astrocyte marker glial fibrillary acidic protein and microglial marker Iba-1 were analyzed by immunohistochemical staining.Apoptotic cell types were detected by immunofluorescence double labeling.Apoptotic cells in the damaged cortex were detected by terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP-biotin nick-end labeling staining.Degenerating neurons in the damaged cortex were detected by Fluoro-Jade B staining.Expression of tissue plasminogen activator was increased at 6 hours,and peaked at 3 days after traumatic brain injury.Neuronal apoptosis and axonal injury were detected after traumatic brain injury.Moreover,neuroserpin enhanced neuronal apoptosis,neuronal injury and axonal injury,and activated microglia and astrocytes.Neuroserpin further deteriorated neurobehavioral function in rats with traumatic brain injury.Our findings confirm that inhibition of endogenous tissue plasminogen activator aggravates neuronal apoptosis and axonal injury after traumatic brain injury,and activates microglia and astrocytes.This study was approved by the Biomedical Ethics Committee of Animal Experiments of Shaanxi Province of China in June 2015.

An update–tissue engineered nerve grafts for the repair of peripheral nerve injuries

Peripheral nerve injuries（PNI） are caused by a range of etiologies and result in a broad spectrum of disability. While nerve autografts are the current gold standard for the reconstruction of extensive nerve damage, the limited supply of autologous nerve and complications associated with harvesting nerve from a second surgical site has driven groups from multiple disciplines, including biomedical engineering, neurosurgery, plastic surgery, and orthopedic surgery, to develop a suitable or superior alternative to autografting. Over the last couple of decades, various types of scaffolds, such as acellular nerve grafts（ANGs）, nerve guidance conduits, and non-nervous tissues, have been filled with Schwann cells, stem cells, and/or neurotrophic factors to develop tissue engineered nerve grafts（TENGs）. Although these have shown promising effects on peripheral nerve regeneration in experimental models, the autograft has remained the gold standard for large nerve gaps. This review provides a discussion of recent advances in the development of TENGs and their efficacy in experimental models. Specifically, TENGs have been enhanced via incorporation of genetically engineered cells, methods to improve stem cell survival and differentiation, optimized delivery of neurotrophic factors via drug delivery systems（DDS）, co-administration of platelet-rich plasma（PRP）, and pretreatment with chondroitinase ABC（Ch-ABC）. Other notable advancements include conduits that have been bioengineered to mimic native nerve structure via cell-derived extracellular matrix（ECM） deposition, and the development of transplantable living nervous tissue constructs from rat and human dorsal root ganglia（DRG） neurons. Grafts composed of non-nervous tissues, such as vein, artery, and muscle, will be briefly discussed.

Selected suitable seed cell, scaffold and growth factor could maximize the repair effect using tissue engineering method in spinal cord injury

Spinal cord injury usually leads to permanent disability, which could cause a huge financial problem to the patient. Up to now there is no effective method to treat this disease. The key of the treatment is to enable the damage zone axonal regeneration and luckily it could go through the damage zone; last a connection can be established with the target neurons. This study attempts to combine stem cell, material science and genetic modification technology together, by preparing two genes modified adipose-derived stem cells and inducing them into neuron direction; then by compositing them on the silk fibroin/chitosan scaffold and implanting them into the spinal cord injury model, seed cells can have features of neuron cells. At the same time, it could stably express the brain-derived neurotrophic factor and neurotrophin-3, both of which could produce synergistic effects, which have a positive effect on the recovery of spinal cord. The spinal cord scaffold bridges the broken end of the spinal cord and isolates with the surrounding environment, which could avoid a scar effect on the nerve regeneration and provide three-dimensional space for the seed cell growth, and at last we hope to provide a new treatment for spinal cord injury with the tissue engineering technique.

Is it time to put traditional cold therapy in rehabilitation of soft-tissue injuries out to pasture?

Cold therapy has been used regularly as an immediate treatment to induce analgesia following acute soft-tissue injuries,however,a prolonged ice application has proved to delay the start of the healing and lengthen the recovery process.Hyperbaric gaseous cryotherapy,also known as neurocryostimulation,has shown the ability to overcome most of the limitations of traditional cold therapy,and meanwhile promotes the analgesic and anti-inflammatory effects well,but the current existing studies have shown conflicting results on its effects.Traditional cold therapy still has beneficial effect especially when injuries are severe and swelling is the limiting factor for recovery after soft-tissue injuries,and therefore no need to be entirely put out to pasture in the rehabilitation practice.Strong randomized controlled trials with good methodological quality are still needed in the future to evaluate the effects of different cryotherapy modalities.

Repairing peripheral nerve injury using tissue engineering techniques

Each year approximately 360,000 people in the United States suffer a peripheral nerve injury （PNI）, which is a leading source of lifelong disability （Kelsey et al., 1997; Noble et al., 1998）. The most frequent cause of PNIs is motor vehicle accidents, while gunshot wounds, stabbings, and birth trauma are also common factors. Patients suffering from disabilities as a result of their PNIs are also burdensome to the healthcare system, with aver- age hospital stays of 28 days each year （Kelsey et al., 1997; Noble et al., 1998）.

Advantages of Ilizarov External Fixation in an Elderly Patient with Pilon Fracture with Severe Soft Tissue Injury and Severe Osteoporosis: A Case Report

Introduction: Pilon fracture in elderly individuals is characterized by senile skin atrophy, poor dermal extensibility, and thin subcutaneous tissue. The use of bulky internal fixation material can thus cause the swelling that accompanies the fracture to induce secondary injury to skin tissue. In addition, initiation of postoperative weight-bearing is delayed due to bone fragility and difficulties with partial weight-bearing, causing a tendency toward prolonged hospitalization. Mean duration of hospitalization after pilon fracture for elderly patients in our department was 79.2 days. Case Presentation: An 80-year-old woman with pilon fracture with soft tissue injury and severe osteoporosis was transferred to our department. The fracture was treated using Ilizarov external fixation. Fourteen days postoperatively, walking with full weight-bearing was permitted. The hospital stay was 28 days. The external fixator of the ankle was removed 87 days postoperatively, at which time the patient was anatomically and functionally recovered and able to walk unaided. Conclusion: Ilizarov external fixation may represent a useful option in elderly patients with pilon fracture showing severe soft tissue injury and severe osteoporosis. The present case provides evidence that this procdure can be successfully applied to the management of such pilon fractures in elderly patients.

Chitosan hydrogel encapsulated with LL-37 peptide promotes deep tissue injury healing in a mouse model

Background:LL-37 peptide is a member of the human cathelicidin family,and has been shown to promote the healing of pressure ulcers.However,the low stability of this peptide within the wound environment limits its clinical use.Chitosan(CS)hydrogel is commonly used as a base material for wound dressing material.Methods:CS hydrogel(2.5%w/v)was encapsulated with LL-37.Cytotoxicity of the product was examined in cultured NIH3 T3 fibroblasts.Effects on immune response was examined by measuring tumor necrosis factor-α(TNF-α)release from RAW 264.7 macrophages upon exposure to lipopolysaccharides.Antibacterial activity was assessed using Staphylococcus aureus.Potential effect on pressure ulcers was examined using a mouse model.Briefly,adult male C57 BL/6 mice were subjected to skin pressure using magnets under a 12/12 h schedule for 21 days.Mice were randomized to receive naked LL-37(20μg),chitosan gel containing 20μg LL-37(LL-37/CS hydrogel)or hydrogel alone under the ulcer bed(n=6).A group of mice receiving no intervention was also included as a control.Results:LL-37/CS hydrogel did not affect NIH3 T3 cell viability.At a concentration of 1–5μg/ml,LL-37/CS inhibited TNF-αrelease from macrophage.At 5μg/ml,LL-37/CS inhibited the growth of Staphylococcus aureus.The area of the pressure ulcers was significantly lower in mice receiving LL-37/CS hydrogel in comparison to all other 3 groups on days 11(84.24%±0.25%),13(56.22%±3.91%)and 15(48.12%±0.28%).Histological examination on days 15 and 21 showed increased epithelial thickness and density of newly-formed capillary with naked LL-37 and more so with LL-37/CS.The expression of key macromolecules in the process of angiogenesis(i.e.,hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor-A(VEGF-A))in wound tissue was increased at both the mRNA and protein levels.Conclusion:Chitosan hydrogel encapsulated with LL-37 is biocompatible and could promote the healing of pressure ulcers.

Reducing intrathecal pressure after traumatic spinal cord injury: a potential clinical target to promote tissue survival

Spinal cord injury （SCI） is an unexpected event that is both devastating and debilitating, resulting in not just motor and sensory loss, but also autonomic dysfunction of the bladder, bowel and sexual organs. Currently, there are no treatments available to improve outcome follow- ing SCI, leaving individuals with permanent and lifelong physical disability. Worldwide it is estimated that more than 500,000 people sustain a SCI each year, with average lifetime cost of paraplegia and quadriplegia estimated at $5 million and $9.5 million respectively. We therefore urgently need effective therapies to improve quality of life following SCI, and this requires a greater understanding of how cell and axonal injury develops after the traumatic event.

A New Model to Study Healing of a Complex Femur Fracture with Concurrent Soft Tissue Injury in Sheep

High energy bone fractures resulting from impact trauma are often accompanied by subcutaneous soft tissue injuries, even if the skin remains intact. There is evidence that such closed soft tissue injuries affect the healing of bone fractures, and vice versa. Despite this knowledge, most impact trauma studies in animals have focussed on bone fractures or soft tissue trauma in isolation. However, given the simultaneous impact on both tissues a better understanding of the interaction between these two injuries is necessary to optimise clinical treatment. The aim of this study was therefore to develop a new experimental model and characterise, for the first time, the healing of a complex fracture with concurrent closed soft tissue trauma in sheep. A pendulum impact device was designed to deliver a defined and standardised impact to the distal thigh of sheep, causing a reproducible contusion injury to the subcutaneous soft tissues. In a subsequent procedure, a reproducible femoral butterfly fracture (AO C3-type) was created at the sheep’s femur, which was initially stabilised for 5 days by an external fixator construct to allow for soft tissue swelling to recede, and ultimately in a bridging construct using locking plates. The combined injuries were applied to twelve sheep and the healing observed for four or eight weeks (six animals per group) until sacrifice. The pendulum impact led to a moderate to severe circumferential soft tissue injury with significant bruising, haematomas and partial muscle disruptions. Posttraumatic measurements showed elevated intra-compartmental pressure and circulatory tissue breakdown markers, with recovery to normal, pre-injury values within four days. Clinically, no neurovascular deficiencies were observed. Bi-weekly radiological analysis of the healing fractures showed progressive callus healing over time, with the average number of callus bridges increasing from 0.4 at two weeks to 4.2 at eight weeks. Biomechanical testing after sacrifice showed in- creasing torsional stiffness between four and eight weeks healing time from 10% to 100%, and increasing ultimate torsional strength from 10% to 64% (relative to the contralateral control limb). Our results demonstrate the robust healing of a complex femur fracture in the presence of a severe soft tissue contusion injury in sheep and demonstrate the establishment of a clinically relevant experimental model, for research aimed at improving the treatment of bone fractures accompanied by closed soft tissue injuries.

Targeted tissue engineering:hydrogels with linear capillary channels for axonal regeneration after spinal cord injury

Spinal cord injury（SCI）frequently results in the permanent loss of function below the level of injury due to the failure of axonal regeneration in the adult mammalian central nervous system（CNS）.The limited intrinsic growth capacity of adult neurons,a lack of growth-promoting factors and the multifactorial inhibitory microenvironment around the lesion site contribute to the lack of axonalregeneration. Strategies such as transplantation of cells,

Advances in spinal cord injury:insights from non-human primates

Spinal cord injury results in significant sensorimotor deficits,currently,there is no curative treatment for the symptoms induced by spinal cord injury.Basic and pre-clinical research on spinal cord injury relies on the development and characterization of appropriate animal models.These models should replicate the symptoms observed in human,allowing for the exploration of functional deficits and investigation into various aspects of physiopathology of spinal cord injury.Non-human primates,due to their close phylogenetic association with humans,share more neuroanatomical,genetic,and physiological similarities with humans than rodents.Therefore,the responses to spinal cord injury in nonhuman primates most likely resemble the responses to traumatism in humans.In this review,we will discuss nonhuman primate models of spinal cord injury,focusing on in vivo assessments,including behavioral tests,magnetic resonance imaging,and electrical activity recordings,as well as ex vivo histological analyses.Additionally,we will present therapeutic strategies developed in non-human primates and discuss the unique specificities of non-human primate models of spinal cord injury.

Advances in extracellular vesicle-based combination therapies for spinal cord injury

Spinal cord injury is a severe insult to the central nervous system that causes persisting neurological deficits.The currently available treatments involve surgical,medical,and rehabilitative strategies.However,none of these techniques can markedly reverse neurological deficits.Recently,extracellular vesicles from various cell sources have been applied to different models of spinal cord injury,thereby generating new cell-free therapies for the treatment of spinal cord injury.However,the use of extracellular vesicles alone is still associated with some notable shortcomings,such as their uncertainty in targeting damaged spinal cord tissues and inability to provide structural support to damaged axons.Therefore,this paper reviews the latest combined strategies for the use of extracellular vesicle-based technology for spinal cord injury,including the combination of extracellular vesicles with nanoparticles,exogenous drugs and/or biological scaffold materials,which facilitate the targeting ability of extracellular vesicles and the combinatorial effects with extracellular vesicles.We also highlight issues relating to the clinical transformation of these extracellular vesicle-based combination strategies for the treatment of spinal cord injury.

Tissue engineering for the repair of peripheral nerve injury

Peripheral nerve injury is a common clinical problem and affects the quality of life of patients. Traditional restoration methods are not satisfactory. Researchers increasingly focus on the field of tissue engineering. The three key points in establishing a tissue engineering material are the biological scaffold material, the seed cells and various growth factors. Understanding the type of nerve injury, the construction of scaffold and the process of repair are necessary to solve peripheral nerve injury and promote its regeneration. This review describes the categories of peripheral nerve injury, fundamental research of peripheral nervous tissue engineering and clinical research on peripheral nerve scaffold material, and paves a way for related research and the use of conduits in clinical practice.

EIGHTY CASES of SOFT TISSUE INJURY TREATED WITH LOCAL CONTRALATERAL NEEDLING METHOD

Contralateral needling method is a common and effective one of acupuncture analgesicmethods.It is used to treat a local pain in the limbs and trunk with a short course of disease,pain and swelling will disappear or be relieved immediately for one or two treatments.Really,there isan effect of relieving pain as soon as the needle is inserted.When the local contralateral needlingmethod is used,the key to improving the effectiveness lies in determining the pain position and its correspondingpoint correctly,the more correct the pain position and its corresponding point are determined,the better the effectiveness will be.In this group,80 cases soft tissue injury were treatedwith local contralateral needling method,the cure rate was 82％,and the total effective rate was100％.

Role of endogenous Schwann cells in tissue repair after spinal cord injury

Schwann cells are glial cells of peripheral nervous system, responsible for axonal myelination and ensheathing, as well as tissue repair following a peripheral nervous system injury. They are one of several cell types that are widely studied and most commonly used for cell transplantation to treat spinal cord injury, due to their intrinsic characteristics including the ability to secrete a variety of neurotrophic factors. This mini review summarizes the recent findings of endogenous Schwann cells after spinal cord injury and discusses their role in tissue repair and axonal regeneration. After spinal cord injury, numerous endogenous Schwann cells migrate into the lesion site from the nerve roots, involving in the construction of newly formed repaired tissue and axonal myelination. These invading Schwann cells also can move a long distance away from the injury site both rostrally and caudally. In addition, Schwann cells can be induced to migrate by minimal insults （such as scar ablation） within the spinal cord and integrate with astrocytes under certain circumstances. More importantly, the host Schwann cells can be induced to migrate into spinal cord by transplantation of different cell types, such as exogenous Schwann cells, olfactory ensheathing cells, and bone marrow-derived stromal stem cells. Migration of endogenous Schwann cells following spinal cord injury is a common natural phenomenon found both in animal and human, and the myelination by Schwann cells has been examined effective in signal conduction electrophysiologically. Therefore, if the inherent properties of endogenous Schwann cells could be developed and utilized, it would offer a new avenue for the restoration of injured spinal cord.

Effects of regenerated tissue extracts after liver injury on the proliferation,differentiation,migration and invasion of SK-HEP1 cells

Objective: To study the effects of regenerated tissue extracts after liver injury on the proliferation, differentiation, migration and invasion of SK-HEP1 cells. Methods: Regenerated tissue extracts after liver injury were used to induce SK-HEP1 cells after enrichment, their effects on the proliferation, differentiation, migration and invasion of SK-HEPI cells were observed through in vitro cell culture, MTT, flow cytometry and transwell assays. Results:In response to the action of regenerated tissue extracts after liver injury, SK-HEP1 cells were blocked in G_0/G_1 phase, their growth rate was distinctly reduced. The number of SK-HEP1^(-fj)colonies decreased. The migration ability of SK-HEPI cells showed a decreased trend on day7 and day 11 after induction. SK-HEPl's invasion ability clearly decreased on days 7 and11 after induction, especially on day 7. Conclusions: To a certain extent, regenerated tissue extracts after liver injury can inhibit the proliferation, differentiation, migration and invasion of hepatoma cells, showing an important potential of being a differentiating agent for the treatment of liver cancer.