Ambystoma mexicanum/immunology* ; Amputation ; Animals ; Biomarkers/metabolism* ; Blastomeres/immunology* ; Cell Lineage/immunology* ; Connective Tissue Cells/immunology* ; Epithelial Cells/immunology* ; Forelimb ; Gene Expression ; High-Throughput Nucleotide Sequencing ; Humans ; Immunity ; Peroxiredoxins/immunology* ; Regeneration/immunology* ; Regenerative Medicine/methods* ; Single-Cell Analysis/methods*

γδ T cells display unique developmental, distributional, and functional patterns and can rapidly respond to various insults and contribute to diverse diseases. Different subtypes of γδ T cells are produced in the thymus prior to their migration to peripheral tissues. γδ T cells are enriched in the liver and exhibit liver-specific features. Accumulating evidence reveals that γδ T cells play important roles in liver infection, non-alcoholic fatty liver disease, autoimmune hepatitis, liver fibrosis and cirrhosis, and liver cancer and regeneration. In this study, we review the properties of hepatic γδ T cells and summarize the roles of γδ T cells in liver diseases. We believe that determining the properties and functions of γδ T cells in liver diseases enhances our understanding of the pathogenesis of liver diseases and is useful for the design of novel γδ T cell-based therapeutic regimens for liver diseases.
Animals ; Cytokines ; immunology ; Humans ; Liver Diseases ; immunology ; Liver Regeneration ; immunology ; Mice ; T-Lymphocytes, Regulatory ; immunology

Currently, the most effective treatment for end-stage liver fibrosis is liver transplantation; however, transplantation is limited by a shortage of donor organs, surgical complications, immunological rejection, and high medical costs. Recently, mesenchymal stem cell (MSC) therapy has been suggested as an effective alternate approach for the treatment of hepatic diseases. MSCs have the potential to differentiate into hepatocytes, and therapeutic value exists in their immune-modulatory properties and secretion of trophic factors, such as growth factors and cytokines. In addition, MSCs can suppress inflammatory responses, reduce hepatocyte apoptosis, increase hepatocyte regeneration, regress liver fibrosis and enhance liver functionality. Despite these advantages, issues remain; MSCs also have fibrogenic potential and the capacity to promote tumor cell growth and oncogenicity. This paper summarizes the properties of MSCs for regenerative medicine and their therapeutic mechanisms and clinical application in the treatment of liver fibrosis. We also present several outstanding risks, including their fibrogenic potential and their capacity to promote pre-existing tumor cell growth and oncogenicity.
Animals ; Cell Differentiation ; Cell Proliferation ; Hepatocytes/immunology/metabolism/pathology/*transplantation ; Humans ; Liver/immunology/metabolism/pathology/physiopathology/*surgery ; Liver Cirrhosis/diagnosis/immunology/metabolism/physiopathology/*surgery ; Liver Regeneration ; *Mesenchymal Stem Cell Transplantation/adverse effects ; *Mesenchymal Stromal Cells/immunology/metabolism/pathology ; Phenotype ; Regenerative Medicine/*methods ; Risk Factors ; Signal Transduction ; Treatment Outcome

There have been many attempts for regeneration of peripheral nerve injury. In this study, we examined the in vivo effects of non-differentiated and neuronal differentiated adipose-derived stem cells (ADSCs) in inducing the neuronal regeneration in the Sprague-Dawley (SD) rats undergoing nerve defect bridged with the PCL nanotubes. Then, we performed immunohistochemical and histopathologic examinations, as well as the electromyography, in three groups: the control group (14 sciatic nerves transplanted with the PCL nanotube scaffold), the experimental group I (14 sciatic nerves with the non-differentiated ADSCs at a density of 7x105 cells/0.1 mL) and the experimental group II (14 sciatic nerves with the neuronal differentiated ADSCs at 7x105 cells/0.1 mL). Six weeks postoperatively, the degree of the neuronal induction and that of immunoreactivity to nestin, MAP-2 and GFAP was significantly higher in the experimental group I and II as compared with the control group. In addition, the nerve conduction velocity (NCV) was significantly higher in the experimental group I and II as compared with the control group (P=0.021 and P=0.020, respectively). On the other hand, there was no significant difference in the NCV between the two experimental groups (P>0.05). Thus, our results will contribute to treating patients with peripheral nerve defects using PCL nanotubes with ADSCs.
Adipose Tissue/cytology ; Animals ; Cell Differentiation ; Electromyography ; Male ; Nanotubes ; *Nerve Regeneration ; Nerve Tissue Proteins/immunology ; Nestin/immunology ; Neural Conduction/physiology ; Peripheral Nerve Injuries/*surgery ; Phosphoprotein Phosphatases/immunology ; Polyesters/*therapeutic use ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve/injuries/surgery ; Stem Cell Transplantation/*methods ; Stem Cells/*cytology ; Tissue Engineering/methods

Mesenchymal stem cells (MSCs) are self-renewing, multipotent progenitor cells with multilineage potential to differentiate into cell types of mesodermal origin, such as adipocytes, osteocytes, and chondrocytes. In addition, MSCs can migrate to sites of inflammation and exert potent immunosuppressive and anti-inflammatory effects through interactions between lymphocytes associated with both the innate and adaptive immune system. Along with these unique therapeutic properties, their ease of accessibility and expansion suggest that use of MSCs may be a useful therapeutic approach for various disorders. In the clinical setting, MSCs are being explored in trials of various conditions, including orthopedic injuries, graft versus host disease following bone marrow transplantation, cardiovascular diseases, autoimmune diseases, and liver diseases. Furthermore, genetic modification of MSCs to overexpress antitumor genes has provided prospects for clinical use as anticancer therapy. Here, we highlight the currently reported uses of MSCs in clinical trials and discuss their efficacy as well as their limitations.
Animals ; Cell Differentiation ; Cell Lineage ; Cell Movement ; Cell Proliferation ; Gene Expression Regulation ; Humans ; *Mesenchymal Stem Cell Transplantation/adverse effects ; Mesenchymal Stromal Cells/immunology/*physiology ; *Regeneration ; Regenerative Medicine/*methods ; Treatment Outcome

OBJECTIVETo observe the change in quantity and morphology of nerve fibers in different periods in granulation tissue in full-thickness burn wound.

METHODSThe granulation tissue samples were harvested from 40 patients with full-thickness burn in our unit at 1st, 2nd, 3rd and 4th post burn week (PBW), 10 samples were obtained at each time point. Donor site tissues from 10 burn patients were used as normal control. Immunofluorescent staining technique with anti-neurofilament (NF) monoclonal antibody was employed to examine the expression of nerve fibers in granulation tissue and normal skin. The morphology of nerve fibers was observed with fluorescence microscope and laser scanning confocal microscope.

RESULTSFluorescence microscopy showed: nerve fibers were short and rare at 1 PBW, the ratio of nerve fibers positive area was (0.14 +/- 0.08)%. Nerve fibers increased slightly and were in single filament without branches, and the positive area ratio of nerve fibers (0.40 +/- 0.09)% was much lower than that of normal control [(0.62 +/- 0.12)%, P < 0.05]. Nerve fibers increased significantly and were arranged like a mesh with more branches and sproutings, and the positive area ratio of nerve fibers was (0.73 +/- 0.16)% at 3 PBW. The quantity of nerve fibers at 4 PBW was similar to that of 3 PBW, and the positive area ratio of nerve fibers was (0.66 +/- 0.13)%. Observations under LSCM: the nerve fibers were short at 1, 2 PBW; was irregular at 3 PBW, among them some were swollen and distorted, and fragmentation and vacuolation were observed. They became aggregated at 4PBW with less branches, similar to that at 3 PBW. The structures of nerve fibers in normal control were intact, without obvious pathological changes.

CONCLUSIONThe change in quantity and morphology of nerve fibers in burn wound is related to the time of granulation tissue development.

Adult ; Burns ; pathology ; Female ; Fluorescent Antibody Technique ; Granuloma ; etiology ; pathology ; Humans ; Male ; Middle Aged ; Nerve Fibers ; metabolism ; pathology ; Nerve Regeneration ; Neurofilament Proteins ; immunology ; Skin ; innervation ; Wound Healing

OBJECTIVE: To examine the biocompatibility of polylactic/glycolic acid (PLGA) and Schwann cells. METHOD: Schwann cells were harvested from rat brachial and sciatic nerves. Schwann cells were cultured with PLGA, observed by phase-contrast microscopy and electron microscopy. RESULT: Schwann cells could attach and proliferate on the surface of the PLGA. CONCLUSION The PLGA has good cellular biocompatibility. It can be used as biomaterial for tissue engineering.
Animals ; Biocompatible Materials ; Cells, Cultured ; Lactic Acid ; Nerve Regeneration ; Polyesters ; Polyglycolic Acid ; Polymers ; Rats ; Rats, Sprague-Dawley ; Schwann Cells ; cytology ; immunology

OBJECTIVENogo-A antibody IN-1 can neutralize Nogo-A, a neurite growth inhibitory protein, promoting axonal regeneration following lesions of the central nervous system (CNS) in adult rats. This study aimed to examine the effect of ventricle injection of Nogo-A antibody on neuronal regeneration in neonatal rats following hypoxic-ischemic brain damage (HIBD).

METHODSA model of neonatal HIBD was prepared by the ligation of the left common carotid artery, followed by 8% hypoxia exposure. Forty HIBD rats were randomly given a ventricle injection of 10 microL Nogo-A antibody IN-1 (IN-1 group) or 10 microL artificial cerebrospinal fluid (artificial CSF group) (n=20 each). Another 20 neonatal rats were sham-operated, without hypoxia-ischemia, and were used as the controls. The levels of Nogo-A and GAP-43 protein in the brain were measured by immunohistochemistry.

RESULTSThe number of immunohistory positive cells of Nogo-A in the brain in the IN-1 group (28.61+/-1.70) was obviously less than that in the artificial CSF (39.52 +/-1.40) and the sham-operated groups (32.78 +/- 1.87) (both P < 0.01). There were significant differences in the Nogo-A protein expression between the artificial CSF and the sham-operated groups (P < 0.01). The GAP-43 protein expression in the IN-1 group (31.14 +/- 1.88) was noticeably higher than that in the artificial CSF group (27.73 +/- 1.43 ) (P < 0.01). Both the IN-1 and the artificial CSF groups showed lower GAP-43 protein levels than the sham-operated groups (33.64 +/- 1.24) (both P < 0.01).

CONCLUSIONSNogo-A antibody can reduce the expression of Nogo-A protein in the brain and thus promote neuronal regeneration in neonatal rats following HIBD. An increased GAP-43 protein expression in the brain after Nogo-A antibody administration shows an enhanced neuronal regeneration in the neonatal rats following HIBD.

Animals ; Animals, Newborn ; Antibodies ; administration & dosage ; Brain Chemistry ; Female ; GAP-43 Protein ; analysis ; Hypoxia-Ischemia, Brain ; metabolism ; physiopathology ; therapy ; Immunohistochemistry ; Injections, Intraventricular ; Male ; Myelin Proteins ; analysis ; antagonists & inhibitors ; immunology ; Nerve Regeneration ; Nogo Proteins ; Rats ; Rats, Sprague-Dawley

BACKGROUND: The hemopoietic stem cells increase in number during the regeneration after chemotherapy or bone marrow transplantation (BMT). Although the proportion of hemopoietic stem cells and their differentiation have been studied by immunophenotyping using the flow cytometry, no substantial research efforts have been directed toward the regenerating marrow. We attempted to discover the proportions of undifferentiated stem cells, committed stem cells, B cell precursors, and myeloid precursors in the regenerating bone marrows during complete remission (CR) and after engraftment of BMT. METHODS: Bone marrow samples from 82 patients with acute leukemia in CR and from 25 patients after BMT engraftment, along with 22 control samples, were used to find the numbers of CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells in the large lymphocyte gate by flow cytometry. We cross-analyzed our results in terms of groups: CR, BMT, and initial diagnosis groups. We performed significance tests on age, relapse, chromosomal abnormalities, clinical outcomes, and initial immunophenotypes of the leukemic cells. RESULTS: The proportions of CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells are more highly distributed in acute B-lymphoblastic leukemia than the normal group and also in the CR than the BMT group. CD19+/CD34+ cells were increased in the relapse group and CD38+/ CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells were increased in the group with chromosomal abnormality. The results were irrelevant to the initial immunophenotype of the leukemic blasts. CONCLUSIONS: The increases of the markers spanned too widely to apply one specific cutoff value to analyze them. They seemed to be the results of normal regeneration, irrelevant to relapse or initial immunophenotype of leukemic blasts.
Acute Disease ; Antigens, CD19/*metabolism ; Antigens, CD34/*metabolism ; Antigens, CD38/*metabolism ; Bone Marrow/physiology ; *Bone Marrow Transplantation ; Flow Cytometry ; Follow-Up Studies ; Granulocyte Colony-Stimulating Factor/therapeutic use ; Hematopoietic Stem Cells/immunology/metabolism ; Humans ; Immunophenotyping ; Leukemia/drug therapy/*metabolism/therapy ; Regeneration ; Remission Induction

OBJECTIVETo evaluate the effects of newborn rat hepatocyte intrasplenic transplantation combined with rat augmenter of liver regeneration (rALR) injection in treating rats with acute hepatic failure.

METHODSAcute hepatic failure (AHF) was induced in rats using D-gal (1.2 g/kg). The rats were then randomly divided into 6 groups. Group I received no further treatment and served as blank controls; group II received 1 ml buffered saline once through intrasplenic injection; group III received 1 ml rALR; group IV received 2 x 10(7)/ml hepatocytes; group V received 2 x 10(7) hepatocytes suspended in 1 ml rALR (50 microg/kg) and group VI received 2 x 10(7) hepatocytes in 1 ml cyclosporine A (10 mg/kg). The rats of the various treated groups were sacrificed at day 1, 5 and at week 2 and their livers and spleens were examined histopathologically. Blood samples of the rats were also obtained to determine the levels of TNF alpha and IL-1 beta.

RESULTSThere were no significant differences in survival between group I, II and III rats. 33.3% of the group IV rats survived for 2 weeks. At week 2, the survival rate of group V rats was significantly higher than that of group IV, but there was of no statistical significant increase when compared to that of group VI rats. Hepatocytes transplanted into spleens survived for 5-7 days in the spleens of group IV and VI rats, but they survived at least 2 weeks in group V. The average serum TNF alpha level in group IV was significantly higher than that in groups V and VI on the first postoperative day, but after four days, only the difference between group IV and group VI was significant (P < 0.05). The average serum level of TNF alpha in group II was higher than that in groups IV, V and VI on the first postoperative day (P < 0.05), but there were no significant differences between those in groups IV, V and VI on the 1st and the 5th postoperative days.

CONCLUSIONNewborn rat hepatocyte intrasplenic transplantation combined with rALR is effective in treating acute hepatic function failure induced by D-gal in rats. The transplanted hepatocytes can survive for at least 2 weeks in the spleens. The rALR mixed with the hepatocytes injected into the spleens may be able to facilitate the hepatocyte regeneration, to inhibit liver cell apoptosis and to suppress the cellular immunity.

Animals ; Cell Transplantation ; methods ; Female ; Hepatocytes ; transplantation ; Liver Failure, Acute ; immunology ; surgery ; Liver Regeneration ; Male ; Proteins ; therapeutic use ; Rats ; Rats, Wistar ; Spleen ; surgery