Macrophages undergo dynamic transitions between M1 and M2 states,exerting profound influences on both inflammatory and regenerative processes.The biocompatible and wound-healing properties of decellularized amniotic m...Macrophages undergo dynamic transitions between M1 and M2 states,exerting profound influences on both inflammatory and regenerative processes.The biocompatible and wound-healing properties of decellularized amniotic membrane(d AM)make it a subject of exploration for its potential impact on the anti-inflammatory response of macrophages.Experimental findings unequivocally demonstrate that d AM promotes anti-inflammatory M2 polarization of macrophage,with its cytokine-rich content posited as a potential mediator.The application of RNA sequencing unveils differential gene expression,implicating the hypoxia inducible factor-1α(HIF-1α)signaling pathway in this intricate interplay.Subsequent investigation further demonstrates that d AM facilitates anti-inflammatory M2 polarization of macrophage through the upregulation of epidermal growth factor(EGF),which,in turn,activates the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway and stabilizes HIF-1α.This cascade results in a noteworthy augmentation of anti-inflammatory gene expression.This study significantly contributes to advancing our comprehension of d AM's immunomodulatory role in tissue repair,thereby suggesting promising therapeutic potential.展开更多
Literature concerning procoagulant activity of the amniotic fluid and pathomechanism of amniotic fluid embolism (AFE) was surveyed and a new concept of its pathogenesis, called the integrated concept of AFE, was prese...Literature concerning procoagulant activity of the amniotic fluid and pathomechanism of amniotic fluid embolism (AFE) was surveyed and a new concept of its pathogenesis, called the integrated concept of AFE, was presented. According to this concept, two components of the amniotic fluid are involved: (i) apoptosis-affected amniotic cells showing a special role in the initiation of disseminated intravascular coagulation (DIC) and (ii) leukotrienes (formerly called slow-reacting substances), inducing bronchial and pulmonary vascular smooth muscle contraction. Although each of these components initiates a different pathogenic pathway, they both lead to the formation of a mechanical barrier on blood flow through the lungs (amniotic debris + microemboli) and/or functional barrier (pulmonary vasoconstriction). An old dilemma, concerning indications for heparin therapy in AFE was recalled in the light of the new concept.展开更多
The main goal of the study was to identify a novel source of human multipotent cells, overcoming ethical issues involved in embryonic stem cell research and the limited availability of most adult stem cells. Amniotic ...The main goal of the study was to identify a novel source of human multipotent cells, overcoming ethical issues involved in embryonic stem cell research and the limited availability of most adult stem cells. Amniotic fluid cells （AFCs） are routinely obtained for prenatal diagnosis and can be expanded in vitro; nevertheless current knowledge about their origin and properties is limited. Twenty samples of AFCs were exposed in culture to adipogenic, osteogenic, neurogenic and myogenic media. Differentiation was evaluated using immunocytochemistry, RT-PCR and Western blotting. Before treatments, AFCs showed heterogeneous morphologies. They were negative for MyoD, Myf-5, MRF4, Myogenin and Desmin but positive for osteocalcin, PPARgamma2, GAP43, NSE, Nestin, MAP2, GFAP and beta tubulin III by RT-PCR. The cells expressed Oct-4, Rex-1 and Runx-1, which characterize the undifferentiated stem cell state. By immunocytochemistry they expressed neural-glial proteins, mesenchymal and epithelial markers. After culture, AFCs differentiated into adipocytes and osteoblasts when the predominant cellular component was fibroblastic. Early and late neuronal antigens were still present after 2 week culture in neural specific media even if no neuronal morphologies were detectable. Our results provide evidence that human amniotic fluid contains progenitor cells with multi-lineage potential showing stem and tissue-specific gene/protein presence for several lineages.展开更多
The transplantation of embryonic stem cells can effectively improve the creeping strength of nerves near an injury site in animals. Amniotic epithelial cells have similar biological properties as em-bryonic stem cells...The transplantation of embryonic stem cells can effectively improve the creeping strength of nerves near an injury site in animals. Amniotic epithelial cells have similar biological properties as em-bryonic stem cells; therefore, we hypothesized that transplantation of amniotic epithelial cells can repair peripheral nerve injury and recover the creeping strength of the brachial plexus nerve. In the present study, a brachial plexus injury model was established in rabbits using the C6root avulsion method. A suspension of human amniotic epithelial cells was repeatedly injected over an area 4.0 mm lateral to the cephal and caudal ends of the C6 brachial plexus injury site （1 × 106 cells/mL, 3μL/injection, 25 injections） immediately after the injury. The results showed that the decrease in stress and increase in strain at 7,200 seconds in the injured rabbit C6 brachial plexus nerve were mitigated by the cell transplantation, restoring the viscoelastic stress relaxation and creep properties of the brachial plexus nerve. The forepaw functions were also signiifcantly improved at 26 weeks after injury. These data indicate that transplantation of human amniotic epithelial cells can effec-tively restore the mechanical properties of the brachial plexus nerve after injury in rabbits and that viscoelasticity may be an important index for the evaluation of brachial plexus injury in animals.展开更多
AIM: To establish an untransfected human corneal epithelial (HCEP) cell line and characterize its biocompatibility with denuded amniotic membrane (dAM). METHODS: The torn HCEP pieces were primarily cultured in DMEM/F1...AIM: To establish an untransfected human corneal epithelial (HCEP) cell line and characterize its biocompatibility with denuded amniotic membrane (dAM). METHODS: The torn HCEP pieces were primarily cultured in DMEM/F12 media (pH 7.2) supplemented with 20% fetal bovine serum and other necessary factors, yielding an HCEP cell line which was its growth performance, chromosome morphology, tumorigenicity and expression of marker proteins analyzed. In addition, the biocompatibility of HCEP cells with dAM was evaluated through histological and immunocytochemistry analyses and with light, electron and slit-lamp microscopies. RESULTS: HCEP cells proliferated to confluence in 3 weeks, which have been subcultured to passage 160. A continuous untransfected HCEP cell line, designated as utHCEPC01, was established with a population doubling time of 45.42 hours as was determined at passage 100. The cells retained HCEP cell properties as were approved by chromosomal morphology and the expression of keratin 3. They, with no tumorigenicity, formed a multilayer epithelium-like structure on dAMs through proliferation and differentiation during air-liquid interface culture, maintained expression of marker proteins including keratin 3 and integrin p 1 and attached tightly to dAMs. The reconstructed HCEP was highly transparent and morphologically and structurally similar to the original. CONCLUSION: An untransfected and non-tumorigenic HCEP cell line was established in this study. The cells maintained expression of marker proteins. The cell line was biocompatible with dAM. It holds the potential of being used for in vitro reconstruction of tissue-engineered HCEP, promising for the treatment of diseases caused by corneal epithelial disorders.展开更多
In this study, we loaded human umbilical cord mesenchymal stem cells onto human amniotic membrane with epithelial cells to prepare nerve conduits, i.e., a relatively closed nerve regeneration chamber. After neurolysis...In this study, we loaded human umbilical cord mesenchymal stem cells onto human amniotic membrane with epithelial cells to prepare nerve conduits, i.e., a relatively closed nerve regeneration chamber. After neurolysis, the injured radial nerve was enwrapped with the prepared nerve conduit, which was fixed to the epineurium by sutures, with the cell on the inner surface of the conduit. Simultaneously, a 1.0 mL aliquot of human umbilical cord mesenchymal stem cell suspension was injected into the distal and proximal ends of the injured radial nerve with 1.0 cm intervals. A total of 1.75 x 107 cells were seeded on the amniotic membrane. In the control group, patients received only neurolysis. At 12 weeks after cell transplantation, more than 80% of patients exhibited obvious improvements in muscular strength, and touch and pain sensations. In contrast, these improvements were observed only in 55-65% of control patients. At 8 and 12 weeks, muscular electrophysiological function in the region dominated by the injured radial nerve was significantly better in the transplantation group than the control group. After cell transplantation, no immunological rejections were observed. These findings suggest that human umbilical cord mesenchymal stem cell-loaded amniotic membrane can be used for the repair of radial nerve injury.展开更多
Survival and differentiation of transplanted cells is closely related to the local microenvironment.The present study cultured human amniotic epithelial cells（HAECs） in a simulated microenvironment in vitro comprisi...Survival and differentiation of transplanted cells is closely related to the local microenvironment.The present study cultured human amniotic epithelial cells（HAECs） in a simulated microenvironment in vitro comprising RPMI 1640 culture medium and the solution extracted from injured brain tissues.Some HAECs were round,triangular in form or irregularly shaped,with extended neuron-like processes;some of the processes were interconnected,representing neuron-like morphology and some HAECs were microtubule-associated protein 2-positive.HAECs survived for at least 4 weeks following transplantation into the center and edges of the trauma focus with traumatic brain injury,and were microtubule-associated protein 2-positive.Moreover,the motor function of rat hind limbs was significantly improved.展开更多
Treatment and functional reconstruction after central nervous system injury is a major medical and social challenge. An increasing number of researchers are attempting to use neural stem cells combined with artificial...Treatment and functional reconstruction after central nervous system injury is a major medical and social challenge. An increasing number of researchers are attempting to use neural stem cells combined with artificial scaffold materials, such as fibroin, for nerve repair. However, such approaches are challenged by ethical and practical issues. Amniotic tissue, a clinical waste product, is abundant, and amniotic epithe- lial cells are pluripotent, have low immunogenicity, and are not the subject of ethical debate. We hypothesized that amniotic epithelial cells combined with silk fibroin scaffolds would be conducive to the repair of spinal cord injury. To test this, we isolated and cultured amniotic epithelial cells, and constructed complexes of these cells and silk fibroin scaffolds. Implantation of the cell-scaffold complex into a rat model of spinal cord injury resulted in a smaller glial scar in the damaged cord tissue than in model rats that received a blank scaffold, or amniotic epithelial cells alone. In addition to a milder local immunological reaction, the rats showed less inflammatory cell infiltration at the trans- plant site, milder host-versus-graft reaction, and a marked improvement in motor function. These findings confirm that the transplantation of amniotic epithelial ceils combined with silk fibroin scaffold can promote the repair of spinal cord injury. Silk fibroin scaffold can provide a good nerve regeneration microenvironment for amniotic epithelial cells.展开更多
Objective To establish the method of isolation, purification, and identification of human amniotic mesenchymal stem cells (hAMSCs). Methods hAMSCs were isolated from human amniotic membrane by trypsin-collagenase dige...Objective To establish the method of isolation, purification, and identification of human amniotic mesenchymal stem cells (hAMSCs). Methods hAMSCs were isolated from human amniotic membrane by trypsin-collagenase digestion, and cultured in Dulbecco's modified Eagle's medinm/F12 medium supplemented with 10% fetal bovine serum. Phenotypic characteristics of these cells were analyzed by means of immunocytochemistry and flow cytometry. Results The cells successfully isolated from human amniotic membrane expressed representative mesenchymal cell surface markers CD44, CD90, and vimentin, but not CD45. Conclusions This study establishes a potential method for isolation of hAMSCs from human amnion, in vitro culture, and identification. The isolated cells show phenotypic characteristics of mesenchymal stem cells.展开更多
AIM:To reconstruct the lamellar cornea using human amniotic epithelial(HAE) cells and rabbit cornea stroma in vitro using tissue engineering technology.·METHODS:Human amnia taken from uncomplicated caesarean sect...AIM:To reconstruct the lamellar cornea using human amniotic epithelial(HAE) cells and rabbit cornea stroma in vitro using tissue engineering technology.·METHODS:Human amnia taken from uncomplicated caesarean sections were digested by collagenase to obtain HAE cells,and the cells were cultured to proliferate.Rabbit corneal epithelial cells were removed by n-heptanol to make lamellar matrix sheets.The second passage of HAE cells were cultured on the corneal stroma sheets for 1 or 2 days,then transferred to an air-liquid interface environment to culture for 2weeks.Tissue engineered lamellar cornea(TELC)morphology was observed by Hematoxylin-eosin(HE)staining;its ultrastructure was observed by transmission electron microscopy(TEM) and scanning electron microscopy(SEM);corneal epithelial cell-specific keratin3 and keratin 12 were detected with immunofluorescence microscopy.·RESULTS:HAE cells grew on the rabbit corneal stroma,forming a monolayer after 1-2 days.About 4-5 layers of epithelial cells developed after 2 weeks of air-liquid interface cultivation,a result similar to normal corneal epithelium.Rabbit corneal stromal cells were significantly reduced after one week,then almost completely disappeared after 2 weeks.TEM showed desmosomes between the epithelial cells;hemidesmosomes formed between the epithelial cells and the basement membrane.SEM revealed that the HAE cells which grew on the lamellar cornea had abundant microvilli.The tissue-engineered cornea expressed keratin 3 and keratin 12,as detected by immunofluorescence assay.·CONCLUSION:Functional tissue-engineered lamellar corneal grafts can be constructed in vitro using HAE cells and rabbit corneal stroma.展开更多
A 10-mm long sciatic nerve injury model was established in fresh normal Chinese patient cadavers. Amniotic membrane was harvested from healthy maternal placentas and was prepared into multilayered,coiled,tubular speci...A 10-mm long sciatic nerve injury model was established in fresh normal Chinese patient cadavers. Amniotic membrane was harvested from healthy maternal placentas and was prepared into multilayered,coiled,tubular specimens.Sciatic nerve injury models were respectively anastomosed using the autologous cadaveric sciatic nerve and human amniotic membrane.Tensile test results showed that maximal loading,maximal displacement,maximal stress,and maximal strain of sciatic nerve injury models anastomosed with human amniotic membrane were greater than those in the autologous nerve anastomosis group.The strain-stress curves of the human amniotic membrane and sciatic nerves indicated exponential change at the first phase,which became elastic deformation curves at the second and third phases,and displayed plastic deformation curves at the fourth phase,at which point the specimens lost their bearing capacity.Experimental findings suggested that human amniotic membranes and autologous sciatic nerves exhibit similar stress-strain curves, good elastic properties,and certain strain and stress capabilities in anastomosis of the injured sciatic nerve.展开更多
AIM： To compare long-term outcome of primary and recurrent pterygium surgery with three different techniques： combined conjunctival autograft and overlay amniotic membrane transplantation （CAT with AMT）, conjuncti...AIM： To compare long-term outcome of primary and recurrent pterygium surgery with three different techniques： combined conjunctival autograft and overlay amniotic membrane transplantation （CAT with AMT）, conjunctival autograft transplantation （CAT） alone and amniotic membrane transplantation （AMT） alone. METHODS： In this retrospective study, 142 eyes of 142 pterygium patients （104 primary, 38 recurrent）who underwent CAT （group A）, AMT （group B） or CAT with AMT （group C） respectively following surgical excision were reviewed and compared based on the recurrences and post-operative complications. RESULTS： The number of recurrence post-surgery were 17 （9 from primary, 8 from recurrent; the same description below）, 18 （10, 8） and 2 （1, 1） in groups A, B, and C respectively; dry eyes were 22 （16, 6）, 27 （18, 9） and 7 （3, 4）; conjunctival inflammations were 30 （17, 13）, 27 （16, 11） and 11 （6, 5）. Patients in group C （either pdmary or recurrent or both） mainly showed significantly better results than those in group A or B （P〈0.05） regarding above-mentioned clinical effects. CONCLUSION： Combined CAT and overly AMT have significantly lower rates of recurrence and postoperative complications for primary and recurrent pterygium surgery than CAT or AMT alone.展开更多
In orthopedics, tissue engineering approach using stem cells is a valid line of treatment for patients with bone defects. In this context, mesenchymal stromal cells of various origins have been extensively studied and...In orthopedics, tissue engineering approach using stem cells is a valid line of treatment for patients with bone defects. In this context, mesenchymal stromal cells of various origins have been extensively studied and continue to be a matter of debate. Although mesenchymal stromal cells from bone marrow are already clinically applied, recent evidence suggests that one may use mesenchymal stromal cells from extra-embryonic tissues, such as amniotic fluid, as an innovative andadvantageous resource for bone regeneration. The use of cells from amniotic fluid does not raise ethical problems and provides a sufficient number of cells without invasive procedures. Furthermore, they do not develop into teratomas when transplanted, a consequence observed with pluripotent stem cells. In addition, their multipotent differentiation ability, low immunogenicity, and anti-inflammatory properties make them ideal candidates for bone regenerative medicine. We here present an overview of the features of amniotic fluid mesenchymal stromal cells and their potential in the osteogenic differentiation process. We have examined the papers actually available on this regard, with particular interest in the strategies applied to improve in vitro osteogenesis. Importantly, a detailed understanding of the behavior of amniotic fluid mesenchymal stromal cells and their osteogenic ability is desirable considering a feasible application in bone regenerative medicine.展开更多
Objective To compare the characterization and myocardial differentiation capacity of arnniotic fluid-derived mesenchymal stromal cells （AF MSCs） and umbilical cord Wharton＇s Jelly-derived mesenchymal stromal cells ...Objective To compare the characterization and myocardial differentiation capacity of arnniotic fluid-derived mesenchymal stromal cells （AF MSCs） and umbilical cord Wharton＇s Jelly-derived mesenchymal stromal cells （WJ MSCs）. Methods The human AF MSCs were cultured from amniotic fluid samples obtained by amniocentesis. The umbilical cord WJ MSCs were obtained from Wharton＇s Jelly of umbilical cords of infants delivered full-term by normal labor. The morphology, growth curves, and analyses by flow cytometry of cell surface markers were compared between the two types of cells. Myocardial genes （GATA-4, c-TnT, a-actin, and Cx43） were detected by real-time PCR and the corresponding protein expressions were detected by Western blot analysis after myocardial induced in AF MSCs and WJ MSCs. Results Our findings revealed AF MSCs and WJ MSCs shared similar morphological characteristics of the fibroblastoid shape. The AF MSCs were easily obtained than the WJ MSCs and had a shorter time to reach adherence of 2.7 ± 1.6 days to WJ MSCs of 6.5 ± 1.8 days. The growth curves by MTT cytotoxic assay showed the AF MSCs had a similar proliferative capacity at passage 5 and passage 10. However, the proliferative capacities ofWJ MSCs were decreased at 5 passage relative to 10 passage. Both AF stem cells and WJ stem cells had the characteristics of mesenchymal stromal cells with some characteristics of embryonic stem cells. They express CD29 and CD105, but not CD34. They were positive for Class I major histocompatibility （MHC I） antigens （HLA-ABC）, and were negative, or mildly positive, for MHC Class II （HLA-DR） antigen. Oct-4 was positive in all the two cells types. Both AF MSCs and WJ MSCs could differentiate along myocardium. The differentiation capacities were detected by the expression of GATA-4, c-TnT, a-actin, Cx43 after myocardial induction. Conclusions Both AF MSCs and WJ MSCs have the potential clinical application for myogenesis in cardiac regenerative therapy.展开更多
Objective： The purpose of this study was to determine the role of Ureaplasma urealyticum-derived lipidassociated membrane proteins （LAMPs） in the host innate immune system, specifically their effect on Toll-like re...Objective： The purpose of this study was to determine the role of Ureaplasma urealyticum-derived lipidassociated membrane proteins （LAMPs） in the host innate immune system, specifically their effect on Toll-like receptors （TLRs）. Methods： LAMPs were derived from U. urea/yticum strains, and human amniotic epithelial cells （HAECs） were isolated from healthy full-term placentas. Cytokine concentrations were determined by enzyme-linked immunosorbent assay （ELISA） and TLR2 mRNA by real-time PCR. Expression of TLR2 was confirmed by Western blotting and immunohistochemistry. Results： LAMPs induced HAECs to produce inflammatory cytokines interleukin （IL）-6, IL-8, and tumor necrosis factor （TNF）-α. Cytokine production was reduced after blocking TLR2 using TLR2 inhibitor （anti-hTLR2-IgA）. Conclusions： LAMPs isolated from U. urealyticum induced TLR2-dependent up-regulation of inflammatory genes and cytokines in HAECs.展开更多
BACKGROUND:Human amniotic epithelial cells(HAECs)can differentiate into neurons,astrocytes and oligodendrocytes.They biologically secrete many active neurotrophins and have the capacity to metabolize dopamine enzymes....BACKGROUND:Human amniotic epithelial cells(HAECs)can differentiate into neurons,astrocytes and oligodendrocytes.They biologically secrete many active neurotrophins and have the capacity to metabolize dopamine enzymes.These features underlie a theoretical basis for the treatment of Parkinson's disease(PD).OBJECTIVE:To investigate the survival and differentiation of transplanted HAECs in the lateral ventricle of PD model rats,and to explore its effect on circling behavior,as well as levels of dopamine(DA),the metabolite homovanillic acid,dihydroxyphenyl acetic acid,5-hydroxyindoleacetic acid,and 5-hydroxytryptamine in the striatum.DESIGN,TIME AND SETTING:A randomized,controlled,animal study was performed at the Institute of Biochemistry and Cell Biology,Shanghai Institute for Biological Sciences,Chinese Academy of Sciences,and Shanghai Celstar Institute of Biotechnology from May 2007 to December 2008.MATERIALS:HAECs were derived from the placental chorion following caesarean delivery at the Shanghai International Matemal and Child Health Hospital.6-hydroxydopamine(6-OHDA),and mouse anti-human Vimentin monoclonal antibody were purchased from Sigma,USA;mouse anti-human nestin and tyrosine hydroxylase(TH)monoclonal antibodies were purchased from Chemicon,USA.METHODS:A total of 114 healthy,adult,Sprague Dawley rats were randomly assigned to two groups:PD model[n=90,stereotactic microinjection of 2μL 6-OHDA(3.5μg/uL)into the striatum]and control(n=24,no treatment).The 51 successful PD model rats were randomly divided into 3 subgroups(n=17):HAEC,PBS,and model.The HAEC and PBS groups were respectively injected with 10μL PBS solution containing 1×10^5/mL HAECs or 10 pL PBS into the lateral ventricle.The model group was not treated.MAIN OUTCOME MEASURES:TH protein expression in the striatum was evaluated by immunohistochemistry 5 weeks after HAEC transplantation.At 10 weeks,HAEC survival in the lateral ventricle was investigated by immunofluorescent staining;differentiation of HAECs in the lateral and third ventricles was examined by TH immunohistochemistry;concentrations of DA,homovanillic acid,dihydroxyphenyl acetic acid,5-hydroxyindoleacetic acid,and 5-hydroxytryptamine in the striatum,as well as DA concentration in the cerebrospinal fluid,were measured with high-performance liquid chromatography-electrochemical detection.Circling behavior of PD model rats was consecutively observed for 10 weeks following intraperitoneal injection of amphetamine 1 week after successful model establishment.RESULTS:tn the HAEC group,the number of TH-positive cells significantly increased in the striatum,and circling behavior significantly decreased,compared with the PBS and model groups(P〈0.01).In addition,monoamine concentrations in the striatum,as well as DA concentrations in the cerebrospinal fluid,significantly increased,compared with the PBS group(P〈0.05-0.01).Moreover,a large number of nestin-,vimentin-,and TH-positive cells were observed in the lateral and third ventricles following HAEC injection.CONCLUSION:HAECs survived for 10 weeks with no overgrowth following transplantation into the lateral ventricle of PD model rats.Moreover,the cells differentiated into dopaminergic neurons,which increased DA secretion.HAEC transplantation improved cycling behavior in PD model rats.展开更多
The amniotic membrane(AM) is the inner layer of the fetal membranes and consist of 3 different layers: the epithelium, basement membrane and stroma which further consists of three contiguous but distinct layers: the i...The amniotic membrane(AM) is the inner layer of the fetal membranes and consist of 3 different layers: the epithelium, basement membrane and stroma which further consists of three contiguous but distinct layers: the inner compact layer, middle fibroblast layer and the outermost spongy layer. The AM has been shown to have anti-inflammatory, anti-fibrotic, anti-angiogenic as well as anti-microbial properties. Also because of its transparent structure, lack of immunogenicity and the ability to provide an excellent substrate for growth, migration and adhesion of epithelial corneal and conjunctival cells, it is being used increasingly for ocular surface reconstruction in a variety of ocular pathologies including corneal disorders associated with limbal stem cell deficiency, surgeries for conjunctival reconstruction, as a carrier for ex vivo expansion of limbal epithelial cells, glaucoma surgeries and sceral melts and perforations. However indiscriminate use of human AM needs to be discouraged as complications though infrequent can occur. These include risk of transmission of bacterial, viral or fungal infections to the recipient if the donors are not adequately screened for communicable diseases, if the membrane is not processed under sterile condi-tions or if storage is improper. Optimal outcomes can be achieved only with meticulous case selection. This review explores the ever expanding ophthalmological indications for the use of human AM.展开更多
Suture and autologous nerve transplantation are the primary therapeutic measures for completely severed nerves. However, imbalances in the microenvironment and adhesion of surrounding tissues can affect the quality of...Suture and autologous nerve transplantation are the primary therapeutic measures for completely severed nerves. However, imbalances in the microenvironment and adhesion of surrounding tissues can affect the quality of nerve regeneration and repair. Previous studies have shown that human amniotic membrane can promote the healing of a variety of tissues. In this study, the right common peroneal nerve underwent a 5-mm transection in rats. Epineural nerve repair was performed using 10/0 non-absorbable surgical suture. The repair site was wrapped with a two-layer amniotic membrane with α-cyanoacrylate rapid medical adhesive after suture. Hindlimb motor function was assessed using footprint analysis. Conduction velocity of the common peroneal nerve was calculated by neural electrical stimulation. The retrograde axoplasmic transport of the common peroneal nerve was observed using fast blue BB salt retrograde fluorescent staining. Hematoxylin- eosin staining was used to detect the pathological changes of the common peroneal nerve sputum. The mRNA expression of axon regeneration-related neurotrophic factors and inhibitors was measured using real-time polymerase chain reaction. The results showed that the amniotic membrane significantly improved the function of the injured nerve;the toe spread function rapidly recovered, the nerve conduction velocity was restored, and the number of fast blue BB salt particles were increased in the spinal cord. The amniotic membrane also increased the recovery rate of the tibialis anterior muscle and improved the tissue structure of the muscle. Meanwhile, mRNA expression of nerve growth factor, growth associated protein-43, collapsin response mediator protein-2, and brain-derived neurotrophic factor recovered to near-normal levels, while Lingo-1 mRNA expression decreased significantly in spinal cord tissues. mRNA expression of glial-derived neurotrophic factor did not change significantly. Changes in mRNA levels were more significant in amniotic-membrane-wrapping-treated rats compared with model and nerve sutured rats. These results demonstrate that fresh amniotic membrane wrapping can promote the functional recovery of sutured common peroneal nerve via regulation of expression levels of neurotrophic factors and inhibitors associated with axonal regeneration. The study was approved by the Committee on Animal Research and Ethics at the Affiliate Hospital of Zunyi Medical University, China (approval No. 112) on December 1, 2017.展开更多
The sciatic nerve is biological viscoelastic solid,with stress relaxation and creep characteristics.In this study,a comparative analysis of the stress relaxation and creep characteristics of the sciatic nerve was cond...The sciatic nerve is biological viscoelastic solid,with stress relaxation and creep characteristics.In this study,a comparative analysis of the stress relaxation and creep characteristics of the sciatic nerve was conducted after simulating sciatic nerve injury and anastomosing with autologous nerve or human amniotic membrane.The results demonstrate that,at the 7 200-second time point,both stress reduction and strain increase in the human amniotic membrane anastomosis group were significantly greater than in the autologous nerve anastomosis group.Our findings indicate that human amniotic membrane anastomosis for sciatic nerve injury has excellent rheological characteristics and is conducive to regeneration of the injured nerve.展开更多
AIM:To investigate the effect of amniotic membrane covering(AMC) on the healing of cornea epithelium and visual acuity for fungal keratitis after debridement.METHODS:Twenty fungal keratitis patients were divided into ...AIM:To investigate the effect of amniotic membrane covering(AMC) on the healing of cornea epithelium and visual acuity for fungal keratitis after debridement.METHODS:Twenty fungal keratitis patients were divided into two groups randomly, the AMC group and the control group, ten patients each group. Both debridement of the infected cornea tissue and standard anti-fungus drugs treatments were given to every patients, monolayer amniotic membrane were sutured to the surface of the entire cornea and bulbar conjunctiva with 10-0 nylon suture for patients in the AMC group.The diameter of the ulcer was determined with slit lamp microscope and the depth of the infiltration was determined with anterior segment optical coherence tomography. Uncorrected visual acuity(UCVA) was tested before surgery and three month after healing of the epithelial layer. The healing time of the cornea epithelium, visual acuity(VA) was compared between the two groups using t- test.RESULTS:There was no statistical difference of the diameter of the ulcer, depth of the infiltration, height of the hypopyon and VA between the two groups beforesurgery(P 】0.05). The average healing time of the AMC group was 6.89 ±2.98 d, which was statistically shorter than that of the control group(10.23±2.78d)(P 【0.05).The average UCVA of the AMC group was 0.138 ±0.083,which was statistically better than that of the control group(0.053±0.068)(P 【0.05).CONCLUSION:AMC surgery could promote healing of cornea epithelium after debridement for fungal keratitis and lead to better VA outcome.展开更多
基金supported by the National Natural Science Foundation of China(No.82302772)Guizhou Basic Research Project(No.ZK[2023]General 201)partially supported by Wuhan Kangchuang Biotechnology Co.,Ltd。
文摘Macrophages undergo dynamic transitions between M1 and M2 states,exerting profound influences on both inflammatory and regenerative processes.The biocompatible and wound-healing properties of decellularized amniotic membrane(d AM)make it a subject of exploration for its potential impact on the anti-inflammatory response of macrophages.Experimental findings unequivocally demonstrate that d AM promotes anti-inflammatory M2 polarization of macrophage,with its cytokine-rich content posited as a potential mediator.The application of RNA sequencing unveils differential gene expression,implicating the hypoxia inducible factor-1α(HIF-1α)signaling pathway in this intricate interplay.Subsequent investigation further demonstrates that d AM facilitates anti-inflammatory M2 polarization of macrophage through the upregulation of epidermal growth factor(EGF),which,in turn,activates the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway and stabilizes HIF-1α.This cascade results in a noteworthy augmentation of anti-inflammatory gene expression.This study significantly contributes to advancing our comprehension of d AM's immunomodulatory role in tissue repair,thereby suggesting promising therapeutic potential.
文摘Literature concerning procoagulant activity of the amniotic fluid and pathomechanism of amniotic fluid embolism (AFE) was surveyed and a new concept of its pathogenesis, called the integrated concept of AFE, was presented. According to this concept, two components of the amniotic fluid are involved: (i) apoptosis-affected amniotic cells showing a special role in the initiation of disseminated intravascular coagulation (DIC) and (ii) leukotrienes (formerly called slow-reacting substances), inducing bronchial and pulmonary vascular smooth muscle contraction. Although each of these components initiates a different pathogenic pathway, they both lead to the formation of a mechanical barrier on blood flow through the lungs (amniotic debris + microemboli) and/or functional barrier (pulmonary vasoconstriction). An old dilemma, concerning indications for heparin therapy in AFE was recalled in the light of the new concept.
文摘The main goal of the study was to identify a novel source of human multipotent cells, overcoming ethical issues involved in embryonic stem cell research and the limited availability of most adult stem cells. Amniotic fluid cells （AFCs） are routinely obtained for prenatal diagnosis and can be expanded in vitro; nevertheless current knowledge about their origin and properties is limited. Twenty samples of AFCs were exposed in culture to adipogenic, osteogenic, neurogenic and myogenic media. Differentiation was evaluated using immunocytochemistry, RT-PCR and Western blotting. Before treatments, AFCs showed heterogeneous morphologies. They were negative for MyoD, Myf-5, MRF4, Myogenin and Desmin but positive for osteocalcin, PPARgamma2, GAP43, NSE, Nestin, MAP2, GFAP and beta tubulin III by RT-PCR. The cells expressed Oct-4, Rex-1 and Runx-1, which characterize the undifferentiated stem cell state. By immunocytochemistry they expressed neural-glial proteins, mesenchymal and epithelial markers. After culture, AFCs differentiated into adipocytes and osteoblasts when the predominant cellular component was fibroblastic. Early and late neuronal antigens were still present after 2 week culture in neural specific media even if no neuronal morphologies were detectable. Our results provide evidence that human amniotic fluid contains progenitor cells with multi-lineage potential showing stem and tissue-specific gene/protein presence for several lineages.
基金financially supported by grants from the Science and Technology Development Plan Program of Jilin Province of China,No.20110492
文摘The transplantation of embryonic stem cells can effectively improve the creeping strength of nerves near an injury site in animals. Amniotic epithelial cells have similar biological properties as em-bryonic stem cells; therefore, we hypothesized that transplantation of amniotic epithelial cells can repair peripheral nerve injury and recover the creeping strength of the brachial plexus nerve. In the present study, a brachial plexus injury model was established in rabbits using the C6root avulsion method. A suspension of human amniotic epithelial cells was repeatedly injected over an area 4.0 mm lateral to the cephal and caudal ends of the C6 brachial plexus injury site （1 × 106 cells/mL, 3μL/injection, 25 injections） immediately after the injury. The results showed that the decrease in stress and increase in strain at 7,200 seconds in the injured rabbit C6 brachial plexus nerve were mitigated by the cell transplantation, restoring the viscoelastic stress relaxation and creep properties of the brachial plexus nerve. The forepaw functions were also signiifcantly improved at 26 weeks after injury. These data indicate that transplantation of human amniotic epithelial cells can effec-tively restore the mechanical properties of the brachial plexus nerve after injury in rabbits and that viscoelasticity may be an important index for the evaluation of brachial plexus injury in animals.
基金Supported by National High Technology Research and Development Program ("863" Program) of China(No. 2006AA02A132)
文摘AIM: To establish an untransfected human corneal epithelial (HCEP) cell line and characterize its biocompatibility with denuded amniotic membrane (dAM). METHODS: The torn HCEP pieces were primarily cultured in DMEM/F12 media (pH 7.2) supplemented with 20% fetal bovine serum and other necessary factors, yielding an HCEP cell line which was its growth performance, chromosome morphology, tumorigenicity and expression of marker proteins analyzed. In addition, the biocompatibility of HCEP cells with dAM was evaluated through histological and immunocytochemistry analyses and with light, electron and slit-lamp microscopies. RESULTS: HCEP cells proliferated to confluence in 3 weeks, which have been subcultured to passage 160. A continuous untransfected HCEP cell line, designated as utHCEPC01, was established with a population doubling time of 45.42 hours as was determined at passage 100. The cells retained HCEP cell properties as were approved by chromosomal morphology and the expression of keratin 3. They, with no tumorigenicity, formed a multilayer epithelium-like structure on dAMs through proliferation and differentiation during air-liquid interface culture, maintained expression of marker proteins including keratin 3 and integrin p 1 and attached tightly to dAMs. The reconstructed HCEP was highly transparent and morphologically and structurally similar to the original. CONCLUSION: An untransfected and non-tumorigenic HCEP cell line was established in this study. The cells maintained expression of marker proteins. The cell line was biocompatible with dAM. It holds the potential of being used for in vitro reconstruction of tissue-engineered HCEP, promising for the treatment of diseases caused by corneal epithelial disorders.
基金the Science and Technology Foundation of Shenyang in China,No.F10-217-1-00
文摘In this study, we loaded human umbilical cord mesenchymal stem cells onto human amniotic membrane with epithelial cells to prepare nerve conduits, i.e., a relatively closed nerve regeneration chamber. After neurolysis, the injured radial nerve was enwrapped with the prepared nerve conduit, which was fixed to the epineurium by sutures, with the cell on the inner surface of the conduit. Simultaneously, a 1.0 mL aliquot of human umbilical cord mesenchymal stem cell suspension was injected into the distal and proximal ends of the injured radial nerve with 1.0 cm intervals. A total of 1.75 x 107 cells were seeded on the amniotic membrane. In the control group, patients received only neurolysis. At 12 weeks after cell transplantation, more than 80% of patients exhibited obvious improvements in muscular strength, and touch and pain sensations. In contrast, these improvements were observed only in 55-65% of control patients. At 8 and 12 weeks, muscular electrophysiological function in the region dominated by the injured radial nerve was significantly better in the transplantation group than the control group. After cell transplantation, no immunological rejections were observed. These findings suggest that human umbilical cord mesenchymal stem cell-loaded amniotic membrane can be used for the repair of radial nerve injury.
基金the Grant from National Basic Research Program of China (973 Program) for Traumatic Projects, No. 2005CB522604
文摘Survival and differentiation of transplanted cells is closely related to the local microenvironment.The present study cultured human amniotic epithelial cells（HAECs） in a simulated microenvironment in vitro comprising RPMI 1640 culture medium and the solution extracted from injured brain tissues.Some HAECs were round,triangular in form or irregularly shaped,with extended neuron-like processes;some of the processes were interconnected,representing neuron-like morphology and some HAECs were microtubule-associated protein 2-positive.HAECs survived for at least 4 weeks following transplantation into the center and edges of the trauma focus with traumatic brain injury,and were microtubule-associated protein 2-positive.Moreover,the motor function of rat hind limbs was significantly improved.
基金supported by the Scientific Research Project Fund of Wuxi Municipal Health and Family Planning Commission,No.MS201402
文摘Treatment and functional reconstruction after central nervous system injury is a major medical and social challenge. An increasing number of researchers are attempting to use neural stem cells combined with artificial scaffold materials, such as fibroin, for nerve repair. However, such approaches are challenged by ethical and practical issues. Amniotic tissue, a clinical waste product, is abundant, and amniotic epithe- lial cells are pluripotent, have low immunogenicity, and are not the subject of ethical debate. We hypothesized that amniotic epithelial cells combined with silk fibroin scaffolds would be conducive to the repair of spinal cord injury. To test this, we isolated and cultured amniotic epithelial cells, and constructed complexes of these cells and silk fibroin scaffolds. Implantation of the cell-scaffold complex into a rat model of spinal cord injury resulted in a smaller glial scar in the damaged cord tissue than in model rats that received a blank scaffold, or amniotic epithelial cells alone. In addition to a milder local immunological reaction, the rats showed less inflammatory cell infiltration at the trans- plant site, milder host-versus-graft reaction, and a marked improvement in motor function. These findings confirm that the transplantation of amniotic epithelial ceils combined with silk fibroin scaffold can promote the repair of spinal cord injury. Silk fibroin scaffold can provide a good nerve regeneration microenvironment for amniotic epithelial cells.
基金Supported by Science and Technology Program of Shenyang (2009-090063, 2011-F10-222-4-00)
文摘Objective To establish the method of isolation, purification, and identification of human amniotic mesenchymal stem cells (hAMSCs). Methods hAMSCs were isolated from human amniotic membrane by trypsin-collagenase digestion, and cultured in Dulbecco's modified Eagle's medinm/F12 medium supplemented with 10% fetal bovine serum. Phenotypic characteristics of these cells were analyzed by means of immunocytochemistry and flow cytometry. Results The cells successfully isolated from human amniotic membrane expressed representative mesenchymal cell surface markers CD44, CD90, and vimentin, but not CD45. Conclusions This study establishes a potential method for isolation of hAMSCs from human amnion, in vitro culture, and identification. The isolated cells show phenotypic characteristics of mesenchymal stem cells.
基金National Natural Science Foundation of China (No.30872808No.81100637)
文摘AIM:To reconstruct the lamellar cornea using human amniotic epithelial(HAE) cells and rabbit cornea stroma in vitro using tissue engineering technology.·METHODS:Human amnia taken from uncomplicated caesarean sections were digested by collagenase to obtain HAE cells,and the cells were cultured to proliferate.Rabbit corneal epithelial cells were removed by n-heptanol to make lamellar matrix sheets.The second passage of HAE cells were cultured on the corneal stroma sheets for 1 or 2 days,then transferred to an air-liquid interface environment to culture for 2weeks.Tissue engineered lamellar cornea(TELC)morphology was observed by Hematoxylin-eosin(HE)staining;its ultrastructure was observed by transmission electron microscopy(TEM) and scanning electron microscopy(SEM);corneal epithelial cell-specific keratin3 and keratin 12 were detected with immunofluorescence microscopy.·RESULTS:HAE cells grew on the rabbit corneal stroma,forming a monolayer after 1-2 days.About 4-5 layers of epithelial cells developed after 2 weeks of air-liquid interface cultivation,a result similar to normal corneal epithelium.Rabbit corneal stromal cells were significantly reduced after one week,then almost completely disappeared after 2 weeks.TEM showed desmosomes between the epithelial cells;hemidesmosomes formed between the epithelial cells and the basement membrane.SEM revealed that the HAE cells which grew on the lamellar cornea had abundant microvilli.The tissue-engineered cornea expressed keratin 3 and keratin 12,as detected by immunofluorescence assay.·CONCLUSION:Functional tissue-engineered lamellar corneal grafts can be constructed in vitro using HAE cells and rabbit corneal stroma.
文摘A 10-mm long sciatic nerve injury model was established in fresh normal Chinese patient cadavers. Amniotic membrane was harvested from healthy maternal placentas and was prepared into multilayered,coiled,tubular specimens.Sciatic nerve injury models were respectively anastomosed using the autologous cadaveric sciatic nerve and human amniotic membrane.Tensile test results showed that maximal loading,maximal displacement,maximal stress,and maximal strain of sciatic nerve injury models anastomosed with human amniotic membrane were greater than those in the autologous nerve anastomosis group.The strain-stress curves of the human amniotic membrane and sciatic nerves indicated exponential change at the first phase,which became elastic deformation curves at the second and third phases,and displayed plastic deformation curves at the fourth phase,at which point the specimens lost their bearing capacity.Experimental findings suggested that human amniotic membranes and autologous sciatic nerves exhibit similar stress-strain curves, good elastic properties,and certain strain and stress capabilities in anastomosis of the injured sciatic nerve.
基金Supported by Natural Science Foundation of Jiangsu Province(No.BK20141346)Nanjing Science and Technology Development Plan(No.201402001)
文摘AIM： To compare long-term outcome of primary and recurrent pterygium surgery with three different techniques： combined conjunctival autograft and overlay amniotic membrane transplantation （CAT with AMT）, conjunctival autograft transplantation （CAT） alone and amniotic membrane transplantation （AMT） alone. METHODS： In this retrospective study, 142 eyes of 142 pterygium patients （104 primary, 38 recurrent）who underwent CAT （group A）, AMT （group B） or CAT with AMT （group C） respectively following surgical excision were reviewed and compared based on the recurrences and post-operative complications. RESULTS： The number of recurrence post-surgery were 17 （9 from primary, 8 from recurrent; the same description below）, 18 （10, 8） and 2 （1, 1） in groups A, B, and C respectively; dry eyes were 22 （16, 6）, 27 （18, 9） and 7 （3, 4）; conjunctival inflammations were 30 （17, 13）, 27 （16, 11） and 11 （6, 5）. Patients in group C （either pdmary or recurrent or both） mainly showed significantly better results than those in group A or B （P〈0.05） regarding above-mentioned clinical effects. CONCLUSION： Combined CAT and overly AMT have significantly lower rates of recurrence and postoperative complications for primary and recurrent pterygium surgery than CAT or AMT alone.
文摘In orthopedics, tissue engineering approach using stem cells is a valid line of treatment for patients with bone defects. In this context, mesenchymal stromal cells of various origins have been extensively studied and continue to be a matter of debate. Although mesenchymal stromal cells from bone marrow are already clinically applied, recent evidence suggests that one may use mesenchymal stromal cells from extra-embryonic tissues, such as amniotic fluid, as an innovative andadvantageous resource for bone regeneration. The use of cells from amniotic fluid does not raise ethical problems and provides a sufficient number of cells without invasive procedures. Furthermore, they do not develop into teratomas when transplanted, a consequence observed with pluripotent stem cells. In addition, their multipotent differentiation ability, low immunogenicity, and anti-inflammatory properties make them ideal candidates for bone regenerative medicine. We here present an overview of the features of amniotic fluid mesenchymal stromal cells and their potential in the osteogenic differentiation process. We have examined the papers actually available on this regard, with particular interest in the strategies applied to improve in vitro osteogenesis. Importantly, a detailed understanding of the behavior of amniotic fluid mesenchymal stromal cells and their osteogenic ability is desirable considering a feasible application in bone regenerative medicine.
文摘Objective To compare the characterization and myocardial differentiation capacity of arnniotic fluid-derived mesenchymal stromal cells （AF MSCs） and umbilical cord Wharton＇s Jelly-derived mesenchymal stromal cells （WJ MSCs）. Methods The human AF MSCs were cultured from amniotic fluid samples obtained by amniocentesis. The umbilical cord WJ MSCs were obtained from Wharton＇s Jelly of umbilical cords of infants delivered full-term by normal labor. The morphology, growth curves, and analyses by flow cytometry of cell surface markers were compared between the two types of cells. Myocardial genes （GATA-4, c-TnT, a-actin, and Cx43） were detected by real-time PCR and the corresponding protein expressions were detected by Western blot analysis after myocardial induced in AF MSCs and WJ MSCs. Results Our findings revealed AF MSCs and WJ MSCs shared similar morphological characteristics of the fibroblastoid shape. The AF MSCs were easily obtained than the WJ MSCs and had a shorter time to reach adherence of 2.7 ± 1.6 days to WJ MSCs of 6.5 ± 1.8 days. The growth curves by MTT cytotoxic assay showed the AF MSCs had a similar proliferative capacity at passage 5 and passage 10. However, the proliferative capacities ofWJ MSCs were decreased at 5 passage relative to 10 passage. Both AF stem cells and WJ stem cells had the characteristics of mesenchymal stromal cells with some characteristics of embryonic stem cells. They express CD29 and CD105, but not CD34. They were positive for Class I major histocompatibility （MHC I） antigens （HLA-ABC）, and were negative, or mildly positive, for MHC Class II （HLA-DR） antigen. Oct-4 was positive in all the two cells types. Both AF MSCs and WJ MSCs could differentiate along myocardium. The differentiation capacities were detected by the expression of GATA-4, c-TnT, a-actin, Cx43 after myocardial induction. Conclusions Both AF MSCs and WJ MSCs have the potential clinical application for myogenesis in cardiac regenerative therapy.
基金Project supported by the Zhejiang Provincial Natural Science Foundation of China(Nos.LY18H040001 and LY16H040003)the Department of Education of Zhejiang Province(No.Y201534723)the Science and Technology Development Project in Hangzhou(No.20160533B13),China
文摘Objective： The purpose of this study was to determine the role of Ureaplasma urealyticum-derived lipidassociated membrane proteins （LAMPs） in the host innate immune system, specifically their effect on Toll-like receptors （TLRs）. Methods： LAMPs were derived from U. urea/yticum strains, and human amniotic epithelial cells （HAECs） were isolated from healthy full-term placentas. Cytokine concentrations were determined by enzyme-linked immunosorbent assay （ELISA） and TLR2 mRNA by real-time PCR. Expression of TLR2 was confirmed by Western blotting and immunohistochemistry. Results： LAMPs induced HAECs to produce inflammatory cytokines interleukin （IL）-6, IL-8, and tumor necrosis factor （TNF）-α. Cytokine production was reduced after blocking TLR2 using TLR2 inhibitor （anti-hTLR2-IgA）. Conclusions： LAMPs isolated from U. urealyticum induced TLR2-dependent up-regulation of inflammatory genes and cytokines in HAECs.
基金Supported by:the Major State Basic Research Development Program of China(973 Program),No.2005CB522604the National Natural Science Foundation of China,No.30271325
文摘BACKGROUND:Human amniotic epithelial cells(HAECs)can differentiate into neurons,astrocytes and oligodendrocytes.They biologically secrete many active neurotrophins and have the capacity to metabolize dopamine enzymes.These features underlie a theoretical basis for the treatment of Parkinson's disease(PD).OBJECTIVE:To investigate the survival and differentiation of transplanted HAECs in the lateral ventricle of PD model rats,and to explore its effect on circling behavior,as well as levels of dopamine(DA),the metabolite homovanillic acid,dihydroxyphenyl acetic acid,5-hydroxyindoleacetic acid,and 5-hydroxytryptamine in the striatum.DESIGN,TIME AND SETTING:A randomized,controlled,animal study was performed at the Institute of Biochemistry and Cell Biology,Shanghai Institute for Biological Sciences,Chinese Academy of Sciences,and Shanghai Celstar Institute of Biotechnology from May 2007 to December 2008.MATERIALS:HAECs were derived from the placental chorion following caesarean delivery at the Shanghai International Matemal and Child Health Hospital.6-hydroxydopamine(6-OHDA),and mouse anti-human Vimentin monoclonal antibody were purchased from Sigma,USA;mouse anti-human nestin and tyrosine hydroxylase(TH)monoclonal antibodies were purchased from Chemicon,USA.METHODS:A total of 114 healthy,adult,Sprague Dawley rats were randomly assigned to two groups:PD model[n=90,stereotactic microinjection of 2μL 6-OHDA(3.5μg/uL)into the striatum]and control(n=24,no treatment).The 51 successful PD model rats were randomly divided into 3 subgroups(n=17):HAEC,PBS,and model.The HAEC and PBS groups were respectively injected with 10μL PBS solution containing 1×10^5/mL HAECs or 10 pL PBS into the lateral ventricle.The model group was not treated.MAIN OUTCOME MEASURES:TH protein expression in the striatum was evaluated by immunohistochemistry 5 weeks after HAEC transplantation.At 10 weeks,HAEC survival in the lateral ventricle was investigated by immunofluorescent staining;differentiation of HAECs in the lateral and third ventricles was examined by TH immunohistochemistry;concentrations of DA,homovanillic acid,dihydroxyphenyl acetic acid,5-hydroxyindoleacetic acid,and 5-hydroxytryptamine in the striatum,as well as DA concentration in the cerebrospinal fluid,were measured with high-performance liquid chromatography-electrochemical detection.Circling behavior of PD model rats was consecutively observed for 10 weeks following intraperitoneal injection of amphetamine 1 week after successful model establishment.RESULTS:tn the HAEC group,the number of TH-positive cells significantly increased in the striatum,and circling behavior significantly decreased,compared with the PBS and model groups(P〈0.01).In addition,monoamine concentrations in the striatum,as well as DA concentrations in the cerebrospinal fluid,significantly increased,compared with the PBS group(P〈0.05-0.01).Moreover,a large number of nestin-,vimentin-,and TH-positive cells were observed in the lateral and third ventricles following HAEC injection.CONCLUSION:HAECs survived for 10 weeks with no overgrowth following transplantation into the lateral ventricle of PD model rats.Moreover,the cells differentiated into dopaminergic neurons,which increased DA secretion.HAEC transplantation improved cycling behavior in PD model rats.
文摘The amniotic membrane(AM) is the inner layer of the fetal membranes and consist of 3 different layers: the epithelium, basement membrane and stroma which further consists of three contiguous but distinct layers: the inner compact layer, middle fibroblast layer and the outermost spongy layer. The AM has been shown to have anti-inflammatory, anti-fibrotic, anti-angiogenic as well as anti-microbial properties. Also because of its transparent structure, lack of immunogenicity and the ability to provide an excellent substrate for growth, migration and adhesion of epithelial corneal and conjunctival cells, it is being used increasingly for ocular surface reconstruction in a variety of ocular pathologies including corneal disorders associated with limbal stem cell deficiency, surgeries for conjunctival reconstruction, as a carrier for ex vivo expansion of limbal epithelial cells, glaucoma surgeries and sceral melts and perforations. However indiscriminate use of human AM needs to be discouraged as complications though infrequent can occur. These include risk of transmission of bacterial, viral or fungal infections to the recipient if the donors are not adequately screened for communicable diseases, if the membrane is not processed under sterile condi-tions or if storage is improper. Optimal outcomes can be achieved only with meticulous case selection. This review explores the ever expanding ophthalmological indications for the use of human AM.
基金supported by Guizhou Province Major Special Projects in Science and Technology of China,No.Qin Ke He Zhong Da Zhuan Xiang Zi [2011]6002the Special Co-operation Funds of the Science and Technology Administration in Provinces and Cities of China,No.Sheng Shi He(2014)59(both to LMY)
文摘Suture and autologous nerve transplantation are the primary therapeutic measures for completely severed nerves. However, imbalances in the microenvironment and adhesion of surrounding tissues can affect the quality of nerve regeneration and repair. Previous studies have shown that human amniotic membrane can promote the healing of a variety of tissues. In this study, the right common peroneal nerve underwent a 5-mm transection in rats. Epineural nerve repair was performed using 10/0 non-absorbable surgical suture. The repair site was wrapped with a two-layer amniotic membrane with α-cyanoacrylate rapid medical adhesive after suture. Hindlimb motor function was assessed using footprint analysis. Conduction velocity of the common peroneal nerve was calculated by neural electrical stimulation. The retrograde axoplasmic transport of the common peroneal nerve was observed using fast blue BB salt retrograde fluorescent staining. Hematoxylin- eosin staining was used to detect the pathological changes of the common peroneal nerve sputum. The mRNA expression of axon regeneration-related neurotrophic factors and inhibitors was measured using real-time polymerase chain reaction. The results showed that the amniotic membrane significantly improved the function of the injured nerve;the toe spread function rapidly recovered, the nerve conduction velocity was restored, and the number of fast blue BB salt particles were increased in the spinal cord. The amniotic membrane also increased the recovery rate of the tibialis anterior muscle and improved the tissue structure of the muscle. Meanwhile, mRNA expression of nerve growth factor, growth associated protein-43, collapsin response mediator protein-2, and brain-derived neurotrophic factor recovered to near-normal levels, while Lingo-1 mRNA expression decreased significantly in spinal cord tissues. mRNA expression of glial-derived neurotrophic factor did not change significantly. Changes in mRNA levels were more significant in amniotic-membrane-wrapping-treated rats compared with model and nerve sutured rats. These results demonstrate that fresh amniotic membrane wrapping can promote the functional recovery of sutured common peroneal nerve via regulation of expression levels of neurotrophic factors and inhibitors associated with axonal regeneration. The study was approved by the Committee on Animal Research and Ethics at the Affiliate Hospital of Zunyi Medical University, China (approval No. 112) on December 1, 2017.
基金the Key Clinical Project from the Ministry of Health in China,No.2007-353
文摘The sciatic nerve is biological viscoelastic solid,with stress relaxation and creep characteristics.In this study,a comparative analysis of the stress relaxation and creep characteristics of the sciatic nerve was conducted after simulating sciatic nerve injury and anastomosing with autologous nerve or human amniotic membrane.The results demonstrate that,at the 7 200-second time point,both stress reduction and strain increase in the human amniotic membrane anastomosis group were significantly greater than in the autologous nerve anastomosis group.Our findings indicate that human amniotic membrane anastomosis for sciatic nerve injury has excellent rheological characteristics and is conducive to regeneration of the injured nerve.
基金Supported by National Natural Science Foundation of China(No.81200661)Nature Science Foundation of Hubei Province(No.2010CDB09802)Wuhan Chenguang Plan Grant(No.201150431124)
文摘AIM:To investigate the effect of amniotic membrane covering(AMC) on the healing of cornea epithelium and visual acuity for fungal keratitis after debridement.METHODS:Twenty fungal keratitis patients were divided into two groups randomly, the AMC group and the control group, ten patients each group. Both debridement of the infected cornea tissue and standard anti-fungus drugs treatments were given to every patients, monolayer amniotic membrane were sutured to the surface of the entire cornea and bulbar conjunctiva with 10-0 nylon suture for patients in the AMC group.The diameter of the ulcer was determined with slit lamp microscope and the depth of the infiltration was determined with anterior segment optical coherence tomography. Uncorrected visual acuity(UCVA) was tested before surgery and three month after healing of the epithelial layer. The healing time of the cornea epithelium, visual acuity(VA) was compared between the two groups using t- test.RESULTS:There was no statistical difference of the diameter of the ulcer, depth of the infiltration, height of the hypopyon and VA between the two groups beforesurgery(P 】0.05). The average healing time of the AMC group was 6.89 ±2.98 d, which was statistically shorter than that of the control group(10.23±2.78d)(P 【0.05).The average UCVA of the AMC group was 0.138 ±0.083,which was statistically better than that of the control group(0.053±0.068)(P 【0.05).CONCLUSION:AMC surgery could promote healing of cornea epithelium after debridement for fungal keratitis and lead to better VA outcome.
