Differentiation Of Endogenous Stem Cells Research Articles

Feather keratin-montmorillonite nanocomposite hydrogel promotes bone regeneration by stimulating the osteogenic differentiation of endogenous stem cells

Bone defects caused by bone trauma, infection, surgery, or other systemic diseases remain a severe challenge for the medical field. To address this clinical problem, different hydrogels were exploited to promote bone tissue regrowth and regeneration. Keratins are natural fibrous proteins found in wool, hair, horns, nails, and feather. Due to their unique characteristics of outstanding biocompatibility, great biodegradability, and hydrophilic, keratins have been widely applicated in different fields. In our study, the feather keratin-montmorillonite nanocomposite hydrogels that consist of keratin hydrogels serving as the scaffold support to accommodate endogenous stem cells and montmorillonite is synthesized. The introduction of montmorillonite greatly improves the osteogenic effect of the keratin hydrogels via bone morphogenetic protein 2 (BMP-2)/phosphorylated small mothers against decapentaplegic homolog 1/5/8 (p-SMAD 1/5/8)/runt-related transcription factor 2 (RUNX2) expression. Moreover, the incorporation of montmorillonite into hydrogels can improve the mechanical properties and bioactivity of the hydrogels. The morphology of feather keratin-montmorillonite nanocomposite hydrogels was shown by scanning electron microscopy (SEM) to have an interconnected porous structure. The incorporation of montmorillonite into the keratin hydrogels was confirmed by the energy dispersive spectrum (EDS). We prove that the feather keratin-montmorillonite nanocomposite hydrogels enhance the osteogenic differentiation of BMSCs. Furthermore, micro-CT and histological analysis of rat cranial bone defect demonstrated that feather keratin-montmorillonite nanocomposite hydrogels dramatically stimulated bone regeneration in vivo. Collectively, feather keratin-montmorillonite nanocomposite hydrogels can regulate BMP/SMAD signaling pathway to stimulate osteogenic differentiation of endogenous stem cells and promote bone defect healing, indicating their promising candidate in bone tissue engineering.

Japanese Laws and the Current Status of Regenerative Medicine in the Tohoku Region.

The aim of this study was to review Japanese laws regarding regenerative medicine and the current status of clinical application of regenerative medicine, to learn about the advantages and problems, and to thereby serve as a reference for measures necessary for the development of regenerative medicine. Regenerative medicine started in 1957 with the transplantation of hematopoietic stem cells, followed by the establishment of embryonic stem cells in 1981 and induced pluripotent stem cells in 2006, and continues to evolve progressively. At the same time, however, problems have emerged due to lax legal regulations, such as the use of treatments that lack scientific evidence. The Japanese government enacted two laws to regulate regenerative medicine: the Law to Ensure the Safety of Regenerative Medicine and the Amend the Pharmaceutical Affairs Law in 2013. These laws were enacted with the aim of providing safe regenerative medicine promptly and smoothly and developing many regenerative medicine products. In these laws, regenerative medicine is defined as medical treatment that restores lost functions of damaged organs and tissues with the help of cellular and tissue-based products. Nowadays, there are two major methods of regenerative medicine. One representative method involves the transplantation of devices that activates self-regenerative ability by introducing living cells into patients' body. The other method is the activation and differentiation of endogenous stem cells with cell growth and differentiation factors. The current status of regenerative medicine in the Tohoku region after the enactment of these laws is described in detail. This clarified the advantages and disadvantages associated with regenerative medicine as it is currently practiced in Japan. Development of regenerative medicine in dentistry will be advanced by learning about its clinical application in medicine.

Bacteria-responsive, Cell-recruitable, and osteoinductive nanocomposite microcarriers for intelligent bacteriostasis and accelerated tissue regeneration

Repairing alveolar bone defects with chronic periodontitis is a major clinical challenge, since the periodontitis causes the intermittent bacteria invasion to irreversibly aggravates the degeneration of alveolar bones. To deal with the challenge, a kind of novel microcarriers with excellent abilities of responding to alkaline phosphatase (ALP) to intelligently eliminate the bacteria and promoting the recruitment and osteogenic differentiation of endogenous stem cells are developed. Mesoporous silica nanocarriers that chemically fix the polyethylene glycol-modified amoxicillin (PEG-AML) through the phosphoester groups and electrostatically adsorb the aspirin (ASP) are first prepared. The nanocarriers are then dispersed in a chitosan solution to prepare the bacteria-responsive, cell-recruitable, and osteoinductive nanocomposite microcarriers. When being stimulated by ALP that are secreted by the bacteria, phosphoester groups of microcarriers are cleaved, and PEG-AML are delivered on demand to well achieve the goal of intelligent bacteriostasis. The microcarriers slowly release the ASP to promote the recruitment and osteogenic differentiation of endogenous stem cells. At 8 weeks post-implantation, the microcarriers facilitate the repair of alveolar bone defects with periodontitis. The blood vessels in the neo-generated alveolar bones grow well. Due to the appropriate degradation time and excellent antibacterial ability of microcarriers, the microcarriers exhibit a better therapeutic efficacy than the standard clinical products (Geistlich Bio-Oss®), providing a new insight into the cure of alveolar bone defects with periodontitis.

Mechanical Loading Promotes the Migration of Endogenous Stem Cells and Chondrogenic Differentiation in a Mouse Model of Osteoarthritis.

Osteoarthritis (OA) is a major health problem, characterized by progressive cartilage degeneration. Previous works have shown that mechanical loading can alleviate OA symptoms by suppressing catabolic activities. This study evaluated whether mechanical loading can enhance anabolic activities by facilitating the recruitment of stem cells for chondrogenesis. We evaluated cartilage degradation in a mouse model of OA through histology with H&E and safranin O staining. We also evaluated the migration and chondrogenic ability of stem cells using in vitro assays, including immunohistochemistry, immunofluorescence, and Western blot analysis. The result showed that the OA mice that received mechanical loading exhibited resilience to cartilage damage. Compared to the OA group, mechanical loading promoted the expression of Piezo1 and the migration of stem cells was promoted via the SDF-1/CXCR4 axis. Also, the chondrogenic differentiation was enhanced by the upregulation of SOX9, a transcription factor important for chondrogenesis. Collectively, the results revealed that mechanical loading facilitated cartilage repair by promoting the migration and chondrogenic differentiation of endogenous stem cells. This study provided new insights into the loading-driven engagement of endogenous stem cells and the enhancement of anabolic responses for the treatment of OA.

Spinal cord decellularized matrix scaffold loaded with engineered basic fibroblast growth factor-overexpressed human umbilical cord mesenchymal stromal cells promoted the recovery of spinal cord injury.

Spinal cord injury (SCI) will lead to irreversible damage of sensory and motor function of central nervous system, which seriously affects patient's quality of life. A variety of nerve engineering materials carrying various stem cells and cell growth factors had used to promote the repair of SCI, but they could not mimic the actual matric niche at spinal cord to promote cell proliferation and differentiation. Thus, developing novel biomaterial providing better niche of spinal cord is a new strategy to treat the severe SCI. In this study, we constructed porcine spinal cord decellularized matrix scaffold (SC-DM) with biocompatibility to load engineered basic fibroblast growth factor-overexpressing human umbilical cord mesenchymal stromal cells (bFGF-HUCMSCs) for treating SCI. The continuously released bioactive bFGF factors from grafted bFGF-HUCMSCs and three-dimensional niche by SC-DM promoted the differentiation of endogenous stem cells into neurons with nerve conduction function, leading a markedly motor function recovery of SCI. These results indicated that the functional bFGF-HUCMSCs/SC-DM scaffold provided more suitable matric niche for nerve cells, that would be a promising strategy for the clinical application of SCI.

Programmed biomolecule delivery orchestrate bone tissue regeneration via MSC recruitment and epigenetic modulation

An on-demand delivery of chemotactic and osteogenic biomolecules to induce the recruitment and osteogenic differentiation of endogenous stem cells for in situ bone regeneration is appealing but challenging. To meet the changing demands at different periods of bone regeneration, a programmed delivery system was successfully fabricated by incorporating the near-infrared (NIR) light-responsive polydopamine-coated hydroxylapatite nanoparticles (nHA@PDA) into the thermo-responsive hydroxybutyl chitosan (HBC) hydrogel to regulate the therapeutic concentrations and time points of chemotactic simvastatin (SIM) and osteogenic pargyline (PGL). This smart hydrogel composite could perform an initial rapid release of SIM to meet the need of endogenous stem cell recruitment at the beginning of bone healing. Meanwhile, a flexible and NIR light-triggered increased release of PGL could promote osteogenic differentiation of migrated cells via a safe and stable epigenetic mechanism. Taken together, a well-orchestrated therapeutic timeline of two drugs was obtained in our programmed delivery system, thus enhancing bone regeneration in a highly effective method. In addition, the small molecular drugs utilized in this study were stable, safe, and easy for translation and clinical applications in bone tissue engineering.

Application of Perinatal Derivatives in Ovarian Diseases.

Reproductive diseases could lead to infertility and have implications for overall health, most importantly due to psychological, medical and socio-economic consequences for individuals and society. Furthermore, economical losses also occur in animal husbandry. In both human and veterinary medicine, hormonal and surgical treatments, as well as assisted reproductive technologies are used to cure reproductive disorders, however they do not improve fertility. With ovarian disorders being the main reproductive pathology in human and bovine, over the past 2 decades research has approached regenerative medicine in animal model to restore normal function. Ovarian pathologies are characterized by granulosa cell and oocyte apoptosis, follicular atresia, decrease in oocyte quality and embryonic development potential, oxidative stress and mitochondrial abnormalities, ultimately leading to a decrease in fertility. At current, application of mesenchymal stromal cells or derivatives thereof represents a valid strategy for regenerative purposes. Considering their paracrine/autocrine mode of actions that are able to regenerate injured tissues, trophic support, preventing apoptosis and fibrosis, promoting angiogenesis, stimulating the function and differentiation of endogenous stem cells and even reducing the immune response, are all important players in their future therapeutic success. Nevertheless, obtaining mesenchymal stromal cells (MSC) from adult tissues requires invasive procedures and implicates decreased cell proliferation and a reduced differentiation capacity with age. Alternatively, the use of embryonic stem cells as source of cellular therapeutic encountered several ethical concerns, as well as the risk of teratoma formation. Therefore, several studies have recently focussed on perinatal derivatives (PnD) that can be collected non-invasively and, most importantly, display similar characteristics in terms of regenerating-inducing properties, immune-modulating properties and hypo-immunogenicity. This review will provide an overview of the current knowledge and future perspectives of PnD application in the treatment of ovarian hypofunction.

Stem Cell‐Recruiting Injectable Microgels for Repairing Osteoarthritis

AbstractThe differentiation potentials and viability of stem cells are often impaired during cell isolation and delivery. Inspired by the phenomenon where islands can recruit seabirds for nesting, “cell island” microgels (MGs), that is, growth factor‐loaded methacrylated hyaluronic acid and heparin blend MGs, which can recruit endogenous stem cells and promote chondrogenic differentiation, are constructed using microfluidic technology and photopolymerization processes, followed by non‐covalently binding platelet‐derived growth factor‐BB (PDGF‐BB) and transforming growth factor‐beta3 (TGF‐β3). The loading efficiency of PDGF‐BB and TGF‐β3 are 96% and 91%, respectively. In vitro and in vivo experiments find that the “cell island” MGs can enhance the migratory capacity of cells and recruit them from their niche via releasing PDGF‐BB. Meanwhile, by using hyaluronic acid, the “cell island” MGs provide a suitable microenvironment for cell attachment and spreading. Furthermore, the “cell island” MGs induce chondrogenic differentiation of the recruited cells via releasing TGF‐β3 and present a promising therapeutic effect for osteoarthritis. In sum, this developed “cell island” MG might serve as a temporary “nest site” to allow the migration, adhesion, and differentiation of endogenous stem cells, which can be a promising candidate rather than the conventional cell‐seeded scaffolds for promoting tissue regeneration.

Generation of niche tuned antifibrotic fibroblasts and non-viral mediated endothelial commitment using adipose stem cells for dermal graft development.

Cell-based skin substitute generation has seen considerable development. Combining synthetic scaffolds with biomimetic fibrin does direct both exogenous and endogenous stem cell differentiation, addressing needs for reliable tissue engineering. However, lack of immediate vasculature within implantable grafts remains critical for its sustenance and integration. Multipotency, high proliferation potential, ability to release multiple growth factors (GFs), and autologous availability highlight the use of human adipose derived mesenchymal stem cells (hADMSCs) in tissue-engineered dermal grafts (TEDG) construction. However, hADMSCs' insufficiency to independently establish angiogenesis within tissue constructs demands improvement of stem cell application for dermal graft survival. Approaches to harness microenvironmentally sensitive paracrine interactions could improve the angiogenic efficiency of hADMSCs within TEDG. This study conceptualized a fibrin-based niche, to direct hADMSCs toward a nonfibrotic fibroblast commitment and incorporation of bioengineered hADMSCs, specifically releasing potent angiogenic factors within TEDG. Coexistence of tuned fibroblast and endothelial lineage committed cells contributed to well-regulated extracellular matrix formation and prevascularization. Adequate cell proliferation; sustained transient release of angiogenic GFs till 20 days; directed dermal, endothelial, fibroblast, and vascular smooth muscle cell differentiation; and favored elastin and collagen deposition were achieved in vitro. In conclusion, specific niche composition and employment of bioengineered hADMSCs favor implantable TEDG construction.

Ginsenoside Rb1/TGF-β1 loaded biodegradable silk fibroin-gelatin porous scaffolds for inflammation inhibition and cartilage regeneration.

Creating a microenvironment with low inflammation and favorable for the chondrogenic differentiation of endogenous stem cells plays an essential role in cartilage repairing. In the present study, we design a novel ginsenoside Rb1/TGF-β1 loaded silk fibroin-gelatin porous scaffold (GSTR) with the function of attenuating inflammation and promoting chondrogenesis. The scaffold has porous microstructure, proper mechanical strength, degradation rate and sustained release of Rb1 and TGF-β1. Rat bone marrow-derived mesenchymal stem cells (rBMSCs) seeded into GSTR scaffolds are homogeneously distributed and display a higher proliferation rate than non-loaded scaffolds (GS). GSTR scaffolds promote the chondrogenic differentiation of rBMSCs and suppress the expression of inflammation genes. Under the stimulation of IL-1β, the inflammation level of the chondrocytes seeded in GSTR scaffolds is also significantly down-regulated. Moreover, GSTR scaffolds implanted into the osteochondral defects in rats effectively promote the regeneration of hyaline cartilage 12weeks after surgery when compared with other groups. It is demonstrated that this scaffold loaded with Rb1 and TGF-β1 can synergistically create a microenvironment favorable for cartilage regeneration by promoting the chondrogenesis and suppressing the inflammation levels in vivo. These results prove it has a great potential to develop this Rb1/TGF-β1 releasing scaffold into a novel and promising therapeutic for cartilage repair.

Effect of Active Ingredients of Chinese Herbal Medicine on the Rejuvenation of Healthy Aging: Focus on Stem Cells.

Stem cells (SCs) are special types of cells with the ability of self-renewal and multidirectional differentiation. As the organism ages, the ability to maintain homeostasis and regeneration deteriorates and the number and activity of stem cells decline. Theoretically, the restoration of stem cells might reverse aging. However, due to their own aging, donor-derived immune rejection, and difficulties in stem cell differentiation control, a series of problems need to be solved to realize the potential for clinical application of stem cells. Chinese herbal medicine is a nature drug library which is suitable for the long-term treatment of aging-related diseases. Modern pharmacological studies have revealed that many active ingredients of Chinese herbal medicines with the effect of promoting stem cells growth and differentiation mainly belong to “reinforcing herbs.” In recent years, exploration of natural active ingredients from Chinese herbal medicines for delaying aging, improving the stem cell microenvironment, and promoting the proliferation and differentiation of endogenous stem cells has attracted substantial attention. This article will focus on active ingredients from Chinese herbs-mediated differentiation of stem cells into particular cell type, like neural cells, endothelial cells, cardiomyocytes, and osteoblasts. We will also discuss the effects of these small molecules on Wnt, Sonic Hedgehog, Notch, eNOS-cGMP, and MAP kinase signal transduction pathways, as well as reveal the role of estrogen receptor α and PPAR γ on selectively promoting or inhibiting stem cells differentiation. This review will provide new insights into the health aging strategies of active ingredients in Chinese herbal medicine in regenerative medicine.

Regenerative Endodontics: A Paradigm Shift in Clinical Endodontics.

ABSTRACTPreserving the natural dentition in its normal form and function is one of the major goals of endodontic treatment. Re-establishing the lost vitality and development of root maturation in immature permanent teeth with pulp necrosis/apical periodontitis is quiet challenging clinically. The very basis of regenerative endodontics relies on the concept of tissue engineering using stem cells, biomimetic scaffold, and bioactive growth factors to regenerate the pulp tissue damaged by microbial infection, injury, or developmental defects. In clinical endodontics, this approach is referred to as a “paradigm shift.” Even though repair instead of true regeneration is achieved with current protocols, it is hoped that further research in the area of stem-cell-based tissue engineering will allow for true regeneration and improved treatment outcomes. The aim of this review is to discuss about the various aspects of regenerative endodontics, current clinical protocols, and the future of pulp regeneration techniques.

New Perspectives of Osteochondral Lesion of the Talus

Osteochondral lesions of the talus(OLT) are the most common articular cartilage defects in the ankle and may cause disability as a result of chronic pain and limited weight-bearing capacity. Numerous treatment strategies for symptomatic OLT have advanced significantly over the past decades.OLT are often managed conservatively for an initial stage before the surgical treatment. However, the conservative management determined solely on symptomatology, and not on the physiological healing. A systematic review for the treatment of OLT demonstrated a 45% success rate of non-operative management. Surgical treatment of OLT is reserved for symptomatic focal lesions that fail to respond to conservative treatments. There are three major operative strategies for OLT, reparative, replacement and regenerative manners.For the reparative modality, arthroscopic bone marrow stimulation(BMS) is widely regarded as the first-line treatment for OLT, as it is a technically undemanding, cost-effective, and minimally invasive procedure with low rates of complication and postoperative pain. Small lesion (<15 mm in diameter or <150mm2 in size) is the ideal candidate for BMS supported by several literatures. And a study reported the long-term follow-up study to date at 8-20 years after BMS in which 78% of patients had an excellent or good functional outcome score.As a replacement strategy, autologous osteochondral transplantation(AOT) is often indicated for symptomatic large, cystic lesions, including those that have failed previous reparative procedures, such as BMS. Osteochondral autograft transfer carries the inherent advantage over osteochondral allograft of being from the host with fresh viable cartilage, most commonly harvested from the ipsilateral knee. Clinical studies have found favorable results with osteochondral autograft techniques, including a recent systematic review of clinical outcomes at mid-term follow-up demonstrating excellent or good outcomes in 87% of patients.In recent years, Scaffold-based regenerative techniques are getting more attention. Matrix-associated chondrocyte implantation(MACI) is a 2-step procedure in which culture-expanded autologous chondrocytes are seeded on a scaffold, which is then secured in the OLT. More recently, 1-step procedures have been developed in which scaffolds and/or orthobiologics, including bone marrow aspirate concentrate(BMAC) and platelet-rich plasma(PRP), and hyaluronic acid(HA) have been used to augment microfracture with the intention of overcoming the 2-step procedures while concurrently promoting chondrogenic differentiation of endogenous stem cells. Matrix-augmented BMS is one such technique that has been reported with good results in case series.In the future, an advanced strategy for tissue engineering with gene therapy may influence the quality of integration and longevity in treatment of OLT.

Gene Delivery into the Inner Ear and Its Clinical Implications for Hearing and Balance.

The inner ear contains many types of cell, including sensory hair cells and neurons. If these cells are damaged, they do not regenerate. Inner ear disorders have various etiologies. Some are related to aging or are idiopathic, as in sudden deafness. Others occur due to acoustic trauma, exposure to ototoxic drugs, viral infections, immune responses, or endolymphatic hydrops (Meniere’s disease). For these disorders, inner ear regeneration therapy is expected to be a feasible alternative to cochlear implants for hearing recovery. Recently, the mechanisms underlying inner ear regeneration have been gradually clarified. Inner ear cell progenitors or stem cells have been identified. Factors necessary for regeneration have also been elucidated from the mechanism of hair cell generation. Inducing differentiation of endogenous stem cells or inner ear stem cell transplantation is expected. In this paper, we discuss recent approaches to hair cell proliferation and differentiation for inner ear regeneration. We discuss the future road map for clinical application. The therapies mentioned above require topical administration of transgenes or drug onto progenitors of sensory cells. Developing efficient and safe modes of administration is clinically important. In this regard, we also discuss our development of an inner ear endoscope to facilitate topical administration.

Effect of icariin in combination with daily sildenafil on penile atrophy and erectile dysfunction in a rat model of bilateral cavernous nerves injury.

The commonly utilized phosphodiesterase type 5 inhibitors do not lead to satisfactory penile erection after radical prostatectomy mainly because of insufficient nitric oxide drive from the damaged cavernous nerves. The aim of this study was to assess the efficacy and mechanisms of icariin in combination with daily sildenafil on neurogenic erectile dysfunction and penile atrophy in a rat model of bilateral cavernous nerves injury. Sixty male Sprague-Dawley rats injected with 5-ethynyl-2-deoxyuridine (50mg/kg) at postnatal day 1 for the purpose of tracking endogenous stem cells in penis. Forty-eight rats of bilateral cavernous nerves injury were randomized equally into gavage feeding of vehicle, sildenafil (10mg/kg), icariin (1.5mg/kg) and sildenafil+icariin, respectively. Twelve sham-operated rats served as control. The intracavernous pressure and mean arterial pressure was measured and mid-penile cross sections were histologically examined 5weeks after surgery. Western blotting of cavernous tissue protein was also performed. Animals treated with sildenafil+icariin had significantly higher mean intracavernous pressure/mean arterial pressure ratio relative to other rats with bilateral cavernous nerves injury (p<0.05). The circumference and mean cross-sectional area of the paired corpus cavernosum were effectively preserved in the sildenafil+icariin. Treatment with sildenafil+icariin significantly increased the cavernous cyclic guanosine monophosphate concentration compared with the icariin group (p<0.05). In addition, the numbers of neuronal nitric oxide synthase-positive nerves and 5-ethynyl-2-deoxyuridine-positive cells co-expressing S100 in the icariin-treated groups were greater compared with the bilateral cavernous nerves injury control group (p<0.05). These data suggest that the combined use of icariin and daily sildenafil holds promise as a potential therapy for neurogenic erectile dysfunction in the future. The underlying mechanisms appears to involve two aspects: (i) icariin promotes differentiation of endogenous stem cells to Schwann cells, which help to repair the damaged neural pathway for erection; (ii) on this basis, sildenafil can further improve penile engorgement through the cyclic guanosine monophosphate-dependent smooth muscle relaxation.

M2 Phenotype Microglia-derived Cytokine Stimulates Proliferation and Neuronal Differentiation of Endogenous Stem Cells in Ischemic Brain.

Microglia play a key role in the immune response and inflammatory reaction that occurs in response to ischemic stroke. Activated microglia promote neuronal damage or protection in injured brain tissue. Extracellular signals polarize the microglia towards the M1/M2 phenotype. The M1/M2 phenotype microglia released pro- and anti-inflammatory cytokines which induce the activation of neural stem/progenitor cells (NSPCs). In this study, we investigated how the cytokines released by microglia affect the activation of NSPCs. First, we treated BV2 cells with a lipopolysaccharide (LPS; 20 ng/ml) for M1 phenotype microglia and interleukin-4 (IL-4; 20 ng/ml) for M2 phenotype microglia in BV2 cells. Mice were subjected to transient middle cerebral artery occlusion (tMCAO) for 1 h. In ex vivo, brain sections containing the subventricular zone (SVZ) were cultured in conditioned media of M1 and M2 phenotype-conditioned media for 3 d. We measured the expression of cytokines in the conditioned media by RT-PCR and ELISA. The M2 phenotype microglia-conditioned media led to the proliferation and neural differentiation of NSPCs in the ipsilateral SVZ after ischemic stroke. The RT-PCR and ELISA results showed that the expression of TGF-α mRNA was significantly higher in the M2 phenotype microglia-conditioned media. These data support that M2 phenotype microglia-derived TGF-α is one of the key factors to enhance proliferation and neural differntiation of NSPCs after ischemic stroke.

Effect of active factor on proliferation and differentiation of endogenous neural stem cells after traumatic brain injury in rats

Objective To investigate the effect of active factor on proliferation and differentiation of neural stem cells(NSCs)in the hippocampal subgranular zone after traumatic brain injury(TBI)in rats. Methods SD rats were divided into sham-injured group (only left dura mater exposed), active factor group (sham injury+ active factor therapy), TBI group (unilateral fluid percussion was used to generate severe TBI), and TBI+ active factor group (TBI+ active factor therapy) according to the random number table (n=10 in each group). Hippocampal homogenates or brain tissues were harvested at 7 d postinjury. Expressions of BrdU and NeuN in the hippocampal subgranular zone were detected by immunofluorescence staining, and five consecutive sections were used to count BrdU and NeuN positive cells. Level of nuclear factor (NF)-κB p65 in hippocampus was detected by Western blot. Results Number of BrdU and NeuN positive cells in TBI group were greatly higher than that in sham-injured group (P 0.05). NF-κBp65 presented over-expression in TBI group (0.81±0.09) while reversed in TBI+ active factor group (1.46±0.13), when compared to sham-injured group (P 0.05). Conclusion Active factor therapy can promote proliferation and neuronal differentiation of NSCs in the hippocampal subgranular zone after TBI in rats and the possible mechanism may relate to activation of the NSCs and inhibition of over-expression of NF-κBp65. Key words: Craniocerebral trauma; Neural stem cells; Active factor

Effects of honey to mobilize endogenous stem cells in efforts intestinal and ovarian tissue regeneration in rats with protein energy malnutrition

Abstract Objective Auto-regeneration of the intestinal and ovarian tissue are experiencing degenerative due to protein energy malnutrition (PEM) through of auto-mobilization, increase of immune response and differentiation of endogenous stem cells. Methods Female rat model of PEM obtained through fasting meals for 5 d and only to drink, causing malnutrition and damage of the intestinal and ovarian tissue. Furthermore, the therapy of honey with a dose of 30% water (T1) and 50% water (T2), respectively for 5 d and compared with the positive control, were fasted without being given honey (T0+) and negative control, not fasted and without being given honey (T0−). Observations on: auto-mobilization, increased immune response, differentiation of stem cells, and regeneration of the intestinal and ovarian tissue with HE staining. Results Auto-mobilization of stem cells based on the expression of CD34+ and CD45+, which is a marker of endogenous stem cells (hematopoietic stem cells/HSCs). Increased immune response is based on Hsp70 expression and PGE2 in intestinal tissue. Differentiation of stem cells into progenitor cells that expected based expression of growth differentiation factor-9 (GDF-9) by immunohistochemistry in ovarian tissue. Conclusions Expression of CD34+ and CD45+, which significantly different in treatment 2 (2). Furthermore, increase of immune response (decrease Hsp70 expression and increased PGE2) in intestinal tissue. Increased immune response causes expression of GDF-9 in ovarian tissue. Decreased of Hsp70 expression, increased PGE2 and increased GDF-9 followed the process of regeneration of the intestinal and ovarian tissue.

AB042. Implications for differentiation of endogenous stem cells: therapeutic effect from icariside II on a rat model of post-prostatectomy erectile dysfunction

Self-renewal and differentiation of endogenous stem cells (SCs) are essential for adult tissue homoeostasis and intrinsic healing capacity. Here, we hypothesize that penis contains a small population of endogenous SCs which might help rejuvenation of damaged erectile function. In this study, 60 newborn male rats were intraperitoneally injected with 5-ethynyl-2-deoxyuridine (EdU; 50 mg) for the purpose of tracking endogenous SCs. Twelve weeks later, 48 rats underwent bilateral cavernous nerves (CN) injury and were randomized into gavage feeding of solvent (vehicle group) or icariside II (ICAII) (0.5, 1.5 and 4.5 mg per day, respectively). Twelve sham-operated rats received vehicle treatment and served as control. The treatments were continued for 4 weeks followed by a washout period of 72 h. Results showed that ICAII treatment significantly restored erectile function and effectively prevented distortion of normal neural anatomy, smooth muscle atrophy and collagen deposition compared to vehicle group. The numbers of label retaining cells (LRCs) co-expressing EdU and differentiated phenotypes (smooth muscle markerα-SMA or Schwann cell marker S100) were significantly higher in three ICAII-treated groups than those in vehicle group in a dose-dependent manner. In addition, the changing trend of p38 mitogen activated protein kinase (MAPK) activity in the penis between groups was same as that of the number of differentiated LRCs. Together, these results suggest that the underlying mechanisms of ICAII in ameliorating erectile function and pathological changes appear to involve enhanced endogenous SCs differentiation, which might be regulated by p38 MAPK signaling pathway.

The effects of functional electrical stimulation on motor function and on the proliferation, migration and differentiation of neural stem cells in the subventricular zone

Objective To investigate the effects of functional electrical stimulation (FES) on motor function and on the expression, proliferation, migration and differentiation of endogenous neural stem cells in the subventricular zone (SVZ) after cerebral ischemia. Methods Middle cerebral artery occlusion (MCAO) was used to induce a model of cerebral ischemia in 108 rats using the modified Zea-Longa method of intraluminal filament occlusion. They were then randomly divided into an FES group, a placebo stimulation group and a control group with 36 cases in each. Superficial FES electrodes were pasted on the paralyzed forelimbs of the rats in the first two groups, though FES treatment was administered only to the FES group beginning on the 3rd day after the MCAO operation. The stimulation was designed to produce extension of the wrist and digits of the paralyzed forelimb. Before, and after 1, 3, 7 and 14 days of the treatment, the neurological deficit was evaluated using modified neurological severity scoring (mNSS). BrdU+ /GFAP+ , BrdU+ /DCX+ and BrdU+ /NeuN+ cells in the SVZ were detected using immunofluorescence technique. Results After 7 and 14 days of treatment, the average motor function of the rats in the FES group had improved significantly when compared with the averages of the other two groups. Compared with the other two groups, the average number of BrdU+ /GFAP+ positive cells in the ischemic SVZ was also significantly greater in the FES group after 7 and 14 days of treatment. After 14 days, BrdU+ /Dcx+ positive cells in the FES group had also increased significantly more, but only a few BrdU+ /NeuN+ cells had appeared in any of the three groups. Conclusion FES can improve motor function after cerebral ischemia, and promote proliferation and differentiation of neural stem cells in the SVZ. Key words: Electrical stimulation; Brain ischemia; Functional electrical stimulation; Neural stem cells; Subventricular zone