RESUMEN
BACKGROUND: Fat grafting is an adjuvant that may improve the quality of radiation-damaged tissue. However, fat grafting for volume restoration in irradiated sites may be less effective because of a poorly vascularized fibrotic recipient bed. External volume expansion has emerged as a potential technique to prepare the recipient sites for improved survival of grafted fat. The authors previously demonstrated increased vasculature with external volume expansion stimulation of irradiated tissues. The authors now hypothesize that external volume expansion's improvements in recipient-site vascularity will increase the volume retention and quality of fat grafts in fibrotic irradiated sites. METHODS: Athymic mice were irradiated until development of chronic radiation injury. Then, the irradiated site was stimulated by external volume expansion (external volume expansion group), followed by subcutaneous fat grafting. Grafts in an irradiated site without external volume expansion stimulation (irradiated control group) and grafts in a healthy nonirradiated (nonirradiated control group) site were used as controls. All grafts were monitored for 8 weeks and evaluated both histologically and by micro-computed tomography for analysis of volume retention. RESULTS: Hyperspectral imaging confirmed a 25 percent decrease in vascularity of irradiated tissue (irradiated control group) compared with nonirradiated tissue (nonirradiated control group). Grafts in the irradiated control group retained 11 percent less volume than grafts in the nonirradiated control group. The experimental external volume expansion group achieved a 20 percent (p = 0.01) increase in retained graft volume compared with the irradiated control group. CONCLUSIONS: External volume expansion stimulation can mitigate the effects of irradiation at the recipient site and in turn help preserve fat graft volume retention. Possible mechanisms include increased vascularity, adipogenic conversion, and increased compliance of a fibrotic recipient site.
Asunto(s)
Traumatismos Experimentales por Radiación/cirugía , Grasa Subcutánea/trasplante , Expansión de Tejido/métodos , Animales , Modelos Animales de Enfermedad , Ratones , Microtomografía por Rayos XRESUMEN
In the prostate, stem and progenitor cell regenerative capacities have been ascribed to both basal and luminal epithelial cells. Here, we show that a rare subset of mesenchymal cells in the prostate are epithelial-primed Nestin-expressing cells (EPNECs) that can generate self-renewing prostate organoids with bipotential capacity. Upon transplantation, these EPNECs can form prostate gland tissue grafts at the clonal level. Lineage-tracing analyses show that cells marked by Nestin or NG2 transgenic mice contribute to prostate epithelium during organogenesis. In the adult, modest contributions in repeated rounds of regression and regeneration are observed, whereas prostate epithelial cells derived from Nestin/NG2-marked cells are dramatically increased after severe irradiation-induced organ damage. These results indicate that Nestin/NG2 expression marks a novel radioresistant prostate stem cell that is active during development and displays reserve stem cell activity for tissue maintenance.
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Antígenos/biosíntesis , Células Epiteliales/metabolismo , Nestina/biosíntesis , Trasplante de Órganos , Próstata/metabolismo , Próstata/trasplante , Proteoglicanos/biosíntesis , Traumatismos Experimentales por Radiación , Tolerancia a Radiación , Células Madre/metabolismo , Animales , Antígenos/genética , Células Epiteliales/patología , Regulación de la Expresión Génica/efectos de la radiación , Masculino , Ratones , Ratones Transgénicos , Nestina/genética , Próstata/patología , Proteoglicanos/genética , Traumatismos Experimentales por Radiación/metabolismo , Traumatismos Experimentales por Radiación/patología , Traumatismos Experimentales por Radiación/cirugía , Células Madre/patologíaRESUMEN
BACKGROUND: Treatment of critical limb ischemia is sometimes difficult because of the patient's condition, and some novel approaches are needed. METHODS: The hindlimbs of Sprague-Dawley rats, after 20-Gy x-ray irradiation and surgical occlusion, were divided into four groups: with a superficial fascial flap, 5.0 × 10 adipose-derived stromal/stem cells, and both combined. The rats were tested for laser tissue blood flow, immunohistologic blood vessel density, and foot paw punch hole wound healing. Green fluorescent protein-tagged Sprague-Dawley rats were used for further investigation by cell tracking for 2 weeks. RESULTS: Laser tissue blood flow demonstrated a significant increase in the combined treatment of flap and adipose-derived stem cells at both 1 and 2 weeks. There were no significant differences between the treatment groups treated with flaps alone and those treated with adipose-derived stem cells alone. Wound healing was significantly increased following combined treatment at 1 week, and there was no wound by 2 weeks except for the no-flap and no-adipose-derived stem cell group. The number of vessels depicted by von Willebrand factor showed a significant increase in the combined treatment group, at both 1 week and 2 weeks. In the cell tracking group, at 2 weeks, the green fluorescent protein-tagged adipose-derived stem cells were significantly more positive in the no-flap group than in the flap group. CONCLUSIONS: Adipose-derived stem cells may be a potent cell source in irradiated and occluded limbs by enhancing tissue blood flow and blood vessel density. Adipose-derived stem cells may play an important role in some difficult ischemic conditions in terms of wound healing.
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Tejido Adiposo/citología , Isquemia/cirugía , Neovascularización Fisiológica , Trasplante de Células Madre , Colgajos Quirúrgicos , Cicatrización de Heridas , Animales , Proteínas Fluorescentes Verdes/análisis , Miembro Posterior/irrigación sanguínea , Miembro Posterior/efectos de la radiación , Isquemia/terapia , Masculino , Traumatismos Experimentales por Radiación/cirugía , Traumatismos Experimentales por Radiación/terapia , Ratas , Ratas Sprague-Dawley , Ratas Transgénicas , Colgajos Quirúrgicos/irrigación sanguínea , Factor de von Willebrand/análisisRESUMEN
Radiation-induced lung injury (RILI) is a fatal condition featured by interstitial pneumonitis and fibrosis. Mesenchymal stem cells (MSCs) have been widely used for treating RILI in rodent models. In the present study, we aimed to investigate whether the therapeutic effects of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) on RILI were in a dose-dependent manner. A total of 100 mice were randomly divided into: a control group (n=25), subject to lung irradiation and injection of phosphate-buffered solution (PBS) via the tail vein; and the hBM-MSC group, subject to lung irradiation followed by injection of a low dose (1x103 hBM-MSCs/g), medium dose (5x103 hBM-MSCs/g) and high dose (1x104 hBM-MSCs/g) of hBM-MSCs in PBS through the tail vein, respectively. After sacrifice, the pulmonary tissues were subject to hematoxylin and eosin (H&E) staining, Masson's trichrome staining and immunohistochemical staining to investigate the pathological changes. Immunofluorescent staining was performed to evaluate the differentiation capacity of hBM-MSCs in vivo by analyzing the expression of SPC and PECAM. hBM-MSCs improved the survival rate and histopathological features in the irradiated mice, especially in the low-dose group. Marked decrease in collagen deposition was noted in the irradiated mice treated using a low dose of hBM-MSCs. In addition, hBM-MSCs attenuated secretion and expression of IL-10 and increased the expression of TNF-α. Furthermore, hBM-MSCs had the potential to differentiate into functional cells upon lung injury. Low-dose hBM-MSCs contributed to functional recovery in mice with RILI.
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Trasplante de Células Madre Mesenquimatosas/métodos , Traumatismos Experimentales por Radiación/cirugía , Neumonitis por Radiación/cirugía , Animales , Células de la Médula Ósea , Diferenciación Celular/fisiología , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Xenoinjertos , Humanos , Ratones , Traumatismos Experimentales por Radiación/complicaciones , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
BACKGROUND: A major area of unmet need is the development of strategies to restore neuronal network systems and to recover brain function in patients with neurological disease. The use of cell-based therapies remains an attractive approach, but its application has been challenging due to the lack of suitable cell sources, ethical concerns, and immune-mediated tissue rejection. We propose an innovative approach that utilizes gut-derived neural tissue for cell-based therapies following focal or diffuse central nervous system injury. RESULTS: Enteric neuronal stem and progenitor cells, able to differentiate into neuronal and glial lineages, were isolated from the postnatal enteric nervous system and propagated in vitro. Gut-derived neural progenitors, genetically engineered to express fluorescent proteins, were transplanted into the injured brain of adult mice. Using different models of brain injury in combination with either local or systemic cell delivery, we show that transplanted enteric neuronal progenitor cells survive, proliferate, and differentiate into neuronal and glial lineages in vivo. Moreover, transplanted cells migrate extensively along neuronal pathways and appear to modulate the local microenvironment to stimulate endogenous neurogenesis. CONCLUSIONS: Our findings suggest that enteric nervous system derived cells represent a potential source for tissue regeneration in the central nervous system. Further studies are needed to validate these findings and to explore whether autologous gut-derived cell transplantation into the injured brain can result in functional neurologic recovery.
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Encéfalo/fisiopatología , Trasplante de Células/métodos , Sistema Nervioso Entérico/fisiología , Células-Madre Neurales/fisiología , Trasplante de Células Madre/métodos , Animales , Encéfalo/efectos de la radiación , Encéfalo/cirugía , Lesiones Encefálicas/cirugía , Lesiones Encefálicas/terapia , Proliferación Celular , Supervivencia Celular , Células Cultivadas , Sistema Nervioso Entérico/citología , Ratones , Ratones Endogámicos C57BL , Células-Madre Neurales/trasplante , Neurogénesis , Traumatismos Experimentales por Radiación/cirugía , Traumatismos Experimentales por Radiación/terapiaRESUMEN
BACKGROUND & AIMS: Radiation-induced liver damage (RILD) is a poorly understood and potentially devastating complication of hepatic radiation therapy (RT) for liver cancers. Previous work has demonstrated that hepatocyte transplantation (HT) can ameliorate RILD in rats. We hypothesized that RT inhibits generation of cellular ATP and suppresses hepatic regeneration. METHODS: To study the metabolic changes that occur in RILD with and without HT, (31)P MRSI data were acquired in rats treated with partial hepatectomy (PH) alone, PH with hepatic irradiation (PHRT) or PHRT with HT (PHRT+HT). RESULTS: Both [γ -ATP] and ATP/Pi (31)P MRSI signal ratio initially decreased and subsequently returned to baseline levels within 2 weeks after PH, which is consistent with other published data. Persistently reduced [γ-ATP] and ATP/Pi (31)P MRSI signal ratio were observed in rats up to 20 weeks after PHRT. However, progressive increases in [γ -ATP] were observed over time in the group of rats receiving PHRT+HT. Normal [γ -ATP] was observed 20 weeks after PHRT+HT (vs. PH alone), although, ATP/Pi levels did not return to normal after PHRT +HT. Ex vivo histological studies were performed to confirm liver repopulation with transplanted hepatocytes and the amelioration of pathologic changes of RILD. CONCLUSIONS: These findings suggest that (31)P MRSI can be used to monitor the progress of RILD and its amelioration using transplanted hepatocytes to simultaneously restore metabolic function while replacing host hepatocytes damaged by RT.
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Metabolismo Energético , Hepatocitos/trasplante , Regeneración Hepática , Hígado/cirugía , Espectroscopía de Resonancia Magnética/métodos , Traumatismos Experimentales por Radiación/cirugía , Adenosina Trifosfato/metabolismo , Animales , Proliferación Celular , Hepatectomía/métodos , Hepatocitos/metabolismo , Hepatocitos/patología , Hígado/metabolismo , Hígado/patología , Hígado/fisiopatología , Masculino , Espectroscopía de Protones por Resonancia Magnética , Traumatismos Experimentales por Radiación/metabolismo , Traumatismos Experimentales por Radiación/patología , Traumatismos Experimentales por Radiación/fisiopatología , Ratas Endogámicas F344 , Recuperación de la Función , Factores de TiempoRESUMEN
Past preclinical studies have demonstrated the capability of using human stem cell transplantation in the irradiated brain to ameliorate radiation-induced cognitive dysfunction. Intrahippocampal transplantation of human embryonic stem cells and human neural stem cells (hNSCs) was found to functionally restore cognition in rats 1 and 4 months after cranial irradiation. To optimize the potential therapeutic benefits of human stem cell transplantation, we have further defined optimal transplantation windows for maximizing cognitive benefits after irradiation and used induced pluripotent stem cell-derived hNSCs (iPSC-hNSCs) that may eventually help minimize graft rejection in the host brain. For these studies, animals given an acute head-only dose of 10 Gy were grafted with iPSC-hNSCs at 2 days, 2 weeks, or 4 weeks following irradiation. Animals receiving stem cell grafts showed improved hippocampal spatial memory and contextual fear-conditioning performance compared with irradiated sham-surgery controls when analyzed 1 month after transplantation surgery. Importantly, superior performance was evident when stem cell grafting was delayed by 4 weeks following irradiation compared with animals grafted at earlier times. Analysis of the 4-week cohort showed that the surviving grafted cells migrated throughout the CA1 and CA3 subfields of the host hippocampus and differentiated into neuronal (â¼39%) and astroglial (â¼14%) subtypes. Furthermore, radiation-induced inflammation was significantly attenuated across multiple hippocampal subfields in animals receiving iPSC-hNSCs at 4 weeks after irradiation. These studies expand our prior findings to demonstrate that protracted stem cell grafting provides improved cognitive benefits following irradiation that are associated with reduced neuroinflammation.
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Trastornos del Conocimiento/etiología , Irradiación Craneana/efectos adversos , Células Madre Pluripotentes Inducidas/trasplante , Traumatismos Experimentales por Radiación/cirugía , Trasplante de Células Madre/métodos , Animales , Trastornos del Conocimiento/cirugía , Xenoinjertos , Hipocampo/cirugía , Humanos , Inmunohistoquímica , Microscopía Confocal , Ratas , Ratas DesnudasRESUMEN
Cutaneous radiation syndrome is the delayed consequence of localized skin exposure to high doses of ionizing radiation. Adipocyte derived stem cells injection may improve tissue regeneration through secreted factors. Thus mesenchymal stem cells secretome optimization, using transient transfection, may represent a new strategy to treat this syndrome. Sonic hedgehog, a secreted protein involved in cell proliferation and angiogenesis, has been chosen as a first candidate. Here preliminary results are reported of the therapeutic potential of transient gene therapy to cure cutaneous radiation syndrome in a minipig model. Adipocyte derived stem cells were transiently transfected by electroporation with a plasmid coding for Sonic Hedgehog. Göttingen minipigs were locally irradiated using a (60)Co gamma source at the dose of 50 Gy and received Phosphate Buffer Salin (controls: n = 8), stem cells (50 × 106 each time, n = 5) or transfected stem cells (25±7 × 106 each time, n = 1). All controls exhibited a homogeneous clinical evolution of cutaneous radiation syndrome with final necrosis (day 91). In stem cell injected minipigs, an ultimate wound healing was observed in four out of five grafted animals (day 130 ± 28, complete in two of them) (historical results). The Sonic hedgehog animal, albeit injected with a lower number of transfected stem cells, presented a very similar evolution of skin healing without necrosis or uncontrolble pain. Globally this preliminary report suggests that local injection of Sonic Hedgehog transfected adipocyte derived stem cells may improve wound healing. Thus work is ongoing to evaluate this therapeutic strategy on a larger number of animals.
Asunto(s)
Terapia Genética/métodos , Traumatismos Experimentales por Radiación/genética , Traumatismos Experimentales por Radiación/terapia , Piel/lesiones , Piel/efectos de la radiación , Porcinos Enanos , Tejido Adiposo/citología , Animales , Modelos Animales de Enfermedad , Estudios de Factibilidad , Proteínas Hedgehog/genética , Traumatismos Experimentales por Radiación/cirugía , Trasplante de Células Madre , Células Madre/citología , Células Madre/metabolismo , Porcinos , TransfecciónRESUMEN
Olfactory ensheathing cells (OECs) and Schwann cells (SCs) share many characteristics, including the ability to promote neuronal repair when transplanted directly into spinal cord lesions, but poor survival and migration when transplanted into intact adult spinal cord. Interestingly, transplanted OECs, but not SCs, migrate extensively within the X-irradiated (40 Gy) adult rat spinal cord, suggesting distinct responses to environmental cues [Lankford et al., (2008) GLIA 56:1664-1678]. In this study, GFP-expressing OECs and SCs were transplanted into juvenile rat brains (hippocampus) subjected to a moderate radiation dose (16 Gy). As in the adult spinal cord, OECs, but not SCs, migrated extensively within the irradiated juvenile rat brain. Unbiased stereology revealed that the number of OECs observed within irradiated rat brains three weeks after transplantation was as much as 20 times greater than the number of cells transplanted, and the cells distributed extensively within the brain. In conjunction with the OEC dispersion, the number of activated microglia in OEC-transplanted irradiated brains was reduced. Unlike in the intact adult spinal cord, both OECs and SCs showed some, but limited, migration within nonirradiated rat brains, suggesting that the developing brain may be a more permissive environment for cell migration than the adult CNS. These results show that OECs display unique migratory, proliferative, and microglia interaction properties as compared with SCs when transplanted into the moderately X-irradiated brain.
Asunto(s)
Movimiento Celular/efectos de la radiación , Proliferación Celular/efectos de la radiación , Mucosa Olfatoria/citología , Mucosa Olfatoria/trasplante , Células de Schwann/citología , Trasplante de Células Madre , Animales , Animales Recién Nacidos , Antígenos , Antígeno CD11b/metabolismo , Células Cultivadas , Femenino , Proteína Ácida Fibrilar de la Glía/metabolismo , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Masculino , Neuroglía/fisiología , Neuroglía/efectos de la radiación , Mucosa Olfatoria/metabolismo , Oligodendroglía/fisiología , Oligodendroglía/trasplante , Proteoglicanos , Traumatismos Experimentales por Radiación/patología , Traumatismos Experimentales por Radiación/cirugía , Ratas , Ratas Sprague-Dawley , Células de Schwann/química , Células de Schwann/metabolismoRESUMEN
The management of proctitis in patients who have undergone very-high-dose conformal radiotherapy is extremely challenging. The fibrosis-necrosis, fistulae, and hemorrhage induced by pelvic overirradiation have an impact on morbidity. Augmenting tissue repair by the use of mesenchymal stem cells (MSCs) may be an important advance in treating radiation-induced toxicity. Using a preclinical pig model, we investigated the effect of autologous bone marrow-derived MSCs on high-dose radiation-induced proctitis. Irradiated pigs received repeated intravenous administrations of autologous bone marrow-derived MSCs. Immunostaining and real-time polymerase chain reaction analysis were used to assess the MSCs' effect on inflammation, extracellular matrix remodeling, and angiogenesis, in radiation-induced anorectal and colon damages. In humans, as in pigs, rectal overexposure induces mucosal damage (crypt depletion, macrophage infiltration, and fibrosis). In a pig model, repeated administrations of MSCs controlled systemic inflammation, reduced in situ both expression of inflammatory cytokines and macrophage recruitment, and augmented interleukin-10 expression in rectal mucosa. MSC injections limited radiation-induced fibrosis by reducing collagen deposition and expression of col1a2/col3a1 and transforming growth factor-ß/connective tissue growth factor, and by modifying the matrix metalloproteinase/TIMP balance. In a pig model of proctitis, repeated injections of MSCs effectively reduced inflammation and fibrosis. This treatment represents a promising therapy for radiation-induced severe rectal damage.
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Trasplante de Células Madre Mesenquimatosas/métodos , Células Madre Mesenquimatosas/citología , Proctitis/patología , Proctitis/cirugía , Traumatismos Experimentales por Radiación/terapia , Animales , Células de la Médula Ósea/citología , Células de la Médula Ósea/metabolismo , Colágeno/metabolismo , Colágeno Tipo I/metabolismo , Factor de Crecimiento del Tejido Conjuntivo/metabolismo , Matriz Extracelular/metabolismo , Matriz Extracelular/patología , Fibrosis/metabolismo , Fibrosis/fisiopatología , Fibrosis/terapia , Humanos , Inflamación/metabolismo , Inflamación/fisiopatología , Inflamación/cirugía , Interleucina-10/metabolismo , Masculino , Células Madre Mesenquimatosas/metabolismo , Membrana Mucosa/diagnóstico por imagen , Membrana Mucosa/metabolismo , Membrana Mucosa/patología , Neovascularización Patológica/metabolismo , Proctitis/etiología , Proctitis/metabolismo , Traumatismos Experimentales por Radiación/metabolismo , Traumatismos Experimentales por Radiación/patología , Traumatismos Experimentales por Radiación/cirugía , Cintigrafía , Recto/diagnóstico por imagen , Recto/metabolismo , Recto/patología , Porcinos , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
This study evaluated the clinical and pathologic effects of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) in the recovery from total body irradiation by comparing it with the effects of granulocyte-colony stimulating factor (G-CSF), an efficacious drug in the treatment of acute bone marrow radiation syndrome. BALB/c mice were treated with G-CSF or hUCB-MSCs after they were irradiated with 7 Gy cobalt-60 γ-rays. Circulating blood counts, histopathologic changes in the bone marrow, and plasma level of Flt-3L and transforming growth factor (TGF-ß1) were monitored in the postirradiation period. Hematologic analysis revealed that the peripheral leukocyte counts were markedly increased in the hUCB-MSCs-treated group, whereas G-CSF-treated mice did not recover significantly. Moreover, differential counts showed that hUCB-MSC treatment has regenerative effects on white blood cells, lymphocytes, and monocytes compared with the irradiated group. Treatment with hUCB-MSCs or G-CSF significantly increased immunoreactivity of Ki-67 until 3 weeks after total body irradiation. However, at 3 weeks, the number of Ki-67 immunoreactive cells significantly increased in the hUCB-MSCs-treated group compared with the G-CSF-treated group. Furthermore, hUCB-MSC treatment significantly modulated plasma levels of the hematopoietic cytokines Flt-3L and TGF-ß1, whereas G-CSF treatment failed to decrease the plasma Flt-3L levels at 2 weeks after irradiation. Based on the differences in circulating blood cell reconstitution and cell density of bone marrow, the authors suggest that MSC treatment is superior to G-CSF treatment for hematopoietic reconstitution following sublethal dose radiation exposure.
Asunto(s)
Síndrome de Radiación Aguda/cirugía , Médula Ósea/patología , Trasplante de Células Madre Mesenquimatosas/métodos , Traumatismos Experimentales por Radiación/cirugía , Síndrome de Radiación Aguda/tratamiento farmacológico , Síndrome de Radiación Aguda/etiología , Animales , Médula Ósea/efectos de los fármacos , Médula Ósea/efectos de la radiación , Células Cultivadas , Femenino , Sangre Fetal/citología , Rayos gamma/efectos adversos , Factor Estimulante de Colonias de Granulocitos/farmacología , Humanos , Antígeno Ki-67/metabolismo , Recuento de Leucocitos , Células Madre Mesenquimatosas/citología , Ratones , Ratones Endogámicos BALB C , Traumatismos Experimentales por Radiación/tratamiento farmacológico , Traumatismos Experimentales por Radiación/etiología , Factores de Tiempo , Factor de Crecimiento Transformador beta1/sangre , Trasplante Heterólogo , Resultado del Tratamiento , Irradiación Corporal Total/efectos adversos , Tirosina Quinasa 3 Similar a fms/sangreRESUMEN
Radiotherapy is a frontline treatment for the clinical management of CNS tumors. Although effective in eradicating tumor cells, radiotherapy also depletes neural stem and progenitor cells in the hippocampus that are important for neurogenesis and cognitive function. Consequently, the use of radiation to control primary and metastatic brain tumors often leads to debilitating and progressive cognitive decrements in surviving patients, representing a serious medical condition that, to date, has no satisfactory, long-term solutions. As a result, we have explored the use of stem cells as therapeutic agents to improve cognition after radiotherapy. Our past work has demonstrated the capability of cranially transplanted human embryonic (hESCs) and neural (hNSCs) stem cells to functionally restore cognition in rats 1 and 4 months after head-only irradiation. We have now expanded our cognitive analyses with hESCs and quantified both survival and differentiated fates of engrafted cells at 1 and 4 months after irradiation. Our findings indicate the capability of hESC transplantation to ameliorate radiation-induced cognitive dysfunction 1 month following cranial irradiation, using a hippocampal-dependent novel place recognition task. Irradiated animals not engrafted with stem cells experienced prolonged and significant cognitive dysfunction. Stereological estimates indicated that 35% and 17% of the transplanted hESCs survived at 1 and 4 months postgrafting, respectively. One month after irradiation and grafting, phenotypic analyses revealed that 26% and 31% of the hESCs differentiated into neurons and astrocytes, while at the 4-month time, neuronal and astrocytic differentiation was 7% and 46%, respectively. Comparison between present and past data with hESCs and hNSCs demonstrates equivalent cognitive restoration and a preference of hNSCs to commit to neuronal versus astrocytic lineages over extended engraftment times. Our data demonstrate the functional utility of human stem cell replacement strategies for ameliorating the adverse effects of cranial irradiation on cognition.
Asunto(s)
Encéfalo/patología , Encéfalo/efectos de la radiación , Irradiación Craneana/métodos , Traumatismos Experimentales por Radiación/cirugía , Trasplante de Células Madre/métodos , Animales , Encéfalo/metabolismo , Diferenciación Celular/fisiología , Diferenciación Celular/efectos de la radiación , Modelos Animales de Enfermedad , Xenoinjertos , Humanos , Masculino , Neuronas/citología , Neuronas/patología , Neuronas/efectos de la radiación , Traumatismos Experimentales por Radiación/etiología , Traumatismos Experimentales por Radiación/patología , Ratas , Ratas DesnudasRESUMEN
Bone marrow transplantation (BMT) is associated with a high risk of mortality, partially because of the harmful effects of the preconditioning myeloablative regimens. We have recently demonstrated increased bone marrow cell survival and proliferation in response to exercise training, which may be attributable to increased quality of the niche. The purpose of the present study was to determine the extent to which exercise preconditioning of recipients could increase the success of BMT. Recipient mice remained sedentary (SED) or were exercise-trained (EX) on a treadmill (3 d/wk for 8 weeks) before reconstitution with green fluorescent protein (GFP)-labeled donor marrow. Recipient survival, both donor-derived and total (donor- and recipient-derived) blood reconstitution were measured by flow cytometry. The first and fourth day after BMT apoptosis, cellularity and donor cell homing were determined in the recipients' bone marrow cavity by flow cytometry. Whereas only 25% of SED mice survived, 82% of EX recipients survived the BMT. Homing of donor-derived marrow cells to the recipients' marrow cavity acutely after BMT was not altered in EX, but EX mice displayed decreased levels (10%; p < 0.05) of activated caspase-3/-7 one day after BMT, leading to a maintenance of marrow cellularity in mice preconditioned with exercise. The acute inhibition of marrow cell apoptosis in EX mice resulted in increased total blood cell reconstitution at 1 and 3.5 months after BMT in EX mice (42% and 43%, respectively; both p < 0.05). Short- and long-term donor-derived engraftment was not different between EX and SED recipients. Exercise training increases recipient survival after BMT with increased total blood cell reconstitution.
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Células de la Médula Ósea/citología , Trasplante de Médula Ósea/rehabilitación , Supervivencia de Injerto , Condicionamiento Físico Animal , Animales , Apoptosis , Células de la Médula Ósea/enzimología , Quimiotaxis de Leucocito , Citocinas/sangre , Femenino , Genes Reporteros , Proteínas Fluorescentes Verdes/análisis , Inflamación/sangre , Masculino , Ratones , Ratones Endogámicos C57BL , Quimera por Radiación , Traumatismos Experimentales por Radiación/sangre , Traumatismos Experimentales por Radiación/patología , Traumatismos Experimentales por Radiación/cirugía , Carrera , Conducta SedentariaRESUMEN
The objective of this study was to explore the protective effects of human bone marrow mesenchymal stem cells (MSC) on hematopoietic organs of irradiated mice. Human bone marrow MSC were isolated, ex vivo expanded, and identified by cell biological tests. Female BALB/c mice were irradiated with (60)Co γ-ray at a single dose of 6 Gy, and received different doses of human MSC and MSC lysates or saline via tail veins. The survival of mice was record daily, and the femurs and spleens were harvested on day 9 and 16 for pathologic examination. The histological changes were observed and the cellularity was scored. The results showed that the estimated survival time of MSC- and MSC lysate-treated mice was comparable to that of controls. The hematopoiesis in the bone marrow of mice that received high-dose (5×10(6)) of MSC or MSC lysates was partially restored on day 9 and the capacity of hemopoietic tissue and cellularity scorings were significantly elevated as compared with that of controls (P < 0.05). Proliferative nudes were also obviously observed in the spleens of mice that received high-dose of MSC or MSC lysates on d 9 after irradiation. The histological structures of the spleen and bone marrow of the mice that received high-doses (5×10(6)) of MSC or MSC lysates were restored to normal, the cell proliferation displayed extraordinarily active. Further, the cellularity scores of the bone marrow were not significantly different between the high-dose MSC and MSC lysate-treated mice. It is concluded that the bone marrow MSC can promote the hematopoietic recovery of the irradiated mice, which probably is associated with the bioactive materials inherently existed in bone marrow cells.
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Hematopoyesis , Trasplante de Células Madre Mesenquimatosas , Traumatismos Experimentales por Radiación/cirugía , Animales , Células de la Médula Ósea/citología , Femenino , Humanos , Células Madre Mesenquimatosas/citología , Ratones , Ratones Endogámicos BALB C , Trasplante HeterólogoRESUMEN
This study was aimed to investigate the effect of human bone marrow mesenchymal stem cells (hBMMSC) on the hematopoietic recovery of sublethally irradiated mice. Female BALb/c mice irradiated with (60)Co γ-ray at a single dose of 6 Gy received graded doses of hBMMSC (1×10(5), 1×10(6) and 5×10(6)) by intravenous infusion. The counts of leukocytes, platelets, erythrocytes and hemoglobin level in peripheral blood, the amount of bone marrow hematopoietic progenitors, and the serum levels of human TPO, SCF and G-CSF as well were evaluated at different time points after transplantation. The results showed that hBMMSC infusion had little protective effect on the survival of irradiated mice. Compared with the control mice, the peripheral blood cell counts of hBMMSC-treated mice were not obviously elevated during 3 weeks after infusion, however, blood cell counts were significantly greater at 4 weeks after cell treatment (P < 0.05). The amount of colony-forming unit of mononuclear cells and granulocyte/monocytes in bone marrow of mice that received middle and high doses of hBMMSC were dramatically greater than that in control mice (P < 0.05). Two days after hBMMSC administration, human G-CSF and SCF could be detected in the sera from hBMMSC-treated mice, and the G-CSF concentration of mice that received high-dose hBMMSC was significantly higher than that in other groups (P < 0.01). Nevertheless, human TPO was undetectable in the sera of all mice tested and serum human G-CSF and SCF could not be detected on days 9 and 16 in all groups. It is concluded that hBMMSC may promote the hematopoietic recovery of irradiated mice, probably by transient secretion of hematopoiesis-associated factors by the implanted cells.
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Hematopoyesis , Trasplante de Células Madre Mesenquimatosas , Traumatismos Experimentales por Radiación/cirugía , Animales , Células de la Médula Ósea/citología , Femenino , Humanos , Células Madre Mesenquimatosas/citología , Ratones , Ratones Endogámicos BALB C , Trasplante HeterólogoRESUMEN
Erythropoiesis-stimulating agents (ESA) are now central to the treatment of renal anemia and are associated with improved clinical outcomes. It is well known that erythropoietin (EPO) is a key regulator of erythropoiesis through its promotion of red blood cell production. In order to investigate the role of ESA on iron metabolism, we analyzed the regulation of the iron regulatory hormone hepcidin by ESA treatment in a bone marrow transplant model in mouse. After treating C57BL/6 mice with continuous erythropoietin receptor activator (C.E.R.A.), recombinant human epoetin-ß (rhEPO), or recombinant human carbamylated epoetin-ß (rhCEPO), we investigated serum hepcidin concentrations and parameters of erythropoiesis. Serum hepcidin concentrations after rhEPO treatment were analyzed in mice subjected to total body irradiation followed by bone marrow transplantation. C.E.R.A. administration caused long-term downregulation of serum hepcidin levels. Serum hepcidin levels in rhEPO-treated mice decreased significantly, whereas there was no change in rhCEPO-treated mice. The reduction in circulating hepcidin levels after rhEPO administration was not observed in irradiated mice. Finally, bone marrow transplantation recovered the response to rhEPO administration that downregulates hepcidin concentration in irradiated mice. These results indicate that ESA treatment downregulates serum hepcidin concentrations, mainly by indirect mechanisms affecting hematopoietic activity in bone marrow cells.
Asunto(s)
Péptidos Catiónicos Antimicrobianos/biosíntesis , Células de la Médula Ósea/efectos de los fármacos , Eritropoyesis/efectos de los fármacos , Eritropoyetina/análogos & derivados , Animales , Péptidos Catiónicos Antimicrobianos/genética , Células de la Médula Ósea/metabolismo , Trasplante de Médula Ósea , Ensayo de Unidades Formadoras de Colonias , Regulación hacia Abajo , Eritropoyesis/genética , Eritropoyetina/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Hepcidinas , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , ARN Mensajero/biosíntesis , Quimera por Radiación , Traumatismos Experimentales por Radiación/metabolismo , Traumatismos Experimentales por Radiación/cirugía , Reacción en Cadena en Tiempo Real de la Polimerasa , Proteínas Recombinantes/farmacología , Organismos Libres de Patógenos Específicos , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Cutaneous radiation syndrome caused by local high dose irradiation is characterized by delayed outcome and incomplete healing. Recent therapeutic management of accidentally irradiated burn patients has suggested the benefit of local cellular therapy using mesenchymal stem cell grafting. According to the proposed strategy of early treatment, large amounts of stem cells would be necessary in the days following exposure and hospitalization, which would require allogeneic stem cells banking. In this context, the authors compared the benefit of local autologous and allogeneic adipocyte-derived stem cell injection in a large animal model. Minipigs were locally irradiated using a 60Co gamma source at a dose of 50 Gy and divided into three groups. Two groups were grafted with autologous (n = 5) or allogeneic (n = 5) adipocyte-derived stem cells four times after the radiation exposure, whereas the control group received the vehicle without cells (n = 8). A clinical score was elaborated to compare the efficiency of the three treatments. All controls exhibited local inflammatory injuries leading to a persistent painful necrosis, thus mimicking the clinical evolution in human victims. In the autologous adipocyte-derived stem cells group, skin healing without necrosis or uncontrollable pain was observed. In contrast, the clinical outcome was not significantly different in the adipocyte-derived stem cell allogeneic group when compared with controls. This study suggests that autologous adipocyte-derived stem cell grafting improves cutaneous radiation syndrome wound healing, whereas allogeneic adipocyte derived stem cells do not. Further studies will establish whether manipulation of allogeneic stem cells will improve their therapeutic potential.
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Adipocitos/citología , Traumatismos Experimentales por Radiación/cirugía , Enfermedades de la Piel/cirugía , Trasplante de Células Madre/métodos , Animales , Femenino , Traumatismos Experimentales por Radiación/fisiopatología , Traumatismos Experimentales por Radiación/terapia , Regeneración , Enfermedades de la Piel/fisiopatología , Enfermedades de la Piel/terapia , Porcinos , Trasplante Homólogo , Resultado del TratamientoRESUMEN
Since 9/11, there have been concerns that terrorists may detonate a radiological or nuclear device in an American city. Aside from several decorporation and blocking agents for use against internal radionuclide contamination, there are currently no medications within the Strategic National Stockpile that are approved to treat the immediate or delayed complications resulting from accidental exposure to radiation. Although the majority of research attention has focused on developing countermeasures that target the bone marrow and gastrointestinal tract, since they represent the most acutely radiosensitive organs, individuals who survive early radiation syndromes will likely suffer late effects in the months that follow. Of particular concern are the delayed effects seen in the lung that play a major role in late mortality seen in radiation-exposed patients and accident victims. To address these concerns, the National Institute of Allergy and Infectious Diseases convened a workshop to discuss pulmonary model development, mechanisms of radiation-induced lung injury, targets for medical countermeasures development, and end points to evaluate treatment efficacy. Other topics covered included guidance on the challenges of developing and licensing drugs and treatments specific to a radiation lung damage indication. This report reviews the data presented, as well as key points from the ensuing discussion.
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Descubrimiento de Drogas , Enfermedades Pulmonares/prevención & control , Pulmón/efectos de la radiación , Modelos Animales , National Institute of Allergy and Infectious Diseases (U.S.)/organización & administración , Guerra Nuclear , Traumatismos Experimentales por Radiación/prevención & control , Protectores contra Radiación/uso terapéutico , Liberación de Radiactividad Peligrosa , Terrorismo , Experimentación Animal/normas , Animales , Biomarcadores , Terapia Combinada , Descubrimiento de Drogas/organización & administración , Descubrimiento de Drogas/normas , Evaluación Preclínica de Medicamentos , Humanos , Enfermedades Pulmonares/tratamiento farmacológico , Enfermedades Pulmonares/etiología , Enfermedades Pulmonares/cirugía , Traumatismos Experimentales por Radiación/tratamiento farmacológico , Traumatismos Experimentales por Radiación/cirugía , Tolerancia a Radiación , Protectores contra Radiación/clasificación , Protectores contra Radiación/provisión & distribución , Proyectos de Investigación , Apoyo a la Investigación como Asunto , Trasplante de Células Madre , Estados Unidos , United States Food and Drug Administration , Poblaciones VulnerablesRESUMEN
Objetivo: Describir el esqueleto de la articulación del hombro en diferentes especies para establecer un modelo experimental de la patología del hombro. Material y metodología: Obtuvimos hombros de diferentes especies: perro (beagle), cerdo (mini-pig), conejo de Nueva Zelanda, liebre, cordero y oveja adulta. Extrajimos las articulaciones escápulo-humerales de ambos lados. Efectuamos una radiografía en cuatro proyecciones (antero-posterior, axial y dos oblicuas) y un TAC con reconstrucción tridimensional. Posteriormente disecamos las piezas para analizar el espacio subacromial y la cavidad glenoidea. Resultados: Ninguna de las especies analizadas posee un acromion desarrollado ni coracoides para definir un espacio subacromial. El cerdo presenta una cápsula posterior más desarrollada que la anterior y un m. subescapular desarrollado y el m. bíceps presenta un tendón en su porción larga pequeño y extraarticular con una corredera bicipital profunda. La oveja presenta un labrum superior desarrollado, con un tendón de la porción larga del m. bíceps en una corredera poco profunda. Conclusión: No hemos encontrado un modelo experimental adecuado para estudiar la patología del espacio subacromial. La oveja y el cerdo pueden ser buenos modelos para estudiar la patología de la glenoides (AU)
Objective: To describe the skeleton of the shoulder joint in different species to establish an experimental model of shoulder pathology. Material and methods: Shoulders were obtained from different species: dog (beagle), pig (minipig), New Zealand rabbit, hare, lamb and adult sheep. The scapulohumeral joints from both sides were extracted. X-ray images were obtained in 4 projections (anteroposterior, axial and two oblique) and a CT scan with 3D reconstruction. We then dissected the pieces to analyze the subacromial space and glenoid cavity. Results: None of the species analyzed possesses a developed acromion or coracoid to define a subacromial space. The pig has a more developed posterior than anterior capsule and a developed subscapularis muscle and the biceps muscle has a tendon on its extraarticular small long portion with a deep bicipital groove. The sheep has a developed superior labrum, with a tendon of the long portion of the biceps muscle in a shallow groove. Conclusion: We did not find an adequate experimental model to study the pathology of the subacromial space. The sheep and the pig may be good model to study the pathology of the glenoid (AU)
Asunto(s)
Animales , Masculino , Femenino , Traumatismos Experimentales por Radiación/diagnóstico , Traumatismos Experimentales por Radiación/cirugía , Modelos Animales , Manguito de los Rotadores/lesiones , Manguito de los Rotadores/cirugía , Manguito de los Rotadores , Articulación del Hombro/anatomía & histología , Articulación del Hombro , Manguito de los Rotadores/anatomía & histología , Manguito de los Rotadores/fisiopatología , Articulación del Hombro/fisiopatologíaRESUMEN
Radiotherapy often provides the only clinical recourse for those afflicted with primary or metastatic brain tumors. While beneficial, cranial irradiation can induce a progressive and debilitating decline in cognition that may, in part, be caused by the depletion of neural stem cells. Given the increased survival of patients diagnosed with brain cancer, quality of life in terms of cognitive health has become an increasing concern, especially in the absence of any satisfactory long-term treatments. To address this serious health concern we have used stem cell replacement as a strategy to combat radiation-induced cognitive decline. Our model utilizes athymic nude rats subjected to cranial irradiation. The ionizing radiation is delivered as either whole brain or as a highly focused beam to the hippocampus via linear accelerator (LINAC) based stereotaxic radiosurgery. Two days following irradiation, human neural stem cells (hNSCs) were stereotaxically transplanted into the hippocampus. Rats were then assessed for changes in cognition, grafted cell survival and for the expression of differentiation-specific markers 1 and 4-months after irradiation. Our cognitive testing paradigms have demonstrated that animals engrafted with hNSCs exhibit significant improvements in cognitive function. Unbiased stereology reveals significant survival (10-40%) of the engrafted cells at 1 and 4-months after transplantation, dependent on the amount and type of cells grafted. Engrafted cells migrate extensively, differentiate along glial and neuronal lineages, and express a range of immature and mature phenotypic markers. Our data demonstrate direct cognitive benefits derived from engrafted human stem cells, suggesting that this procedure may one day afford a promising strategy for the long-term functional restoration of cognition in individuals subjected to cranial radiotherapy. To promote the dissemination of the critical procedures necessary to replicate and extend our studies, we have provided written and visual documentation of several key steps in our experimental plan, with an emphasis on stereotaxic radiosurgey and transplantation.