Endogenous Production Of Growth Factors Research Articles

Versatile hydrogel facilitating angiogenesis and cell recruitments for enhanced calvarial bone regeneration

The repair of large-size cranial bone defects caused by traumatic brain injury (TBI) remains a substantial clinical challenge. On one hand, traditional bone implants with intrinsic brittle and poor recovery features hinder their immediate implantation for cranioplasty applications. On the other hand, using exogenous growth factors to enhance the osteo-bioactivity of bone implants often leads to efficacy, safety, and cost concerns. Thus far, the authors develop a growth factor-free pliable hydrogel with multiple functions for mediating endogenous growth factor production and stem cell functions in cranioplasty. The pliable hydrogels are based on GelMA networks, in which the mechanical properties and protein affinity were strengthened by the crosslinked poly (ethylene glycol) disuccinimidyl succinate (PEG-(SS)2), while the antioxidant capability and osteoinductivity were remarkedly enhanced through the decoration of magnesium-seamed C-propylpyrogallol[4]arene cages (PgC3Mg). In vitro and in vivo results confirmed that the versatile hydrogel with excellent biocompatibility and biodegradability can improve osteogenic differentiation and cranial bone regeneration by facilitating growth factor production, endogenous cell recruitment and angiogenesis. These findings indicate that the versatile hydrogels represent a potential avenue for developing growth factor-free pliable scaffolds in cranioplasty after TBI.

A Milled Microdevice to Advance Glia-Mediated Therapies in the Adult Nervous System.

Neurodegenerative disorders affect millions of adults worldwide. Neuroglia have become recent therapeutic targets due to their reparative abilities in the recycling of exogenous neurotoxins and production of endogenous growth factors for proper functioning of the adult nervous system (NS). Since neuroglia respond effectively to stimuli within in vivo environments on the micron scale, adult glial physiology has remarkable synergy with microscale systems. While clinical studies have begun to explore the reparative action of Müller glia (MG) of the visual system and Schwann Cells (ShC) of the peripheral NS after neural injury, few platforms enable the study of intrinsic neuroglia responses to changes in the local microenvironment. This project developed a low-cost, benchtop-friendly microfluidic system called the glia line system, or gLL, to advance the cellular study needed for emerging glial-based therapies. The gLL was fabricated using elastomeric kits coupled with a metal mold milled via conventional computer numerical controlled (CNC) machines. Experiments used the gLL to measure the viability, adhesion, proliferation, and migration of MG and ShC within scales similar to their respective in vivo microenvironments. Results illustrate differences in neuroglia adhesion patterns and chemotactic behavior significant to advances in regenerative medicine using implants and biomaterials, as well as cell transplantation techniques. Data showed highest survival and proliferation of MG and ShC upon laminin and illustrated a four-fold and two-fold increase of MG migration to dosage-dependent signaling from vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF), respectively, as well as a 20-fold increase of ShC migration toward exogenous brain-derived neurotrophic factor (BDNF), compared to media control. The ability to quantify these biological parameters within the gLL offers an effective and reliable alternative to photolithography study neuroglia in a local environment ranging from the tens to hundreds of microns, using a low-cost and easily fabricated system.

The double inhibition of endogenously produced BMP and Wnt factors synergistically triggers dorsal telencephalic differentiation of mouse ES cells.

Embryonic stem (ES) cells are becoming a popular model of in vitro neurogenesis, as they display intrinsic capability to generate neural progenitors that undergo the known steps of in vivo neural development. These include the acquisition of distinct regional fates, which depend on growth factors and signals that are present in the culture medium. The control of the intracellular signaling that is active at different steps of ES cell neuralization, even when cells are cultured in chemically defined medium, is complicated by the endogenous production of growth factors. However, this endogenous production has been poorly investigated so far. To address this point, we performed a high-throughput analysis of the expression of morphogens during mouse ES cell neuralization in minimal medium. We found that during their neuralization, ES cells increased the expression of members of Wnt, Fibroblast Growth Factor (FGF), and BMP families. Conversely, the expression of Activin/Nodal and Shh ligands was low in early steps of neuralization. In this experimental condition, neural progenitors and neurons generated by ES cells expressed a gene expression profile that was consistent with a midbrain identity. We found that endogenous BMP and Wnt signaling, but not FGF signaling, synergistically affected ES cell neural patterning, by turning off a profile of dorsal/telencephalic gene expression. Double BMP and Wnt inhibition allowed neuralized ES cells to sequentially activate key genes of cortical differentiation. Our findings are consistent with a novel synergistic effect of Wnt and BMP endogenous signaling of ES cells in inhibiting a cortical differentiation program.

Simple signaling molecules for inductive bone regenerative engineering.

With greater than 500,000 orthopaedic procedures performed in the United States each year requiring a bone graft, the development of novel graft materials is necessary. We report that some porous polymer/ceramic composite scaffolds possess intrinsic osteoinductivity as shown through their capacity to induce in vivo host osteoid mineralization and in vitro stem cell osteogenesis making them attractive synthetic bone graft substitutes. It was discovered that certain low crystallinity ceramics partially dissociate into simple signaling molecules (i.e., calcium and phosphate ions) that induce stem cells to endogenously produce their own osteoinductive proteins. Review of the literature has uncovered a variety of simple signaling molecules (i.e., gases, ions, and redox reagents) capable of inducing other desirable stem cell differentiation through endogenous growth factor production. Inductive simple signaling molecules, which we have termed inducerons, represent a paradigm shift in the field of regenerative engineering where they can be utilized in place of recombinant protein growth factors.

Properties of dehydrated human amnion/chorion composite grafts: Implications for wound repair and soft tissue regeneration

PURION(®) processed dehydrated human amnion/chorion membrane (dHACM; MiMedx Group, Marietta, GA) tissue products were analyzed for the effectiveness of the PURION(®) process in retaining the native composition of the amniotic membrane and preserving bioactivity in the resulting products. dHACM was analyzed for extracellular matrix (ECM) composition through histological staining and for growth factor content via multiplex ELISA arrays. Bioactivity was assessed by evaluating endogenous growth factor production by human dermal fibroblasts in response to dHACM and for thermal stability by mechanical tests and in vitro cell proliferation assays. Histology of dHACM demonstrated preservation of the native amnion and chorion layers with intact, nonviable cells, collagen, proteoglycan, and elastic fibers distributed in the individual layers. An array of 36 cytokines known to regulate processes involved in inflammation and wound healing were identified in dHACM. When treated with dHACM extracts, bioactivity was demonstrated through an upregulation of basic fibroblast growth factor, granulocyte colony-stimulating factor, and placental growth factor biosynthesis, three growth factors involved in wound healing, by dermal fibroblasts in vitro. After conditioning at temperatures ranging from -78.7 to +73.5°C, dHACM retained its tensile strength and ability to promote proliferation of dermal fibroblasts in vitro. Elution experiments demonstrated a soluble fraction of growth factors that eluted from the tissue and another fraction sequestered within the matrix. The PURION(®) process retains the native composition of ECM and signaling molecules and preserves bioactivity. The array of cytokines preserved in dHACM are in part responsible for its therapeutic efficacy in treating chronic wounds by orchestrating a "symphony of signals" to promote healing.

Atypical Hypertrophic Osteoarthropathy as a Presenting Complaint in a Non-Smoker

A 70-year-old non-smoker initially presented to accident and emergency with a painful left foot. This was an atypical presentation of hypertrophic osteoarthropathy (HPOA) secondary to grade 4 adenocarcinoma of the lung , highlighting the importance of routine chest x-ray in clinical assessment of atypical bone pain . Diagnosis was made using an isotope bone scan, the most sensitive method for detecting HPOA . The tumour and HPOA symptoms responded well to the first line chemotherapy regimen of cisplatin and pemetrexed. This supports the current understanding that treatment of the primary lung lesion will treat HPOA. The mechanism of this action is hypothesised to be by reduction of endogenous growth factor production, either by reduction in cell numbers producing the factor or direct inhibition of factor production: VEGF is emerging as the predominant growth factor causing HPOA. We believe the mechanism of cisplatin action on HPOA symptoms may be two fold; firstly, by direct reduction in VEGF secreting tumour bulk and secondly, by reduction of VEGF transcription. After disease progression of the lung tumours and recurrence of HPOA symptoms the patient received second line docetaxel. HPOA symptoms were unchanged but the chemotherapy stabilised the tumours. This lack of effect on HPOA symptoms may be due to a lack of tumour reduction or more speculatively the lack of suppression of VEGF transcription. HPOA symptoms were significantly improved when the patient was started on octreotide and current studies suggest that VEGF inhibition may be the mechanism of octretotide ’ s effect. doi: http://dx.doi.org/10.4021/jmc1057w

Atypical Hypertrophic Osteoarthropathy as a Presenting Complaint in a Non-Smoker

A 70-year-old non-smoker initially presented to accident and emergency with a painful left foot. This was an atypical presentation of hypertrophic osteoarthropathy (HPOA) secondary to grade 4 adenocarcinoma of the lung, highlighting the importance of routine chest x-ray in clinical assessment of atypical bone pain. Diagnosis was made using an isotope bone scan, the most sensitive method for detecting HPOA. The tumour and HPOA symptoms responded well to the first line chemotherapy regimen of cisplatin and pemetrexed. This supports the current understanding that treatment of the primary lung lesion will treat HPOA. The mechanism of this action is hypothesised to be by reduction of endogenous growth factor production, either by reduction in cell numbers producing the factor or direct inhibition of factor production: VEGF is emerging as the predominant growth factor causing HPOA. We believe the mechanism of cisplatin action on HPOA symptoms may be two fold; firstly, by direct reduction in VEGF secreting tumour bulk and secondly, by reduction of VEGF transcription. After disease progression of the lung tumours and recurrence of HPOA symptoms the patient received second line docetaxel. HPOA symptoms were unchanged but the chemotherapy stabilised the tumours. This lack of effect on HPOA symptoms may be due to a lack of tumour reduction or more speculatively the lack of suppression of VEGF transcription. HPOA symptoms were significantly improved when the patient was started on octreotide and current studies suggest that VEGF inhibition may be the mechanism of octretotide’s effect. J Med Cases. 2013;4(4):204-207 doi: https://doi.org/10.4021/jmc1057w

Induced chronic hypoxia negates the pro-angiogenic effect of surface immobilized heparin in a polyurethane porous scaffold

Porous scaffolds are frequently utilized in tissue regeneration. We have developed a polyurethane (PU) scaffold with a freely interconnecting porosity that can be modified with a covalently linked heparinized surface. The ability of this surface functionality to stimulate vessel and cellular growth into the PU scaffold has been evaluated by subcutaneous implantation of discs in the rat under normoxia and chronic hypoxia (hypobaric chamber) for 10 days. The heparinized surface alone was able to significantly increase vascularization and cellularization under normoxia (p < 0.05), but this response was negated by hypoxia. Addition of vascular endothelial growth factor to heparinized discs resulted in increased vascularity and cellularization under both conditions (p < 0.05). This suggests that endogenous growth factor production was limiting under chronic hypoxia but that an angiogenic response could still occur with exogenous delivery of factors.

Immunohistochemical study of the growth factors, aFGF, bFGF, PDGF-AB, VEGF-A and its receptor (Flk-1) during arteriogenesis

Growth factors are viewed as main arteriogenic stimulators for collateral vessel growth. However, the information about their native expression and distribution in collateral vessels is still limited. This study was designed to profile expression of acidic and basic FGF, platelet-derived growth factor (PDGF-AB) and vascular endothelial growth factor (VEGF-A) and its receptor, fetal liver kinase-1 (Flk-1) during arteriogenesis by confocal immunofluorescence in both dog ameroid constrictor model and rabbit arteriovenous shunt model of arteriogenesis. We found that: (1) in normal arteries (NA) in dog heart, aFGF, bFGF, and PDGF-AB all were mainly expressed in endothelial cells (EC) and media smooth muscle cells (SMC), but the expression of aFGF was very weak, with those of the other two being moderate; (2) in collateral arteries (CAs), aFGF, bFGF, and PDGF-AB all were significantly upregulated (P < 0.05); they were present in all the layers of the vascular wall and were 2.1, 1.7, and 1.9 times higher than that in NA, respectively; and (3) in NA in rabbit hind limb, VEGF-A was absent, Flk-1 was only weakly present in endothelial cells, but in one week CAs VEGF-A and Flk-1 were significantly increased in both shunt and ligation sides; this was more evident in the shunt-side CAs, 2.3, and 2 times higher than that in the ligation side, respectively. In conclusion, our data demonstrate for the first time that growth factors, aFGF, bFGF, and PDGF-AB are significantly upregulated in collateral vessels in dog heart, and enhanced VEGF-A and its receptor, Flk-1, are associated with rapid and lasting increased shear stress. These findings suggest that endogenous production of growth factors could be an important factor promoting collateral vessel growth.

Effects of FGF-2 and EGF removal on the differentiationof mouse neural precursor cells

Cell therapy for neurological disorders has advanced, and neural precursor cells (NPC) may become the ideal candidates for neural transplantation in a wide range of diseases. However, additional work has to be done to determine either the ideal culture environment for NPC expansion in vitro, without altering their plasticity, or the FGF-2 and EGF mechanisms of cell signaling in neurospheres growth, survival and differentiation. In this work we evaluated mouse neurospheres cultured with and without FGF-2 and EGF containing medium and showed that those growth factors are responsible for NPC proliferation. It is also demonstrated that endogenous production of growth factors shifts from FGF-2 to IGF-1/PDGFb upon EGF and FGF-2 withdrawal. Mouse NPC cultured in suspension showed different patterns of neuronal localization (core versus shell) for both EGF and FGF-2 withdrawal and control groups. Taken together, these results show that EGF and FGF-2 removal play an important role in NPC differentiation and may contribute to a better understanding of mechanisms of NPC differentiation. Our findings suggest that depriving NPC of growth factors prior to grafting might enhance their chance to effectively integrate into the host.

VEGF and odontoblast-like cells: Stimulation by low frequency ultrasound

Objective Vascular endothelial growth factor (VEGF) has been implicated in the regulation of dental pulp and dentine repair. Therapeutic ultrasound was shown to be effective for fracture repair. We investigated whether low frequency ultrasound influences the production of VEGF by odontoblast-like cells. Moreover, we examined the direct effects of VEGF on odontoblast-like cell proliferation. Design MDPC-23, an established odontoblast-like cell line, was exposed to increasing intensities of 30 kHz ultrasound using an ultrasonic tip probe. Results After 24 h cell culture, WST-1 analysis of cell viability and number showed a dose-dependent decrease in the number of viable cells with increasing ultrasound power. However, the relative concentration of VEGF as analysed by ELISA and normalised to cell number was significantly increased in the culture supernatants indicating an ultrasound-induced stimulation of odontoblastic VEGF secretion. Analysis of VEGF gene expression by sqRT-PCR revealed the expression of the main VEGF isoforms in the MDPC-23 cells, i.e. VEGF 120 and VEGF 164 as well as to a minor extent VEGF 188. Low power ultrasound increased gene expression of all VEGF isoforms. Addition of recombinant VEGF to the cell cultures significantly stimulated cell proliferation. Gene expression of the VEGF receptors Flt1/VEGFR1 and KDR/VEGFR2 was detected in the MDPC-23, suggesting the possibility that VEGF may act on the odontoblast-like cells in an autocrine manner. Conclusions Our results indicate that ultrasound promoted VEGF expression and production by odontoblast-like cells and that VEGF may have autocrine effects on these cells. It is proposed that ultrasound may influence odontoblast activity and dentine repair by modulating production of endogenous growth factors in the dentine-pulp complex.

Aortic rings stimulate inflammatory angiogenesis in a subcutaneous implant in vivo model

Rat or mouse aortic rings produce angiogenic outgrowths in vitro through endogenous production of growth factors and inflammatory cytokines. To further investigate this process in vivo, collagen-Gelfoam constructs containing aortic rings were implanted subcutaneously in syngeneic animals. Aortic rings stimulated a prominent angiogenic response characterized by peri- and intra-aortic accumulation of florid granulation tissue. Conversely, implants without rings elicited a non-specific inflammatory reaction without significant angiogenesis. The angiogenic response to the rings peaked at day 14 and was followed by regression of neovessels, which were mostly reabsorbed by day 28. Gene expression studies showed upregulated expression of angiogenic growth factors and cytokines in implants with rings. Tracking experiments with LacZ expressing ROSA26 transgenic mice demonstrated that both the aorta and the host contributed to the angiogenic response. These studies show that the angiogenic properties of the rodent aorta can be studied in the live animal under conditions that can be monitored and quantified. This in vivo assay can be used to study the molecular mechanisms by which the arterial wall and its proangiogenic cytokines regulate formation of granulation tissue during wound healing.

The use of heparan sulfate to augment fracture repair in a rat fracture model

Fracture healing is a complex process regulated by numerous growth and adhesive factors expressed at specific stages during healing. The naturally occurring, cell surface-expressed sugar, heparan sulfate (HS), is known to bind to and potentiate the effects of many classes of growth factors, and as such, may be a potential candidate therapy for enhancing bone repair. This study investigated the local application of bone-derived HS in the repair of rat femoral fractures. After 2 weeks, there was a significant increase in the callus size of rats administered with 5 microg HS compared to the control and 50 microg HS groups, presumably due to increased trabecular bone volume rather than increased cartilage production. In addition, 5 microg HS increased the expression of ALP, Runx2, FGF-1, IGF-II, TGF-beta1, and VEGF. It is hypothesized that these increases resulted from changes in HS-mediated receptor/ligand interactions that increase local growth factor production to augment bone formation. The findings of this study demonstrate the anabolic potential of HS in bone repair by recruiting and enhancing the production of endogenous growth factors at the site of injury.

Gene Therapy Approaches for Intervertebral Disc Degeneration

Review of the literature concerning the development of gene therapy approaches for the treatment of intervertebral disc degeneration. To provide an overview of gene therapy principles, summarize the results of completed gene therapy studies, and discuss considerations for the direction of future research. Intervertebral disc degeneration is characterized by progressive loss of the proteoglycan matrix in the nucleus pulposus. Exogenous growth factors have been shown to transiently increase matrix synthesis. Gene therapy offers exciting potential to induce and sustain endogenous production of growth factors within the intervertebral disc and thus possibly alter the degenerative course. Published and presented scientific literature was examined. Several in vitro and in vivo studies have documented the capacity of gene therapy to favorably modify the biologic functions of intervertebral disc cells with the delivery of the cDNA for various growth factors. Currently, investigators are exploring the efficacy and safety of gene therapy in animal models of degeneration. With promising initial results and an immense potential clinical impact, gene therapy approaches for treatment of intervertebral disc degeneration will continue to receive dedicated research efforts.

Mouse embryonic stem cells and preimplantation embryos require signaling through the phosphatidylinositol 3-kinase pathway to suppress apoptosis

Whereas most mammalian cells require extracellular signals to suppress apoptosis, preimplantation embryos can survive and develop to the blastocyst stage in defined medium without added serum or growth factors. Since cells of these embryos are capable of undergoing apoptosis, it has been suggested that their lack of dependence upon exogenous growth factors results from the production of endogenous growth factors that suppress apoptosis by an autocrine signaling mechanism. In the present study, we have examined the growth factor requirements and intracellular signaling pathways that suppress apoptosis in both mouse preimplantation embryos and embryonic stem (ES) cells, which are derived from the blastocyst inner cell mass. Cultured ES cells, in contrast to intact embryos, required serum growth factors to prevent apoptosis. Suppression of ES cell apoptosis by serum growth factors required the phosphatidylinositol 3-kinase (PI 3-kinase) signaling pathway, since apoptosis was rapidly induced by inhibition of PI 3-kinase with LY294002. In contrast, inhibition of MEK/ERK signaling with U0126 or of mTOR with rapamycin had no detectable effect on ES cell survival. Thus, like most mammalian cells, the survival of ES cells is mediated by growth factor stimulation of PI 3-kinase signaling. Treatment with LY294002 (but not with U0126 or rapamycin) similarly induced apoptosis of mouse blastocysts in serum-free medium, indicating that intact preimplantation embryos are also dependent upon PI 3-kinase signaling for survival. These results demonstrate that PI 3-kinase signaling is required to suppress apoptosis of both ES cells and intact preimplantation embryos, consistent with the hypothesis that survival of preimplantation embryos is maintained by endogenous growth factors that stimulate the PI 3-kinase pathway.

Vascular Growth Factor Expression in a Rat Model of Severe Limb Ischemia

Background. In ischemic tissue hypoxia induces production of vascular growth factors, especially VEGF, which initiate local angiogenesis. Collateralization—or arteriogenesis—occurs at a distance from the ischemic tissue and depends on different growth factors such as FGF-2. A spatial discrepancy in endogenous growth factor production in limb ischemia may have implications for therapeutic angiogenesis. The present study elucidates if such spatial and temporal variation occurs. Materials and methods. A two-staged procedure was performed to generate severe long-lasting limb ischemia in 60 rats. At 1, 7, 28, and 56 days, subgroups were subjected to perfusion assessment with laser Doppler imaging and angiography. Muscle samples and foot skin were gathered to measure growth factor expression and signs of angiogenesis using immunohistochemistry. Results. There was an early twofold increase ( P < 0.05) in both VEGF and FGF-2 levels in distal muscle from the ischemic leg, but no significant rise in the thigh. The concentrations decreased over time with an exception for VEGF in soleus and FGF-2 in anterior tibial muscle, which remained high. An increased capillarity was noted ( P < 0.05) in soleus after 28 days, and the number of BrdU-positive ECs was elevated in all ischemic samples at 56 days. Collateral arteries were observed on the angiograms after 7 days. Conclusions. The results suggest that in limb ischemia any major increase in vascular growth factor production is limited to ischemic tissue. The spatial and temporal distribution patterns of growth factor production are complex and to a great extent influenced by inflammation.

The temporal course of adenovirus-mediated gene transfer in the rat larynx.

Polypeptide growth factors have important influences on wound-healing and scar tissue formation. Specific growth factors or their inhibitors may potentially decrease scar tissue formation and prevent subglottic stenosis. Gene transfer using recombinant adenovirus may be an ideal method to mediate endogenous production of growth factors to inhibit fibrosis. The study incorporated adenovirus-mediated transduction of normal and stenotic rat larynges and histologic analysis of the sequential expression of a beta-galactosidase marker gene over time. The study was conducted at the animal care facility of an academic children's hospital. We report successful transduction in normal and injured rat larynx with peak expression of beta-galactosidase at 2 days after transduction and almost complete disappearance by 7 days. There appeared to be an early inflammatory response to the viral injection, but at 7 and 14 days after injection (transduction) the uninjured rat larynges resumed a normal histologic appearance. All distant sites stained negative for beta-galactosidase. Recombinant adenovirus-mediated gene transfer is feasible in the rat larynx with transient duration and limited toxicity. Adenovirus-mediated gene transfer has the potential to deliver growth factors that modulate wound healing and inflammation in the larynx by inhibiting fibrosis.

Role of the renin-angiotensin system in vascular diseases: expanding the field.

The renin-angiotensin system (RAS) has emerged as one of the essential links in the pathophysiology of vascular disease. Angiotensin (Ang) II, the main peptide of the RAS, was considered as a vasoactive hormone, but in the past years, this view has been modified to a growth factor that regulates cell proliferation/apoptosis and fibrosis. Recently, this view has been enlarged with a novel concept: Ang II participates in the inflammatory response, acting as a proinflammatory mediator. In resident vascular cells, Ang II produces chemokines, cytokines, and adhesion molecules, which contribute to the migration of inflammatory cells into the tissue injury. Ang II is also a chemotactic and mitogenic factor for mononuclear cells. The molecular mechanisms of Ang II-induced vascular damage are mediated by the activation of transcription factors, redox signaling systems, and production of endogenous growth factors. In addition, other components of the RAS could also be involved in the pathogenesis of cardiovascular diseases. The Ang II degradation product Ang III shares some of its properties with Ang II, including chemotaxis and production of growth factors and chemokines. All these data clearly demonstrate that Ang II is a true cytokine, show the complexity of the RAS in pathological processes, and provide some mechanistic responses of the beneficial effects of the treatment with RAS blockers in cardiovascular diseases.

Apoptosis in Chronic Myelogenous Leukemia: Studies of Stage-Specific Differences

In this study we compared rates of apoptosis, survival and metabolic activity from CML peripheral blood neutrophils with peripheral blood and bone marrow neutrophils from healthy volunteer donors and studied the influence of the disease stage and of cytokines including G-CSF, GM-CSF and IL-1β on these parameters. Quantification of apoptosis by morphology, diphenylamine DNA fragmentation assay and by gel electrophoresis showed similar rates of apoptosis in chronic phase CML and normal peripheral blood neutrophils when the cells were incubated in RPMI + FCS or in serum-free medium. However, there were lower rates of apoptosis in accelerated and blast phase CML neutrophils (p <. 001) and in normal bone marrow neutrophils (p <. 001) compared to normal peripheral blood neutrophils (incubated in RPMI + FCS). Survival among neutrophils from chronic phase or accelerated/blast phase CML patients was significantly longer (p < 0.002 and p < 0.001, respectively) than among normal peripheral blood neutrophils but neutrophils purified from normal bone marrow had a survival rate which fell between that of normal peripheral blood and chronic phase CML patients. When the cells were incubated in RPMI + autologous sera, neutrophils from chronic phase CML patients showed markedly lower rates of apoptosis (p < 0.001) and maintained higher metabolic activities (p < 0.002) compared to normal neutrophils. G-CSF and GM-CSF were found to considerably decrease the rate of apoptosis in chronic phase CML neutrophils (p < 0.001 and p = 0.008 respectively) while IL-1β did not show any antiapoptotic effect. It is suggested that the endogenous production of growth factors may therefore participate in delaying apoptotic cell death, during the progression of CML.

In vivo localization of the insulin-like growth factors I and II (IGF I and IGF II) gene expression during human lung development.

The insulin-like growth factors I and II (IGF I and IGF II) are synthesized in many organs during human development and are involved in the growth and differentiation of tissues. Correlations between lung growth and maturation and the local production of IGFs have been poorly explored in humans. Using in situ hybridization we localized the synthesis of IGFs in the human fetal respiratory tract over an extended period of the gestation and we demonstrated time dependent changes. IGF mRNAs were expressed throughout gestation with a clear predominance of IGF II and a decreasing expression of both IGFs after the 20th week of gestation. They were mainly detected in the mesodermal-derived components of the respiratory tract, especially in the undifferentiated mesenchyme of the lung buds up to 20 weeks of gestation. At this time the local production of collagen and the proliferation of adjacent epithelial cells were predominant features. Later, mesenchymal hybridization decreased. Weak epithelial hybridization was observed during the first stages of growth and progressively decreased when the epithelium underwent maturation: early in the trachea, later in the distal lung buds. A consistent expression of IGF II, but not IGF I, in the endothelium, throughout gestation, was also observed. The IGFs may act on the near epithelial cell proliferation in both autocrine and paracrine ways. They may also stimulate the maturation of the connective tissue. This endogenous production of growth factors may play a role in the somatic growth during prenatal life.