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Fatigue debonding of the roughened stem-cement interface: effects of surface roughness and stem heating conditions.

The aim of this study was to determine the effects of cyclic loading on the debond process of a roughened stem-cement interface used in total hip arthroplasty. The specific goals were to assess the effects of two surgeon-controlled variables (stem heating and degree of stem surface roughness) and to determine if an independent finite element-based fracture mechanics model could be used to predict the debond response. A clamped cantilever beam geometry was used to determine the fatigue debond response of the stem-cement interface and was created using an experimental mold that simulated in vivo cementing conditions. A second experiment was performed using a torsion-loading model representative of the stem-cement-bone composite. For both experiments, two stem heating (room temperature and 50 degrees C) and surface roughness conditions (grit blasted: Ra = 2.3 and 5.1 microm) were used. Finally, a finite element model of the torsion experiment with provision for crack growth was developed and compared with the experimental results. Results from both experiments revealed that neither stem preheating nor use of a stem with a greater surface roughness had a marked effect on the fatigue debond response. There was substantial variability in the debond response for all cases; this may be due to microscopic gaps at the interface for all interface conditions. The debond rate from the finite element simulation (10(-7.31) m/cycle) had a magnitude similar to the experimental torsion model (10(-(6.77 +/- 1.25)) m/cycle). This suggests that within the context of the experimental conditions studied here that the debond response could be assessed using a linear elastic fracture mechanics-type approach.

Biocompatible Materials↗

Migration of cemented stem and restrictor after total hip arthroplasty: a radiostereometry study of 25 patients with Lubinus SP II stem.

The aim of this study was to investigate the migration pattern of the Lubinus SP II prosthesis stem. Migration at the stem-cement and at the bone-cement interface was assessed. Twenty-five patients were studied by radiostereometry during a period of 2 years. The migration of the head, of 2 points on the collar, of the stem tip, and also of the cement restrictor, was measured. The collar and the head were found stable at 2 years' follow-up, whereas an anterior migration occurred at the stem tip (median, 0.3 mm; 25th percentile, 0.04 mm; 75th percentile, 1.27 mm). Virtually no subsidence or axial rotation of the stems was observed. An anterior migration of the restrictor marker was also observed, compatible with a rigid sagittal plane rotation of the stem and of the cement mantle about the collar. Therefore, stem tip migration occurred at the bone-cement interface. Key words: total hip arthroplasty, anatomic stem, radiostereometry, subsidence, bone-cement interface.

Aged↗

Flower Formation in Excised Tobacco Stem Segments: III. Deoxyribonucleic Acid Content in Stem Tissue of Vegetative and Flowering Tobacco Plants.

A method has been developed that extracts DNA from stem tissue of flowering tobacco plants, Nicotiana tabacum cv. Wis. 38. The DNA content of stem tissue from a flowering tobacco plant is correlated with its capacity to flower in vitro. Stem segments known to form 100% floral buds contain 10 times more DNA per gram fresh weight than segments that form 5% floral buds and 95% vegetative buds, and in the uppermost 28 centimeters of flowering tobacco plant stems the DNA content decreases roughly in parallel with the floral gradient.((3)H)-Thymidine incorporation into DNA in the internodes is inhibited by attached leaves in flowering tobacco plants but promoted in vegetative plants. Low indoleacetic acid concentrations (2.8 and 5.7 mum) inhibit DNA synthesis in internodes of defoliated stems of flowering tobacco plants to the same extent as attached leaves, whereas the same concentrations of indoleacetic acid promote DNA synthesis in internodes of defoliated stems of vegetative plants. The optimal concentration (11.4 mum) of indoleacetic acid supplied to defoliated stems of vegetative plants increased the rate of DNA synthesis to 4 times the rate in defoliated stems without indoleacetic acid treatment. This increase more than compensates for the promotion of DNA synthesis by the young leaves. Thus, the opposite effects of young leaves on DNA synthesis in internodes of flowering and vegetative tobacco plants may be related to their auxin content.

Journal Article↗

Femoral cement pressurization during total hip arthroplasty. The role of different femoral stems with reference to stem size and shape.

Aseptic loosening is the most common cause of femoral component failure following total hip arthroplasty (THA). Improvements have been made in cementing technique, but little attention has been directed toward the effect of different femoral components on bone cement pressurization. The effect of three different femoral stem sizes in one femoral canal and of eight different femoral stem designs on bone cement extrusion pressures was investigated on autopsy specimens. In a given femur significant increases in bone cement pressurization were achieved about the distal two-thirds of the femoral component in progressing from a small to a medium to a large stem. Similarly, femoral stems with rounded, rectangular cross sections generated greater bone cement pressurization about the distal two-thirds of the stems than stems trapezoidal and diamond-shaped in cross section due to their increased stem volume. The results of this study favor the use of the largest femoral component stem, preferably one with a rounded, rectangular cross section, that will fit comfortably in a given femoral canal and still allow a surrounding cement envelope such that bone cement pressurization will be enhanced with implant introduction.

Cadaver↗

Isolation and cloning of multipotential stem cells from the embryonic human CNS and establishment of transplantable human neural stem cell lines by epigenetic stimulation.

Stem cells that can give rise to neurons, astroglia, and oligodendroglia have been found in the developing and adult central nervous system (CNS) of rodents. Yet, their existence within the human brain has not been documented, and the isolation and characterization of multipotent embryonic human neural stem cells have proven difficult to accomplish. We show that the developing human CNS embodies multipotent precursors that differ from their murine counterpart in that they require simultaneous, synergistic stimulation by both epidermal and fibroblast growth factor-2 to exhibit critical stem cell characteristics. Clonal analysis demonstrates that human C NS stem cells are multipotent and differentiate spontaneously into neurons, astrocytes, and oligodendrocytes when growth factors are removed. Subcloning and population analysis show their extensive self-renewal capacity and functional stability, their ability to maintain a steady growth profile, their multipotency, and a constant potential for neuronal differentiation for more than 2 years. The neurons generated by human stem cells over this period of time are electrophysiologically active. These cells are also cryopreservable. Finally, we demonstrate that the neuronal and glial progeny of long-term cultured human CNS stem cells can effectively survive transplantation into the lesioned striatum of adult rats. Tumor formation is not observed, even in immunodeficient hosts. Hence, as a consequence of their inherent biology, human CNS stem cells can establish stable, transplantable cell lines by epigenetic stimulation. These lines represent a renewable source of neurons and glia and may significantly facilitate research on human neurogenesis and the development of clinical neural transplantation.

Animals↗

The happy destiny of frozen haematopoietic stem cells: from immature stem cells to mature applications.

Forty years ago, van Putten described in the European Journal of Cancer (see this issue) quantitative studies on the optimal storage techniques of mouse and monkey bone marrow suspensions. Survival of the animals after irradiation following injection with stored bone marrow cell suspensions was the endpoint. He observed some species differences, but based on the data obtained considered a careful trial of the glycerol-polyvinylpyrrolide (PVP) combination for storage of marrow in man was indicated. In spite of this, dimethyl sulphoxide has become the 'standard' cryopreservant for human marrow stem cells. Over the last 40 years, there has been a tremendous increase in knowledge about haematopoietic stem cells and their use in the clinic. Haematopoietic stem cells are now known to travel between the bone marrow and peripheral blood and are the best-characterised adult stem cells. These cells are currently widely used for transplantations in the clinic and are obtained from a wide variety of sources. These include the bone marrow, peripheral blood, cord blood, autologous as well as allogeneic stem cells from related or unrelated donors. Increasingly, data has become available that adult haematopoietic stem cells can generate differentiated cells belonging to other cell types, a process called "developmental plasticity". Thus, they may contribute to non-haematopoietic tissue repair in multiple organ systems. This has created a whole new potential therapeutic armamentarium for the application of haematopoietic stem cells outside of the area of malignancies and haematopoietic disorders.

Adult↗

Hypothesis: cell plasticity, linking embryonal stem cells to adult stem cell reservoirs and metastatic cancer cells?

Embryonal stem (ES) cells are the earliest ontogenetically identifiable stem cells of the embryo proper for all subsequent mesenchymal stem cells and for highly specialized differentiated cells. This review characterizes, in a working hypothesis, the role of reversible EMT/MET (epithelialmesenchymal transition) as a manifestation of cell plasticity 1) in the development of ES cells to adult stem cells (hematopoietic stem cells) and 2) in metastasizing cancer cells. Animal studies support the concept that EMT/MET is a key manifestation of cell plasticity in the development of ES cells to adult stem cells, and in conversion of localized to metastasizing cancer cells. In fact, ES cells may persist to postnatal life, in cytologically verifiable form and/or within the frame of EMT/MET, as ultimate reservoir for adult stem cells. Furthermore, EMT could possibly serve as a conceptional link between physiologic and pathologic signaling pathways. Clonal confirmation in humans is necessary.

Adult↗

Stem cell sources for clinical islet transplantation in type 1 diabetes: embryonic and adult stem cells.

Lifelong immunosuppressive therapy and inadequate sources of transplantable islets have led the islet transplantation benefits to less than 0.5% of type 1 diabetics. Whereas the potential risk of infection by animal endogenous viruses limits the uses of islet xeno-transplantation, deriving islets from stem cells seems to be able to overcome the current problems of islet shortages and immune compatibility. Both embryonic (derived from the inner cell mass of blastocysts) and adult stem cells (derived from adult tissues) have shown controversial results in secreting insulin in vitro and normalizing hyperglycemia in vivo. ESCs research is thought to have much greater developmental potential than adult stem cells; however it is still in the basic research phase. Existing ESC lines are not believed to be identical or ideal for generating islets or beta-cells and additional ESC lines have to be established. Research with ESCs derived from humans is controversial because it requires the destruction of a human embryo and/or therapeutic cloning, which some believe is a slippery slope to reproductive cloning. On the other hand, adult stem cells are already in some degree specialized, recipients may receive their own stem cells. They are flexible but they have shown mixed degree of availability. Adult stem cells are not pluripotent. They may not exist for all organs. They are difficult to purify and they cannot be maintained well outside the body. In order to draw the future avenues in this field, existent discrepancies between the results need to be clarified. In this study, we will review the different aspects and challenges of using embryonic or adult stem cells in clinical islet transplantation for the treatment of type 1 diabetes.

Adult↗

Use of fluorescence in situ hybridization to detect minimal residual disease in hematopoietic stem cell assays from peripheral blood stem cells of 2 patients with trisomy 8 acute myeloid leukemia.

Fluorescence in situ hybridization (FISH) was used to analyze peripheral blood stem cells (PBSC) and stem cell assays (SCA) derived from them in 2 patients with acute myeloid leukemia (AML) with trisomy 8 as the sole chromosome abnormality prior to undergoing autologous stem cell transplantation. In both cases, the demonstration of cells containing trisomy 8 in the stem cell product led to significant changes in the patients' treatment. In the initial PBSC collections from each patient, trisomy 8 was found in aspirated granulocyte-monocyte (GM) colonies or aspirated GM clusters but not entire cell populations of SCA dish or uncultured PBSC (one patient). FISH analyses for specific cytogenetic abnormalities in hematopoietic stem cell cultures may be a more useful means of assessing the quality of the stem cell product in patients being considered for autologous stem cell transplantation.

Adolescent↗

Genetic selection of mouse male germline stem cells in vitro: offspring from single stem cells.

Spermatogenesis originates from a small population of spermatogonial stem cells. These cells are believed to divide infinitely and support spermatogenesis throughout life in the male. In this investigation, we examined the possibility of deriving transgenic offspring from single spermatogonial stem cells. Spermatogonial stem cells were transfected in vitro with a plasmid vector containing a drug resistant gene. Stably transfected stem cell clones were isolated by in vitro drug selection; these clones were expanded and used to produce transgenic progeny following spermatogonial transplantation into infertile recipients. An average of 49% of the offspring carried the transgene, and the recipient mice continued to produce monoclonal transgenic progeny a year after transplantation. Thus, a somatic cell-based genetic approach can be used to modify and select clones of spermatogonial stem cells in a manner similar to embryonic stem cells. The feasibility of genetic selection using postnatal spermatogonial stem cells demonstrates their extensive proliferative potential and provides the opportunity to develop new methods for generating stable animal transgenics or for germline gene therapy.

Animals↗

Blood stem cells compared with bone marrow as a source of hematopoietic cells for allogeneic transplantation. IBMTR Histocompatibility and Stem Cell Sources Working Committee and the European Group for Blood and Marrow Transplantation (EBMT).

Peripheral blood cells are increasingly used in place of bone marrow as a source of hematopoietic stem cells for allogeneic transplantation. The relative efficacy of these 2 approaches is unknown. This retrospective multivariate analysis compared results of 288 HLA-identical sibling blood stem cell transplantations with results of 536 HLA-identical sibling bone marrow transplantations. No transplants were T-cell depleted. Median follow-up was 12 months, and analyses focused on 1-year outcomes. Recipients of blood stem cell transplants had more rapid recovery of neutrophils to at least 0.5 x 10(9)/L (median time to recovery, 14 days, compared with 19 days for marrow transplants; P <.001) and of platelets to at least 20 x 10(9)/L (median time, 18 days, compared with 25 days for marrow transplants; P <.001). There was no significant difference in the incidence of grades II to IV acute graft versus host disease (GVHD). The incidence of chronic GVHD was significantly higher after blood stem cell transplantation (1-year probability [95% confidence interval], 65% [56%-72%] compared with 53% [47%-59%]; P =.02) Relapse incidence in the 2 transplant groups did not differ significantly. Treatment-related mortality rates were lower and leukemia-free survival rates were higher with blood stem cell transplants in patients with advanced leukemia (acute leukemia in second remission or chronic myelogenous leukemia in accelerated phase) but not in early leukemia (acute leukemia in first remission or chronic myelogenous leukemia in chronic phase). The median time from transplantation to hospital discharge was 23 days after blood stem cell transplantation and 28 days after bone marrow transplantation (P =.003). Further study with longer follow-up is necessary to definitively establish the role of blood stem cells for allogeneic transplantation, especially in patients with good-risk disease. (Blood. 2000;95:3702-3709)

Acute Disease↗

[Methodologies for isolation, expansion and differentiation of somatic stem cells(tissue stem cells)].

Recent studies have shown that pluriponten stem cells are also found in adult tissue. These adult stem cells has a great advantage over other stem cells sources, human embryos or human fetal tissue, that trouble many people on ethical grounds. While the adult stem cells have great potential for use in the cell therapy, we must pass numerous technical hurdles to reach a goal. Here we describe a subset of problems encountered in isolation and expansion of somatic stem cells for clinical application. As it is necessary to obtain the enough number of cells to supply the defect tissue and organ, we required purification and ex vivo expansion of stem cells, overcoming senescence. We also refer to the argument about plasticity of adult stem cells.

Adult↗

The role of stem cell source in autologous hematopoietic stem cell transplantation for patients with myelodysplastic syndromes.

BACKGROUND AND OBJECTIVES: Intensive chemotherapy followed by autologous hematopoietic stem cell transplantation (HSCT) is a curative treatment option for patients with myelodysplastic syndromes (MDS). Peripheral blood (PB) HSCT was introduced in 1992 and PB has become the source of choice of autologous stem cells worldwide. Autologous PB stem cells result in faster hematopoietic recovery, but may be associated with a higher risk of relapse. DESIGN AND METHODS: We analyzed the data of 336 patients transplanted after 1992 with either bone marrow (BM) (n=104) or PB (n=232). RESULTS: Various factors had an impact on event-free survival in univariate analysis: age hazard ratio [HR]=1.1 per 10 years; p=0.12), source of stem cells (HR=1.2, p=0.22), interval between diagnosis and transplantation (HR=1.0 per month; p=0.87), and therapy-related vs primary disease (HR=0.5; p=0.002). In the multivariate Cox model, the event-free survival was not different after PB or BM HSCT with a HR of 0.93 (95% confidence interval of 0.67 - 1.30; p=0.67). The relapse risk after transplantation with stem cells from either source was similar with a HR of 1.1. A significant interaction (p=0.02) between age and the source of stem cells indicated a more favorable potential of autologous PB HSCT in young age groups. INTERPRETATION AND CONCLUSIONS: Autologous PB and BM HSCT result in equivalent outcomes. Therefore, given the more rapid hematopoietic recovery PB is the preferred source of stem cells.

Analysis of Variance↗

Stem cell factor enhances the survival of murine intestinal stem cells after photon irradiation.

Recombinant rat stem cell factor (SCF) has been shown to decrease lethality in mice exposed to total-body irradiation (TBI) in the lower range of lethality through radioprotection of hematopoietic stem cells and acceleration of bone marrow repopulation. This study evaluates the effect of SCF on the survival of the intestinal mucosal stem cell after TBI. This non-hematopoietic stem cell is clinically relevant. Gastrointestinal toxicity is common during and after abdominal and pelvic radiation therapy and limits the radiation dose in these regions. As observed with bone marrow, the administration of SCF to mice prior to TBI enhanced the survival of mouse duodenal crypt stem cells. The maximum enhancement of survival was seen when 100 micrograms/kg of SCF was given intraperitoneally 8 h before irradiation. This regimen increased the survival of duodenal crypt stem cells after 12.0 Gy TBI from 22.5 +/- 0.7 per duodenal cross section for controls to 30.0 +/- 1.7 after treatment with SCF (P = 0.03). The TBI dose producing 50% mortality at 6 days (LD50/6) was increased from 14.9 Gy for control mice to 19.0 Gy for mice treated with SCF (dose modification factor = 1.28). These findings demonstrate that SCF has radioprotective effects on a non-hematopoietic stem cell population and suggest that SCF may be of clinical value in preventing radiation injury to the intestine.

Animals↗

Stem cells from midguts of Lepidopteran larvae: clues to the regulation of stem cell fate.

Previously, we showed that isolated stem cells from midguts of Heliothis virescens can be induced to multiply in response to a multiplication protein (MP) isolated from pupal fat body, or to differentiate to larval types of mature midgut cells in response to either of 4 differentiation factors (MDFs) isolated from larval midgut cell-conditioned medium or pupal hemolymph. In this work, we show that the responses to MDF-2 and MP in H. virescens stem cells decayed at different time intervals, implying that the receptors or response cascades for stem cell differentiation and multiplication may be different. However, the processes appeared to be linked, since conditioned medium and MDF-2 prevented the action of MP on stem cells; MP by itself appeared to repress stem cell differentiation. Epidermal growth factor, retinoic acid, and platelet-derived growth factor induced isolated midgut stem cells of H. virescens and Lymantria dispar to multiply and to differentiate to mature midgut cells characteristic of prepupal, pupal, and adult lepidopteran midgut epithelium, and to squamous-like cells and scales not characteristic of midgut tissue instead of the larval types of mature midgut epithelium induced by the MDFs. Midgut stem cells appear to be multipotent and their various differentiated fates can be influenced by several growth factors.

Animals↗

Characterization of a population of cells in the bone marrow that phenotypically mimics hematopoietic stem cells: resting stem cells or mystery population?

We have identified a population of cells in murine bone marrow that has many of the phenotypic characteristics attributed to resting hematopoietic stem cells but does not reconstitute irradiated mice. These cells express high levels of Sca-1, H-2K and CD38 and low levels of Thy-1.1, but do not express CD34 nor any of the lineage markers including CD3, CD4, CD5, CD8 NK1.1, I-A, B220, Ig(MGA), CD40, kappa, Mac-1, Gr-1 or Ter119. In addition, this population can be found at normal frequency in nu/nu as well as rag-1-/- mice. These cells incorporate only low levels of Rh123, are resistant to the cytotoxic effects of 5-fluorouracil and, consistent with their resting phenotype, less than 2% of these cells are in the S/G2/M phases of the cell cycle. The only phenotypic characteristic that distinguishes these cells from the lineage- Sca-1+, Thy-1.1low long-term reconstituting hematopoietic stem cell population is their lack of c-kit expression. Here we have explored the possibility that these cells represent a truly resting population of hematopoietic stem cells. We found that the lineage-, Sca-1+, c-kit- cells do not respond to hematopoietic growth factors in vitro, either alone or in combination with stromal layers. Furthermore, these cells do not form in vivo spleen colonies nor do they have the ability to reconstitute irradiated mice. Thus, this population may represent either a population of resting stem cells for which we lack the appropriate activating stimulus, or simply represent a "mystery population" that phenotypically mimics most of the physical properties of resting stem cells. Given the close phenotypic similarity of the c-kit- mystery population cells to the c-kit+ long-term reconstituting stem cells, investigators must be rigorous to exclude their effects from other stem cell assays.

ADP-ribosyl Cyclase↗

Survival of mammary stem cells in suspension culture: implications for stem cell biology and neoplasia.

There is increasing evidence that a variety of neoplasms including breast cancer may result from transformation of normal stem and progenitor cells. In the past, isolation and characterization of mammary stem cells has been limited by the lack of suitable culture systems able to maintain these cells in an undifferentiated state in vitro. We have recently described a culture system in which human mammary stem and progenitor cells are able to survive in suspension and produce spherical colonies composed of both stem and progenitor cells. Recent observation that adult stem cells from other tissues may also retain the capacity for growth under anchorage independent conditions suggests a common underlying mechanism. We propose that this mechanism involves the interaction between the canonical Wnt signal pathway and E-cadherin. The Wnt pathway has been implicated in normal stem cell self-renewal in vivo. Furthermore, there is evidence that deregulation of this pathway in the mammary gland and other organs may play a key role in carcinogenesis. Thus, the development of in vitro suspension culture systems not only provides an important new tool for the study of mammary cell biology, but also may have important implications for understanding key molecular pathways in both normal and neoplastic stem cells.

Animals↗

Abnormal brain-stem function (brain-stem auditory evoked response) correlates with acoustic cry features in term infants with hyperbilirubinemia.

We hypothesized that changes in brain-stem auditory evoked responses related to bilirubin would be associated with changes in cry because of the anatomic proximity in the brain stem of cranial nerves 8 (auditory) and 9 to 12 (vagal complex, which controls cry). Brain-stem auditory evoked responses and computerized cry analysis were used to study the concurrent effects of moderate hyperbilirubinemia on auditory function and cry. Fifty term infants were divided equally into two groups on the basis of serum bilirubin concentrations: low (less than 8 mg/dl; 136) mumol/L and moderate (10 to 20 mg/dl, 170 to 342 mumol/L). Forty-three infants had successful tracings of brain-stem auditory evoked responses recorded with a Cadwell model 5200A evoked response unit during two successive trials, and a cry recording of each infant was analyzed by computer. The moderate serum bilirubin group had an increase in percent cry phonation (p less than 0.02) and an increase in the variability of the first formant (p less than 0.04) in comparison with the low serum bilirubin group. Serum bilirubin values correlated positively with brain-stem conduction time (r = 0.36, p less than 0.01), percent phonation (r = 0.42, p less than 0.004), and variability of the first formant (r = 0.39, p less than 0.02). Percent phonation, the voiced component produced by increased neural control, correlated with the interpeak of waves latencies I to III (r = 0.32, p less than 0.03) and brain-stem conduction time (wave I to V) (r = 0.35, p less than 0.01). We conclude that hyperbilirubinemia affects adjoining areas of the brain stem that control hearing and cry production.

Brain Stem↗