Application of Mesenchymal Stem Cells for repair and regeneration of cartilage and bone

Australian efforts to provide orthopaedic surgeons with living, load-bearing scaffolds suitable for current joint (knee and hip) replacement surgery, non-union fracture repair, and miniscal and growth plate cartilage regeneration are being lead by teams at the Institute for Medical and Veterinary Science and Women's and Children's Hospital in Adelaide; the Peter MacCallum and St Vincent's Medical Research Institutes in Melbourne; and the Mater Medical Research Institute and new Institute for Health and Biomedical Innovation at QUT, Brisbane. In each case multidisciplinary teams are attempting to develop autologous living tissue constructs, utilising mesenchymal stem cells (MSC), with the intention of effecting seamless repair and regeneration of skeletal trauma and defects. In this article we will briefly review current knowledge of the phenotypic properties of MSC and discuss the potential therapeutic applications of these cells as exemplified by their use in cartilage repair and tissue engineering based approaches to the treatment of skeletal defects.

Spatial/surveying programs repositioning within new STEM Faculty at QUT

Many of the undergraduate and postgraduate programs of the former Faculty of Built Environment and Engineering PLUS Faculty of Sciences and Technology are changing as a result of merging these two large organisations, with some disciplines relocating to faculties of Creative Industries and Health respectively. The new STEM precinct under construction has begun rising from the proverbial hole-in-the-ground. Existing Surveying and Spatial Sciences programs, assets and staff are being repositioned with the newly formed School of Earth, Environment and Biological Sciences.2011. Golden graduates morning tea organised by QUT Alumni. Technology upgrades to the Mapping Sciences lab benefits 3-D learning experiences. Second and third-year students are undertaking Work Integrated Learning (WIL) over the summer vacation period. Final year students recently presented capstone project presentations at mini-conference in the Gibson Rooms overlooking a vibrant Southbank and sparkling Brisbane River. Discussion on end of year graduation ceremony held at QPAC.

Perceived gender differences in STEM learning in the middle school

Women are underrepresented in science, technology, engineering and mathematics (STEM) university coursework, reflecting long-standing gender issues that have existed in core middle-school STEM subject areas. Using data from a survey and written responses, we report on findings following the introduction of engineering education in middle school classes across three schools (grade level 7, n=122). The engineering experiences fused science, technology and mathematics concepts. The survey revealed higher percentages for girls than boys in 13 of the 24 items; however there were six items with a 20% difference in their perceptions about learning in STEM. For instance, despite girls recording that they have been provided equal or more opportunities than boys in STEM, they believed they do not do as well as boys (80% boys, 48% girls) or want to seek a career in STEM (39% boys, 17% girls). The written responses revealed gender differences across a number of themes in the students’ responses, including resources, group work, the nature and type of learning experiences, content knowledge, and teachers’ instructional style. Exposing students to STEM education facilitates an awareness of their learning and may assist girls to consider studying STEM subjects or STEM careers.

A brief history of haemotopoietic stem cell Ex vivo expansion

Haematopoiesis is the process by which a hierarchy of mature and progenitor blood cells are formed. These cell populations are all derived from multipotent haematopoietic stem cells (HSC), which reside in the bone marrow ‘niche’ of adult humans. Over the lifetime of a healthy individual, this HSC population replenishes between 1010-1011 blood cells on a daily basis. Dysregulation of this system can lead to a number of haematopoietic diseases, including aplastic anaemias and leukaemias, which result in, or require for disease resolution, bone marrow cell depletion. In 1956, E. Donnall Thomas demonstrated that haematopoiesis could be restored by transplanting bone marrow-derived cells from one man into his identical twin brother, who was suffering from advanced leukaemia. His success drew significant interest in academic research and medicine communities, and 12 years later, the first successful allogeneic transplant was performed. To this day, HSCs remain the most studied and characterised stem cell population. In fact, HSCs are the only stem cell population routinely utilised in the clinic. As such, HSCs function as a model system both for the biological investigation of stem cells, as well as for their clinical application. Herein, we briefly review HSC transplantation, strategies for the ex vivo cultivation of HSCs, recent clinical outcomes, and their impact on the future direction of HSC transplantation therapy.

Engaging parents in STEM: Coteaching and cogenerative dialoguing in a Queensland high school

Diminished student interest in science, technology, engineering and mathematics (STEM) is recognised by educators, researchers and public policy makers as a concerning global trend. Inviting stakeholders like scientists and industry specialists to discuss their work is one means schools use to facilitate student engagement in the sciences. However, these visits generally comprise one-off sessions with minimal relevance to students’ particular and ongoing learning needs. This case study investigated coteaching and cogenerative dialoguing with parents in teaching a Year-8 multidisciplinary unit with science and technology foci. Two parents cotaught alongside the resident teacher and researcher over eight months. This paper concentrates on one parent, a medical scientist by profession. Data sources included video and audio recordings of cogenerative dialogues and classroom interactions, student work samples and journal entries. Data were interrogated using the sociological constructs of fields and capitals and the dialectic of structure|agency. The findings reveal how (a) the parent’s science and technology knowledge was tailored to the students’ needs initially and continually and (b) student-generated data indicated enhanced engagement in science and technology. The research speaks to schools and governments about enhancing STEM education by furthering collaborative relationships with relevant stakeholders.

The world of adolescence and the world of STEM : are they irrevocably separated?

Adolescents are indeed bothered by the complexities of the present and future and are concerned with making sense out of the multiple demands of parents, teachers, and peers while trying to develop identities as autonomous individuals. In this confused world, contemporary school science does not fit their view of desirable world as evident in the findings of the ROSE study. However, there are bright spots where teachers, community, parents and youth do engage with STEM. This paper will report on initiatives drawn from a decade of research in schools that have attempted to reconcile the interests of youth and the contemporary world of science. The aim is to identify those factors that do stimulate student interest. These case studies were conducted generally using both qualitative and quantitative data and findings analysed in terms of program outcomes and student engagement. The key finding is that the formation of relationships and partnerships in which students have high degree of autonomy and sense of responsibility is paramount to positive dispositions towards STEM. The findings raise some hope that innovative schools and partnerships can foster innovation and connect youth with the real world.

Teachers' conceptions of how to engage students through inquiry learning : foundations for STEM

The advocacy for inquiry-based learning in contemporary curricula assumes the principle that students learn in their own way by drawing on direct experience fostered by the teacher. That students should be able to discover answers themselves through active engagement with new experiences was central to the thinking of eminent educators such as Pestalozzi, Dewey and Montessori. However, even after many years of research and practice, inquiry learning as a referent for teaching still struggles to find expression in the average teachers' pedagogy. This study drew on interview data from 20 primary teachers. A phenomenographic analysis revealed three conceptions of teaching that support inquiry learning in science in the primary years of schooling: (a) The Experience-centred conception where teachers focused on providing interesting sensory experiences to students; (b) The Problem-centred conception where teachers focused on challenging students with engaging problems; and (c) The Question-centred conception where teachers focused on helping students to ask and answer their own questions. Understanding teachers' conceptions of teaching has implications for both the enactment of inquiry teaching in the classroom as well as the uptake of new teaching behaviours during professional development, with enhanced outcomes for engaging students in STEM.

Proteomics Approaches in the Identification of Molecular Signatures of Mesenchymal Stem Cells

Mesenchymal stem cells (MSCs) are undifferentiated, multi-potent stem cells with the ability to renew. They can differentiate into many types of terminal cells, such as osteoblasts, chondrocytes, adipocytes, myocytes, and neurons. These cells have been applied in tissue engineering as the main cell type to regenerate new tissues. However, a number of issues remain concerning the use of MSCs, such as cell surface markers, the determining factors responsible for their differentiation to terminal cells, and the mechanisms whereby growth factors stimulate MSCs. In this chapter, we will discuss how proteomic techniques have contributed to our current knowledge and how they can be used to address issues currently facing MSC research. The application of proteomics has led to the identification of a special pattern of cell surface protein expression of MSCs. The technique has also contributed to the study of a regulatory network of MSC differentiation to terminal differentiated cells, including osteocytes, chondrocytes, adipocytes, neurons, cardiomyocytes, hepatocytes, and pancreatic islet cells. It has also helped elucidate mechanisms for growth factor–stimulated differentiation of MSCs. Proteomics can, however, not reveal the accurate role of a special pathway and must therefore be combined with other approaches for this purpose. A new generation of proteomic techniques have recently been developed, which will enable a more comprehensive study of MSCs. Keywords

Biological responses of human bone marrow mesenchymal stem cells to Sr-M-Si (M = Zn, Mg) silicate bioceramics

Strontium (Sr), Zinc (Zn), magnesium (Mg), and silicon (Si) are reported to be essential trace elements for the growth and mineralization of bone. We speculated that the combination of these bioactive elements in bioceramics may be effective to regulate the osteogenic property of boneforming cells. In this study, two Sr-containing silicate bioceramics, Sr2ZnSi2O7 (SZS) and Sr2MgSi2O7 (SMS), were prepared. The biological response of human bone marrow mesenchymal stem cells (BMSCs) to the two bioceramics (in the forms of powders and dense ceramic bulks) was systematically studied. In powder form, the effect of powder extracts on the viability and alkaline phosphatase (ALP) activity of BMSCs was investigated. In ceramic disc form, both direct and indirect coculture of BMSCs with ceramic discs were used to investigate their biological response, including attachment, proliferation, ALP activity, and bone-related genes expression. Beta-tricalcium phosphate (b-TCP) and akermanite (Ca2MgSi2O7, CMS) were used as control materials. The results showed that the Sr, Zn, and Si (or Sr, Mg, and Si)-containing ionic products from SZS and SMS powders enhanced ALP activity of BMSCs, compared to those from b-TCP. Both SZS and SMS ceramic discs supported the growth of BMSCs, and most importantly, significantly enhanced the ALP activity and bone-related genes expression of BMSCs as compared to b-TCP. The results suggest that the specific combination of bioactive ions (Sr, Zn, Si, e.g.) in bioceramics is a viable way to improve the biological performance of biomaterials, and the form of materials and surface properties were nonnegligible factors to influence cell response.

ATF5, a possible regulator of osteogenic differentiation in human adipose-derived stem cells

The regulatory pathways involved in maintaining the pluripotency of embryonic stem cells are partially known, whereas the regulatory pathways governing adult stem cells and their "stem-ness" are characterized to an even lesser extent. We, therefore, screened the transcriptome profiles of 20 osteogenically induced adult human adipose-derived stem cell (ADSC) populations and investigated for putative transcription factors that could regulate the osteogenic differentiation of these ADSC. We studied a subgroup of donors' samples that had a disparate osteogenic response transcriptome from that of induced human fetal osteoblasts and the rest of the induced human ADSC samples. From our statistical analysis, we found activating transcription factor 5 (ATF5) to be significantly and consistently down-regulated in a randomized time-course study of osteogenically differentiated adipose-derived stem cells from human donor samples. Knockdown of ATF5 with siRNA showed an increased sensitivity to osteogenic induction. This evidence suggests a role for ATF5 in the regulation of osteogenic differentiation in adipose-derived stem cells. To our knowledge, this is the first report that indicates a novel role of transcription factors in regulating osteogenic differentiation in adult or tissue specific stem cells. © 2012 Wiley Periodicals, Inc.

Chemopreventive effect of PSP through targeting of prostate cancer stem cell-like population

Recent evidence suggested that prostate cancer stem/progenitor cells (CSC) are responsible for cancer initiation as well as disease progression. Unfortunately, conventional therapies are only effective in targeting the more differentiated cancer cells and spare the CSCs. Here, we report that PSP, an active component extracted from the mushroom Turkey tail (also known as Coriolus versicolor), is effective in targeting prostate CSCs. We found that treatment of the prostate cancer cell line PC-3 with PSP led to the down-regulation of CSC markers (CD133 and CD44) in a time and dose-dependent manner. Meanwhile, PSP treatment not only suppressed the ability of PC-3 cells to form prostaspheres under non-adherent culture conditions, but also inhibited their tumorigenicity in vivo, further proving that PSP can suppress prostate CSC properties. To investigate if the anti-CSC effect of PSP may lead to prostate cancer chemoprevention, transgenic mice (TgMAP) that spontaneously develop prostate tumors were orally fed with PSP for 20 weeks. Whereas 100% of the mice that fed with water only developed prostate tumors at the end of experiment, no tumors could be found in any of the mice fed with PSP, suggesting that PSP treatment can completely inhibit prostate tumor formation. Our results not only demonstrated the intriguing anti-CSC effect of PSP, but also revealed, for the first time, the surprising chemopreventive property of oral PSP consumption against prostate cancer.

3D mesenchymal stem/stromal cell osteogenesis and autocrine signalling

Mesenchymal stem/stromal cells (MSC) are rapidly becoming a leading candidate for use in tissue regeneration, with first generation of therapies being approved for use in orthopaedic repair applications. Capturing the full potential of MSC will likely require the development of novel in vitro culture techniques and devices. Herein we describe the development of a straightforward surface modification of an existing commercial product to enable the efficient study of three dimensional (3D) human bone marrow-derived MSC osteogenic differentiation. Hundreds of 3D microaggregates, of either 42 or 168 cells each, were cultured in osteogenic induction medium and their differentiation was compared with that occurring in traditional two dimensional (2D) monolayer cultures. Osteogenic gene expression and matrix composition was significantly enhanced in the 3D microaggregate cultures. Additionally, BMP-2 gene expression was significantly up-regulated in 3D cultures at day 3 and 7 by approximately 25- and 30-fold, respectively. The difference in BMP-2 gene expression between 2D and 3D cultures was negligible in the more mature day 14 osteogenic cultures. These data support the notion that BMP-2 autocrine signalling is up-regulated in 3D MSC cultures, enhancing osteogenic differentiation. This study provides both mechanistic insight into MSC differentiation, as well as a platform for the efficient generation of microtissue units for further investigation or use in tissue engineering applications.

Implantation of osteogenic differentiated donor mesenchymal stem cells causes recruitment of host cells

The interaction between host and donor cells is believed to play an important role in osteogenesis. However, it is still unclear how donor osteogenic cells behave and interact with host cells in vivo. The purpose of this study was to track the interactions between transplanted osteogenic cells and host cells during osteogenesis. In vitro migration assay was carried out to investigate the ability of osteogenic differentiated humanmesenchymal stemcells (O-hMSCs) to recruit MSCs. At the in vivo level, O-hMSCs were implanted subcutaneously or into skull defects in severe combined immunodeficient (SCID) mice. New bone formation was observed bymicro-CT and histological procedures. In situ hybridization (ISH) against human Alu sequences was performed to distinguish donor osteogenic cells from host cells. In vitro migration assay revealed an increased migration potential of MSCs by co-culturing with O-hMSCs. In agreement with the results of in vitro studies, ISH against human Alu sequences showed that host mouse MSCs migrated in large numbers into the transplantation site in response to O-hMSCs. Interestingly, host cells recruited by O-hMSCs were the major cell populations in newly formed bone tissues, indicating that O-hMSCs can trigger and initiate osteogenesis when transplanted in orthotopic sites. The observations fromthis study demonstrated that in vitro induced O-hMSCs were able to attract hostMSCs in vivo andwere involved inosteogenesis togetherwith host cells,whichmay be of importance for bone tissue-engineering applications.