An in vitro comparison of the incorporation, growth, and chondrogenic potential of human bone marrow versus adipose tissue mesenchymal stem cells in clinically relevant cell scaffolds used for cartilage repair

Aim. To compare the incorporation, growth, and chondrogenic potential of bone marrow (BM) and adipose tissue (AT) mesenchymal stem cells (MSCs) in scaffolds used for cartilage repair. Methods. Human BM and AT MSCs were isolated, culture expanded, and characterised using standard protocols, then seeded into 2 different scaffolds, Chondro-Gide or Alpha Chondro Shield. Cell adhesion, incorporation, and viable cell growth were assessed microscopically and following calcein AM/ethidium homodimer (Live/Dead) staining. Cell-seeded scaffolds were treated with chondrogenic inducers for 28 days. Extracellular matrix deposition and soluble glycosaminoglycan (GAG) release into the culture medium was measured at day 28 by histology/immunohistochemistry and dimethylmethylene blue assay, respectively. Results. A greater number of viable MSCs from either source adhered and incorporated into Chondro-Gide than into Alpha Chondro Shield. In both cell scaffolds, this incorporation represented less than 2% of the cells that were seeded. There was a marked proliferation of BM MSCs, but not AT MSCs, in Chondro-Gide. MSCs from both sources underwent chondrogenic differentiation following induction. However, cartilaginous extracellular matrix deposition was most marked in Chondro- Gide seeded with BM MSCs. Soluble GAG secretion increased in chondrogenic versus control conditions. There was no marked difference in GAG secretion by MSCs from either cell source. Conclusion. Chondro-Gide and Alpha Chondro Shield were permissive to the incorporation and chondrogenic differentiation of human BM and AT MSCs. Chondro-Gide seeded with BM MSCs demonstrated the greatest increase in MSC number and deposition of a cartilaginous tissue.

An investigation of primary human cell sources and clinical scaffolds for articular cartilage repair

Damage to articular cartilage of the knee can be debilitating because it lacks the capacity to repair itself and can progress to degenerative disorders such as osteoarthritis. The current gold standard for treating cartilage defects is autologous chondrocyte implantation (ACI). However, one of the major limitations of ACI is the use of chondrocytes, which dedifferentiate when grown in vitro and lose their phenotype. It is not clear whether the dedifferentiated chondrocytes can fully redifferentiate upon in vivo transplantation. Studies have suggested that undifferentiated mesenchymal stem or stromal cells (MSCs) from bone marrow (BM) and adipose tissue (AT) can undergo chondrogenic differentiation. Therefore, the main aim of this thesis was to examine BM and AT as a cell source for chondrogenesis using clinical scaffolds. Initially, freshly isolated cells were compared with culture expanded MSCs from BM and AT in Chondro-Gide®, Alpha Chondro Shield® and Hyalofast™. MSCs were shown to grow better in the three scaffolds compared to freshly isolated cells. BM MSCs in Chondro-Gide® were shown to have increased deposition of cartilage specific extracellular matrix (ECM) compared to AT MSCs. Further, this thesis has sought to examine whether CD271 selected MSCs from AT were more chondrogenic than MSCs selected on the basis of plastic adherence (PA). It was shown that CD271+MSCs may have superior chondrogenic properties in vitro and in vivo in terms of ECM deposition. The repair tissue seen after CD271+MSC transplantation combined with Alpha Chondro Shield® was also less vascularised than that seen after transplantation with PA MSCs in the same scaffold, suggesting antiangiogenic activity. Since articular cartilage is an avascular tissue, CD271+MSCs may be a better suited cell type compared to the PA MSCs. Hence, this study has increased the current understanding of how different cell-scaffold combinations may best be used to promote articular cartilage repair.

Re-engineering materials management: a case study on an Indian refinery

Materials management function is always a major concern to the management of any industrial organisation as high inventory and an inefficient procurement process affect the profitability to a great extent. Problems multiply due to a very current business environment in India. Hence, existing materials planning and procurement processes and inventory management systems require a re-look with respect to a changing business environment. This study shows a radical improvement in materials management function of an Indian petroleum refinery through business process re-engineering (BPR) by analysing current processes, identifying key issues, deriving paradigm shifts and developing re-engineered processes through customer value analysis. BPR has been carried out on existing processes of “materials planning and procurement” and “warehousing and surplus disposal”. The re-engineered processes for materials management function trigger a few improvement projects that were identified by the group of executives who took part in the re-engineering exercise. Those projects were implemented in an integrated framework with the application of the state of art information technology tools.

Enabling a simulation capability in the organisation

Enabling a Simulation Capability in the Organisation addresses the application of simulation modelling techniques in order to enable better informed decisions in business and industrial organisations. The book’s unique approach treats simulation not just as a technical tool, but within as a support for organisational decision making, showing the results from a survey of current and potential users of simulation to suggest reasons why the technique is not used as much as it should be and what are the barriers to its further use. By incorporating an evaluation of six detailed case studies of the application of simulation in industry by the author, the book will teach readers: •the role of simulation in decision making; •how to introduce simulation as a tool for decision making; and •how to undertake simulation studies that lead to change in the organisation. Enabling a Simulation Capability in the Organisation provides an introduction to the state of the art in simulation modelling for researchers in business studies and engineering, as well a useful guide to practitioners and managers in business and industry.

Managing projects in fast track:a case of public sector organisation in India

Completing projects faster than the normal duration is always a challenge to the management of any project, as it often demands many paradigm shifts. Opportunities of globalization, competition from private sectors and multinationals force the management of public sector organizations in the Indian petroleum sector to take various aggressive strategies to maintain their profitability. Constructing infrastructure for handling petroleum products is one of them. Moreover, these projects are required to be completed in faster duration compared to normal schedules to remain competitive, to get faster return on investment, and to give longer project life. However, using conventional tools and techniques of project management, it is impossible to handle the problem of reducing the project duration from a normal period. This study proposes the use of concurrent engineering in managing projects for radically reducing project duration. The phases of the project are accomplished concurrently/simultaneously instead of in a series. The complexities that arise in managing projects are tackled through restructuring project organization, improving management commitment, strengthening project-planning activities, ensuring project quality, managing project risk objectively and integrating project activities through management information systems. These would not only ensure completion of projects in fast track, but also improve project effectiveness in terms of quality, cost effectiveness, team building, etc. and in turn overall productivity of the project organization would improve.

Process re-engineering for effective implementation of projects

Time, cost and quality are the prime objectives of any project. Unfortunately, today’s project management does not always ensure the realisation of these objectives. The main reasons of project non-achievement are changes in scope and design, changes in Government policies and regulations, unforeseen inflation, under-estimation and mis-estimation. An overall organisational approach with the application of appropriate management philosophies, tools and techniques can only solve the problem. The present study establishes a methodology for achieving success in implementing projects using a business process re-engineering (BPR) framework. Internal performance characteristics are introspected through condition diagnosis that identifies and prioritises areas of concern requiring attention. Process re-engineering emerges as a most critical area for immediate attention. Project process re-engineering is carried out by eliminating non-value added activities, taking up activities concurrently by applying information systems rigorously and applying risk management techniques throughout the project life cycle. The overall methodology is demonstrated through applications to cross country petroleum pipeline project organisation in an Indian scenario.

Physical and biological properties of a novel siloxane adhesive for soft tissue applications

The aim of this study was to investigate the adhesive properties of an in-house amino-propyltrimethoxysilane-methylenebisacrylamide (APTMS-MBA) siloxane system and compare them with a commercially available adhesive, n-butyl cyanoacrylate (nBCA). The ability of the material to perform as a soft tissue adhesive was established by measuring the physical (bond strength, curing time) and biological (cytotoxicity) properties of the adhesives on cartilage. Complementary physical techniques, X-ray photoelectron spectroscopy, Raman and infrared imaging, enabled the mode of action of the adhesive to the cartilage surface to be determined. Adhesion strength to cartilage was measured using a simple butt joint test after storage in phosphate-buffered saline solution at 37°C for periods up to 1 month. The adhesives were also characterised using two in vitro biological techniques. A live/dead stain assay enabled a measure of the viability of chondrocytes attached to the two adhesives to be made. A water-soluble tetrazolium assay was carried out using two different cell types, human dermal fibroblasts and ovine meniscal chondrocytes, in order to measure material cytotoxicity as a function of both supernatant concentration and time. IR imaging of the surface of cartilage treated with APTMS-MBA siloxane adhesive indicated that the adhesive penetrated the tissue surface marginally compared to nBCA which showed a greater depth of penetration. The curing time and adhesion strength values for APTMS-MBA siloxane and nBCA adhesives were measured to be 60 s/0.23 MPa and 38 min/0.62 MPa, respectively. These materials were found to be significantly stronger than either commercially available fibrin (0.02 MPa) or gelatin resorcinol formaldehyde (GRF) adhesives (0.1 MPa) (P <0.01). Cell culture experiments revealed that APTMS-MBA siloxane adhesive induced 2% cell death compared to 95% for the nBCA adhesive, which extended to a depth of approximately 100-150 μm into the cartilage surface. The WST-1 assay demonstrated that APTMS-MBA siloxane was significantly less cytotoxic than nBCA adhesive as an undiluted conditioned supernatant (P <0.001). These results suggest that the APTMS-MBA siloxane may be a useful adhesive for medical applications. © VSP 2005.