Chizu Nakamoto | PubMed Health

PubMed2006

Osteogenic progenitors in bone marrow aspirates from smokers and nonsmokers.

UNLABELLED: Bone marrow aspirates contain connective tissue progenitor cells which can proliferate to form cell colonies that express connective tissue phenotypes. The number of osteogenic connective tissue progenitor cells can be estimated by counting colonies that express alkaline phosphatase, an early marker of osteoblastic differentiation. Because tobacco use is associated with decreased bone mass and fracture nonunion, we tested the hypothesis that current or previous tobacco use is an independent determinant of marrow cellularity or prevalence of osteogenic connective tissue progenitor cells among marrow-derived cells. Marrow aspirates were obtained from the anterior iliac crest of 62 patients who were grouped as never smoked, past smokers, or current smokers. The number of nucleated cells per aspirate was determined. Cells were cultured for 6 days in osteogenic media. The prevalence of osteogenic connective tissue progenitor cells was determined by counting colony forming units. The area of each colony was assessed using quantitative image analysis. Cellularity of bone marrow was found to decrease with age. We observed no relationship between smoking status and marrow cellularity, colony prevalence, or colony area. This suggests that tobacco use is not associated with a change in prevalence of osteogenic connective tissue progenitor cells in bone marrow, or their intrinsic biologic capacity to undergo early osteoblastic differentiation. LEVEL OF EVIDENCE: Diagnostic Study, Level I (testing of previously developed diagnostic criteria on consecutive patients--with universally applied reference "gold" standard). See the Guidelines for Authors for a complete description of levels of evidence.

Adult↗

PubMed2004

Engineering principles of clinical cell-based tissue engineering.

Tissue engineering is a rapidly evolving discipline that seeks to repair, replace, or regenerate specific tissues or organs by translating fundamental knowledge in physics, chemistry, and biology into practical and effective materials, devices, systems, and clinical strategies. Stem cells and progenitors that are capable of forming new tissue with one or more connective tissue phenotypes are available from many adult tissues and are defined as connective tissue progenitors. There are four major cell-based tissue-engineering strategies: (1) targeting local connective tissue progenitors where new tissue is desired, (2) transplanting autogenous connective tissue progenitors, (3) transplanting culture-expanded or modified connective tissue progenitors, and (4) transplanting fully formed tissue generated in vitro or in vivo. Stem cell function is controlled by changes in stem cell activation and self-renewal or by changes in the proliferation, migration, differentiation, or survival of the progeny of stem cell activation, the downstream progenitor cells. Three-dimensional porous scaffolds promote new tissue formation by providing a surface and void volume that promotes the attachment, migration, proliferation, and desired differentiation of connective tissue progenitors throughout the region where new tissue is needed. Critical variables in scaffold design and function include the bulk material or materials from which it is made, the three-dimensional architecture, the surface chemistry, the mechanical properties, the initial environment in the area of the scaffold, and the late scaffold environment, which is often determined by degradation characteristics. Local presentation or delivery of bioactive molecules can change the function of connective tissue progenitors (activation, proliferation, migration, differentiation, or survival) in a manner that results in new or enhanced local tissue formation. All cells require access to substrate molecules (oxygen, glucose, and amino acids). A balance between consumption and local delivery of these substrates is needed if cells are to survive. Transplanted cells are particularly vulnerable. Theoretical calculations can be used to explore the relationships among cell density, diffusion distance, and cell viability within a graft and to design improved strategies for transplantation of connective tissue progenitors. Rational strategies for tissue engineering seek to optimize new tissue formation through the logical selection of conditions that modulate the performance of connective tissue progenitors in a graft site to produce a desired tissue. This increasingly involves strategies that combine cells, matrices, inductive stimuli, and techniques that enhance the survival and performance of local or transplanted connective tissue progenitors.

Cell Culture Techniques↗

PubMed2003

Practical Modeling Concepts for Connective Tissue Stem Cell and Progenitor Compartment Kinetics.

Stem cell activation and development is central to skeletal development, maintenance, and repair, as it is for all tissues. However, an integrated model of stem cell proliferation, differentiation, and transit between functional compartments has yet to evolve. In this paper, the authors review current concepts in stem cell biology and progenitor cell growth and differentiation kinetics in the context of bone formation. A cell-based modeling strategy is developed and offered as a tool for conceptual and quantitative exploration of the key kinetic variables and possible organizational hierarchies in bone tissue development and remodeling, as well as in tissue engineering strategies for bone repair.

Journal Article↗