Definition of TE Why TE has developed Applications of TE Challenges The future

Creation of a functional biological substitute using living cells and a matrix to maintain, improve or restore damage to tissues and organs (Atala, A. Engineering tissues, organs

and cells. 2007 J Tissue Eng Regen Med 1: 83-96)

Bringing together the fields of medicine, biology, engineering and biotechnology

Tissue Engineering is an interdisciplinary field that applies the principles of engineering and the life sciences toward the development of biological substitutes that restore, maintain, or improve tissue function
Langer and Vacanti. Science 260, 920 (1993)

Prior to 1950 references to the concept of tissue engineering may be found in the literature
In the 1970s and 1980s research on what we now call tissue engineering emerges In 1987 the term tissue engineering was coined

In 1988 the first meeting called tissue engineering was held at Lake Tahoe
In the 1990s research accelerates and an industry begins to emerge

Failing tissues and organs Shortfalls of current options Autologous tissues Allogeneic tissues Xenogeneic tissues Synthetic materials

To create products that improve tissue function or heal tissue defects. Replace diseased or damaged tissue Because Donor tissues and organs are in short supply We want to minimize immune system response by using our own cells or novel ways to protect transplant.

Tissue from the same patient The ideal option Biocompatible, no immune response Natural Decreasing availability of healthy tissue from patients with disease

Tissue from another person More practical than autologous harvest Severe shortage of donors Increased immune response to foreign material (Santos, T. et al. TERMIS, NA 2007) Immunosuppressant drugs required Unpleasant side effects Expensive

Tissue harvested from animals Potentially readily available Immune response from host to foreign material (Santos, T. et al. TERMIS, NA 2007) Risk of disease transmission from animal to human e.g. prions Significant ethical considerations

Biocompatible made with the patients own cells Engineered to fill the exact role required Degradation rate, composition, size, mechanical properties Off-the-shelf availability

e Cell Source
Embryonic stem cells Adult stem cells Progenitor cells

Signals
Growth factors Drugs Mechanical forces

ECM
Metals Ceramics Synthetic polymers Natural polymers

Allow cell attachment and migration Deliver and retain cells and biochemical factors Enable diffusion of vital cell nutrients and expressed products Exert certain mechanical and biological influences to modify the behaviour of the cell phase

Various textures and materials Encourage cells to grow Allow nutrients to permeate Wont harm the patient

Polymers Collagen Laminin Fibrin Matrigel Decellularized matrix Ceramics Hydroxyapatite Calcium phosphate Bioglass

Perfusion-decellularized matrix: using nature's platform to engineer a bioartificial heart. Ott, et al.

3-dimensional Cross-linked Porous Biodegradable Proper surface chemistry Matching mechanical strength Biocompatible Promotes natural healing Accessibility Commercial Feasibility

Able to cause change and affect structure and function of a graft Examples: -Skin model: fibroblasts and keratinocytes -Vascular construct: smooth muscle and endothelial cells -Cartilage: chondrocytes Stem cells much potential but difficult to direct differentiation and achieve sufficient cell numbers

Growth factors Directing cellular activity Bioreactors Designed to expose cells to physical stimuli and/or maintain desired conditions Examples Bladder urothelium: cyclic strain Vascular graft: pulsatile flow

ADVANTAGES This approach seeks to overcome the problem of limited implant lifetimes by regenerating tissue in culture. Alleviates problem of donor shortage by using a few culture cells to regenerate entire tissue

DISADVANTAGES
May face same rejection issues as transplanted tissues Long term stability of tissues generated in this manner is still unknown Approaches to generate complex tissue types still very experimental

ADVANTAGES
Seeks to solve all of the potential problems of the other approaches The bodys own cells or growth factors are used for regeneration, so no autograft of transplanted tissue is needed The implant will regenerate natural tissue and the components of the guiding scaffold are biodegradable so long term stability is not an issue

DISADVANTAGES Faithful regeneration of complex tissue is an unsolved problem In vivo synthesis of skin successfully recovers the overall structure of the organ but fails to regenerate organelles such as the sweat glands and hair follicles Proper scaffolds for many cell types (e.g. nerve) have not been fully developed yet, primarily because the biological cues necessary for regeneration are not well understood

FIRST APPROACH Seeding of cells into various gels (collagen gels, fibrin and other component) The true integration of cells into gels allow them to reorganize the surrounding matrix.

FIRST APPROACH
Drawback: Week mechanical resistance of the obtained substitutes. The structural integrity may be sufficient for skin, but not so for substitutes in vascular or orthopaedic systems This problem has been addressed with glycation and magnetic alignment of the collagen fibres

SECOND APPROACH
The seeding of cells into scaffolds The cells thrive in the porous material and secrete various amount of extracellular matrix depending on their nature. (developed by Robert Langers group MIT)

Advantage: of the scaffold approach is the immediate creation of a three-dimensional structure that already has significant structural properties Drawback: The intrinsic nature of most of these polymers, which are suture materials, entails slow degradation with an ensuing lowering of pH of surrounding tissues. This leads to a slow but rather protracted low-level inflammatory process.

THIRD APPROACH
In this approach various types of cells, mostly of mesencymal origin, are grown in such a fashion within a culture flask that they literally embedded themselves in their very oven extracellular matrix. Among many factors, the addition of sodium ascorbate allows the significant appearance of the various components of the extracellular matrix. These sheets are then either stacked or rolled to obtain various tissue substitutes.

Advantage: The absence of extraneous collagens and any synthetic material Drawback: This approach is time consuming

In surgery Transplantation of failing tissues/organs Aiding tissues in the healing process In the laboratory Observing immunological, pathological and healing changes in human tissue without harming patients Drug therapies: efficacy and side effects of drugs

INITIAL STAGE

CAP STAGE

BELL STAGE

CROWN FORMATION

Obtain patients bone cells From the hip Seed onto scaffold Implant scaffold Use body as incubator All appropriate growth factors available Remove and implant in proper location

Osteoinductive Demineralized freeze-dried bone allograft (DFDBA) Partially pure proteins (BMP) BMP-2 BMP-4 BMP-7 BMP-9

Osteoconductive Freeze-dried bone Autograft Ceramics Bioglasses Coral-derived Deproteinized bovine bone Polylactic acid (PLA)/polyglycolic acid (PGA)

Osteogenic Mesenchymal stem cells (MSC) Marrow Platelet-rich plasma (PRP) PRP + white blood cells (WBC) Emdogain Gene therapy Fibroblast growth factor (FGF) Peptide TP508 Peptide P15 Platelet-derived growth factor (PDGF

Limited Success: full regeneration of tissues that do not regenerate spontaneously has not yet been achieved A lot of success with bone Skin has no glands and hair Engineered cartilage is not articular

How much is enough? We do not really know. Pain relief, partial function and aesthetic improvement are often achieved using incomplete tissues Do we need (always, in some cases) full regeneration? If so, is it needed immediately or over time? What are the minimal requirements for engineered tissue grafts at the time of implantation?

Widely used in 10 Yrs

Human skin Artificial blood, kidney, liver, nerve cells, synthetic bone Cells for targeted detoxification

Ten years & beyond

Molecular pharming of wide range of large therapeutic molecules (proteins, lipids, enzymes, etc.) For wide range of diseases (cancer, heart, stroke, diabetes) In-body drug manufacture

Probiotics, prebiotics Nutraceuticals in food

Unforeseen hurdles in the creation of multicellular constructs

In the laboratory
Supply of nutrients Removal of waste Size Mechanical stability

In the patient
Availability Evidence of efficacy Safety; graft rejection Cost

Micromechanical effects Cell differentiation and growth (especially in load-bearing tissues) can be affected by micromechanical stresses transmitted by the scaffold Cell function deterioration Cross-application to other areas (gene therapy, drug delivery) Multicellular tissues and organs Complex, multicomponent structures (vascularized tissues) Regeneration-inducing factors (proteins)

Vascularization :The approach of stimulating the ingrowths of blood vessels into solid organs has not been successful. Such organs rapidly, within hours or even minutes, demanded blood irrigation for survival and proper function. Innervation

Some tissues already in clinical use Improvements needed to increase availability and safety For widespread use, reduced cost is essential Further work should focus on: vascularisation of new tissue; maintaining nutrient supply to cells in matrix with increasing size Achieving full potential of stem cells to differentiate into desired cell types