Biomaterials - II

BME 379/385, CHE 379, Spring 2003 (Schmidt)

Goals:
By the end of this lecture, you should be able to: • List and describe some of the common natural materials used in TE • Describe how cells interact with the surrounding ECM • Describe how these interactions can be modulated • Define "biomimetic" • Define "Vroman effect" • Describe the biological responses to implanted materials • Describe common methods to assess biocompatibility

Outline:
I. II. III. IV. Natural Materials in Tissue Engineering Cell Interactions with Materials Controlling Cell Interactions - "Biomimetic" Materials Analysis of Material Biocompatibility

BME 379/385, CHE 379: Biomaterials II -- 2

I.

Natural Materials in Tissue Engineering

Last time, we focused predominantly on synthetic materials. Examples of some of the common synthetic polymers used in tissue engineering include: PLA, PGA, and PLGA. Natural Materials are playing a larger role in TE applications. Examples of Natural Materials include:

BME 379/385, CHE 379: Biomaterials II -- 3

II.

Cell Interactions with Materials

SURFACE vs. BULK properties

Function Bioactivity

Structure Support Shape

Surface Properties governed by:

Interactions:

Material

ECM

Cell

BME 379/385, CHE 379: Biomaterials II -- 4

Cell Interactions/Cell Adhesion Modulate:

How do cells interact with their environment?

BME 379/385, CHE 379: Biomaterials II -- 5

Integrins are the main class of receptors that mediate binding of cells to the ECM (e.g., fibronectin, laminin, collagen)...

Example of Cell Shape Changes and Binding:

• Cell shape changes induced via binding of adrenaline, thrombin, or ADP to their respective G-Protein Coupled Receptors (GPCRs). • Activation of the cell surface platelet-specific integrin αIIb β3 also occurs. • Only activated cell surface αIIb β3 binds to fibrinogen, fibronectin, and von Willebrand factor, leading to adherence to the vessel wall and formation of a platelet plug.

> In tissue engineering applications, cell adhesion and migration are critical for tissue formation <

BME 379/385, CHE 379: Biomaterials II -- 6

III.

Controlling Cell Interactions - "Biomimetic" Materials

Upon implantation, all materials are coated by serum proteins (e.g., fibronectin, vitronectin), and are eventually encapsulated in fibrous tissue, as described in Section IV of these notes. Thus, many techniques are geared at reducing non-specific adhesion "programming in" only specific cell adhesion sites for interaction. Biomimetic materials are thus defined as: This can be controlled through:

Other ways to control cell shape and adhesion:

BME 379/385, CHE 379: Biomaterials II -- 7

Geometry can be used to monitor cell migration...

PNAS 98:5992-5996, 2001

Cell shape can be used to control cell viability...

Science 276:1425-1428, 1997

BME 379/385, CHE 379: Biomaterials II -- 8

Cell shape and viability/function continued....

20:40=20 µm circles separated by 40 µm 5:10= 5 µm circles separated by 10 µm 3:6= 3 µm circles separated by 6 µm

Science 276:1425-1428, 1997

BME 379/385, CHE 379: Biomaterials II -- 9

Definitions for types of cell migration behavior:
Haptotaxis:

Chemotaxis:

Chemoaffinity:

Many migration assays and models exist to study migration behavior of cells on various materials.

BME 379/385, CHE 379: Biomaterials II -- 10

IV.

Analysis of Material Biocompatibility

Tissue Response to Materials
Key Cells Involved in Tissue Response: Blood Cells (identified using a blood smear) are key players.

Blood cells consist of white blood cells (leukocytes) and red blood cells. Granulocytes (eosinophils, basophils, neutrophils), monocytes (macrophages), and lymphocytes together comprise white cells.

Red blood cells are biconcave discs that lack a nucleus and carry hemoglobin which is responsible for oxygen transport. What are the functions of neutrophils (polymorphonuclear leukocytes or PMNs), macrophages, and lymphocytes?

Platelets are cell fragments involved in clotting mechanisms to prevent excessive bleeding when tissue is injured.

BME 379/385, CHE 379: Biomaterials II -- 11

Response to a Material Implant: Consider what happens to the biomaterial upon implantation into a vascularized tissue of an animal or patient. Answer the following: What is the first event that occurs?

Define the Vroman Effect.

How can the event above be controlled? Give some examples of how this can be minimized or modulated.

What other factors may affect this event?

BME 379/385, CHE 379: Biomaterials II -- 12

Now let's consider the broader host response to the implanted biomaterial. Define and distinguish the following responses. What factors and/or cells are important in each case? Acute Inflammation:

Chronic Inflammation:

Granulation Tissue:

Foreign Body Reaction:

Fibrosis and Fibrous Encapsulation:

BME 379/385, CHE 379: Biomaterials II -- 13

Inflammation: The four signs associated with an inflammatory response: 1) Red (rubor) appearance at the injury site due to the presence of RBCs just under the skin. 2) The site becomes swollen (tumor) due to the fluid that came with the blood . 3) Site is warm (calor), which is the result of the warm blood going to a cooler tissue site. 4) Painful (dolor) due to impingement on and damage to local nerve networks. Healing in the presence of a biomaterial: Define biocompatibility? When a material is not biocompatible, the inflammatory response continues as a chronic inflammatory response and progresses to giant cells and granulomas. The host tries to neutralize the foreign object (implant). Formation of the fibrous capsule is an indication that the material is biocompatible and will occur as an early step in healing. Infection: Presence of infection will prevent the resolution of inflammation, and an undesired chronic inflammatory response will occur. What would happen if bacteria are involved during the process of wound healing?

BME 379/385, CHE 379: Biomaterials II -- 14

Immune response: A specific antigen-antibody based immune response will occur if cells from another species or patient are implanted. Allergic reactions are specific immune responses against nonbiological entities. These reactions are not desired and can have detrimental side effects (and must be considered in device design).

Why are degradation products from the materials of an implant an important concern?

Biocompatibility testing: An essential question to consider when a biomaterial is used in a biological system is: "will this material stimulate the appropriate biological response for the intended use?". Sterilization is crucial to the use of the material in vivo. (see Handout) In vitro tests are common tests performed using tissue or cell cultures. In vivo tests to assess possible chronic inflammation are also common. Tests are designed for cytotoxicity, stimulation of immune response, irritation to tissues, provocation of chronic inflammation, effects on blood and blood components, and effects on genetic factors including mutations and tumor formation. In implantation of a nerve guidance channel composed of a polymer based material, what are some of the biocompatibility tests involved? Provide some broad and specific tests.