FUNDAMENTAL PRINCIPLES OF

RADIOBIOLOGY

I. LAW OF BERGONIÉ AND

TRIBONDEAU (1906)
Principle

Radiosensitivity of living tissue varies with maturation and metabolism.

Radiation dose in GYt (rad)

Key Points

Tissues are more radiosensitive when they:

1.​ Contain stem cells

-​ Stem cells (and organs) are radiosensitive; mature cells are radioresistant
2.​ Are younger
3.​ Have high metabolic activity
4.​ Have high proliferation rate
5.​ Have high growth rate​

Mature, specialized cells are more radioresistant.

Important Concept

The response of tissue is determined by the amount of energy deposited per unit mass
(radiation dose).

Even under controlled conditions, tissue responses may vary due to modifying factors.
II. FACTORS AFFECTING
RADIOSENSITIVITY

A. PHYSICAL FACTORS
1. Linear Energy Transfer (LET)

Definition

LET = Rate at which radiation transfers energy to soft tissue.

Unit

keV/µm (kiloelectron volts per micrometer)

Importance

• Expresses radiation quality​

• Determines radiation weighting factor (WR)​
• Higher LET → denser ionization → greater biologic damage

Diagnostic X-rays

Approximate LET = 3 keV/µm

2. Relative Biologic Effectiveness (RBE)

Definition

Compares biologic effectiveness of test radiation to standard radiation (usually diagnostic

x-rays).

Formula
RBE=Dose of standard radiationDose of test radiationRBE = \frac{\text{Dose of standard
radiation}}{\text{Dose of test radiation}}RBE=Dose of test radiationDose of standard
radiation​
Important Facts

• Diagnostic x-rays → RBE = 1​

• Higher LET → Higher RBE​
• Lower LET → Lower RBE

As LET increases:

●​ RBE increases
●​ Reaches maximum
●​ Then decreases (Overkill effect)​

3. Protraction and Fractionation

Protraction

Delivering dose slowly over a long time.

Fractionation

Dividing total dose into smaller doses separated by time.

Effect

Both reduce biologic damage because:​

• Sublethal damage repair occurs​
• Repopulation occurs

Longer exposure time → higher total dose required to produce the same effect.

B. BIOLOGIC FACTORS

1. Oxygen Effect
Tissue is more sensitive when irradiated in oxygenated (aerobic) state than in
hypoxic/anoxic state.
Oxygen Enhancement Ratio (OER)
OER = Dose under anoxic conditions/Dose under aerobic conditions

Important Points

• OER is higher for low-LET radiation​

• OER decreases as LET increases​
• Oxygen enhances indirect effect​
• Hyperbaric oxygen increases tumor radiosensitivity

2. Age
• Most sensitive: Before birth (in utero)​
• Sensitivity decreases until maturity​
• Slight increase again in old age

Most resistant period: Young adulthood

3. Recovery
If cell survives initial radiation:

• Sublethal damage repair occurs​

• Repopulation occurs

❖ Interphase death occurs when the cell dies before replicating

❖ Repair & Repopulation - the combined processes of intracellular repair and repopulation
contribute to recovery from radiation damage

High dose → tissue shrinkage → Atrophy

4. Chemical Agents
Must be present at time of irradiation.

Radiosensitizers (Increase sensitivity)

• Halogenated pyrimidines​
• Methotrexate​
• Actinomycin D​
• Hydroxyurea

Radioprotectors (Decrease sensitivity)

• Sulfhydryl compounds​
• Cysteine​
• Cysteamine

Not widely used in humans due to toxicity.

5. Hormesis
Concept: Very low radiation dose may stimulate beneficial biological responses.

“A little bit of radiation is good for you” (Not universally accepted.)

III. RADIATION DOSE–RESPONSE

RELATIONSHIPS
Applications
1.​ Cancer therapy planning
2.​ Studying low-dose radiation effects​
Two Characteristics
1.​ Linear or Nonlinear
2.​ Threshold or Nonthreshold​

TYPES OF EFFECTS
A. Deterministic Effects
• High dose​
• Early response​
• Has threshold​
• Severity increases with dose

Examples:

●​ Skin burn
●​ Cataract​

B. Stochastic Effects
• Low dose​
• Late response​
• No threshold​
• Incidence increases with dose

Examples:

●​ Cancer
●​ Leukemia
●​ Genetic effects​
Linear Dose–Response Relationships
Definition

A linear dose–response relationship means that the biologic response is directly

proportional to the radiation dose.

If the radiation dose is doubled, the biologic response is also doubled.

Dose–Response Curve Types

1. Linear, Nonthreshold (LNT) Model

Characteristics

●​ Response is directly proportional to dose.

●​ No threshold dose.
●​ Any radiation dose, no matter how small, is expected to produce some effect.
●​ Commonly used for stochastic effects (e.g., cancer, genetic mutations).​

Important Concept: Natural Response

In some linear, nonthreshold curves (commonly labeled Curve A):

●​ At zero radiation dose, a response is still observed.

●​ This is called the Natural Response.
●​ It represents the background occurrence of the effect (e.g., cancer) even without
radiation exposure.

This means radiation adds to an already existing baseline risk.

2. Linear, Threshold Model

Characteristics

●​ Response is directly proportional to dose.

●​ A threshold dose (DT) must be exceeded before any response is observed.
●​ Below the threshold → no measurable effect.​

Quick Comparison
Feature Linear Nonthreshold (LNT) Linear Threshold

Threshold present? No Yes

Any dose causes effect? Yes No

Commonly associated Stochastic effects Some tissue

with reactions

3. Nonlinear Nonthreshold
• Exponential or exaggerated small-dose response
4. Nonlinear Threshold (Sigmoid)
• No response below DT​
• Response increases​
• Reaches inflection point​
• After inflection → less effective increase

Inflection Point – when the curves stops bending up and begins bending down

Common for deterministic effects.

❖ No radiation responses have been observed after radiation doses less than 100 mGyt (10
rad)

IV. MOLECULAR RADIOBIOLOGY

A. Irradiation of Macromolecules
Macromolecules are more radiosensitive in vivo than in vitro.

In vitro requires a higher dose for measurable effect.

●​ The human body is composed of approximately 80% water.​

●​ Because of this high water content, most radiation interactions occur with water
molecules, not directly with DNA.​

2. Principal Molecular Interaction in the Body

The primary molecular interaction of ionizing radiation in the body is with water.

This process is called: Radiolysis of Water

When radiation interacts with water:

 ●​ Water molecules become ionized.​

●​ Free radicals are produced.​

●​ These free radicals are highly reactive and unstable.

Effects in Solution (In Vitro)

1.​ Main-chain scission​
→ Backbone break​
→ Decreased viscosity​

2.​ Cross-linking​
→ Molecules join​
→ Increased viscosity​

3.​ Point lesions​

→ Single bond disruption​
→ Not directly detectable​
→ May cause malfunction​

These may be repaired intracellularly.

B. DNA – THE CRITICAL TARGET

DNA is the most radiosensitive molecule.

Controls:​
• Cellular metabolism​
• Reproduction​
• Growth

Located in nucleus as chromosomes.

DNA Structure
Backbone:​
• Deoxyribose​
• Phosphate

Bases:​
• Adenine​
• Guanine​
• Thymine​
• Cytosine

Nucleotide = Base + Sugar + Phosphate

Effects of Radiation on Macromolecules In Vitro

1.​ Main-Chain Scission​

○​ Occurs when the backbone of a long-chain macromolecule is broken.​

○​ Results in long single molecules breaking into smaller molecules.​

○​ Reduces the viscosity of the solution.​

2.​ Cross-Linking​

○​ Can increase the viscosity of the solution.​

○​ Involves interactions between different macromolecular chains.​

3.​ Point Lesions​

○​ Involves disruption of single chemical bonds.​

○​ Usually not detectable.​

○​ Causes minor modifications of the molecule.​

○​ May lead to cellular malfunction.​

Reversibility of Effects:
 ●​ Effects can be repaired through intracellular repair mechanisms and cellular
recovery processes.​

Macromolecular Synthesis

Energy Flow in Cells:

●​ Catabolism → Energy → Synthesis → Anabolism

Protein and Nucleic Acid Synthesis:

●​ Essential for cell survival and reproduction.

●​ Proteins are synthesized via translation:​

○​ mRNA (messenger RNA) carries the genetic information from DNA.

○​ tRNA (transfer RNA) reads the code from mRNA to assemble proteins.​

●​ DNA is the primary molecule containing the genetic blueprint.​

○​ Located in the nucleus as part of chromosomes.

○​ Chromosomes control cell growth, development, and functions.
○​ Ultimately determine the characteristics of the organism.

S Phase (DNA Synthesis)

●​ DNA replicates so the cell can divide.​

●​ Chromatid-to-centromere ratio changes:​

○​ Before S phase: 1 chromatid : 1 centromere​

○​ After S phase: 2 chromatids (sister chromatids) : 1 centromere​

●​ Centromeres stay the same, only DNA is doubled.​

●​ Prepares the cell for mitosis so each daughter cell gets the full set of DNA.​
Radiation Effects on DNA
1. Chromosome Aberrations
Types:​
• Terminal deletion​
• Dicentric formation​
• Ring formation

2. Point Mutations
• Change in base​
• Incorrect genetic code​
• Passed to daughter cells

3. Outcomes
• Recovery​
• Cell death​
• Deterministic effect​
• Stochastic effect​
• Genetic abnormalities

V. RADIOLYSIS OF WATER (Indirect

Effect)
Humans are ~80% water.

Principal radiation interaction in body occurs with water.

Process
H₂O → Ionization → Ion Pair

Produces:​
• H⁺​
• OH⁻​
• H• (Hydrogen radical)​
• OH• (Hydroxyl radical)

Free Radicals

●​ Definition: Highly unstable molecules with an unpaired electron.

●​ Lifetime: Very short – less than 1 millisecond.
●​ Behavior:​

○​ Can diffuse through the cell.

○​ Can interact with molecules far from where they were formed.​

●​ Energy: Contain excess energy that can be transferred to other molecules.​

●​ Effects on Macromolecules:​

○​ Can disrupt chemical bonds.

○​ Can produce point lesions (minor damage to molecules).​

●​ Mechanism: Damage can occur at a distance from the original ionizing event.​

Free Radicals from Water Radiolysis

Two Primary Free Radicals:

●​ HO* (Hydroxyl radical)​

●​ H* (Hydrogen radical)​
Radiolysis of Water (H₂O)

1.​ Radiation splits water molecules, producing free radicals:​

○​ H₂O → HO* + H*​

2.​ Hydrogen Peroxide (H₂O₂)​

○​ Considered the principal damaging product of water radiolysis.​

○​ Formed when free radicals combine or react with oxygen.​

3.​ Hydroperoxyl Radical (HO₂*) Formation​

○​ H* can react with O₂ → HO₂*

○​ HO* + HO* → H₂O₂ (hydrogen peroxide)
○​ HO₂* can also produce H₂O₂ and O₂​

Summary of Reactions

●​ H₂O → HO* + H*
●​ H* + O₂ → HO₂*
●​ HO* + HO* → H₂O₂
●​ HO₂* → H₂O₂ + O₂​

Key Point:

●​ H₂O₂ and HO₂* are highly reactive and damaging, causing cellular injury even far
from the initial radiation site.

Hydroxyl radical (OH•)​

→ Most damaging

Hydrogen peroxide (H₂O₂)​

→ Toxic to cells

Hydroperoxyl radical (HO₂•)​

→ Major damaging product
VI. DIRECT VS INDIRECT EFFECT
Direct Effect
• Radiation hits DNA directly​
• More common with high-LET radiation

Indirect Effect
• Radiation ionizes water​
• Free radicals formed​
• Free radicals damage DNA

Most common mechanism in humans.

Correct answer to Question #6:​

Indirect Effect

VII. CELLULAR RADIOBIOLOGY

A. Target Theory
●​ Basic Idea:
○​ For a cell to die after radiation, its critical target molecule (usually DNA)
must be inactivated.​

●​ Interaction:
○​ The interaction between radiation and cellular molecules is random.​
Intracellular Molecular Movement

●​ Hit:
1.​ When radiation actually interacts with the target, a “hit” occurs.​

●​ Types of Hits:
1.​ Direct Hit
■​ Radiation interacts directly with the critical target molecule.​

2.​ Indirect Hit

■​ Radiation interacts with other molecules, producing free radicals
that damage the target molecule.​

Key Point:

●​ Both direct and indirect hits can lead to cell death, but indirect hits are more
common in water-rich cells due to radiolysis of water.​

B. Cell Survival Kinetics

Cell Cloning After Radiation

●​ Surviving Cells:
○​ When cells are irradiated, some survive the damage.​

●​ Proliferation:
○​ Single surviving cells can divide repeatedly, forming colonies.​

Key Point:

●​ This process is used to study cell survival and radiation effects in vitro.

1. Single-Target, Single-Hit Model

• One target per cell​
• One hit → death​
• Exponential survival curve​
• Common with high-LET radiation

2. Multi-Target, Single-Hit Model

• Multiple targets​
• Several hits required​
• Shoulder region present​
• Represents sublethal damage repair

Formula
S = N/N₀ = 1 (1 - e^D/D₀)

Where:

S/N₀ = Surviving fraction​

D = Dose​
D₀ = Mean lethal dose​
n = Extrapolation number

Smaller D₀ → Greater radiosensitivity​

Larger n → Wider shoulder → More repair capacity
VIII. CELL CYCLE AND
RADIOSENSITIVITY
Cell cycle phases:​
G₁ – S – G₂ – M

Most radiosensitive: M phase​

Most radioresistant: Late S phase

Average human cell cycle: ~24 hours

IX. MODIFICATION OF RADIATION

EFFECTS
1.​ Recovery
2.​ Oxygen
3.​ LET

High LET → Dense ionization → Less repairable damage​

Low LET → More indirect effect → Oxygen dependent