The insect cuticle: (2) hierarchical structure

This post is part of a series on the insect cuticle as a biological material that can inspire novel engineered materials. The characteristics of the cuticle, setting it apart from most synthetic/engineered materials, will be discussed in this series. The introduction to the series can be found here.

Structural hierarchy, and thus strength, is intrinsic

The insect cuticle can have great strength and flexibility, much greater than if it were just made up of stacked sheets of chitin and protein. The cuticle also exhibits energy absorption properties. This is all due to its ingenious structure. It has a complex hierarchical micro-architecture that spans multiple length scales from the nanoscale to the macroscale, which exhibit a remarkable combination of stiffness, ability for crack deflection, low weight and strength. The cuticle’s special self-assembly characteristics, as those of other biological materials, have attracted interest from materials scientists for the development of laminated composite materials and from bioengineers interested in molecular scale self-assembly.

Many biological materials exhibit structural hierarchy, and some have been studied in more detail than insect cuticle: nacre (mother-of-pearl) and bone. There is often hierarchy within hierarchy, at the nano-, micro-, all the way to the meso-level.  Hierarchical structure is primarily due to the outcome of developmental pathways of biological systems that create the material. Having hierarchy within a biological material is intrinsic. The only forces acting during material synthesis are intermolecular, and are thus very weak and work on only a short range.

This can result in very beneficial properties.

  • areas and layers that are softer than the rest can greatly affect the failure properties of a material because these interfaces can stop cracks, or divert them.
  • The hierarchical material’s microstructures can exhibit dramatic increases in compressive strength compared with that of solids of similar density with conventional structure.

Since human-made materials are often made using a top down approach, by forcing structure and shape onto a material, we do not often see structural hierarchies in engineered materials, or only to a limited extend.

Characteristic Figure from "Insects Did It 1st" book by Akre, Paulson and Catts. Illustration by E. Paul Catts.

Characteristic figure from “Insects Did It 1st” book by Akre, Paulson and Catts. This illustration by E. Paul Catts accompanied the chapter on plywood/cuticle. Plywood is a human-made material that is layered – if not quite in a hierarchical manner.

The mechanical properties of biological materials, such as the insect cuticle, may give insights into the design of more advanced engineered composites.

The layers

Meso-level hierarchy

Strictly speaking the insect’s integument is comprised of the epidermal cell layer (epidermis) and the cuticle. The Integument itself is build upon a basement membrane (basal lamina).

The insect integument. Let's call this meso-level hierarchy. The integument is comprised of an epidermal cell layer and cuticle.

The insect integument. Let’s call this meso-level hierarchy. The integument is comprised of an epidermal cell layer and cuticle, all on top of basement membrane. (Drawing by Marianne Alleyne)

The basement membrane is only 0.5 μm thick and separates the epidermal cells from the circulating hemolymph (=insect blood).  The membrane can some proteins through, and nerves and trachea also penetrate it, but when this membrane gets breached alarm bells go off in the insect’s body. Often an immune response is mounted by blood cells in response to the breach and the epidermal cells will help prepare it.

The epidermis is comprised of living cells arranged in a single layer. The cells can be modified into structures such as dermal glands and sensory receptors.

The cuticle is secreted by the epidermal cells and can be incredibly thin (e.g. larval endoparasitoids) or incredibly thick (e.g. adult rhinoceros beetles). The cuticle is non-living. It lines the external surface of the body as well as the lining of the trachea and the anterior and posterior sections of the alimentary canal and even parts of the reproductive system. This means that often even these structures undergo a re-lining during a molt.

Meso/Micro-level hierarchy

Several horizontal divisions of the cuticle are obvious, when using an electron microscope, giving the cuticle at this micro/meso-level also a hierarchical structure. These divisions came about because the sublayers were produced in a certain sequence.

The insect integument. Let's call this meso-level hierarchy. The integument is comprised of an epidermal cell layer and cuticle, all on top of basement membrane. (Drawing by Marianne Alleyne)

The meso/micro-level hierarchy of the insect integument. The cuticle is comprised of the epicuticle and procuticle. The procuticle is in turn subdivided into the endo-, meso- and exocuticle. (Drawing by Marianne Alleyne)

  1. There is a thick inner PROCUTICLE. This is the only layer that contains chitin, it also contains proteins.  As the molting cycle progresses the procuticle becomes horizontally subdivided.
    a)    Exocuticle – Is the first portion of procuticle to become synthesized, and then is pushed outward and becomes the outer layer of the procuticle.  Contains heavily cross-linked (insoluble) proteins and chitin.  This layer cannot be reused and is shed during the molt.
    b)    Endocuticle – This layer is formed just above the epidermis.  Consists of several lamellar layers of protein and chitin.  In soft-bodied insects and in areas of flexibility it is this layer that comprises most of the cuticle.  It is flexible because there is little cross-linking of proteins.  It can also be reabsorbed.
    c)    Mesocuticle – This layer cannot always be identified.  It appears to be a transitional layer.  Proteins are yet untanned (like endocuticle) but impregnated with lipids and proteins (like exocuticle), may also contain chitin.
  2. Thin outer EPICUTICLE consists of several layers, which are produced by the epidermal cells and dermal glands.  This layer does not contain chitin.  Since it is only 1 to 4 um thick it has been incredibly difficult to study. It is lipids in the wax layer on top of the epicuticle that play protective and communicative roles in the life of insects.

Nano/Micro-level hierarchy

The cuticle is a composite consisting of chitin fibers within proteinaceous matrix, all arranged in a layered structure. Several horizontal divisions can be observed. Each layer (or lamina) consists of a layer of parallel chitin chains but the laminae are stacked in such a way that the chitin molecules are arranged in an antiparallel manner.

So what does this mean? Lets see if we can build a cuticle graphically from the nano- to micro-scale.

  • Chitin is one of the two major components of the procuticle, it can make up almost half of the total dry weight of the cuticle – but it probably most likely to be less than 20% for some of the most familiar insects, such as caterpillars. Chitin is a very stable molecule: insoluble in water, dilute acids, concentrated alkali, alcohol, and organic solvents. These features make the molecule a great biological building block, but very difficult to study. Proteins are the other major component of the insect cuticle. It is the interaction between proteins and chitin that provides the mechanical function of the cuticle – giving it strength. (In the future I will come back to interesting proteins, such as resilin.)
  • Chitin is very similar to cellulose. It is an acetylated polysaccharide. A ribbon-like chitin chain is created when N-acetyl-D-glucosamines and glucosamines link together.
A portion of a chitin chain. In most insect cuticles these chains are arranged in an anti-parallel manner.

A portion of a chitin chain. In most insect cuticles these chains are arranged in an anti-parallel manner.

  • Adjacent chains of chitin are linked together through hydrogen bonds. Most commonly the orientation of the chitin chains relative to each other is anti-parallel. This arrangement (already at this nano-level hierarchical scale) allows for tighter packaging and contributes to the strength and stability of the cuticle. The grouping of 18-25 chitin molecule chains then creates chitin microfibrils that are about 3nm thick. These microfibrils are wrapped in protein.

Nanofibrils are created when groups of chitin molecules link by way of hydrogen bonds and are wrapped in protein.

  • At the next hierarchical level the nanofibrils cluster together into long chitin-protein fibers.
Nanofibrils clustered together to form chitin-protein fibers.

Nanofibrils cluster together to form chitin-protein fibers.

  • The microfibrils are laid down in an almost parallel pattern within a single layer. This creates a network or matrix of chitin and proteins, and other components such as minerals. The chitin microfibrils are covalently linked to the proteins.
Chitin-protein fibers are woven into a matrix.

Chitin-protein fibers are woven into a matrix.

  • These layers are stacked on top each other in successive layers that are rotated by a constant angle to produce a helicoidal arrangement that further contributes to the strength of the overall cuticle.
The helicoidal stacking of chitin-protein layers (matrix) creates a twisted arrangement. This creates a arching pattern in the cuticle as a whole. (Pictures by Marianne Alleyne)

The helicoidal stacking of chitin-protein layers (matrix) creates a twisted arrangement. This creates an arching pattern in the cuticle as a whole. (Here layers were created by stacking sushi mats in a twisted manner. Pictures by Marianne Alleyne)

It is the hierarchical structure created at the nano-scale, when chitin molecules link together and form chains in close association with proteins, all the way up to the meso-scale, where different cuticular layers are stacked in non-random orientations, that makes the insect cuticle such an inspiring biological material. It is because of this hierarchical structure that the insect cuticle can take on all the different functions that it has. And ultimately the cuticle is one of the main reasons why insects can be found to be thriving in so many different habitats – air, water, land, even inside horse guts. Modern analysis of small-scale composites and self-assembly-type manufacturing techniques will enable us to make materials equally as versatile as the insect cuticle.

Further reading:

Design and mechanical properties of insect cuticle. J. F. V. Vincent and U. G. K. Wegst (2004) Arthropod Structure and Development V33 pp-187-199. DOI: 10.1016/j.asd.2004.05.006


Side note about those sushi mats:

The insect cuticle: (1) multi-functionality

This post is part of a series on the insect cuticle as a biological material that can inspire novel engineered materials. The characteristics of the cuticle, setting it apart from most synthetic/engineered materials, will be discussed in this series. The introduction to the series can be found here.

Cellular and acellular layers make up the insect cuticle. With the most interior layer being comprised of living epidermal cells that secret the outside layers, which then completely cover the insect – even some of the interior surfaces such as the trachea, foregut and hindgut – are lined with cuticle. (The hierarchical characteristics of these layers will be discussed in the next post.). One of the characteristics that makes the insect cuticle, an “inspirational” biological material is its multi-functionality – something that is rarely seen in engineered materials.

Living materials, including the insect cuticle, often exhibit novel properties that are difficult to incorporate (all at once) into engineered materials. Unique physical and chemical interactions of the biomolecules that make up the cuticle (building blocks) at the nanometer scale convey characteristics such as high strength, energy absorption, and flexibility. Currently multi-functionality in engineered materials is limited to different functions due to a hybrid of several distinct phases the material can attain.

Below I have sketched out some of the functions of the insect cuticle. (If I forgot some important functions please leave a comment below and I will add them to the diagram)

insect cuticle functions

Insect Cuticle Functions – probably not complete – click on diagram to enlarge.

Insect Cuticle Functions

Protective Barrier

The insect’s exoskeleton/cuticle/integument is doing the functions of both our skin and our bones. The cuticle completely covers the insect (~armored skin), while at the same time serving as a supportive skeleton (~bones).

The protective covering creates a barrier. Precious water and ions are prevented from freely moving outward, while pathogens, parasites and dangerous chemicals are prevented from moving inward. This function is not at all trivial for insects. Since insects are relatively small they present a large surface area to the outside environment so that loss of water is a greater problem than it is for larger animals such as mammals.

Structure and Form

The insect’s exoskeleton gives the insect structure and form. And over an individual’s lifetime that form can change. In the case of holometabolous insects, such as flies, wasps, bees, beetles, butterflies and moths, this form change is striking. As an immature caterpillar a moth has a cuticle that stretches and is relatively soft, as a pupa the same individual (using the same building blocks, or biomolecules) has a cuticle that is extremely tough and does not change shape easily. Then as an adult moth the cuticle, including the wings, has yet other features that make the insect successful.

The change in structure and form seen in holometabolous insects, and to some extend the growth strategies employed by ametabolous and hemimetabolous insects, enables the animal to exploit different habitats and diets – even during its lifetime.

This is one of the most striking things about the cuticle. During the separate life stages the cuticle has different functions, it therefore has distinct characteristics and appearance. Yet, the biomolecules that make up the cuticle are pretty much the same, and one individual can synthesize all these seemingly very different types of cuticle.

How different is the cuticle from life stage to life stage? I asked family, friends and colleagues to describe, in non-scientific terms, what the cuticle of each form of the hornworm (=moth) feels like. Here are their responses.

Caterpillar or Larva


Caterpillar (Drawing by Marianne Alleyne)

Soft, rubbery, squishy, velvety, muscular, cold. Feels like pleather, like a writhing rubber pickle, like play dough that I rolled in my hands to make a snake.



Pupa (Drawing by Marianne Alleyne)

Hard, smooth, leathery, acrylic, slick. Feels weird & dead.


Adult Moth (Drawing by Marianne Alleyne)

Adult Moth (Drawing by Marianne Alleyne)

Fuzzy, hairy, soft. Feels like perfumed talc from my grandmother’s vanity. Holding it will probably feel like a fluttering beakless bird but since I haven’t held a fluttering bird, with or without a beak, I can’t be sure.


The rigidity that the exoskeleton exhibits makes it possible that insects can make rather precise muscle movements since those are due to the insertion of muscles to the integument wall. It is often the cuticle that has important biomechanical features that enables an insect to run, jump, dig, fly or swim. These precise movements are also essential for respiration (creating flow of air in and out of the tracheal system), food manipulation, excretion and osmoregulation.

Sensing the Environment

Maybe surprisingly a rigid integument is not necessarily limiting awareness of the surroundings. The cuticle has been modified in many insects into structures that can sense most of the modalities that we can sense with our skin. Some examples are the trichoid sensilla, the campaniform sensilla and chordotonal organs. Note that all three of these sensors use the cuticle as an integral part of their structure.

The functionality of the trichoid sensilla - a mechanosensory hair - is dependent on the rigidity of the cuticle.

The functionality of the trichoid sensilla – mechanosensory hairs – is dependent on the rigidity of the cuticle. (Drawing by Marianne Alleyne)

The campaniform sensilla is a mechanoreceptor found in insects. When the exoskeleton bends the resulting strain stimulates the sensilla.

The campaniform sensilla are mechanoreceptors found in insects. When the exoskeleton bends the resulting strain stimulates the sensillum. (Drawing by Marianne Alleyne)

The chordotonal organ is a stretch receptor that senses to what degree the cuticle is being deformed. This deformation can then give information about movement of body parts, gravity (proprioception) and vibrations of the surrounding air (hearing).

The chordotonal organ is a stretch receptor that senses to what degree the cuticle is being deformed. This deformation can then give information about movement of body parts, gravity (proprioception) and vibrations of the surrounding air (hearing).

Energy Storage

For many insects the cuticle also represents a temporary food store. The basic building blocks, to some extend, can be withdrawn during times of nutritive stress. Having to molt to be able to grow in size is one of the drawbacks of having an exoskeleton. Molting consumes time, energy and metabolic resources, and makes the insect vulnerable against pathogens, predators and water loss.  Reabsorbing much of the cuticle during molting minimizes some of these costs. (The closed-loop cycle characteristics of the cuticle will be discussed in part 8 of this series.)

Behavior Modulation

The single layer of epidermal cells that secretes the cuticle also secretes and deposits within or on the cuticle hydrocarbons that are involved in behavioral sequences that are important in recognition and mating. Pheromones and pigments are also deposited. The cuticle may also have modified structures that are important in mating or other behavioral processes – such as bumps,  hairs, nano-scale structures that create structural colors.

By further studying the insect cuticle it is my hope that new materials can be created, using similar “manufacturing” steps as employed by insects, that can provide increased function through integrated or further-integrated systems. Insects do it, we should too.

Next: the structural hierarchy of the insect cuticle.

(My apologies to dumpy grey-brown moths everywhere. That is a terrible drawing of an adult moth. Oh, well)

Insects have advanced degrees in Material Science and Manufacturing

There is much to learn from nature about biological materials – materials produced by a biological system. Just think about the materials in nature that you are most familiar with: wood, wool, cotton, shells, skin….consider:

  • Animals and plants are constructed from materials that are readily at hand – no dangerous mining endeavors in geo-politically sensitive areas. Sea shells are made from materials available in the sea water. Bone is made from components available in our (and our mother’s) diet. Insect cuticle is synthesized from maternal sources and from the individual’s diet.
  • The materials found in nature have many of the same attributes as engineered materials, yet avoid the heat-beat-and-treat manufacturing methods that are so costly to us and our environment. We manufacture materials by applying large amounts of energy so we can first homogenize materials, then add even more energy, and dangerous chemicals, to impose the structure that we want.
  • The periodic table categorizes more than 100 elements. Although around 25 types of elements can be found in biomolecules, only six elements are most common. These are carbon, hydrogen, nitrogen, oxygen, phosphorus, and sulfur. Few metals are used by nature, although iron, zinc, and manganese are essential in various ways.

PeriodicTable6If you create and then overlay* Ashby plots of biological and engineered materials you will learn that nature has many more tricks up its sleeve than we do. Insect cuticle is very similar to skin and bone, and some of the engineered materials, it is also ubiquitous, yet we know still so very little about it.

Collection of live and preserved insects hanging out in the Alleyne-lab. All insects have a cuticle with the same basic building blocks. But the cuticle of different species, or even different life stages, differ in hardness, flexibility. permeability, color, etc. (Photo: Marianne Alleyne)

At first glance the structure that is a characteristic of insects and other arthropods, the cuticular exoskeleton, seems such a simple yet sophisticated material. Upon closer inspection it is actually maddeningly complex and enigmatic, just ask any researcher working on it (needless to say, there aren’t too many). The chemical analysis of the insect cuticle is a frustrating undertaking.  Perform any manipulation to isolate it and “voila” the structure in which you were interested has been changed. The cuticle has been described by biochemists as “intractable.” It is possible to (partially) dissolve cuticle in various solvents and analyze the results, but the results are quantitative, both because it is not easy to break apart the assembled materials, but also because chemical transformations can occur during the process of dissolution. In the future multidisciplinary (including molecular) approaches may give us more insights into the exact composition of different types of cuticle. Maybe we will soon learn ‘why some bugs go “squish” when you step on them, but others go “crunch” even though they made out of same material’

As an old seasoned insect physiologist I am still amazed by the insect cuticle since it can have varied structure at the meso-level (allowing insects as a group to live out many life histories), yet the basic (nano-, micro-) structure and components of the cuticle are not numerous or terribly exotic. The insect cuticle is all around us, it is what makes the insects so successful.

The cuticle is produced at atmospheric pressure and at temperatures that are not extreme. Simple building blocks, mainly chitin, proteins and water, make up the insect cuticle. It can have many different features and characteristics just because the building blocks occur in different ratios or orientations. Large amounts of the cuticle are recycled by the individual insect itself, and the remainder can easily be broken down by other life forms.

Can we humans manufacture such an amazingly versatile material in a similar manner?

To manufacture anything close to the insect cuticle we need to understand the characteristics of the material. The characteristics of biological materials, that set them apart from most synthetic/engineered materials, were recently defined and listed by Meyers et al. (1). (The closed-loop cycle characteristic of the cuticle is rather unique to arthropod cuticle and should be added to the list when considering the insect cuticle).

  1. Multi-functionality
  2. Hierarchical structure
  3. Self-assembly
  4. Hydration
  5. Mild synthesis conditions
  6. Self-healing capabilities
  7. Closed-loop cycle
  8. Evolutionary and ecological constraints

Over the next few days/posts I will discuss each characteristic as it relates to the insect cuticle. At the conclusion of the series I will discuss how they all combine and result in the interesting mechanical properties of the insect cuticle, and how cuticle-inspired materials might become useful.

*Feel free to contact me for Bioinspiration learning module using Ashby plots.


  1. Meyers, M. A., J. McKittrick & P-Y Chen (2013) Structural Biological Materials: Critical Mechanics-Materials. Science V339, pp 773-779. DOI: 10.1126/science.1220854

Next: the multi-functionality of the insect cuticle.