Transcribed by Leslie Afable Basic Tissues Lecture 17 Continuation of Bone I by Dr.

Wishe

February 3, 2014

Slide 12 Gross Structure of a Long Bone Dr. Harvey I. Wishe This was the last slide that we looked at. It was a summary slide where we had gone over the various components of a long bone, the types of bone, and we discussed periosteal and endosteal surfaces. This was more or less of a gross-type look at bone. Slide 13 G&H Text Fig. 7-10 Compact & Spongy Bone Dr. Harvey I. Wishe Now we are into the histological discussion of your typical long bone. This happens to represent a longitudinal section and as you look at the longitudinal section we are best doing this in an organized fashion. Youll see this blue structure over here, this is a connective tissue layer called a periosteum. It surrounds the bone, not at the articulating surface but rather it is surrounding shaft. It consists of two layers, an outer fibrous layer and an inner osteogenic layer. Very similar to what we discussed in terms of hyaline cartilage. There is a great deal of similarity between the different hard tissues. Then as you look at this blue region, you see little lines, those little lines represent collagen fibers called SHARPEYS FIBERS. Sharpeys fibers attach the periosteum to the underlying bone. You have to realize that once bone forms, you cant put in fibers. So the fibers have to be there to begin with then the bone forms on top of and around the fibers. Then this area right in here that is labeled, represents the outer circumferential lamellae or you can use the term PERIOSTEAL LAMELLAE, which makes sense because it is underneath the periosteum. Thats actual bone tissue. As we get more internal to the bone, you are going to find a certain organization where the functional unit of your compact bone is the picture that you see on the top and thats your OSTEON or HAVERSIAN SYSTEM. There are many of them in the compact bone. Then we get towards the inside of the bone. Again we see some bone lamellae and what doesnt really show up is your ENDOSTEUM. But all this region in here looks like your household sponge. There is a connective tissue layer that lines all the internal spaces. So I am just going to put this red mark here and thats called your endosteum. So the endosteum is lining all the trabeculae, all the partitions of spongy bone. In addition, it will line this haversian canal. So the inside of the bone has an endosteal surface with which the endosteum is associated with. So weve covered so far, 3 types of lamellae. Periosteal, which is on the outside. Endosteal which is on the inside. Then I really didnt mention it, but youll notice around the osteon there are also more lamellae and they encircle the haversian canal. Around this haversian canal you will also find an endosteum. Those lamellae are called HAVERSIAN LAMELLAE, they are concentric rings. Initially what an osteon is forming, thats the haversian canal and we will just put in one layer of bone. When you are looking at a mature osteon, thats the haversian canal and now youll have many layers of bone. In fact, you can have anywhere from 5-20 layers.. 5-20 haversian lamellae. The big difference between peri and endosteum is the number of layers making up each. Periosteum has 2 layers-- outer fibers, inner osteogenic. Whereas the endosteum only has one layer, an osteogenic layer. In a few minutes we will see a picture illustrating all of that. Then as we look at this region, this is sort of like you just pulled 2 straws out of a 1

Transcribed by Leslie Afable

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box. What we are really doing, there is the haversian canal here and then each set of haversian lamellae are now being pulled up as well. And the reason you can see the different lamellae depends on the arrangement of the collagen. So what you really see, here is your collagen going in this direction and then it goes this way and that way so you really have a preferential arrangement of collagen fibers and they extend in different directions. As a result, the various lamellaes can be distinguished one from the other. This is best seen on a ground section which I will show you what that is shortly. The other thing that you should be noticing is all these little spinal like structures. You can see the same thing up here (top right photo) while there is a dark space thats called your lacuna. Maybe this will be a good spot for me to draw something. Diagram Drawn By Wishe During Lecture

Dr. Harvey I. Wishe So what I am essentially drawing is an osteoblast. An osteoblast has cytoplasmic processes. Then the cell produces your organic matrix, and as a result you find the whole cell and the processes becoming entrapped. The organic matrix has been deposited outside the cell, outside the processes. The cell and the processes are completely enclosed by the organic matrix. What I am drawing in red represents a space called the lacuna. So between the organic matrix and the cell itself, you are going to get a space called the lacuna. The same thing happened with your chondroblast producing cartilage matrix and gets trapped in the lacuna. You will see the same thing with cementum. So they all have similar type of scenarios. So the dark yellow color is surrounding the cytoplasmic processes. In essence, it is a space. But that is NOT called a lacuna, its a canaliculus. So the space that surrounds the cell body of the osteoblast is your lacuna, and the space that surrounds the cytoplasmic processes is referred to as a canaliculus. The lacuna and canaliculi are continuous with one another and they are filled with extracellular fluid. And

Transcribed by Leslie Afable

February 3, 2014

because of this arrangement, material can be transported back and forth. I will show you some pictures of that shortly as well. Slide 13 G&H Text Fig. 7-10 Compact & Spongy Bone 3rd Ed. Dr. Harvey I. Wishe Now other than the haversian canal, I really havent mentioned anything about blood vessels. The haversian canal does have blood vessels, nerves, lymphatic vessels. When it comes to compact bone, thats the only place where you will find vascularity. There are no blood vessels any place else. Now here is a blood vessel on the outside of the long bone and it passes through the periosteum into the inside of the bone through canals. You can see a canal here and another, and same thing here. Those are called Volkmanns canals. So volkmanns canals are interconnecting canals and they interconnect haversian canals. Haversian canals, essentially, are parallel to the long axis of the bone and volkmanns canal are more or less perpendicular to the long axis of the bone. In addition to blood vessels coming in through the periosteal surface, lets look at this part of the compact bone. What do we find in these spaces? Anyone remember? Well, surrounding the little pieces of bone is the endosteum but whats in the space? So you have pieces of spongy bone but there are spaces in between. BONE MARROW. So in the bone marrow there are plenty of blood vessels. So youll have coming from the bone marrow, moving into the compact bone. Let me just put in here for instance, another canal and I will make that a volkmanns canal. So blood vessels can enter from 2 different sources, the periosteal surface and the endosteal surface. So thats how compact bone gets all the blood for bringing in nutrients and getting rid of waste products and having respiratory exchange. If we look at the spongy bone, you dont have any blood vessels in the spongy. But what surrounds the spongy bone is the bone marrow. Thats where the blood vessels are. So the blood vessels come up to the spongy bone and between the two you now have an interchange of nutrients, waste products and respiratory gases. If we go back to our haversian canal, so you are having all sorts of good stuff being brought in and bad stuff taken out. This material diffuses from the blood vessel internally and it goes through these little spinal like lines, which really represent the canaliculi which are not blood vessels but merely channel ways. Think of the haversian canal as first avenue and think of the canaliculi as 24th street and 25th street. Because of this arrangement, you have a tremendous interconnection between all parts of the compact bone. As you look at this picture, you can sort of get that feeling. But some of the upcoming pictures show it a lot better and you will understand how well structured the compact bone actually is. Slide 14 G&H Atlas Plate 4-3 Fig. 2 Dr. Harvey I. Wishe This is a decalcified section of compact bone. The previous slide picture was just diagrammatic. But here you can see the osteons (Os) or haversian systems. There is always a canal in the middle, its called a Haversian Canal (HC) and some people call it a central canal. Surrounding the canal are haversian lamellae, thats essentially what we are talking about. In the haversian lamellae you will find spaces called lacuna. What you are seeing in the spaces are the dark staining structures which are really the nuclei of your osteocytes. When an osteoblast traps 3

Transcribed by Leslie Afable

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itself in the matrix (we can refer to the picture he drew) it is no longer an osteoblast, it is now an osteocyte. The same thing with cartilage, chondroblast traps itself in the matrix and it is now a chondrocyte. The cell is sitting in the lacuna. You will find the same story with cementum. Cementoblast -- cementocyte. Dentin is similar but it is different and I dont think its a good idea right now to get involved with that. VC is a Volkmanns canal, so here is one haversian canal and here is another one and they are linked together by a Volkmanns canal. Here is another Volkmanns canal and another built into here. Now blood is coming into the compact bone via a Volkmanns canal and the blood passes down into the haversian canals and then we have a diffusion process that takes place distributing the various important ingredients and removing the bad stuff etc. You see the 2 arrows, and there are 2 more arrows over here, and they are pointing to a basophilic line. Those are called cement lines. There are 2 types of cement lines, arrest and reversal. An arrest line means bone formation is occurring, and thats me walking and all of a sudden it stops for some reason and then it starts up again. So you temporarily halted the process. The reversal line has a different implication and thats when you change processes. So you can have bone formation followed by bone resorption followed by bone formation again. So youre reversing the process. Breakdown of bone with formation of bone. What appears between the old and new bone happens to be a reversal line. You dont have to worry about identifying which of these is arrest or reversal. Slide 15 G&H Text, Fig. 7-21 Dr. Harvey I. Wishe Another fairly decent picture. There are some nice osteons over here, and you can see the haversian lamella going around the canal. What do you think I just drew? A haversian canal? No. A pizza pie with 8 slices. When bone resorption occurs, lets say you eat 7 slices of pie. As a result, you have 1 slice left over. So you got rid of most of the slices and you got rid of the haversian canal and you are left with this one slice. This one slice is equivalent to the last type of lamella called interstitial lamella. We can see a little something over here or in here. So interstitial lamella are groups of lamella that are the remnants of bone resorption and we will go into that a little more deeply.

Transcribed by Leslie Afable Slide 16 G&H Text, Fig. 7.5 Compact Bone

February 3, 2014

Dr. Harvey I. Wishe Here is a rather high power of your compact bone. It is showing you a really nice haversian canal. Here is your space and these cells labeled Op means osteoprogenitor cells. So in order to form bone in compact bone, you have to have osteoprogenitor cells coming in from some place. They wind up lining all the haversian canals. They are originally mesenchyme cells which sort of differentiate a little bit and become predestined to become osteoblasts and then you have bone forming. One of the earlier pictures that I drew with a large haversian canal, then it becomes a small haversian canal.. Well how did that canal fill up? Because of the osteoprogenitor cells laying down more layers. Having said that, 1.. 2.. Which is the oldest bone? 1 or 2? 1, because it is furthest on the outside. 2 is closest to the osteoprogenitor cells so that is the one that is more recently formed. You will notice all these basophilic lines that are cement lines. You do see lacuna with your osteocyte sitting in it. In reality, you are only seeing the nucleus. Slide 17 G&H Atlas Plate 4-3 Fig. 4 Compact Bone

Dr. Harvey I. Wishe This is a ground section compared to the previous slide which was decalcified. That was one of the questions on the quiz, the difference between hard and soft tissues. Do you have to decalcify soft tissues before you can cut it? No. But bone you do otherwise you cant cut it. You try to get rid of the calcium such that the bone now becomes sort of soft and it is easier to cut. So decalcification is important in preparation of hard tissues for cutting for you to stain and look at. Ground section means I walked up to one of you with a hammer and chisel and I 5

Transcribed by Leslie Afable

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need a knife to slit open the skin with various layers and get to the bone and I take the hammer and chisel and get a little chip of bone. I put it on a slide and this is what you see. This is not stained. That is your natural intact bone. But you can still see the osteon very nicely and the haversian canal. When you look at ground bone, anything that was a space comes out dark. Thats why you are seeing the canal dark as opposed to light. So theres the haversian canal. This is also part of a haversian canal as well as this and roughly this area would be you volkmanns canal interlinking 2 different systems. This has a perfect example right here of my interstitial lamella. The only thing is that it is a square pizza as opposed to the normal triangular slice. Slide 18 G&H Plate 4-4 Fig. 1 Canaliculi Dr. Harvey I. Wishe Very high power of an osteon with a canal. Again, all these dark structures in here represent lacuna, NOT cells. Cells fall out of the ground substance. Then you can see all these various radiating lines, those are the canaliculi. Then you start to follow it.. So here is the haversian canal in the center and there are canaliculi going to this lacuna and canaliculi passing to the next one and you have a continuous flow system throughout the osteon and thats how you have the transport of various things through compact bone. Slide 19 G&H Text Fig. 7-11 Dr. Harvey I. Wishe Another picture again. You can see the canaliculi much better and this gives you a good impression of the interconnection of all the layers of your haversian lamella. Slide 20 Microscopic Structure of Bone Dr. Harvey I. Wishe So in terms of a summary, we have a periosteum which has 2 layers. Outer fibrous and inner osteogenic. And dont forget Sharpeys fibers that attach the periosteum and endosteum to the underlying tissue. By the way, Sharpeys fibers also attach to cementum but that we will deal with at the appropriate time. So youre PDL actually extends from the cementum of a tooth to your alveolar bone and into both hard tissues you will get your Sharpeys fibers which represents your anchorage point. The endosteum is a single layer with osteogenic potential. Here we have 4 types of lamellae, periosteal endosteal. There are other terms for this and if you look at an earlier picture you will see that periosteal is also known as your outer circumferential lamellae and it goes around, thats what circumferential means. Outer means its on the outside surface, periosteal surface. You have the same type of arrangement for the endosteal but there it is called inner circumferential lamellae. Then we have the haversian lamellae which surrounds all the haversian canals and the interstitial lamellae which represents remnants of bone resorption. We will go into that a little more deeply. So the functional unit of bone is an osteon, haversian system, and the volkmanns canal from the outside and inside bring blood vessels into the compact bone. Then we talked about cement lines.

Transcribed by Leslie Afable

February 3, 2014

Slide 21 Osteon/Haversian System Dr. Harvey I. Wishe These are the components of your osteon. A haversian or central canal, the haversian lamellae which go around the canal. Each lamellae has lacuna and each lacuna will be an osteocyte. Then there are cement lines generally surrounding an osteon. Slide 22 Types of Bone Dr. Harvey I. Wishe Types of bone. We briefly alluded to this, but we have primary immature woven spongy bone. This is the first bone that forms no matter what. If you are first forming a bone, if you are repairing a bone, if you have a bone tumor then woven bone will be the first type of bone formed. It is very cellular and because it is so cellular, you will have less ground substance and less mineral. There is collagen present but it is completely disorganized. Because of that, you will not see bone lamellae. That gets resorbed and is now replaced by your secondary mature trabeculae bone. There is another word in here called lamellated and some folks like to use that but I DONT. Lamellated means it has lamellae, but compact bone also has lamellae. So if you say lamellated bone, I dont know which one you are discussing. So I would tend to leave lamellated out of any discussion. So when you are dealing with mature spongy bone, you will see lamellae. All that means is that the collagen has been organized in a specific fashion so that the collagen fibers are lined up in a preferential arrangement. Then some of this mature secondary spongy bone will become compact bone. Slide 23 G&H Atlas Plate 4-4 Fig. 2 Woven Bone Dr. Harvey I. Wishe Here we have a picture essentially showing us woven bone. What you are seeing are pieces of bone. T stands for trabeculae. Its just a piece of bone. Here we have a little guy and I could put S in there and that means spicule. What is the difference between spicule and trabeculae? One is small, that is a spicule. Trabeculae are just a larger piece of bone. As you look at this bone you see a lot of cells. They are all in lacuna. This area and down in this region. So once you start getting so many cells you know that its not a mature bone and it has to be woven bone. What is this tissue in here (lighter colored area in between the whole regions labelled T)? Anyone have any idea? It will become bone marrow, that is true, but in this stage of the game what is it? It is embryonic connective tissue and what do we call embryonic connective tissue? Mesenchyme. Its not mucus because mucus would have big gaps and the gaps are where Whartons jelly used to be but were dissolved out. So all this tissue is mesenchyme. Notice also the number of blood vessels. That is another key to being mesenchyme. Then you will notice Hc which stands for haversian canal. You dont really go from woven bone to compact bone. Usually the woven bone becomes a mature spongy bone and some of the mature spongy bone then becomes compact bone. You will find a lot of this happening from the internal surface, the endosteal surface. More so than at the periosteal surface. So this shows us actually how compact bone could form. There has to be a blood vessel and each one of these has a blood vessel. So I technically could start forming bone around this or that (medium pink areas), if theres no blood vessel then you wont get an osteon. By the way, cartilage does not have blood 7

Transcribed by Leslie Afable

February 3, 2014

vessels which we mentioned last week. It doesnt need blood vessels. So if there are no blood vessels, which tissue will form first? Cartilage or bone? It has to be cartilage because in order for bone to form, you require vascularity. You require oxygen. So bone has a high oxygen tension and cartilage has a low oxygen tension. So that determines how bone is going to form and we will get to intramembranous vs endochondrial shortly. Those are the 2 methods of bone formation. Slide 24 A. Fig. 10-1 Alveolar Bone; Tooth PDL Alveolar Bone Dr. Harvey I. Wishe Just to show you that bone does exist in terms of the mandible. This happens to be the mandible because look at how thick this bone is so this is cortical bone right over here wrapping around the mandible. This is referred to as your interdental bone in between 2 teeth same thing here. And then all the tissue in this area is really part of the PDL. Technically we are looking at alveolar bone which will have lamellated bone and some spongy bone and even possibly some compact bone. Slide 25 A. Fig. 12-5 Alveolar Bone Dr. Harvey I. Wishe This is a very nice slide. Again, there is one tooth and there is a second tooth and there is your alveolar bone and these are all collagen fibers making up the PDL. Now you will notice going into the alveolar bone, you can see the fibers. The part of the collagen fiber that is going into the alveolar bone, what do we call that? Sharpeys fibers. The same thing could happen at this end, the sharpeys fibers would extend into the cementum of the tooth. You dont have to know this now, but these fibers go from the cementum of one tooth to the cementum of the other tooth, those are you interdental fibers. This is the alveolar crest and the fibers inserting here are your alveolar crest fibers. These fibers seem to be at an angle and we call those oblique fibers. Slide 26 A. Fig. 11-2 Alveolar Bone Dr. Harvey I. Wishe In this picture these fibers are more or less straight, so they are horizontal fibers. There are a lot of different fibers that go into making up the PDL. I remember we use the word gomphosis to describe this as a joint. Its more of a cushioning type scenario and the fibers respond to the pressure that is brought to bear upon the tooth. Slide 27 Bone Matrix Dr. Harvey I. Wishe Matrix. Inorganic and organic matrix. So in terms of the inorganic matrix, we are talking about calcium which is organized into hydroxyapatite crystals. And 65% of the bone is inorganic in nature. Dentin is 70% and enamel is 96%. Most of the hydroxyapatite, the calcium is in combination with phosphate, calcium phosphate. Thats about 85%. 10% calcium combines with carbonate. The remaining 5% is calcium in combination with various other ingredients. The hydroxyapatite crystals are slender crystalline structures and they tend to be deposited on top of, around, along side your collagen fibers. Hydration shell -- calcium or the hydroxyapatite crystal does bind to water. As it does that, the outside of the crystal has a shell of water. Thats your hydration shell but the water 8

Transcribed by Leslie Afable

February 3, 2014

is held onto the crystal. Hydroxyapatite which we abbreviate as HAP is responsible for the hardness of the tissue. But something very hard isn't necessarily 100% protective. Something hard could fracture very easily. Use your concept of a porcelain plate, that is pretty hard but if you drop it on the floor it cracks to smithereens and you have to pick up all the pieces. Now for mineralization to occur, the HAP has to attach to something and thats what we call the nucleation core. It was once believed that nucleation core were actually your collagen fibers. But then it was found out that it wasnt the collagen fibers but rather these NCPs noncollagenous proteins. When mineralization occurs you tend to lose some fluid but you dont lose much collagen. Now sometimes it is important to measure the degree of calcification and I have just listed 2 particular lets call them diseases. One we will talk about again, osteomalacia. This is the adult form of rickets which children have and it relates to a deficiency in vitamin D. If you dont have enough vitamin D you wont have the calcium binding as it should. You will be deficient not only in D but also in calcium. There is a technique you can use to determine the measurement of calcification. It turns out in osteomalacia, you have NOT a 65% mineralization rate but only like 35%. So the tissue is not as hard as it should be. This could be temporary scenario. Pregnant women, a lot of the calcium goes to support the early developing child. So she could be just temporarily having this condition. The second, osteitis fibrosa cystica is a situation where you have too much bone resorption via a cell known as an osteoclast. As a result, you lose bone and mineralization and the bone begins to break down. So those are just 2 examples of where it would be to the doctors advantage to measure the degree of calcification. Now we come to organic matrix and the organic matrix makes up 35% of the bone. Essentially we are dealing with type I collagen which is roughly 85% of the 35%. The other 15% of the 35% happens to be your NCPs. Then I listed type V collagen. Well if we wanted to say that collagen is 100%, type I is 97% of that. Type V is 3%. You dont have to know the particular numbers, they are just there for illustration. But you should know that the organic matrix is 35% organic in nature with type I making up most of it and now the NCPs make up the rest. It turns out that your osteoblast produces your type I collagen and it produces your NCPs. About half the amount of time is devoted to producing type I and half the time is devoted to producing NCPs. By the way, collagen is a much larger molecule and NCPs are roughly 1/10 the size of your collagen. Slide 28 Noncollagenous Proteins Dr. Harvey I. Wishe This lists the various NCPs. Bone attachment molecules, proteoglycans, osteocalcin, growth factors, alkaline phosphatase and osteonectin. There are 4 bone attachment molecules. So these chemical entities play an important role in attachment of your various components in your bone. All but one will bind to calcium as well. The one that doesnt bind to calcium happens to be your fibronectin. It is meant strictly for attachment and spreading of cells. The others are involved in attachment and NOT spreading of cells but binding to calcium. Then we have proteoglycans which was mentioned in connective tissue. What is a proteoglycan molecule mostly made up of? Carbohydrates! Look at the suffix glycan that means carbohydrates. So in reality, carbohydrates make up maybe 9

Transcribed by Leslie Afable

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70% and the protein makes up roughly 30%. I think theres probably a conference that I mentioned a description of your proteoglycan. You can think of it as a tree and the stem of the tree is the protein core and then all the branches and leaves coming off represent your carbohydrates. What is the name of those carbohydrates? GAG molecules. Another example was a test tube brush, finally I gave you the idea of a caterpillar. Where the body of a caterpillar is your protein core and all the little itsy bitsy feet represent your GAG molecules. Mostly GAGs have sulfur in them. Chondroitin sulfate or heparin sulfate. But some dont have sulfate. These 3 substances (versican, biglycan, decorin) represent your protein core under different circumstances. The first one, the versican is when the bone is first forming. The biglycan is when the bone has got a good way into forming but its not complete. And finally the last one, decorin, is the protein core when you have your fully formed bone. And they are all linked to GAG moelcules like chondroitin sulfate. The heparin sulfate is believed to be linked to the membranes of the cell and maybe it facilitates/makes it easier for osteoblasts to interact with the extracellular matrix. So we are up to osteocalcin. Thats a relative to glutamic acid. Most of these factors, by the way, are produced by the osteoblast. Theres a relationship between the amount of vitamin D on the amount of the factors that are produced. The osteocalcin is a very strong calcium binding agent. It probably leads to the most binding of calcium of all the factors. Then we have the growth factors and this could be something like fibroblast growth factor or insulin-like growth factor, transforming growth factor, all of them play a role in bone formation. Alkaline phosphatase is an enzyme which you should realize on your own and when you go to a physician and he does a blood test, and by the way, youre entitled to get a copy of your blood test and you should. This way you can see what your values are. Some people dont give a hoot about it, doctor says youre OK, fine I dont have to know anything more. But the fact that you are in the science field and you are doing dentistry, you really have to know this type of information about your patient. What kind of condition the patient has, is he or she producing too much bone or not enough bone? This could effect the treatment. So the fact that you have to know this information about your patient, you might as well know it about yourself as well. Alkaline phosphatase is a direct determinant of the amount of bone that is produced. You get alkaline phosphatase, you know bone is being made. If it is too much, that means something is wrong. If it is not enough, that means something is wrong as well. Alkaline phosphatase plays some sort of role in regulating your mineralization as well. And finally the last item, the osteonectin, is another fairly strong calcium binding molecule. It has a very high affinity for your calcium and its probably the most abundant NCP that is around. So there are a lot of molecules here that bind calcium. This is just not a simplistic matter.

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