Limited success in regenerating large bone defects has been achieved by bridging them with osteoconductive materials. These substitutes lack the osteogenic and osteoinductive properties of bone autograft. A direct approach would be to stimulate osteogenesis in these biomaterials by the addition of fresh bone-marrow cells (BMC). We therefore created osteoperiosteal gaps 2 cm wide in the ulna of adult rabbits and either bridged them with coral alone (CC), coral supplemented with BMC, or left them empty. Coral was chosen as a scaffold because of its good biocompatibility and resorbability. In osteoperiosteal gaps bridged with coral only, the coral was invaded chiefly by fibrous tissue. It was insufficient to produce union after two months. In defects filled with coral and BMC an increase in osteogenesis was observed and the bone surface area was significantly higher compared with defects filled with coral alone. Bony union occurred in six out of six defects filled with coral and BMC after two months. An increase in the resorption of coral was also observed, suggesting that resorbing cells or their progenitors were present in bone marrow and survived the grafting procedure. Our findings have shown that supplementation of coral with BMC increased both the resorption of material and osteogenesis in defects of a clinical significance

1. In two-month-old rabbits the femoral heads were made necrotic by transecting the ligament of the femoral head and applying a ligature around the femoral neck. The animals were killed at different periods, from six hours to twenty-one weeks after the operation. The changes in the femoral heads were studied histologically, microradiographically and radiographically. 2. In the first three weeks the necrotic bone marrow was penetrated by granulation tissue in which cellular differentiation gradually developed. Subsequently large quantities of new bone were deposited on the dead trabeculae. This led to an increase in the bone volume at the expense of the marrow volume; this increase coincided with an increase in the radiographic density (sclerosis) of the femoral head. The new bone tissue was attached to the necrotic trabeculae by a specific cement line and showed the features of woven bone. At a later stage lamellar bone was deposited. From six weeks on a normal bone-marrow ratio was gradually restored with concomitant radiographic loss of sclerosis. 3. It is suggested that mechanical weakening of the femoral head is the consequence of this late post-operative restoration of the normal pre-operative bone-to-marrow ratio, the new bone trabeculae being mechanically inferior because of the presence of woven bone and cement lines. This weakness may initiate collapse and deformation of the revascularised femoral head

1 . The extensor digitorum longus of the rabbit was partly denervated by section of one of its two nerve branches and examined histologically for evidence of sprouting of new fibres. 2. Sections from material fixed two and three days after operation showed terminal bundles in which varying numbers of axons and motor end-plates have degenerated. This supports the concept that the motor unit is not confined to single groups of neighbouring muscle fibres, but innervates fibres scattered throughout the muscle. 3. New fine fibres branching from intact intramuscular axons to reinnervate denervated muscle fibres were observed as early as four days after operation. 4. Such new fibres were most numerous in the early weeks after operation and their numbers then declined. Two months after operation no small fibres or simple end-plates were seen. 5. No new fibres were seen in areas of the muscle containing only denervated nerve fibres. The new fibres were formed only under the stimulus of proximity to the degenerating ones. 6. The relationship of these findings to the mechanism of recovery of human muscle affected by poliomyelitis is discussed

We studied the cellular response to physeal distraction in the growth plates of skeletally immature rabbits. We used a new method of labelling and detection of proliferating cells with bromodeoxyuridine (BUdR) and an anti-BUdR antibody. The application of an external fixator but no distraction force produced no changes in the growth plates. After five days of distraction at a maximum force of 20 N, the growth plate became thicker, mainly because of an increase in the number of hypertrophic chondrocytes, but there was no evidence of increased cell proliferation. Recent fractures were seen at the junction of growth plate and metaphysis but the increase in bone length was insignificant. After ten days of distraction at the same maximum force, the chondrocyte columns had become disorganised and cell proliferation was significantly decreased. There was an increase in bone length due to distraction of the fracture gap. In this model, physeal distraction did not stimulate cell proliferation, but actually inhibited it. The apparent increase in growth-plate thickness produced by distraction is not due to increased cell production, but results from inhibition of endochondral ossification and the consequent accumulation of hypertrophic chondrocytes. Any growth after distraction depends on the ability of growth-plate chondrocytes to divide. The decrease in proliferative activity which we found after ten days of distraction suggests the need for caution in the use of such procedures in young children

Chondrocytes at the lower zone of the growth plate must be eliminated to facilitate longitudinal growth; this is generally assumed to involve apoptosis. We attempted to provide definitive electron-microscopic evidence of apoptosis in chondrocytes of physes and chondroepiphyses in the rabbit. We were, however, unable to find a single chondrocyte with the ultrastructure of ‘classical’ apoptosis in vivo, although such a cell was found in vitro. Instead, condensed chondrocytes had a convoluted nucleus with patchy chromatin condensations while the cytoplasm was dark with excessive amounts of endoplasmic reticulum. These cells were termed ‘dark chondrocytes’. A detailed study of their ultrastructure combined with localisation methods in situ suggested a different mechanism of programmed cell death. In addition, another type of death was identified among the immature chondrocytes of the chondroepiphysis. These cells had the same nucleus as dark chondrocytes, but the lumen of the endoplasmic reticulum had expanded to fill the entire non-nuclear space, and all cytoplasm and organelles had been reduced to dark, worm-like inclusions. Since these cells appeared to be ‘in limbo’, they were termed ‘paralysed’ cells. It is proposed that ‘dark chondrocytes’ and ‘paralysed cells’ are examples of physiological cell death which does not involve apoptosis. It is possible that the confinement of chondrocytes within their lacunae, which would prevent phagocytosis of apoptotic bodies, necessitates different mechanisms of elimination

Aims

Femoroacetabular impingement (FAI) is a potential cause of hip osteoarthritis (OA). The purpose of this study was to investigate the expression profile of matrix metalloproteinases (MMPs) in the labral tissue with FAI pathology.

1. When cortisone is administered to rabbits there is early rapid resorption of bone and a partial inhibition of new bone formation. After a few days the effect becomes less obvious, so that, if observations are made at later stages, the results may be ascribed then to simple inhibition of bone growth. 2. The effect of mechanical stress has been studied in the jaw. When tooth movement is induced mechanically there is, in ordinary circumstances, a resorption of bone on the side to which the tooth is moving (the "pressure" side) and bone formation on the opposite side (the "tension" side). After administration of cortisone there is increased resorption on the "pressure" side and there is greater resorption of connective tissues here. On the "tension" side there is resorption and inhibition of bone formation. 3. In the areas of stress, when cortisone is administered, collagen fibres are no longer in apposition, being separated by spaces presumably filled with altered ground substance; this kind of change may be responsible for many of the observed phenomena. 4. A.C.T.H. does not produce a demonstrable resorptive effect on bone or connective tissue until it has been administered for periods longer than is required for cortisone (three weeks); even then the change is not pronounced. 5. In the guinea pig there is slight delay in bone formation with large doses of both cortisone and A.C.T.H., but no significant bone resorption occurs

Aims

The success of anterior cruciate ligament reconstruction (ACLR) depends on osseointegration at the graft-tunnel interface and intra-articular ligamentization. Our aim was to conduct a systematic review of clinical and preclinical studies that evaluated biological augmentation of graft healing in ACLR.

Aims

Cigarette smoking has a negative impact on the skeletal system, causes a decrease in bone mass in both young and old patients, and is considered a risk factor for the development of osteoporosis. In addition, it disturbs the bone healing process and prolongs the healing time after fractures. The mechanisms by which cigarette smoking impairs fracture healing are not fully understood. There are few studies reporting the effects of cigarette smoking on new blood vessel formation during the early stage of fracture healing. We tested the hypothesis that cigarette smoke inhalation may suppress angiogenesis and delay fracture healing.

Lesions within the articular cartilage layer of synovial joints do not heal spontaneously. Some repair cells may appear, but their failure to become established may be related to problems of adhesion to proteoglycan-rich surfaces. We therefore investigated whether controlled enzymatic degradation of surface proteoglycan molecules to a depth of about 1 μm, using chondroitinase ABC, would improve coverage by repair cells. We created superficial lesions (1.0 × 0.2 × 5 mm) in the articular cartilage of mature rabbit knees and treated the surfaces with 1 U/ml of chondroitinase ABC for four minutes. The defects were studied by histomorphometry and electron microscopy at one, three and six months. At one month, untreated lesions were covered to a mean extent of 28% by repair cells; this was enhanced to a mean of 53% after enzyme treatment. By three months, the mean coverage of both control and chondroitinase-ABC-treated defects had diminished dramatically to 0.2% and 13%, respectively, but at six months both untreated and treated lesions had a similar coverage of about 30%, not significantly different from that achieved in untreated knees at one month. These findings suggest that, with time, chondrocytes near the surface of the defect may compensate for the loss of proteoglycans produced by enzyme treatment, thereby restoring the inhibitory properties of the matrix as regards cell adhesion. This supposition was confirmed by electron microscopy. Our results have an important bearing on attempts made to induce healing responses by transplanting chondrogenic cells or by applying growth factors

Objectives

Staphylococcus aureus (S. aureus) is the most commonly implicated organism in septic arthritis, a condition that may be highly destructive to articular cartilage. Previous studies investigating laboratory and clinical strains of S. aureus have demonstrated that potent toxins induced significant chondrocyte death, although the precise toxin or toxins that were involved was unknown. In this study, we used isogenic S. aureus mutants to assess the influence of alpha (Hla)-, beta (Hlb)-, and gamma (Hlg)-haemolysins, toxins considered important for the destruction of host tissue, on in situ bovine chondrocyte viability.

1. The antigenicity of cancellous bone has been investigated in ninety-seven rabbits. 2. The immune responses of lymph nodes draining fresh homografts of cancellous bone (Burwell and Gowland 1961b) has been used as a histological indicator of the antigenicity of components of fresh homologous cancellous bone and also of the antigenicity of homologous bone subjected to a variety of physical or chemical treatments. 3. The principal antigenic component of a fresh homograft of iliac cancellous bone is the nucleated cells of the red marrow. 4. Homologous marrow-free cancellous bone does not usually produce cytological evidence of an immune response in the lymph node draining the graft, unless new homograft bone formation occurs. 5. The treatment of marrow-containing cancellous bone by boiling, freezing at - 20 degrees Centigrade, freeze-drying, irradiation or by merthiolate solution impairs the transplantation antigenicity of the tissue as a homograft. 6. The immersion of cancellous bone in a glycerol-serum-Ringer solution which is then slowly cooled to - 79 degrees Centigrade, stored for one week and then rapidly thawed, allows considerable preservation of the antigenicity of the red marrow. 7. Knowledge concerning the antigenicity of fresh and treated homologous bone is discussed. 8. Evidence is presented to show that the large and medium lymphoid cell response of lymph nodes draining homografts is due principally to the T-antigens, rather than H-antigens, of the grafts. 9. The changes which occur in the first regional lymph nodes draining tissue homografts may provide another test system to assess the transplantation antigenicity of foreign tissues or extracts of foreign tissues other than bone

1. The effects of the insertion of pieces of fresh cancellous bone into the subcutaneous tissues of the ear upon lymph nodes and spleens have been investigated in seventy rabbits. 2. The main immunological response is found to occur in the first regional nodes draining the sites of insertion of homografts of bone, which show a considerable increase in weight compared with nodes draining autografts of bone. 3. An increased number of large and medium lymphoid cells occurs principally in the first regional node of the homografted animals, as Scothorne and McGregor (1955) observed using skin as the homografted tissue. 4. The large and medium lymphoid cell response is found in both the cortex and the medulla of the lymph nodes. In the cortex a sectoral distribution of the cellular response is observed and the name reactive cortex is given to these sectors. Evidence is presented to show that the sectoral pattern of reactivity is probably determined by the localised entry into the node of iso-antigens through lymphatic vessels draining the bed of the graft. 5. We have made a quantitative analysis of the large and medium lymphoid cell response in the reactive parts of the diffuse lymphoid tissue of the cortex. The mean maximal large and medium lymphoid cell response occurs five days after the insertion of bone homografts. 6. The origin and fate of the large and medium lymphoid cells and their role in the production of antibodies is reviewed in the light of recent work. 7. A correlation is made between the maximal production of large and medium lymphoid cells in the first regional lymph node, the invasion of the graft bed with small lymphocytes and the inhibition of new bone formation in the homografts

1. The synovial membrane and capsule in osteoarthritis of the hip have been studied in twenty-five cases. Dissections have been made on fresh cadavers to establish the normal structure and function of these tissues at different ages. 2. Fragments of bone and cartilage were found beneath the synovial surface in twenty-three cases of the twenty-five cases of osteoarthritis. 3. The source of these fragments is the degenerate articular surfaces. 4. The fibrosis of the synovial membrane and capsule follows the synovial hyperplasia which accompanies the phagocytosis of these fragments. 5. A similar histological picture has been produced by injecting fragmented cartilage into the knee joints of rabbits. The injected fragments are found beneath the surface, and synovial hyperplasia is followed by subsynovial fibrosis. 6. The greatest amount of this joint debris is found in the lowest part of the joint cavity. 7. The joint capsule is particularly sensitive to traction. 8. All parts of the capsule are tight in extension, which is the weight-bearing position. 9. Fibrotic shortening of the capsule in the lowest part of the joint cavity explains many of the symptoms and signs of the disease: pain is caused by an attempt to stretch the capsule; muscle spasm occurs in the muscles supplied by the sensory nerves of this part of the capsule; extension, medial rotation and abduction, which tighten this area, are lost first; progressive shortening causes deformity in the opposite direction, namely flexion, lateral rotation and adduction; the loss of extension causes a more rapid wearing of articular cartilage on weight bearing; subperiosteal new bone is formed on the under-surface of the neck of the femur. 10. The symptomatology is discussed

Aternatives to autogenous bone graft for spinal fusion have been investigated for many years. It has been shown that osteoconductive materials alone do not give a rate of fusion which is comparable to that of autogenous bone graft. We analysed the effectiveness of porous ceramic loaded with cultured mesenchymal stem cells as a new graft material for spinal fusion in an animal model. Posterolateral fusion was carried out at the L4/L5 level in 40 White New Zealand rabbits using one of the following graft materials: porous ceramic granules plus cultured mesenchymal stem cells (group I); ceramic granules plus fresh autogenous bone marrow (group II); ceramic granules alone (group III); and autogenous bone graft (group IV). The animals were killed eight weeks after surgery and the spines were evaluated radiographically, by a manual palpation test and by histological analysis. The rate of fusion was significantly higher in group I compared with group III and higher, but not significantly, in group I compared with groups II and IV. In group I histological analysis showed newly formed bone in contact with the implanted granules and highly cellular bone marrow between the newly formed trabecular bone. In group II, thin trabeculae of newly formed bone were present in the peripheral portion of the fusion mass. In group III, there was a reduced mount of newly formed bone and abundant fibrous tissue. In group IV, there were thin trabeculae of newly formed bone close to the decorticated transverse processes and dead trabecular bone in the central portion of the fusion mass. In vitro cultured mesenchymal stem cells may be loaded into porous ceramic to make a graft material for spinal fusion which appears to be more effective than porous ceramic alone. Further studies are needed to investigate the medium- to long-term results of this procedure, its feasibility in the clinical setting and the most appropriate carrier for mesenchymal stem cells

1. Transplantations of autografts and of Kiel bone to the iliac bone and to muscle tissue were performed in rabbits. Through labelling with two tetracycline compounds which have different fluorescent colours in ultraviolet light, bone formation between the labelling periods could be followed. 2. It was shown that bone formation between the fifth and the tenth day after transplantation to bone took place in about 50 per cent of the fresh autografts. Storage of the transplants in saline for one hour before replacement had little adverse effect, whereas exposure to air for one hour seemed to reduce the osteogenic effect of the grafts. Bone formation was not observed in grafts of Kiel bone during this period. 3. The fact that new bone formation in fresh autografts could be demonstrated even during the first four days after transplantation to bone indicates that osteogenic cells from the fresh autografts continue their activity under favourable conditions. This is supported by microradiographic and histological evidence. 4. The amount of callus which developed in close contact with the grafts during the first ten days after transplantation to bone was more pronounced both in fresh autografts and in autografts kept in saline than in autografts exposed to air for one hour. Callus developing at a later stage showed no significant difference between the various grafts, including those of Kiel bone. 5. In fresh autografts transplanted to muscle tissue callus formation could be demonstrated in most cases by the tenth day, indicating either survival of osteoblasts or the transformation of more primitive cells from the graft or from the host bone into osteogenic cells. No bone formation was observed when Kiel bone was embedded in muscle tissue

1. An experimental study of the effects of nerve and muscle lesions upon the growth of bone has been made. In each case animals were subjected to unilateral lesions in the hind limb, the other limb serving as a control. The growth of the tibia was measured by calculating the difference between the length of the bone on a radiograph at the beginning of the experiment and the length of the dried bone after necropsy. The weights of the dried bones were compared. 2. In the young rabbit simple exposure of the common peroneal nerve, or division of the sural nerve, produced no change in the growth rate of the tibia. Division of both peroneal nerves, producing paralysis of the muscles below the knee, led to lengthening of the affected tibia, and this lengthening persisted until maturity several months later. A similar lengthening was seen after division of the tendons around the ankle. In spite of this lengthening the tibia on the side of the operation was almost always lighter than its fellow. 3. In the puppy division of the anterior nerve roots supplying the hind limb produced a significant lengthening of the tibia of the affected limb three months after operation. No significant changes in limb length occurred after lumbar sympathectomy in the puppy. 4. The significance of these experimental nerve lesions has been considered together with recent observations upon the growth of bone in the presence of lower motor neurone lesions in the child. From this analysis it is suggested that the initial effect of paralysis is to produce lengthening of the affected bone. This lengthening is probably due to the hyperaemia of disuse. In the presence of persistent paralysis the growth of the limb is ultimately depressed. This depression is rarely seen in the experimental animal because the growing period is relatively short. The possible causes of this secondary depression of bone growth have been considered

We implanted bone harvest chambers (BHCs) bilaterally in ten mature male New Zealand white rabbits. Polyethylene particles (0.3 ± 0.1 −m in diameter, 6.4×10. 12. particles/ml) were implanted for two, four or six weeks bilaterally in the BHCs, with subsequent removal of the ingrown tissue after each treatment. In addition to the particles, one side also received 1.5 −g of recombinant transforming growth factor ß1 (TGFβ1). At two weeks, the bone area as a percentage of total area was less in chambers containing TGFβ compared with those with particles alone (7.8 ± 1.3% v 16.9 ± 2.7% respectively; 95% confidence interval (CI) for difference -14.0 to -4.30; p = 0.002). At four weeks, the percentage area of bone was greater in chambers containing TGFβ compared with those with particles alone (31.2 ± 3.4% v 22.5 ± 2.0% respectively; 95% CI for difference 1.0 to 16.4; p = 0.03). There were no statistical differences at six weeks, despite a higher mean value with TGFβ treatment (38.2 ± 3.9% v 28.8 ± 3.5%; 95% CI for difference -4.6 to 23.3; p = 0.16). The number of vitronectin-receptor-positive cells (osteoclast-like cells) was greater in the treatment group with TGFβ compared with that with particles alone; most of these positive cells were located in the interstitium, rather than adjacent to bone. TGFβ1 is a pleotropic growth factor which can modulate cellular events in the musculoskeletal system in a time- and concentration-dependent manner. Our data suggest that there is an early window at between two and six weeks, in which TGFβ may favourably affect bone ingrowth in the BHC model. Exogenous growth factors such as TGFβ may be a useful adjunct in obtaining osseointegration and bone ingrowth, especially in revisions when there is compromised bone stock and residual particulate debris

Bone is one of the most highly adaptive tissues in the body, possessing the capability to alter its morphology and function in response to stimuli in its surrounding environment. The ability of bone to sense and convert external mechanical stimuli into a biochemical response, which ultimately alters the phenotype and function of the cell, is described as mechanotransduction. This review aims to describe the fundamental physiology and biomechanisms that occur to induce osteogenic adaptation of a cell following application of a physical stimulus. Considerable developments have been made in recent years in our understanding of how cells orchestrate this complex interplay of processes, and have become the focus of research in osteogenesis. We will discuss current areas of preclinical and clinical research exploring the harnessing of mechanotransductive properties of cells and applying them therapeutically, both in the context of fracture healing and de novo bone formation in situations such as nonunion.

Cite this article: Bone Joint Res 2019;9(1):1–14.