Temporal profile of microvascular disturbances in rat tibial periosteum following closed soft tissue trauma

Background and aims Bone devascularization due to impaired periosteal perfusion following fracture with severe soft tissue trauma has been proposed to precede and underlie perturbed bone healing. The extent and temporal relationship of periosteal microcirculatory deteriorations after severe closed soft tissue injury (CSTI) are not known. We hypothesized that periosteal microcirculation is adversely affected and the manifestation of trauma-initiated microvascular impairment in periosteum is substantially prolonged following CSTI.Material and methods Using the controlled-impact injury device, we induced standardized CSTI in the tibial compartment of 35 isoflurane-anesthetized rats. Following the trauma the rats were assigned to five groups, differing in time of analysis (2 h, 24 h, 48 h, 1 and 6 weeks). Non-injured rats served as controls. Before the metaphyseal/diaphyseal periosteum was surgically exposed, intramuscular pressure within tibial compartment was measured. Using intravital fluorescence microscopy (IVM) we studied the microcirculation of the tibial periosteum. We calculated the edema index (EI) by measuring the skeletal muscle wet-to-dry weight ratio (EI = injured limb/contralateral limb).Results Microvascular deteriorations of periosteal microhemodynamics caused by isolated CSTI were reflected by persistent decrease in nutritive perfusion, markedly prolonged increase in microvascular permeability associated with increasingly sustained leukocyte rolling and adherence throughout the entire study period, mostly pronounced 48 h after the trauma. Peak level in capillary leakage coincided with the maximum leukocyte adherence, tissue pressure, and edema. Microcirculation of tibial periosteum in control rats demonstrated a homogeneous perfusion with no capillary or endothelial dysfunction.Conclusion Isolated CSTI in absence of a fracture exerts long-lasting disturbances in periosteal microcirculation, suggesting a delayed temporal profile in manifestation of CSTI-induced periosteal microvascular dysfunction and inflammation. These observations may have therapeutic implications in terms of preserving periosteal integrity and considering the interaction of skeletal muscle damage and periosteal microvascular injury during management of musculoskeletal trauma.The paper was presented at the 2nd Musculoskeletal Symposium Significance of Musculo-Skeletal Soft Tissue on Pre-Pperative Planning, Surgery and Healing, 13–14 February 2003, Berlin, Germany     

Mechanics of Avian Fibrous Periosteum: Tensile and Adhesion Properties During Growth

We report the results of direct mechanical tests of the fibrous periosteum from the tibiotarsi of white leghorn chicks at 4, 6, 8, 9, 10, 11, 12, and 14 weeks of age using a newly developed sample isolation technique. Additionally, this technique allows the determination of the apparent in vivo load on the fibrous periosteum. The periosteum has a highly nonlinear stress-strain relationship at all ages. For loading below the in vivo level, the periosteum is pliant and mean tensile modulus is 3.35 MPa (±1.84 SD, n = 75). For loading above the in vivo level, tensile stiffness is nearly two orders of magnitude greater. In the region of high stiffness, mean modulus is 229.5 MPa (±89.6, n = 72). In vivo, the periosteum is loaded at the transition between these two stiffness regions. We interpret this as indicating that, in vivo, the collagen fibers of the periosteum are aligned, but subject to minimal loading. Stress levels in the periosteum corresponding to in vivo conditions indicate modest loading, and mean apparent in vivo stress levels are 0.92 MPa (±0.37 SD, n = 67). A second technique demonstrated that the adhesion of the periosteum in the diaphyseal region (1–6 weeks of age) is minimal, but is substantial in the metaphyseal region. The metaphyseal adhesion will affect the transmission of load between the physes. These studies suggest that growth of the fibrous periosteum follows the longitudinal growth of the bone, rather than the periosteum having a direct mechanical influence on growth plate activity. Comparison of tensile properties over the course of growth indicates a substantial increase in periosteal stiffness in the early portion of the growth period, which reaches a maximum at approximately 9 weeks posthatching. There is also a marked decline in periosteal stiffness as growth rate declines in the latest stages of growth (14 weeks). This suggests that the basic properties of periosteal collagen may undergo a transition during the course of this tissue’s brief functional lifetime; that is, during long bone growth.     

Determinants of bone size and strength in 13-year-old South African children: the influence of ethnicity, sex and pubertal maturation

We have previously shown ethnic differences in bone mass between pre-pubertal black and white children using DXA. To investigate these ethnic differences further, using pQCT, and to determine the influence of sex and pubertal development, we measured appendicular bone variables in 13-year-old children using pQCT. We collected pQCT data on a cohort of 471 black and white children at age 13years. Black boys and girls were shorter and had less lean mass than their white peers, and black boys were lighter than white boys at an earlier stage of pubertal development. Metaphyseal (4%) radial trabecular density was greater in the black girls than their white peers (239.5±49.5 vs. 222.7±34.2 mg/cm(3); p<0.05). Bone strength index was not different between the ethnic groups. All metaphyseal measures were 3-41% greater in boys than girls, after adjusting for height where appropriate. Diaphyseal (38%) tibial values, including total area, endosteal diameter, tibial diameter, periosteal circumference and polar strength-strain index were 4-22% greater in the black than white children and in boys than in girls. Cortical density was greater in black than white boys (1079.0±39.4 vs. 1058.7±34.5 mg/mm(3); p<0.001) and greater in the girls than boys (black: 1129.3±33.7 vs. 1079.0±39.4 mg/mm(3); p<0.001; white: 1126.8±28.3 vs. 1058.7±34.5mg/mm(3); p<0.001). Cortical thickness was less in the black groups. Lower leg muscle cross-sectional area (MCSA) was higher in white than black children, and forearm MCSA was higher in white than black boys. There was no difference in fat cross-sectional area between the ethnic groups. In conclusion, ethnic and sex differences in both metaphyseal and diaphyseal bone parameters exist during puberty, which are not accounted for by differences in body size or skeletal maturity. South African black children have wider diaphyseal regions of appendicular bones with greater measures of bone strength.     

Lower trabecular volumetric BMD at metaphyseal regions of weight‐bearing bones is associated with prior fracture in young girls

Understanding the etiology of skeletal fragility during growth is critical for the development of treatments and prevention strategies aimed at reducing the burden of childhood fractures. Thus we evaluated the relationship between prior fracture and bone parameters in young girls. Data from 465 girls aged 8 to 13 years from the Jump‐In: Building Better Bones study were analyzed. Bone parameters were assessed at metaphyseal and diaphyseal sites of the nondominant femur and tibia using peripheral quantitative computed tomography (pQCT). Dual‐energy X‐ray absorptiometry (DXA) was used to assess femur, tibia, lumbar spine, and total body less head bone mineral content. Binary logistic regression was used to evaluate the relationship between prior fracture and bone parameters, controlling for maturity, body mass, leg length, ethnicity, and physical activity. Associations between prior fracture and all DXA and pQCT bone parameters at diaphyseal sites were nonsignificant. In contrast, lower trabecular volumetric BMD (vBMD) at distal metaphyseal sites of the femur and tibia was significantly associated with prior fracture. After adjustment for covariates, every SD decrease in trabecular vBMD at metaphyseal sites of the distal femur and tibia was associated with 1.4 (1.1–1.9) and 1.3 (1.0–1.7) times higher fracture prevalence, respectively. Prior fracture was not associated with metaphyseal bone size (ie, periosteal circumference). In conclusion, fractures in girls are associated with lower trabecular vBMD, but not bone size, at metaphyseal sites of the femur and tibia. Lower trabecular vBMD at metaphyseal sites of long bones may be an early marker of skeletal fragility in girls. © 2011 American Society for Bone and Mineral Research.     

Anatomical effects of periosteal elevation.

Research that involves harvesting the periosteum is common. The exact technique of harvesting is rarely described; however, it may be of vital importance because techniques may vary in their ability to raise the osteogenic cambial layer, which is reported to be tightly adherent to the underlying cortex. This study was performed to define how the cambial and fibrous layers of the periosteum are affected by different techniques of stripping. The periosteum was raised from the tibia and the humerus of adult rabbits with four stripping techniques. The stripped bone surface was examined histologically and with a scanning electron microscope to determine whether the fibrous and cambial layers of the periosteum had been removed and whether there had been damage to the underlying cortex. The results from the two anatomical sites were the same. Raising the periosteum with cortical bone chips (shingling) or with a periosteal elevator removed both layers of the periosteum and caused considerable damage to the surface of the cortex. Raising the periosteum with a sharp scalpel or by simply pulling it off removed the fibrous layer but left the osteogenic layer intact adherent to the cortex. We conclude that some techniques of periosteal elevation fail to harvest the osteogenic layer and therefore may lead to unexpected experimental results. We suggest that authors describe the exact technique of periosteal stripping that was employed.     

Osteoclasts differentiate from resident precursors in an in vivo model of synchronized resorption: a temporal and spatial study in rats

Osteoclasts differentiate from mononucleated precursors expressing monocyte markers, which gradually evolve to preosteoclasts expressing the osteoclast phenotype. Although the role of osteogenic cells in these changes has been well documented in vitro, their contribution in vivo has not been established. In this study, a synchronized wave of resorption was activated along the mandibular periosteum. The periosteum adjacent to the bone surface studied was separated by a computer-assisted technique into an osteogenic alkaline phosphatase-positive compartment and an outer nonosteogenic compartment. Specific markers (nonspecific esterase [NSE], tartrate-resistant acid phosphatase [TRAP], and ED1 antibody, a marker of the monocyte-macrophage lineage) were used to follow osteoclast differentiation quantitatively as a function of time after activation of resorption, from day 0 to day 4 (peak of resorption in this model). Local cell proliferation was assessed in parallel. Between day 0 and day 3, the thickness of the osteogenic compartment decreased by 50% (p < 0.0002). In the osteogenic compartment, proliferating cell numbers fell by 80% at 12 day, NSE(+) cells (located farthest from the bone surface) increased 3. 9-fold on day 4 vs. day 0 (p < 0.005), ED1(+) cells decreased between day 0 and day 2 (p < 0.02) before returning to their initial value, and TRAP(+) cells increased 2.7-fold between day 1 and day 3 (p < 0.0005). Resorption was absent in the site studied on day 0, but on day 4 there were 20.5 osteoclast nuclei per millimeter of bone surface. The cell ratio changed from 30.3 NSE(+) and ED1(+) (some of which were also TRAP(+)) cells per millimeter on day 0 to 37.6 mononucleated cells plus 20.5 osteoclast nuclei on day 4. In the nonosteogenic compartment, an entry of ED1(+)/NSE(-) was observed on 12 day (+23 cells, p < 0.02 vs. day 0). This was followed by a return of ED1(+) cell numbers to the control level on day 1, and a transient increase in NSE(+) cells (+47% on day 2 vs. day 1, p < 0.02). TRAP(+) cells were never seen in this compartment. Proliferating cell numbers did not change throughout the study. Our results strongly suggest that the osteoclasts present on day 4 differentiated from the pool of TRAP(+), ED1(+), and NSE(+) cells present at the site on day 0. The osteogenic compartment was gradually replenished by cells migrating from the nonosteogenic compartment, which was supplemented by ED1(+) cells recruited from the circulation early after activation. Moreover, osteogenic cells appeared to be as crucial in vivo for the acquisition of the TRAP phenotype as previously shown in vitro.     

Lengthening osteotomy in the metaphysis and diaphysis. An experimental study in the ovine tibia

In 20 sheep, aged seven to eight months, a tibial lengthening osteotomy was performed to compare the process of repair of the metaphyseal and diaphyseal regions of the bone. Analogous to clinical lengthening, two frame configurations of a bilateral external fixation device were used to obtain adequate fixation of the bone segments in the metaphyseal and diaphyseal lengthening osteotomies. A daily distraction rate of 1 mm for 4.5 weeks yielded an average length increase of 2.6 cm (12.5%). After death at 4.5 weeks postdistraction, the elongated bones were tested mechanically with the contralateral tibiae as controls. No significant difference in relative torsional strength of the elongated tibiae was found between the two groups. Inferior mechanical stability of the bone segments in metaphyseal compared with diaphyseal lengthening (due to differences in frame rigidity and distribution of muscular moments) influenced healing to such an extent that any superior biologic, osteogenic potential in the metaphyseal bone region was nullified. With the clinical use of two configurations of a given external device necessary for fixation of the bone segments in a metaphyseal or diaphyseal lengthening osteotomy, the empirically accepted idea that metaphyseal healing is superior may not be correct.     

Effects of lesion between bone, periosteum and muscle on fracture healing in rats

We assessed the effects of periosteal detachment from bone and musculature on the healing of diaphyseal fracture. In 30 male Wistar rats we produced a partial osteotomy, which was manually broken in the middiaphysis of the left femur. All fractures were reamed and stabilized with an 1.6 mm steel pin. The animals were randomly assigned to 3 groups. In group 1, a subperiosteal detachment between cortex and periost was created in the middle third of the diaphysis. An extraperiosteal detachment between periost and the surrounding musculature was performed in group 2. In group 3, the periosteum was isolated from the musculature by an extraperiosteal detachment and application of an e-PTFE sheath (Gore-Tex ® expanded polytetrafluoroethylene) around the shaft between the periost and the surrounding muscles. The rats were killed after 4 weeks and callus formation and mechanical characteristics were measured. All fractures healed by production of external callus. The callus area was significantly less in the group where periost was mechanically isolated from the surrounding muscles compared to the other groups. Bending moment, bending rigidity and fracture energy were less in this group than in groups 1 and 2. No differences were detected between the sub- and extraperiosteal groups, either in callus formation or in mechanical measurements. Our findings underline the importance of the muscle-periosteal connection for periosteal healing of diaphyseal fractures.     

Chondroclasts and Osteoclasts in Bones of Young Rats: Comparison of Ultrastructural and Functional Features

The aim of the present study was to characterize cells involved in resorption during endochondral bone formation. We investigated whether the cells involved in cartilage breakdown at the epiphyseal/metaphyseal border, i.e., chondroclasts, share the characteristics of bone/cartilage-resorbing osteoclasts at the metaphyseal/diaphyseal border regarding ultrastructural features and functional activity. Morphometric evaluation showed that chondroclasts do not form ruffled borders and clear zones, i.e., well-known resorption characteristics, to the same extent as osteoclasts, present at the lower metaphysis. Instead, chondroclasts tend to express an undifferentiated surface adjacent to the matrix, not structurally different from the basolateral plasma membrane. Tartrate-resistant acid phosphatase (TRAP) was used as a marker for functional activity. Immunohistochemical staining by light microscopy was strong in both chondroclasts and in osteoclasts. Furthermore, in situ hybridization revealed large amounts of TRAP mRNA in chondroclasts as well as in osteoclasts. Ultrastructural immunohistochemistry suggests extensive secretion of the TRAP enzyme in the ruffled border area of both chondroclasts and osteoclasts. Intracellular accumulation was seen particularly in chondroclasts, possibly as a consequence of a relative disinclination to develop a ruffled border. Thus, semiquantitative estimation of TRAP distribution showed an inverse relationship between extracellular and intracellular TRAP in chondroclasts and osteoclasts. These results indicate that chondroclasts and osteoclasts differ, not only with respect to location but possibly also by mode of action. The observed differences may reflect the maturation sequence of these multinucleated cells when associated with different metaphyseal trabecular surfaces. Received: 22 January 1998 / Accepted 8 April 1998     

Role of fracture hematoma and periosteum during fracture healing in rats: interaction of fracture hematoma and the periosteum in the initial step of the healing process

To study the mechanisms of fracture healing, we investigated the interaction between fracture hematoma and periosteum during the early phase of fracture healing in rats. Experimentally induced fractures of the tibia in untreated rats were compared histologically with such fractures in rats in which either the bone marrow or the periosteum had been removed. The extent of periosteal cell proliferation and chondrogenesis in the fracture hematoma was evaluated on experimental days 3, 6, 10, and 14. On day 3, periosteal cell proliferation at the tibial fracture site was decreased in the bone marrow-removed rats compared with the proliferation in untreated rats. Little chondrogenesis in the fracture hematoma was seen through day 6 in the periosteum-removed rats. These results suggest that the periosteum is important for mediating the primary steps of chondrogenesis and enchondral ossification in the fracture hematoma and that the fracture hematoma may be essential for periosteal cell proliferation during fracture healing. Received for publication on April 5, 1999; accepted on July 21, 1999     

Aged mice have enhanced endocortical response and normal periosteal response compared with young‐adult mice following 1 week of axial tibial compression

With aging, the skeleton may lose its ability to respond to positive mechanical stimuli. We hypothesized that aged mice are less responsive to loading than young‐adult mice. We subjected aged (22 months) and young‐adult (7 months) BALB/c male mice to daily bouts of axial tibial compression for 1 week and evaluated cortical and trabecular responses using micro–computed tomography (µCT) and dynamic histomorphometry. The right legs of 95 mice were loaded for 60 rest‐inserted cycles per day to 8, 10, or 12 N peak force (generating mid‐diaphyseal strains of 900 to 1900 µε endocortically and 1400 to 3100 µε periosteally). At the mid‐diaphysis, mice from both age groups showed a strong anabolic response on the endocortex (Ec) and periosteum (Ps) [Ec.MS/BS and Ps.MS/BS: loaded (right) versus control (left), p < .05]. Generally, bone formation increased with increasing peak force. At the endocortical surface, contrary to our hypothesis, aged mice had a significantly greater response to loading than young‐adult mice (Ec.MS/BS and Ec.BFR/BS: 22 months versus 7 months, p < .001). Responses at the periosteal surface did not differ between age groups (p > .05). The loading‐induced increase in bone formation resulted in increased cortical area in both age groups (loaded versus control, p < .05). In contrast to the strong cortical response, loading only weakly stimulated trabecular bone formation. Serial (in vivo) µCT examinations at the proximal metaphysis revealed that loading caused a loss of trabecular bone in 7‐month‐old mice, whereas it appeared to prevent bone loss in 22‐month‐old mice. In summary, 1 week of daily tibial compression stimulated a robust endocortical and periosteal bone‐formation response at the mid‐diaphysis in both young‐adult and aged male BALB/c mice. We conclude that aging does not limit the short‐term anabolic response of cortical bone to mechanical stimulation in our animal model. © 2010 American Society for Bone and Mineral Research     

The chondrogenic potential of periosteum decreases with age.

Periosteum contains undifferentiated mesenchymal stem cells that possess the potential for chondrogenesis during cartilage repair and in fracture healing. With aging, the chondrogenic potential of periosteum declines significantly. An organ-culture model was used to investigate the relationship between the chondrogenic potential of periosteum and aging. A total of 736 periosteal explants from the proximal medial tibiae of 82 rabbits, aged 2 weeks to 2 years, were cultured in agarose suspension conditions conductive for chondrogenesis. and analyzed using histomorphometry, collagen typing, wet weight measurement, 3H-thymidine and 35S-sulfate uptake, autoradiography, and PCNA immunostaining. The rabbits were skeletally mature by 6 months and stopped increasing in weight by 12 months. Chondrogenesis declined significantly with age (P < 0.0001) and was maximal in the 1.5-2 month-old rabbits. Explants from the 6 month-old rabbits formed 50% less cartilage. and by 12 months chondrogenesis reached a steady state minimal level. In parallel with this decrease in chondrogenic potential similar decreases were measured in 3H-thymidine uptake (P < 0.0001). 35S-sulfate uptake (P = 0.0117), as well as the thickness (P < 0.0001) and the total number of cells in the cambium layer of the periosteum (P < 0.0001). Autoradiography with 3H-thymidine and PCNA immunostaining confirmed the measured decrease in proliferative activity in the cambium layer where the chondrocyte precursors reside, although the percentage of proliferating cells did not change significantly with age. The most dramatic change was the marked decrease (87%) in the thickness and total cell number in the cambium layer of the perisoteum between the 2 and 12 month-old rabbits (P < 0.05). These data confirm a decline in the chondrogenic potential of periosteum with aging. Thus, one possibility for improving cartilage formation by periosteal transplantation after skeletal maturity would be to stimulate an increase in the total number of cells in the chondrocyte precursor pool early during chondrogenesis.     

Effect of periosteal stripping on cortical bone perfusion: A laser doppler study in sheep

The goals of internal fixation are an accurate reduction and stable fixation in the presence of adequate bony vascularity. This can be achieved by a variety of means including plate fixation. A certain amount of periosteal stripping is necessary for proper open reduction of a fracture and for proper plate application. With displaced diaphyseal fractures, cortical bone perfusion (CBP) is already compromised. Further damage, in terms of periosteal stripping for plate fixation, may not be acceptable. Little information is available as to what extent the periosteum contributes to cortical bone perfusion. The purpose of this study was to determine the acute effects of periosteal stripping on cortical bone perfusion in a sheep tibia model. Twenty-three sheep were operated on and had the medial aspect of their right tibia exposed. Cortical bone perfusion measurements were obtained using laser Doppler flowmetry prior to periosteal stripping and after periosteal stripping. The results of this study show that the cortical bone perfusion significantly decreased by 20% after periosteal stripping over the entire length of the tibia. We therefore conclude that the periosteum contributes to diaphyseal bone perfusion and that it is important to preserve this source with fractures where blood supply is already significantly compromised.     

Mechanisms responsible for longitudinal growth of the cortex: coalescence of trabecular bone into cortical bone

BACKGROUND: The purpose of the present study was to determine whether longitudinal growth of the cortex occurs through intramembranous bone formation involving the periosteum or through endochondral bone formation involving the growth plate and to explore the cellular and biochemical mechanisms responsible for this process. METHODS: Cortical bone formation was studied in the metaphyses of growing New Zealand White rabbits by means of (1) oxytetracycline labeling and fluorescence microscopy, (2) computer-assisted histomorphometry, (3) osteoblast culture and [(3) H]-thymidine incorporation in the presence of periosteum or periosteum-conditioned medium, and (4) surgical insertion of membranes between the periosteum and the underlying spongiosa. RESULTS: Within the metaphyseal cortex, oxytetracycline labeling produced fluorescent closed curves outlining enlarging trabeculae derived from coalescing endochondral trabecular bone. In this region of coalescing trabeculae close to the periosteum, osteoblast surface was increased compared with trabeculae farther from the periosteum (p < 0.001). The osteoclast surface did not differ. In vitro, osteoblast proliferation was increased in the presence of periosteum (p < 0.001) or periosteum-conditioned medium (p < 0.001). Surgical insertion of permeable or impermeable membranes between the periosteum and the spongiosa did not prevent cortex formation. CONCLUSIONS: These observations demonstrate that metaphyseal cortical bone is formed by coalescence of endochondral trabecular bone. This coalescence is associated with increased osteoblast surface in the peripheral spongiosa. The increased osteoblast surface could be due to inductive effects of periosteum; in the present study, periosteum stimulated osteoblast proliferation in vitro but was not required for metaphyseal cortical bone formation in vivo. Clinical Relevance: Understanding metaphyseal cortical growth may help to elucidate the pathophysiology of osseous growth disorders in children.     

Reversibility of nontraumatic disuse osteoporosis during its active phase

The potential for the recovery of bone lost during the active phase of disuse osteoporosis, both in the diaphyseal compacta and metaphyseal spongiosa was tested in young adult and old Beagle dogs. Immobilization for up to 60 weeks was achieved by placing the forelimb in a spica cast and remobilization by removing it. Bone volume was estimated in the third metacarpus, radius, ulna and humerus at the mid-diaphysis and at the level of distal metaphyseal spongiosa in both forelimbs by radiography and histomorphometry.

Measurements carried out on animals remobilized showed considerable recovery of the original bone loss. In both age groups, the residual deficits increased, however, with the duration of immobilization and were similar in the metaphyseal spongiosa and in the diaphyseal compacta. The old dogs which began the study with 10% less bone than the younger dogs, showed smaller proportional losses than the younger dogs but greater residual deficits, most evident in the diaphysis. In both age groups the distal, weight-bearing bones tended to show greater losses and also greater recovery both in diaphyseal compacta and the metaphyseal spongiosa. Thus, 28 weeks after cast removal following 32 weeks of immobilization the following findings were noted: In the third metacarpal diaphyseal compacta in the younger dogs, a 53.6% loss (mostly from the periosteal envelope) decreased to 16.3% (a 70% recovery) while in the older dogs a 37.6% loss (mostly from the endosteal envelope) decreased to 23% (a 40% recovery). In metacarpal metaphyseal spongiosa in young adult dogs, a 50% loss reduced to 16.8% (a 66% recovery) while in the older dogs a 47% loss reduced to 19% (a 60% recovery).

These observations apply only to the effect of remobilization on recovery of bone loss incurred during the active phase of disuse osteoporosis. The same potential for recovery may not exist later in the inactive phase of established disuse osteoporosis. The permanent losses, however, could be prevented by appropriate measures taken during the active phase of osteoporosis.     

Early muscle-periosteal lesion inhibits fracture healing in rats

We assessed the effects of muscular detachment from the periosteum on fracture healing, focusing on a muscle-periosteal lesion in the initial healing process. In 30 male Wistar rats we produced a partial osteotomy in the mid-diaphysis of the left femur which was then manually broken. All fractures were reamed and stabilized with a 1.6 mm steel pin. The animals were randomly assigned to 3 groups. In group 1, an extraperiosteal detachment between muscle and periosteum was created in the middle third of the diaphysis. In group 2, an extraperiosteal detachment was created with application of an ePTFE sheath (Gore-Tex expanded polytetrafluoroethylene) around the shaft between muscle and periosteum during the first 2 weeks following fracture. In group 3, the dissection was identical, while the ePTFE sheath was installed after 2 weeks. The rats were killed after 4 weeks, and their bones were evaluated radiographically and mechanically by the three-point bending test. The fractures healed by production of external callus, and radiographs revealed various degrees of periosteal callus with a radiolucent fracture line, most evident after early muscle-periosteal isolation. The callus area was significantly smaller after early muscle isolation, compared to extraperiosteal dissection alone and later tissue isolation. Bending moment and stiffness were also less in this group than in groups 1 and 3, while fracture energy was less than in group 1. No differences in mechanical properties were detected between extraperiosteal dissection alone and late-tissue isolation. This animal study underlines the importance of early muscle-periosteal apposition for fast periosteal healing of diaphyseal fractures.     

Early muscle-periosteal lesion inhibits fracture healing in rats

We assessed the effects of muscular detachment from the periosteum on fracture healing, focusing on a muscle-periosteal lesion in the initial healing process. In 30 male Wistar rats we produced a partial osteotomy in the mid-diaphysis of the left femur which was then manually broken. All fractures were reamed and stabilized with a 1.6 mm steel pin. The animals were randomly assigned to 3 groups. In group 1, an extraperiosteal detachment between muscle and periosteum was created in the middle third of the diaphysis. In group 2, an extraperiosteal detachment was created with application of an e-PTFE sheath (Gore-Tex® expanded polytetrafluoro-ethylene) around the shaft between muscle and periosteum during the first 2 weeks following fracture. In group 3, the dissection was identical, while the e-PTFE sheath was installed after 2 weeks. The rats were killed after 4 weeks, and their bones were evaluated radiographically and mechanically by the three-point bending test. The fractures healed by production of external callus, and radiographs revealed various degrees of periosteal callus with a ra-diolucent fracture line, most evident after early muscle-periosteal isolation. The callus area was significantly smaller after early muscle isolation, compared to extraperiosteal dissection alone and later tissue isolation. Bending moment and stiffness were also less in this group than in groups 1 and 3, while fracture energy was less than in group 1. No differences in mechanical properties were detected between extraperiosteal dissection alone and late-tissue isolation. This animal study underlines the importance of early muscle-periosteal apposition for fast periosteal healing of diaphyseal fractures.     

Testing of a new one-stage bone-transport surgical procedure exploiting the periosteum for the repair of long-bone defects

BACKGROUND: A recently proposed one-stage bone-transport surgical procedure exploits the intrinsic osteogenic potential of the periosteum while providing mechanical stability through intramedullary nailing. The objective of this study was to assess the efficacy of this technique to bridge massive long-bone defects in a single stage. METHODS: With use of an ovine femoral model, an in situ periosteal sleeve was elevated circumferentially from healthy diaphyseal bone, which was osteotomized and transported over an intramedullary nail into a 2.54-cm (1-in) critical-sized diaphyseal defect. The defect-bridging and bone-regenerating capacity of the procedure were tested in five groups of seven animals each, which were defined by the absence (Group 1; control) or presence of the periosteal sleeve alone (Group 2), bone graft within the periosteal sleeve (Groups 3 and 5), as well as retention of adherent, vascularized cortical bone chips on the periosteal sleeve with or without bone graft (Groups 4 and 5). The efficacy of the procedure was assessed qualitatively and quantitatively. RESULTS: At sixteen weeks, osseous bridging of the defect was observed in all twenty-eight experimental sheep in which the periosteal sleeve was retained; the defect persisted in the remaining seven control sheep. Among the experimental groups 2 through 5, significant differences were observed in the density of the regenerated bone tissue; the two groups in which vascularized bone chips adhered to the inner surface of the periosteal sleeve (Groups 4 and 5) showed a higher mean bone density in the defect zone (p < 0.02) than did the other groups. In these two groups with the highest bone density, the addition of bone graft was associated with a significantly lower callus density than that observed without bone graft (p < 0.05). The volume of regenerate bone (p < 0.02) was significantly greater in the groups in which the periosteal sleeve was retained than it was in the control group. Among the experimental groups (groups 2 through 5), however, with the numbers studied, no significant differences in the volume of regenerate bone could be attributed to the inclusion of bone graft within the sleeve or to vascularized bone chips remaining adherent to the periosteum. CONCLUSIONS: The novel surgical procedure was shown to be effective in bridging a critical-sized defect in an ovine femoral model. Vascularized bone chips adherent to the inner surface of the periosteal sleeve, without the addition of morselized cancellous bone graft within the sleeve, provide not only a comparable volume of regenerate bone and composite tissue (callus and bone) but also a superior density of regenerate bone compared with that after the addition of bone graft.     

Humeral head circle‐fit method greatly increases reliability and accuracy when measuring anterior–posterior radiographs of the proximal humerus

Measurements made on routine A–P radiographs can predict strength/quality of the proximal humerus, as shown in terms of two easy‐to‐measure parameters: Cortical index (CI) and mean‐combined cortical thickness (MCCT). Because of high variability inherent when using established methods to measure these parameters, we describe a new orientation system. Using digitized radiographs of 33 adult proximal humeri, five observers measured anatomical reference locations in accordance with: (i) Tingart et al. (2003) method, (ii) Mather et al. (2013) method, and (iii) our new humeral head Circle‐Fit method (CFM). The Tingart and Mather methods measure CI and MCCT with respect to upper and lower edges of 20 mm tall rectangles fit to a proximal diaphyseal location where endosteal (Tingart) or periosteal (Mather) cortical margins become parallel. But high intra‐ and inter‐observer variability occurs when placing the rectangles because of uncertainty in identifying cortical parallelism. With the CFM an adjustable circle is fit to the humeral head articular surface, which reliably and easily establishes a proximal metaphyseal landmark (M1) at the surgical neck. Distal locations are then designated at successive 10 mm increments below M1, including a second metaphyseal landmark (M2) followed by diaphyseal (D) locations (D1, D2 ⋯D6). D1 corresponds most closely to the proximal edges of the rectangles used in the other methods. Results showed minimal inter‐observer variations (mean error, 1.5 ± 1.1 mm) when the CFM is used to establish diaphyseal locations for making CI and MCCT measurements when compared to each of the other methods (mean error range, 10.7 ± 5.9 to 13.3 ± 6.7 mm) (p < 0.001). © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:2313–2322, 2017.

     

Periosteum: biology, regulation, and response to osteoporosis therapies

Periosteum contains osteogenic cells that regulate the outer shape of bone and work in coordination with inner cortical endosteum to regulate cortical thickness and the size and position of a bone in space. Induction of periosteal expansion, especially at sites such as the lumbar spine and femoral neck, reduces fracture risk by modifying bone dimensions to increase bone strength. The cell and molecular mechanisms that selectively and specifically activate periosteal expansion, as well as the mechanisms by which osteoporosis drugs regulate periosteum, remain poorly understood. We speculate that an alternate strategy to protect human bones from fracture may be through targeting of the periosteum, either using current or novel agents. In this review, we highlight current concepts of periosteal cell biology, including their apparent differences from endosteal osteogenic cells, discuss the limited data regarding how the periosteal surface is regulated by currently approved osteoporosis drugs, and suggest one potential means through which targeting periosteum may be achieved. Improving our understanding of mechanisms controlling periosteal expansion will likely provide insights necessary to enhance current and develop novel interventions to further reduce the risk of osteoporotic fractures.