Cancellous Bone Healing Research Articles

Guided cortical and cancellous bone formation using a minimally invasive technique of BMSC- and BMP-2-laden visible light-cured carboxymethyl chitosan hydrogels

A cavity defect inside the bone is formed by deformed cancellous bone from the fixation of the cortical bone, and consequently, abnormal bone healing occurs. Therefore, repairing cancellous bone defects is a remarkable topic in orthopedic surgery. In this study, we prepared bone marrow-derived stem cell (BMSC)-laden and bone morphogenetic protein-2 (BMP-2)-laden visible light-cured carboxymethyl chitosan (CMCS) hydrogels for cortical and cancellous bone healing. Proton nuclear magnetic resonance (1H NMR) analysis confirmed the methacrylation of CMCS (CMCSMA), resulting in 55 % of substitution. The higher concentration of CMCSMA hydrogel resulted in the lower swelling ratio, the larger viscosity, the slower degradation behavior, and the stronger compressive strength. The 5 w/v% hydrogel exhibited a controlled BMP-2 release for 14 days, while the 7 and 10 w/v% hydrogels displayed a controlled BMP-2 release for 28 days. Results of in vitro cytotoxicity and cell proliferation assays revealed the biocompatibility of the samples. In vivo animal tests demonstrated that BMSC- and BMP-2-laden 7 w/v% CMCSMA (CMCSMA+Cell+BMP-2) improved bone formation in the defected cortical and cancellous bones of the femur, as analyzed by micro-computed tomography (micro-CT) and histological evaluations. Consequently, we suggested that CMCSMA+Cell+BMP-2 can be a valuable scaffold for restoring cortical and cancellous bone defects.

Reconstruction of anterior frontal wall and contours defects using bone substitute

Aim: The aim of this article is to report on the safety and long-term efficacy of Cerament® BoneVoid Filler bone substitute for repairing craniofacial bone defects. Post-traumatic cranioplasty is a complex and challenging procedure for all maxillo-craniofacial surgeons and neurosurgeons, especially when repairing large areas. The standard criterion for repairing small cranial defects is the use autogenous bone from the iliac crest or split calvarial grafts. Autogenous grafts may result in donor-site morbidity, increased surgical time, reabsorption, blood loss, and longer recovery time . Alloplastic materials used for bone repair, such as methyl methacrylate, hydroxyapatite, titanium, or porous polyethylene, are expected to have optimal properties, including easy adaptation, biocompatibility, ingrowth of new tissue, stability of shape, and low rate of reabsorption. A cranial implant should be easily shaped and positioned, allowing easy tissue growth. In very wide cranium defects the new technology is a custom made cranial implant constructed three-dimensionally with different types of materials. However, this procedure is very expensive with various infection rates depending on the kind of material used and on the chemicophysical composition of the implant. Methods: The authors report the case of a 50-year-old man with a severe deformity of the forehead-supra orbital area as a result of a previous complex fronto-facial trauma treated in an emergency Unit. Secondary correction and reconstruction of the residual deformities were performed by using Cerament® Bone Void Filler, an alloplastic biphasic material, composed of 40% hydroxyapatite, 60% calcium sulfate and the radio-contrast agent iohexol. The unique ratio of hydroxyapatite and calcium sulfate is designed to enable Cerament to resorb at the same rate that bone forms. Calcium sulfate acts as a resorbable carrier for hydroxyapatite which is highly osteoconductive, promoting bone ingrowth.It seems to be a promising bone graft substitute in the management of bony irregularities in the fronto-orbital area. Conclusion: The patient was first hospitalized as the result of a serious craniofacial trauma. One year after the first emergency cranio-orbital reconstructive operation, a marked deformity of the frontal region appeared with a “grid effect” due to the inadequate plate-bony fixation of the fractures applied during the first bony recomposition and because it was not as rigid as it should have been . A secondary surgery for deformity correction was performed. The hardware was totally removed and the bony deformity smoothed, reshaped, covered and filled using Cerament® Bone Void Filler, a biomaterial. The patient recovered with a satisfactory cranium-forehead shape, no complications, and complete disappearance of a frowning look of the fronto-orbital region. Recently, increased use of bone substitutes in the reconstruction of bone defects has been fuelled by donor site complications associated with autologous bone harvesting. Cerament® BoneVoid Filler is a biphasic and injectable bone substitute that has a highly compressive strength and the ability to promote cancellous bone healing

The combined effects of teriparatide and anti-RANKL monoclonal antibody on bone defect regeneration in ovariectomized mice.

The purpose of this study was to investigate the combined effects of teriparatide (TPTD) and anti-murine receptor activator of nuclear factor-κB ligand monoclonal antibody (anti-RANKL Ab) on both cancellous and cortical bone healing in ovariectomized mice. Thirteen-week-old mice were divided into the sham-operated group (n=11) or the ovariectomized group (n=44). At 1 month post-operation, all mice underwent bone defect surgery on the left femoral metaphysis (cancellous bone healing model) and right femoral mid-diaphysis (cortical bone healing model). After creating the bone defects, all ovariectomized mice were assigned to one of four groups to receive 1) saline (5 times a week; CNT group), 2) TPTD (40μg/kg 5 times a week; TPTD group), 3) anti-RANKL Ab (5mg/kg once; Ab group), or 4) a combination of TPTD and anti-RANKL Ab (COMB group). The following analyses were performed: Time-course microstructural analysis of healing in both cancellous and cortical bone in the bone defect, measuring the volumetric bone mineral density and the cortical bone thickness of the tibia as a representative of whole body bone with the use of micro-computed tomography, and histological analysis. Regeneration of cancellous bone volume in the COMB group was the highest among the four groups, and combined treatment accelerated the formation of medullary callus during the early phase of bone regeneration. On the other hand, there were no significant differences in the regeneration of cortical bone volume during the early phase of bone regeneration among the four groups. Furthermore, lamellar bone was not well identified in the all four groups. Volumetric bone mineral density in the tibia in the COMB group was significantly higher compared with that in the CNT and TPTD groups and tended to be higher compared with that in the Ab group. The mean values of cortical bone thickness in the TPTD and COMB groups were significantly higher than that in the CNT group. In a mouse model of postmenopausal osteoporosis, combination therapy of TPTD and anti-RANKL Ab accelerates regeneration of cancellous bone more effectively than either agent alone during the early phase of bone regeneration.

Depletion of cytotoxic (CD8+) T cells impairs implant fixation in rat cancellous bone.

As cytotoxic (CD8+ ) T cells seem to impair shaft fracture healing, we hypothesized that depletion of CD8+ cells would instead improve healing of cancellous bone. Additionally, we also tested if CD8-depletion would influence the healing of ruptured Achilles tendons. Rats received a single injection of either anti-CD8 antibodies or saline and put through surgery 24 h later. Three different surgical interventions were performed as follows: (1) a drill hole in the proximal tibia with microCT (BV/TV) to assess bone formation; (2) a screw in the proximal tibia with mechanical evaluation (pull-out force) to assess fracture healing; (3) Achilles tendon transection with mechanical evaluation (force-at-failure) to assess tendon healing. Furthermore, CD8-depletion was confirmed with flow cytometry on peripheral blood. Flow cytometric analysis confirmed depletion of CD8+ cells (p < 0.001). Contrary to our hypothesis, depletion of CD8+ cells reduced the implant pull-out force by 19% (p < 0.05) and stiffness by 34% (p < 0.01), although the bone formation in the drill holes was the same as in the controls. Tendon healing was unaffected by CD8-depletion. Our results suggest that CD8+ cells have an important part in cancellous bone healing. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.

Cyclin-dependent kinase 8/19 inhibition suppresses osteoclastogenesis by downregulating RANK and promotes osteoblast mineralization and cancellous bone healing.

Cyclin-dependent kinase 8 (CDK8) is a mediator complex-associated transcriptional regulator that acts depending on context and cell type. While primarily under investigation as potential cancer therapeutics, some inhibitors of CDK8-and its paralog CDK19-have been reported to affect the osteoblast lineage and bone formation. This study investigated the effects of two selective CDK8/19 inhibitors on osteoclastogenesis and osteoblasts in vitro, and further evaluated how local treatment with a CDK8/19 inhibitor affects cancellous bone healing in rats. CDK8/19 inhibitors did not alter the proliferation of neither mouse bone marrow-derived macrophages (BMMs) nor primary mouse osteoblasts. Receptor activator of nuclear factor κΒ (NF-κB) ligand (RANKL)-induced osteoclastogenesis from mouse BMMs was suppressed markedly by inhibition of CDK8/19, concomitant with reduced tartrate-resistant acid phosphatase (TRAP) activity and C-terminal telopeptide of type I collagen levels. This was accompanied by downregulation of PU.1, RANK, NF-κB, nuclear factor of activated T-cells 1 (NFATc1), dendritic cell-specific transmembrane protein (DC-STAMP), TRAP, and cathepsin K in RANKL-stimulated BMMs. Downregulating RANK and its downstream signaling in osteoclast precursors enforce CDK8/19 inhibitors as anticatabolic agents to impede excessive osteoclastogenesis. In mouse primary osteoblasts, CDK8/19 inhibition did not affect differentiation but enhanced osteoblast mineralization by promoting alkaline phosphatase activity and downregulating osteopontin, a negative regulator of mineralization. In rat tibiae, a CDK8/19 inhibitor administered locally promoted cancellous bone regeneration. Our data indicate that inhibitors of CDK8/19 have the potential to develop into therapeutics to restrict osteolysis and enhance bone regeneration.

Effects of Povidone-Iodine or Ethanol Exposure on Bone Formation at the Osteotomy Site of the Proximal Tibia in Rats

Background: Povidone-iodine (PVI) irrigation is currently used to decrease the frequency of postoperative surgical site infections. Ethanol (EtOH) is sometimes applied to prevent local recurrence after curettage of benign bone tumors. However, the effects of PVI and EtOH on surrounding soft tissue and on bone union are unclear. The purpose of this study was to determine whether PVI or EtOH adversely affects the cancellous bone healing of the osteotomy site at the proximal tibia in rats. Methods: A cancellous bone osteotomy was performed at the right proximal tibia in 4-month-old, female, Sprague Dawley rats. Vehicle, 10% PVI, or 95% EtOH-soaked gauze was inserted into the osteotomy site and maintained for 6 minutes. The rats were euthanized 2 or 4 weeks after the osteotomy. Results: Two weeks after treatment, the bone union rate was significantly higher in the vehicle group than in the PVI group and the EtOH group (p < 0.001). However, the bone union rate was not significantly different between the PVI and EtOH groups. There was no significant difference among the three groups in the bone union rate 4 weeks after treatment. Conclusion: PVI or EtOH delayed bone union of the cancellous bone osteotomy site of the proximal tibia in the early phase (2 weeks), but not at 4 weeks, in rats.

Different composition of leucocytes in cortical and cancellous bone healing in a mouse model.

ObjectivesCortical and cancellous bone healing processes appear to be histologically different. They also respond differently to anti-inflammatory agents. We investigated whether the leucocyte composition on days 3 and 5 after cortical and cancellous injuries to bone was different, and compared changes over time using day 3 as the baseline.MethodsTen-week-old male C56/Bl6J mice were randomized to either cancellous injury in the proximal tibia or cortical injury in the femoral diaphysis. Regenerating tissues were analyzed with flow cytometry at days 3 and 5, using panels with 15 antibodies for common macrophage and lymphocyte markers. The cellular response from day 3 to 5 was compared in order to identify differences in how cancellous and cortical bone healing develop.ResultsBetween day 3 and 5, the granulocytes increased in the cancellous model, whereas the lymphocytes (T cells, B cells, NK cells) and monocytes (CD11b+, F4/80+, CD206+, CD14+) increased in the cortical model.ConclusionThese results suggest an acute type of inflammation in cancellous bone healing, and a more chronic inflammation in cortical healing. This might explain, in part, why cancellous healing is faster and more resistant to anti-inflammatory drugs than are diaphyseal fractures.Cite this article: L. Tätting, O. Sandberg, M. Bernhardsson, J. Ernerudh, P. Aspenberg. Different composition of leucocytes in cortical and cancellous bone healing in a mouse model. Bone Joint Res 2018;7:620–628. DOI: 10.1302/2046-3758.712.BJR-2017-0366.R2.

Guided tissue engineering for healing of cancellous and cortical bone using a combination of biomaterial based scaffolding and local bone active molecule delivery

A metaphyseal bone defect due to infection, tumor or fracture leads to loss of cancellous and cortical bone. An animal model separating the cancellous and cortical healing was used with a combination of a macroporous gelatin-calcium sulphate-hydroxyapatite (Gel-CaS-HA) biomaterial as a cancellous defect filler, and a thin collagen membrane (CM) guiding cortical bone regeneration. The membrane was immobilized with bone morphogenic protein-2 (rhBMP-2) to enhance the osteoinductive properties. The Gel-CaS-HA cancellous defect filler contained both rhBMP-2 and a bisphosphonate, (zoledronate = ZA) to prevent premature callus resorption induced by the pro-osteoclast effect of rhBMP-2 alone. In the first part of the study, the CM delivering both rhBMP-2 and ZA was tested in a muscle pouch model in rats and the co-delivery of rhBMP-2 and ZA via the CM resulted in higher amounts of bone compared to rhBMP-2 alone. Secondly, an established tibia defect model in rats was used to study cortical and cancellous bone regeneration. The defect was left empty, filled with Gel-CaS-HA alone, Gel-CaS-HA immobilized with ZA or Gel-CaS-HA immobilized with rhBMP-2+ZA. Functionalization of the Gel-CaS-HA scaffold with bioactive molecules produced significantly more bone in the cancellous defect and its surroundings but cortical defect healing was delayed likely due to the protrusion of the Gel-CaS-HA into the cortical bone. To guide cortical regeneration, the cortical defect was sealed endosteally by a CM with or without rhBMP-2. Subsequently, the cancellous defect was filled with Gel-CaS-HA containing ZA and rhBMP-2+ZA. In the groups where the CM was doped with rhBMP-2, significantly higher number of cortices bridged. The approach to guide cancellous as well as cortical bone regeneration separately in a metaphyseal defect using two bioactive molecule immobilized biomaterials is promising and could improve the clinical care of patients with metaphyseal defects.

Use of a biphasic cement bone substitute in the management of metaphyseal fractures

ObjectivesIn recent years, the increase in utilisation of bone substitutes in the reconstruction of bone defects has been fuelled by donor site complications associated with autologous bone harvesting. However the ability of bone substitute to stimulate bone union while maintaining fracture reduction has been a topic of debate. Cerament Bone Void Filler (CBVF) is a novel biphasic and injectable ceramic bone substitute that has high compressive strength and the ability to promote cancellous bone healing. Materials and MethodThis is a retrospective study to evaluate the surgical outcome of utilising CBVF in the treatment of depressed metaphyseal bone fractures over a two year period. The patients were followed up for at least six months after surgery and clinical parameters such as wound site complications were collated. Radiographic imaging was evaluated to determine loss of fracture reduction and rate of cement resorption. ResultsThirteen patients with depressed metaphyseal fractures were enrolled, which included: (i) one proximal humerus fracture; (ii) three tibial plateau fractures; and (iii) nine calcaneal fractures. None of the patients showed significant collapse in fracture reduction after six months of follow up. Cement resorption was noted in one patient as early as three weeks after surgery. There were no cases of cement leak or wound site complications. ConclusionCerament Bone Void Filler (CBVF) is a promising bone graft substitute in the management of depressed metaphyseal bone fractures, with the ability to maintain fracture reduction despite cement resorption.

Effects of combined therapy of alendronate and low-intensity pulsed ultrasound on metaphyseal bone repair after osteotomy in the proximal tibia of glucocorticoid-induced osteopenia rats

ObjectivesGlucocorticoid (GC) treatment inhibits activation of runt-related transcription factor 2 (Runx2), which is essential for osteoblast differentiation from stem cells. As a result, GC treatment results in bone loss, GC-induced osteoporosis (GIO), elevated fracture risk, and delayed bone healing. Bisphosphonates such as alendronate (ALN) are recommended for treating or preventing GIO, and low-intensity pulsed ultrasound (LIPUS) facilitates fracture healing and maturation of regenerated bone. Combined therapy with ALN and LIPUS may stimulate cancellous bone healing in GIO rats. Here, we examined the effect of ALN and LIPUS on cancellous bone osteotomy repair in the proximal tibia of GIO rats. MethodsPrednisolone (10 mg/kg body weight/day) was administered for 4 weeks to induce GIO in 6-month-old female Sprague-Dawley rats. Tibial osteotomy was then performed and daily subcutaneous injection of ALN (1-μg/kg body weight) was subsequently administered alone or in combination with LIPUS (20 min/day) for 2 or 4 weeks. ResultsALN significantly increased bone mineral density (BMD) at 2 and 4 weeks, and ALN + LIPUS significantly increased BMD at 4 weeks. Bone union rates were significantly increased after 2 and 4 weeks ALN and ALN + LIPUS treatment. Lastly, ALN and ALN + LIPUS significantly increased the proportion of Runx2 positive cells at 4 weeks. ConclusionsALN monotherapy and combined ALN and LUPUS treatment augmented BMD and stimulated cancellous bone repair with increased Runx2 expression at the osteotomy site in GIO rats. However, the combined treatment had no additional effect on cancellous bone healing compared to ALN monotherapy.

Temporal role of macrophages in cancellous bone healing

Macrophages are important phagocytosing and cytokine producing cells with effects on fracture healing. We used clodronate-containing liposomes to reduce the number of macrophages, in order to study their role in the early phases of cancellous bone healing.Holes were drilled bilaterally into the cancellous bone of the proximal metaphysis of the tibia of 60 mice. A screw was inserted in the hole in the right tibia. The day of surgery was day 0. Clodronate-containing liposomes were injected intraperitoneally as a single injection either day −4 or −1 (before surgery) or day 1 or 3 (after surgery). A control group underwent surgery as above, but received no clodronate. The mice were killed day 7. The mechanical quality of the new formed cancellous bone holding the screw was evaluated by a pull-out test. The contents of the drill hole in the left tibia was analyzed by microCT.Another set of 20 mice received a drill hole in the metaphysis of the right tibia, and were given either clodronate or saline injections days −3 and −2. The animals were killed day 1 and 3. Flow cytometry was used to analyze the composition of macrophage subpopulations in the regenerating tissue.Flow cytometry showed that clodronate injections day −3 and −2 led to a decrease in mature monocytes day 1 together with an increase in immature monocytes. On day 3 this effect had mostly disappeared, suggesting that the effect of the injections lasted 3 to 5days.Mechanical testing revealed that the injections prior to surgery decreased the strength of the new formed bone, holding the screw, by about half. Bone density in the drill hole was similarly reduced. In contrast, the injections given day 1 and 3 had smaller and statistically insignificant effects.Since their depletion at later time points failed to produce a significant effect, it seems that the role of macrophages in cancellous bone is most crucial during the first two days after trauma.

No effect of teriparatide on migration in total knee replacement

Background and purpose — Aseptic loosening is a main cause of late revision in total knee replacement (TKR). Teriparatide, a recombinant parathyroid hormone (PTH), stimulates osteoblasts and has been suggested to improve cancellous bone healing in humans. This might also be relevant for prosthesis fixation. We used radiostereometric analysis (RSA) to investigate whether teriparatide influences prosthesis fixation. Early migration as measured by RSA can predict future loosening.Patients and methods — In a randomized controlled trial with blind evaluation, 50 patients with osteoarthritis of the knee were allocated to a teriparatide treatment group (Forsteo, 20 μg daily for 2 months postoperatively) or to an untreated control group. RSA was performed postoperatively and at 6 months, 12 months, and 24 months. The primary effect variable was maximal total point motion (MTPM) from 12 to 24 months.Results — Median maximal total point motion from 12 to 24 months was similar in the 2 groups (teriparatide: 0.14 mm, 10% and 90% percentiles: 0.08 and 0.24; control: 0.13 mm, 10% and 90% percentiles: 0.09 and 0.21). [Authors: this is perhaps better than using "10th" and "90th", which looks ugly in print./language editor] The 95% confidence interval for the difference between group means was −0.03 to 0.04 mm, indicating that no difference occurred.Interpretation — We found no effect of teriparatide on migration in total knee replacement. Other trials using the same dosing have suggested a positive effect of teriparatide on human cancellous fracture healing. Thus, the lack of effect on migration may have been due to something other than the dose. In a similar study in this issue of Acta Orthopaedica, we found that migration could be reduced with denosumab (Ledin et al. 2017). The difference in response between the anabolic substance teriparatide and the antiresorptive denosumab suggests that resorption has a more important role during the postoperative course than any deficit in bone formation.

Inter-trabecular bone formation: a specific mechanism for healing of cancellous bone

Background and purpose — Studies of fracture healing have mainly dealt with shaft fractures, both experimentally and clinically. In contrast, most patients have metaphyseal fractures. There is an increasing awareness that metaphyseal fractures heal partly through mechanisms specific to cancellous bone. Several new models for the study of cancellous bone healing have recently been presented. This review summarizes our current knowledge of cancellous fracture healing.Methods — We performed a review of the literature after doing a systematic literature search.Results — Cancellous bone appears to heal mainly via direct, membranous bone formation that occurs freely in the marrow, probably mostly arising from local stem cells. This mechanism appears to be specific for cancellous bone, and could be named inter-trabecular bone formation. This kind of bone formation is spatially restricted and does not extend more than a few mm outside the injured region. Usually no cartilage is seen, although external callus and cartilage formation can be induced in meta­physeal fractures by mechanical instability. Inter-trabecular bone formation seems to be less sensitive to anti-inflammatory treatment than shaft fractures.Interpretation — The unique characteristics of inter-trabecular bone formation in metaphyseal fractures can lead to differences from shaft healing regarding the effects of age, loading, or drug treatment. This casts doubt on generalizations about fracture healing based solely on shaft fracture models.

Earlier effect of alendronate in mouse metaphyseal versus diaphyseal bone healing.

Healing of injured cancellous bone is characterized by a transient stage of rapid bone formation throughout the traumatized bone volume, often followed by similarly rapid resorption. This is different from the slower diaphyseal healing via an external callus. We, therefore, hypothesized that antiresorptive treatment might have an earlier positive effect in cancellous bone healing than in diaphyseal fractures. One hundred and twenty-three male C57bl6 mice received either an internally stabilized diaphyseal osteotomy of the femur or a screw inserted into the tibial metaphysis. The mice were randomized to daily alendronate injections (200 μg/kg/day), or control injections, and killed for mechanical testing after 14, 21, or 28 days. The hypothesis was tested by a three-way Anova (time, site, and drug). The ultimate force was increased by bisphosphonate treatment in both models. There was a significant interaction between time, site, and drug (p < 0.001) so that the full positive effect of alendronate was evident in the metaphysis at 14 days, but first after 28 days in the diaphysis. While the early effect in the metaphysis might be translated into earlier healing, the late effect in the diaphysis was due to delayed remodeling of the callus, which might have less clinical importance. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:793-799, 2017.

Experimental models for cancellous bone healing in the rat

Background and purpose — Cancellous bone appears to heal by mechanisms different from shaft fracture healing. There is a paucity of animal models for fractures in cancellous bone, especially with mechanical evaluation. One proposed model consists of a screw in the proximal tibia of rodents, evaluated by pull-out testing. We evaluated this model in rats by comparing it to the healing of empty drill holes, in order to explain its relevance for fracture healing in cancellous bone. To determine the sensitivity to external influences, we also compared the response to drugs that influence bone healing.Methods — Mechanical fixation of the screws was measured by pull-out test and related to the density of the new bone formed around similar, but radiolucent, PMMA screws. The pull-out force was also related to the bone density in drill holes at various time points, as measured by microCT.Results — The initial bone formation was similar in drill holes and around the screw, and appeared to be reflected by the pull-out force. Both models responded similarly to alendronate or teriparatide (PTH). Later, the models became different as the bone that initially filled the drill hole was resorbed to restore the bone marrow cavity, whereas on the implant surface a thin layer of bone remained, making it change gradually from a trauma-related model to an implant fixation model.Interpretation — The similar initial bone formation in the different models suggests that pull-out testing in the screw model is relevant for assessment of metaphyseal bone healing. The subsequent remodeling would not be of clinical relevance in either model.

Cancellous bone healing around strontium-doped hydroxyapatite in osteoporotic rats previously treated with zoledronic acid.

Bisphosphonates (BPs) are potent anti-osteoporotic agents. Strontium-doped hydroxyapatite (HA) (SrHA) has been reported to increase bone density and improve trabecular microarchitecture in osteoporotic animals. But information about the effect of SrHA on the surrounding bone tissue in osteoporotic animals previously on BPs treatment is limited. We hypothesize that SrHA will induce increased bone density in the vicinity of the material when compared to HA, even in osteoporotic animals previously treated with BPs. HA and 10%SrHA (HA with 10 mol % calcium substituted by strontium) implants were prepared and characterized by scanning electronic microscopy (SEM), X-ray photoemission spectroscopy (XPS), and X-ray diffraction (XRD). Osteoporotic animal model was established by bilateral ovariectomy. Twelve weeks later, all OVX rats accepted subcutaneous injection of zoledronic acid (ZOL) at the dose of 1.5 μg/kg weekly for another twelve weeks. Subsequently, rod-shaped HA and SrHA implants were inserted in the distal femur of the OVX animals previously treated with ZOL. Eight weeks after implantation, specimens were harvested for histological and micro-computed tomography (micro-CT) analysis. Compared to HA, 10%SrHA raised the percent bone volume by 32.7%, the mean trabecular thickness by 36.5%, the mean trabecular number by 34.3%, the mean connectivity density by 38.4%, while the mean trabecular separation showed no significant difference. 10%SrHA also increased the bone area density by 36.3% in histological analysis. Results from this study indicated that 10%SrHA increased bone density and improved trabecular microarchitecture around implants in osteoporotic animals previously treated with ZOL when compared to HA.

Autologous resurfacing.

There is a discrepancy between the interest in joint-reconstructions and the current knowledge about the healing-processes involved. Major reconstructions are performed with osteosynthesized allografts and fresh allografts for cartilage. The main question to be answered is: what do we know about metaphyseal and epiphyseal cancellous bone healing, contact healing of the subchondral bone and its influence on cartilage healing? Can we achieve healing of all four compartments in contact? The purpose is to systematically investigate through animal testing the healing processes of metaphyseal and epiphyseal bone, including the subchondral bone and the healing of cartilage of press-fit-inserted grafts, considering nondemineralized high-resolution histology. Primary cancellous-bone healing of osteosynthesized hemi-osteotomies was studied in 13 canine tibial heads, the contact healing was investigated in 7 dogs and 18 giant-rabbits comparing contact-healing of press-fit-inserted autologs cylindrical grafts with empty defects applying the wet-grinding diamond-technology. Bench-experiments on the epiphyseal bones of swine including pullout-tests of cylindrical grafts formed the basis for validation of that press-fit diamond technology. Primary metaphyseal and epiphyseal contact healing, including hyaline cartilage, was found in all compartments of the meta-and epiphysis in the precisely performed experiments. The press-fit principle, which employs cylindrical grafts and diamond instrumentation featuring a difference of 15/100 mm between graft and recipient bed, achieved high loads between 73.48 and 178.95 N (mean value 118.16 and standard deviation 32.79) in the pullout tests. Autologous press-fit grafting with alignment of the bony baseplate using wet-grinding precision has attained promising histo-morphological results.

A sheep model for cancellous bone healing.

Appropriate well-characterized bone defect animal models remain essential for preclinical research. This pilot study demonstrates a relevant animal model for cancellous bone defect healing. Three different defect diameters (8, 11, 14 mm) of fixed depth (25 mm) were compared in both skeletally immature (18-month-old) and aged sheep (5-year-old). In each animal, four defects were surgically created and placed in the cancellous bone of the medial distal femoral and proximal tibial epiphyses bilaterally. Animals were euthanized at 4 weeks post-operatively to assess early healing and any biological response. Defect sites were graded radiographically, and new bone formation quantified using μCT and histomorphometry. Fibrous tissue was found within the central region in most of the defects with woven bone normally forming near the periphery of the defect. Bone volume fraction [bone volume (BV)/TV] significantly decreased with an increasing defect diameter. Actual BV, however, increased with defect diameter. Bone ingrowth was lower for all defect diameters in the aged group. This pilot study proposes that the surgical creation of 11 mm diameter defects in the proximal tibial and distal femoral epiphyses of aged sheep is a suitable large animal model to study early healing of cancellous bone defects. The refined model allows for the placement of four separate bone defects per animal and encourages a reduction in animal numbers required for preclinical research.

Bone Healing of an Unfixed Bone Fragment of the Distal Segment in Sagittal Split Ramus Osteotomy

In bilateral sagittal split ramus osteotomy for the correction of asymmetry of the mandible, yawing movement of the distal segment can displace the proximal segment. This displacement can be minimized through osteotomy of the posterior part of the distal segment (ie, distal cutting). This free bone fragment is usually removed because of the difficulty of fixation. No previous studies have examined whether union of the bone fragment after distal cutting could join and consequently strengthen the thinned mandibular posterior border. This study used CT imaging to evaluate bone union of the unfixed bone fragment at 3 months postoperatively. The location of the bone fragment and the morphology of bone healing were evaluated in 2D and 3D. The amount of cancellous bone healing between the free bone segment and the proximal segment averaged 63.69%. There was no correlation between the size of the bone gap and the degree of bone union. In most cases, the free bone fragment was located between the distal and proximal segments and tended to dislocate in an anterior-superior direction. Because the postoperative follow-up period was only 3 months, a longer-term study of the changes in bone volume after remodeling is necessary.

Effects of combined therapy of alendronate and low-intensity pulsed ultrasound on metaphyseal bone repair after osteotomy in the proximal tibia of aged rats

Bisphosphonates and low-intensity pulsed ultrasound (LIPUS) are both known to maintain or promote callus formation during diaphyseal fracture healing. However, the effect of these treatments on the repair of metaphyseal fractures has not been elucidated. To evaluate the effects of bisphosphonates and/or LIPUS on cancellous bone healing, an osteotomy was performed on the proximal tibial metaphysis of 9-month-old Sprague-Dawley rats (n = 64). Treatment with alendronate (1 μg/kg/day), LIPUS (20 min/day), or a combination of both was administered for 2 or 4 weeks, after which changes in bone mineral density (BMD), bone histomorphometric parameters, and the rate of cancellous bony bonding were measured. Alendronate suppressed bone resorption parameters at 2 weeks (p = 0.019) and increased bone volume and BMD at 4 weeks (p = 0.034 and p = 0.008, respectively), without affecting bony bonding. LIPUS had no significant effect on any of the histomorphometric parameters at 2 or 4 weeks, but significantly increased in BMD at 4 weeks (p = 0.026) as well as the percentage of bony bonding at both 2 and 4 weeks (p < 0.01). The combined therapy also showed significantly increased BMD compared with the control group at 4 weeks (p = 0.010) and showed a trend toward increased bony bonding. In conclusion, alendronate and LIPUS cause an additive increase in BMD at the affected metaphysis: alendronate increases the bone volume at the osteotomy site without interrupting metaphyseal repair, whereas LIPUS promotes metaphyseal bone repair, without affecting bone histomorphometric parameters.