Fresh-frozen human bone allograft in vertical ridge augmentation: clinical and tomographic evaluation of bone formation and resorption

The aim of the current study is to evaluate fresh-frozen human bone allografts (FHBAs) used in vertical ridge augmentation clinically and by computed tomography, and to analyze the resulting bone formation and graft resorption. Sixteen FHBAs were grafted in the maxillae and mandibles of 9 patients. The FHBAs, which were provided by the Musculoskeletal Tissue Bank of Marilia Hospital (Unioss), were frozen at -80°C. After 7?months, dental implants were placed and bone parameters were evaluated. Vertical bone formation was measured by computerized tomography before (T0) and at 7?months (T1) after the surgical procedure. Bone graft resorption was measured clinically from a landmark screw head using a periodontal probe. The results were analyzed by Student's t-test. Significant differences existed in the bone formation values at T0 and T1, with an average change of 4.03?±?1.69?mm. Bone graft resorption values were 1.0?±?0.82?mm (20%). Implants were placed with varying insertion torque values (35-45?Ncm), and achieved primary stability. This study demonstrates that FHBAs promote satisfactory vertical bone formation with a low resorption rates, good density, and primary implant stability.     

Reconstruction of the edentulous mandible with fresh frozen bone grafts and implants: a 4-year report of a prospective clinical study

Allogen bones from tissue bank are often used in dentistry although the data analyzing the long-term success in mandible are scarce. This study evaluated by computed tomography scans (CTS) the bone resorption around the implants installed on fresh frozen bone (FFB) previously grafted, after 4 years of occlusal rehabilitation. Six subjects were grafted with blocks in posterior mandible using FFB. After 6 months, 27 implants were placed and after further 4 months the prostheses were delivered. Following 4 years of the final rehabilitation procedures, another CTS was done in order to measure the resorption in periimplant bone crest at the proximal implant surfaces. It was observed a 100 % survival rate of the implants after 4 years of the fixture installation. The marginal bone resorption after 48 months was 2.82 ± 1.63 mm and no statistical significant difference was observed along the region where the implants were fixed when compared with the interimplantar space. In addition there was no significant correlation regarding the length of the implant used and the amount of marginal bone resorption. The conclusion is that grafted areas with FFB are suitable to implant installation in the posterior mandible.     

Evaluation of bone height in osseous free flap mandible reconstruction: an indirect measure of bone mass

Osseous free flaps have become the preferred method of mandibular reconstruction after oncologic surgical ablation. To elucidate the long-term effects of free flap mandibular reconstruction on bone mass, maintenance or reduction in bone height over time was used as an indirect measure of preservation or loss in bone mass. Factors potentially influencing bone mass preservation were evaluated; these included site of reconstruction (central, body, ramus), patient age, length of follow-up, adjuvant radiotherapy, and the delayed placement of osseointegrated dental implants. A retrospective analysis of patients undergoing osseous free flap mandible reconstruction for oncologic surgical defects between 1987 and 1995 was performed. Postoperative Panorex examinations were used to evaluate bone height and bony union after osteotomy. Fixation hardware was used as a reference to eliminate magnification as a possible source of error in measurement. There were 48 patients who qualified for this study by having at least 24 months of follow-up. There were 27 male and 21 female patients, with a mean age of 45 years (range, 5 to 75 years). Mandibular defects were anterior (24) and lateral (24). Osseous donor sites included the fibula (35), radius (6), scapula (4), and ilium (3). There were between zero and four segmental osteotomies per patient (excluding the ends of the graft). Nineteen percent of all patients had delayed placement of osseointegrated dental implants. Initial Panorex examinations were taken between 1 and 9 months postoperatively (mean, 2 months). Follow-up Panorex examinations were taken 24 to 104 months postoperatively (mean, 47 months). The bony union rate after osteotomy was 97 percent. Bone height measurements were compared by site and type of reconstruction. The mean loss in fibula height by site of reconstruction was 2 percent in central segments, 7 percent in body segments, and 5 percent in ramus segments. The mean loss in bone height after radial free flap mandible reconstruction was 33 percent in central segments and 37 percent in body segments; ramus segments did not lose height. The central and body segments reconstructed with scapular free flaps did not lose height, but one ramus segment lost 20 percent of height. There was no loss in bone height in mandibular body reconstruction with the ilium free flap. Fibula free flaps did not significantly lose bone height when evaluated with respect to age, follow-up, radiation therapy, or dental implant placement. The retention in bone height demonstrated in this study suggests that bone mass is preserved after osseous free flap mandible reconstruction. The greatest amount of bone loss was seen after multiply osteotomized radial free flaps were used for central mandibular reconstruction. The ability of the fibula free flap to maintain mass over time, coupled with its known advantages, further supports its use as the "work horse" donor site for mandible reconstruction.     

Predicting bone remodeling around tissue- and bone-level dental implants used in reduced bone width

The objective of this study was to predict time-dependent bone remodeling around tissue- and bone-level dental implants used in patients with reduced bone width. The remodeling of bone around titanium tissue-level, and titanium and titanium–zirconium alloy bone-level implants was studied under 100 N oblique load for one month by implementing the Stanford theory into three-dimensional finite element models. Maximum principal stress, minimum principal stress, and strain energy density in peri-implant bone and displacement in x- and y- axes of the implant were evaluated. Maximum and minimum principal stresses around tissue-level implant were higher than bone-level implants and both bone-level implants experienced comparable stresses. Total strain energy density in bone around titanium implants slightly decreased during the first two weeks of loading followed by a recovery, and the titanium–zirconium implant showed minor changes in the axial plane. Total strain energy density changes in the loading and contralateral sides were higher in tissue-level implant than other implants in the cortical bone at the horizontal plane. The displacement values of the implants were almost constant over time. Tissue-level implants were associated with higher stresses than bone-level implants. The time-dependent biomechanical outcome of titanium–zirconium alloy bone-level implant was comparable to the titanium implant.     

Effect of bovine fetal growth plate as a new xenograft in experimental bone defect healing: radiological,histopathological and biomechanical evaluation

Bone grafting is used to enhance healing in osteotomies, arthrodesis, and multifragmentary fractures and to replace bony loss resulting from neoplasia or cysts. They are source of osteoprogenitor cells and induce bone formation and provide mechanical support for vascular and bone ingrowth. Autografts are used commonly but quantity of retrieved bone is limit. This study was designed to evaluate autograft and new xenograft (Bovine fetal growth plate) effects on bone healing process. Twenty male White New Zealand rabbits were used in this study. In autograft group the defect was filled by fresh autogenous cortical graft, in xenograft group the defect was filled by a segment of bovine fetal growth plate and was fixed by cercelage wire. Radiological, histopathological and biomechanical evaluations were performed blindly and results scored and analyzed statistically. Statistical tests did not support significant differences between two groups at the 14th and 28th postoperative day radiographically (P > 0.05). There was a significant difference for remodeling at the 42nd postoperative radiologically (P < 0.05). Xenograft was superior to autograft at the 56th postoperative day for radiological bone formation (P < 0.03). Histopathological and biomechanical evaluation revealed no significant differences between two groups. The results of this study indicate that satisfactory healing occurred in rabbit radius defect filled with calf fetal growth plate. Complications were not identified and healing was faster than cortical autogenous grafting. It was concluded that the use of calf fetal growth plate as a new xenograft is an acceptable alternative to cortical autogenous graft and could reduce the morbidity associated with harvesting autogenous graft during surgery.     

Finite element analysis of peri-implant bone volume affected by stresses around Morse taper implants: effects of implant positioning to the bone crest

Abstract

Objectives: The purpose of the present study was to evaluate the distribution and magnitude of stresses through the bone tissue surrounding Morse taper dental implants at different positioning relative to the bone crest. Materials and Methods: A mandibular bone model was obtained from a computed tomography scan. A three-dimensional (3D) model of Morse taper implant-abutment systems placed at the bone crest (equicrestal) and 2?mm bellow the bone crest (subcrestal) were assessed by finite element analysis (FEA). FEA was carried out on axial and oblique (45°) loading at 150 N relatively to the central axis of the implant. The von Mises stresses were analysed considering magnitude and volume of affected peri-implant bone. Results: On vertical loading, maximum von Mises stresses were recorded at 6-7?MPa for trabecular bone while values ranging from 73 up to 118?MPa were recorded for cortical bone. On oblique loading at the equiquestral or subcrestal positioning, the maximum von Mises stresses ranged from 15 to 21?MPa for trabecular bone while values at 150?MPa were recorded for the cortical bone. On vertical loading, >99.9vol.% cortical bone volume was subjected to a maximum of 2?MPa while von Mises stress values at 15?MPa were recorded for trabecular bone. On oblique loading, >99.9vol.% trabecular bone volume was subjected to maximum stress values at 5?MPa, while von Mises stress values at 35?MPa were recorded for >99.4vol.% cortical bone. Conclusions: Bone volume-based stress analysis revealed that most of the bone volume (>99% by vol) was subjected to significantly lower stress values around Morse taper implants placed at equicrestal or subcrestal positioning. Such analysis is commentary to the ordinary biomechanical assessment of dental implants concerning the stress distribution through peri-implant sites.     

Effects of fixation and decalcification on the immunohistochemical localization of bone matrix proteins in fresh-frozen bone sections

To examine the stability of bone matrix proteins for crystal dislocation, the immunolocalization of type I collagen, bone sialoprotein, and osteopontin was investigated during different stages of fixation and decalcification. Four-week-old rat femurs were rapidly frozen, and were sectioned without fixation or decalcification. Thereafter, following or bypassing fixation in 4% paraformaldehyde, these sections were decalcified in 5% EDTA for 0-5 min. Before decalcification, marked radiopacity of bone matrix was observed in contact microradiography (CMR) images, and electron probe microanalysis (EPMA) demonstrated intense localization for phosphorus and calcium. In fixed and unfixed sections without decalcification, immunolocalization of bone matrix proteins were almost restricted to osteoid. After 1 min of decalcification, reduced radiopacity was apparent in the CMR images, and less phosphorus and calcium was observed by EPMA, which completely disappeared by 5 min decalcification. After 3-5 min of decalcification, unfixed sections showed that these proteins were immunolocalized in bone matrix, but were not detectable in osteoid. However, fixed sections demonstrated that these were found in both bone matrix and osteoid. The present findings suggest that bone matrix proteins are embedded in calcified matrix which is separated from the aqueous environment and that they hardly move, probably due to firm bonding with each other. In contrast, matrix proteins in osteoid are subject to loss after decalcification because they may be bound to scattered apatite crystals, not to each other.     

Incidence of bone metastases and survival after a diagnosis of bone metastases in breast cancer patients

Background: Bone is the most common metastatic site associated with breast cancer. Using a database of women with breast cancer treated at Guy's Hospital, London 1976–2006 and followed until end 2010, we determined incidence of and survival after bone metastases. Methods: We calculated cumulative incidence of bone metastases considering death without prior bone metastases as a competing risk. Risk of bone metastases was modelled through Cox-regression. Survival after bone metastases diagnosis was calculated using Kaplan–Meier methodology. Results: Of the 7064 women, 589 (22%) developed bone metastases during 8.4 years (mean). Incidence of bone metastases was significantly higher in younger women, tumour size >5 cm, higher tumour grade, lobular carcinoma and ≥four positive nodes, but was not affected by hormone receptor status. Median survival after bone metastases diagnosis was 2.3 years in women with bone-only metastases compared with <1 year in women with visceral and bone metastases. There was a trend for decreased survival for patients who developed visceral metastases early, and proportionately fewer patients in this group. Interpretation: Incidence of bone metastases has decreased but bone metastases remain a highly relevant clinical problem due to the large number of patients being diagnosed with breast cancer.     

Effects of physical training on bone mineral density and bone metabolism

The purpose of this study was to examine the influences of long-term walking training and walking and jumping training on bone mineral density (BMD) and bone metabolism. Data from 28 healthy premenopausal women was assessed. The subjects were divided into the walking group (WG; 17 women mean+/-SE age 35+/-2 years), and the walking and jumping group (WJG; 11 women mean+/-SE age 39+/-1 years). BMD was measured in the lumbar spine and proximal femur using dual energy X-ray absorptiometry (DXA). As markers of bone metabolism, this study was to measure bone formation markers, bone-alkaline phosphatase (B-ALP: measured by enzyme immunoassay/EIA) and osteocalcin (BGP: by radioimmunoassay/RI) as well as bone resorption markers, parathyroid hormone (PTH: measured by/RI) and type I collagen cross-linked N-telopeptides (NTx: by EIA). Despite the significant decrease in body weight (p<0.05), no corresponding decrease in BMD was observed. Moreover, no significant difference in bone markers BGP, PTH, and NTx was observed. B-ALP was significantly increased (p<0.05) after one year, and the rate of this increase was greater in the WJG than in the WG. It is thus concluded that walking training for one year is beneficial for the promotion of bone formation, and that jumping stimulus maintain BMD effectively.     

A new bone banking technique to maintain osteoblast viability in frozen human iliac cancellous bone

The aim of this study was to develop a new cryopreservation technique to maintain the osteoblast viability in frozen iliac bone and to prove cell viability using cell culture techniques.Human iliac cancellous bones were frozen with and without 10% Me(2)SO at -80 degrees C. The tubes were kept in a -80 degrees C freezer for at least 2 days. After the storage period, the frozen bone was thawed by placing the tube in a 37 degrees C water bath. A serial enzymatic digestion technique using 0.2% collagenase was employed to isolate osteoblast-like cells from the bone. The cells that were released were inoculated into tissue culture flasks containing DMEM supplemented with 10% FCS. They were incubated at 37 degrees C in a humidified atmosphere of 95% air and 5% CO(2). Cells of the second passage were plated at a density of 5 x 10(3)cells/cm(2) in a 24-well plate and used for characterization. For characterization, WST-1 assay, determination of alkaline phosphatase, Type I collagen assay, osteocalcin assay, and von Kossa staining were used. The assays were performed at 3, 6, 9, and 12 days after plating the cells. Based on the results of this study, we conclude that the osteoblast-like cells in the frozen bone can survive, only when the bone is frozen with cryoprotectants to prevent injury during freezing and thawing.     

Modulation of bone ingrowth of rabbit femur titanium implants by in vivo axial micromechanical loading.

Titanium implants commonly used in orthopedics and dentistry integrate into host bone by a complex and coordinated process. Despite increasingly well illustrated molecular healing processes, mechanical modulation of implant bone ingrowth is poorly understood. The objective of the present study was to determine whether micromechanical forces applied axially to titanium implants modulate bone ingrowth surrounding intraosseous titanium implants. We hypothesized that small doses of micromechanical forces delivered daily to the bone-implant interface enhance implant bone ingrowth. Small titanium implants were placed transcortically in the lateral aspect of the proximal femur in 15 New Zealand White rabbits under general anesthesia and allowed to integrate with the surrounding bone for 6 wk. Micromechanical forces at 200 mN and 1 Hz were delivered axially to the right femur implants for 10 min/day over 12 consecutive days, whereas the left femur implants served as controls. The average bone volume 1 mm from mechanically loaded implants (n = 15) was 73 +/- 12%, which was significantly greater than the average bone volume (52 +/- 21%) of the contralateral controls (n = 15) (P < 0.01). The average number of osteoblast-like cells per endocortical bone surface was 55 +/- 8 cells/mm(2) for mechanically loaded implants, which was significantly greater than the contralateral controls (35 +/- 6 cells/mm(2)) (P < 0.01). Dynamic histomorphometry showed a significant increase in mineral apposition rate and bone-formation rate of mechanically stressed implants (3.8 +/- 1.2 microm/day and 2.4 +/- 1.0 microm(3).microm(-2).day(-1), respectively) than contralateral controls (2.2 +/- 0.92 microm/day and 1.2 +/- 0.60 microm(3).microm(-2).day(-1), respectively; P < 0.01). Collectively, these data suggest that micromechanical forces delivered axially on intraosseous titanium implants may have anabolic effects on implant bone ingrowth.     

Treatment of large complex cranial bone defects by using hydroxyapatite ceramic implants

Hydroxyapatite ceramic implants were used in the reconstruction of very large and complex-form cranial bone defects in nine patients. The bone defects were the result of craniectomy after infections and other complications such as severe brain edema, after neurosurgery, and as a result of trauma, subdural hemorrhage, and surgery for brain tumor. The size, shape, and curvature of the hydroxyapatite ceramic implants were determined based on high-precision, full-scale models fabricated through a laser lithographic molding method by using computed tomographic data. The use of this method allowed the fabrication of hydroxyapatite ceramic implants of shapes that accurately matched the area of bone defect, allowing for a minimum of adjustment during the operation even with a complex-form implantation. Not only were good cranial contour reconstructed and aesthetically satisfactory results obtained in the cases treated by incorporating this series of techniques, but neurologic conditions present in some cases were also improved to some extent. The postoperative course has been steady for all nine patients, with no blood transfusions required during or after the operations and no implants requiring removal because of infection or other postoperative complications. The average length of postoperative hospitalization for the nine cases was 11.7 days, remarkably short considering the clinical conditions.     

The process of bone regeneration from devitalization to revitalization after pedicle freezing with immunohistochemical and histological examination in rabbits

The pedicle freezing procedure by liquid nitrogen is a method for the reconstruction of tumor-bearing bone after malignant tumor resection. However, the regenerative mechanism of bone after the pedicle freezing procedure is unclear. We investigated the complete process from devitalization to revitalization of bone after the pedicle freezing procedure in 13 rabbits. After osteotomy the 5 mm distal femurs were immersed in liquid nitrogen, and the specimens were divided into frozen area and sub-frozen area. The bilateral femurs were harvested for evaluation of bone regeneration by histological and immunohistochemical examination (VEGF, CD31, BMP-2 and Runx2) from 1 week to 52 weeks. The diameter of operating femurs was compared with contralateral femurs from 6 weeks to 52 weeks.No viable cells could be found from 1 to 8 weeks in the frozen area, and a mean 1.83 cm necrotic range were detected in the sub-frozen area. The periosteal reaction, massive fibrous tissue and immature bone matrix invaded from the normal area to the necrotic area from 12 weeks. Subsequently, the necrotic bone was gradually replaced by newly formed bone by creeping substitution, with endochondral and intramembrane bone formation. The diameter of frozen femurs was significantly larger than the contralateral femur at the same period from 8 weeks to 52 weeks (P < 0.01). All immunohistochemical factors were positively expressed in both areas at different time points. The active osteoblasts and microvessel migrated from marrow cavity and periosteum into dead bone. This study suggested that the frozen bone not only provides a scaffold but also possesses excellent osteoinductive properties.     

自体颏部骨移植并种植体修复前牙槽骨缺损的应用研究

目的:评价自体颏部骨移植方法在前牙槽骨缺损植骨及种植的效果。方法:对16例前牙骨缺损的患者进行自体颏部骨移植,分别于植骨12～16周后植入种植体,共植入18枚种植体,术后16～20周完成修复治疗。结果:16例自体骨移植患者中,1例出现术后感染,经局部冲洗换药后后愈合良好,其余患者移植骨均已成活,修复后种植体至今均无脱落,局部外形良好。结论:自体颏部骨移植能有效恢复局部牙槽外形,为前牙美学修复提供良好的基础。     

Comparison of frozen and freeze-dried particulate bone allografts

Freeze-dried and frozen particulate bone allografts are used interchangeably on the assumption that the biologic behavior of these grafts is similar. Dissimilarities in biologic behavior and differences in the rate and extent of bone incorporation of freeze-dried and frozen particulate grafts were demonstrated in a comparative study using a non-human primate model. Freeze-dried particulate allografts induced new bone formation and healing of the osseous defects much faster than the frozen allografts.     

Low areal bone mineral density values in adolescents and young adult turner syndrome patients increase after long-term transdermal estradiol therapy

OBJECTIVE: To study the effects of long-term estradiol therapy on areal bone mineral density (aBMD) values in young adult Turner syndrome patients. METHODS: The effects of 2-year transdermal estradiol administration on lumbar, L2-L4, aBMD values were evaluated in 12 Turner syndrome patients, 15.41-21.85 years old, who had reached adult height and had low aBMD values. Puberty was induced in all at a chronological age above 12 years and menarche appeared between 13.82 and 15.40 years. The patients were on oral estrogen/gestagen therapy from then until the start of the study. Adhesive patches of 17-beta-estradiol designed to be worn for 72 h and deliver 100 microg of estradiol per day, which results in a steady mean serum estradiol level of 75 pg/ml, were used for 21 days. From day 11 to day 21, 10 mg of oral didrogesterone were also added. Nutritional and physical activity habits were evaluated at the beginning, after 1 year and at the end of the study. RESULTS: aBMD values significantly increased from 0.910 +/- 0.065 to 1.005 +/- 0.086 g/cm2 (10.06 +/- 3.37%) and the z-score from -2.38 +/- 0.63 to -1.54 +/- 0.71 (0.81 +/- 0.30 z-score). No significant differences were observed in body mass index, calcium intake and physical activity habits at the start, during and at the end of the study. CONCLUSION: In summary, our results underline the importance of estrogens for bone mass peaking and suggest that this therapeutic protocol may be useful in the therapy of Turner syndrome patients with low bone mass.     

Utilisation of bone allograft by orthopaedic surgeons in Scotland

The use of bone allograft in orthopaedic surgery has been predicted to increase, particularly in joint revision surgery. This has led to a potential problem with supply. Questionnaires were distributed to all 146 Consultant Orthopaedic surgeons working in Scotland in 2000. They were asked to indicate their current usage of bone and tissue allograft, any problems encountered with supply and if alternatives to allograft, such as processed bone, might be used. The questions asked were very similar to those asked in a previous study in 1995 to enable comparisons to be made. Replies were received from 125 Consultants (87%) of whom 93 reported using bone allograft. Forty-one consultants (46%) predicted an increase in their requirement for bone allograft, and 23 (26%) felt they could currently use more bone if this was available. Sixty percent of surgeons would consider using processed bone as an alternative. In comparison with figures from 1995, an increasing number of surgeons are prepared to use processed bone as an alternative to fresh frozen allograft. This revised version was published online in July 2006 with corrections to the Cover Date.     

HLA-typing analysis following allogeneic bone grafting for sinus lifting

According to the Brazilian Association of Organ Transplants, in 2015, 19,408 bone transplants were performed in Brazil, over 90% by Dental Surgeons. The surgical technique itself has a respectable number of reports regarding its clinical efficacy, as measured by long-term survival of dental implants in grafted areas. Uncertainty remains, however, as to whether fresh frozen grafts from human bone donors remain immunologically innocuous in the body of the host. Six male with no previous medical history of note, including systemic diseases, surgery or blood transfusion were selected. These patients underwent reconstructive procedures (sinus lifting) using fresh frozen human bone from a tissue bank. All patients had venous blood samples collected prior to surgery and 6 months after the procedure. Anti-HLA analysis for the detection of HLA (human leukocyte antigen) antibodies was performed using methods such as the LABScreen PRA Class I and Class II, LABScreen Single Antigen Class I and Class II, Luminex Platform. Reactive individuals to the screening tests (LABScreen PRA) were further investigated to determine the specificity of the antibodies detected (LABScreen Single Antigen) with a cutoff value of median fluorescence intensity ≥500. As a result, it was observed that two patients (33%) were positive in screening tests, one presenting with anti-HLA Class I and II sensitization and the other with anti-HLA class II. The specificity analysis showed that the patients sensitized to HLA class II presented 4 specificities, 3 of which immunologically relevant. In the second individual, 23 specificities were identified, 6 of which immunologically important for HLA class I and 4 specificities for HLA class II, 3 of these were immunologically important. All specificities detected had average fluorescence. These findings are suggestive that sinus-lifting procedures with allogeneic bone can induce immunological sensitization.     

Similarity in the fatigue behavior of trabecular bone across site and species

Within the context of improving knowledge of the structure-function relations for trabecular bone for cyclic loading, we hypothesized that the S-N curve for cyclic compressive loading of trabecular bone, after accounting for differences in monotonic strength behavior, does not depend on either site or species. Thirty-five cores of fresh-frozen elderly human vertebral trabecular bone, harvested from nine donors (mean+/-S.D., age=74+/-17 years), were biomechanically tested in compression at sigma/E(0) values (ratio of applied stress to pre-fatigue elastic modulus) ranging from 0.0026 to 0.0070, and compared against literature data (J. Biomech. Eng. 120 (1998) 647-654) for young bovine tibial trabecular bone (n=37). As reported for the bovine bone, the number of cycles to failure for the human vertebral bone was related to sigma/E(0) by a power-law relation (r(2)=0.54, n=35). Quantitative comparison of these data against those reported for the bovine bone supported our hypothesis. Namely, when the differences in mean monotonic yield strain between the two types of bone were accounted for, a single S-N curve worked well for the pooled data (r(2)=0.75, n=72). Since elderly human vertebral and young bovine tibial trabecular bone represent two very different types of trabecular bone in terms of volume fraction and architecture, these findings suggest that the dominant failure mechanisms in trabecular bone for cyclic loading occur at the ultrastructural level.     

Collection,processing and freezing of equine bone marrow cells

There is no consensus on aspects of equine bone marrow collection and processing. The study aimed to describe the collection of large volumes of bone marrow from horses of advanced age, with emphasis on bone marrow mononuclear cells (BMMCs) recovery and viability after cryopreservation. Fourteen horses, aged 3–24 years, were divided into three experiments. E1 studied the feasibility of collecting 200 mL from the sternums of horses of advanced age; E2 examined the number of cells obtained from the first and last syringe of each puncture; and E3 investigated the influence of heparin concentration on the prevention of cell aggregation, and cell viability after freezing in liquid nitrogen. Bone marrow aspirations were done with syringes pre-filled with Iscove's modified Dulbecco's medium and different concentrations of sodium heparin. BMMCs were counted, cell viability was determined, and samples were frozen. Bone marrow collection from the sternum is safe, even at large volumes and from horses of advanced age, and the number of cells recovered decreases with successive aspirations (p < 0.0001). Heparin concentration influenced cell aggregation, and recovered cells continued to be commercially viable after 150 days in frozen storage.