Effects of autogenous cancellous bone on healing of homogenous cortical bone grafts

A comparative clinical and histological study was made of the effects of autogenous cancellous grafts on the rate of healing of homogenous cortical bone grafts in dogs: their right foreleg received homogenous bone grafts and their left foreleg received pre-drilled homogenous bone grafts filled with autogenous cancellous bone. Histologically, the use of homogenous cortical grafts with pre-drilled holes and autogenous cancellous bone appeared to accelerate reorganization and development of these grafts. This was seen particularly 1–2 months post-operatively and at nine months when the graft in the left leg was made up of spongy bone marrow; whereas the right leg graft still contained compact dead bone and the marrow was filled with fibrous tissue.     

Effects of gelatin sponge implantation in cancellous bone defects in dogs

Reports from oral surgeons suggested that packing bone defects with cellulose or collagen stimulated cancellous bone replacement. The study reported here was designed to evaluate a similar method for augmenting the remodeling of cancellous bone defects and graft donor sites. Gelatin sponge was implanted into bony defects created in the proximal humeral metaphysis of 5 adult Beagles after collection of cancellous bone. A similar defect created in the contralateral humerus was left unpacked. Four months after creation of the defect, the dogs were euthanatized, and the surgical sites were evaluated by use of correlated microradiographic and histologic examination. Unpacked sites had filled completely with loosely woven trabecular bone. Remodeling had not taken place in sites packed with gelatin sponge. Gelatin sponge did not elicit any reaction in the surrounding bone, but remained in situ and obstructed new bone formation. Gelatin sponge blocked rather than augmented cancellous bone replacement.     

Comparison of bone healing by demineralized bone matrix and autogenous cancellous bone in horses

OBJECTIVE: The purpose of this study was to compare bone healing induced by equine demineralized bone matrix (DBM) to autogenous cancellous bone graft (ACB) or no graft (control) in a rib-defect model in horses. STUDY DESIGN: The osteogenic properties of ACB and DBM were evaluated in bilateral 19-mm circular defects created in the outer cortex of the 6th and 8th ribs of each horse. ANIMALS OR SAMPLE POPULATION: Eight mature horses. METHODS: Three rib defects in each horse were randomly treated with each of the 3 treatment groups, and the fourth rib defect received a random treatment. Rib sections, including the defects, were harvested 56 days after implantation and examined for bone mineral density, percent ash and calcium and graded for signs of radiographic and histological healing. RESULTS: All ribs were fractured at the defect site and were classified as nonunion fractures 56 days after implantation. There were no significant differences among groups in bone mineral density and signs of radiographic or histological healing. There was an increased volume of bone in control and ACB-treated sites compared with DBM-treated sites. Rib defects treated with ACB were significantly higher in percent ash and calcium than those treated with DBM. DBM elicited no inflammatory reaction, and remodeling occurred around the periphery and within vascular channels of the decalcified particles. CONCLUSION: DBM particles remodel from the periphery, which may explain the significantly lower percent ash, calcium, and bone when compared with ACB, because 2- to 4-microL pieces of DBM may act as space-occupying masses until completely mineralized. There was no evidence of enhanced healing associated with the use of DBM in this model. CLINICAL RELEVANCE: Particles of 2 to 4 mm DBM should not be used as an aid to fracture repair because particles of this size interfere with normal mineralization. However, our model of nonunion fracture healing may be useful in future studies.     

Effects of a fixed compression load on the osteogenic effect of autogenous cancellous bone grafts in dogs.

A standardized cortical defect was created on the caudal cortex of the proximal portion of each ulna in 5 adult mixed-breed dogs. One gram of autogenous cancellous bone graft (ACBG) was obtained from the greater tubercle of the ipsilateral humerus. The cortical defect in the ulna of 1 limb was filled with 1 g of ACBG that had been compressed with 2-MPa pressure for 30 seconds. One gram of noncompressed ACBG was placed into the contralateral ulnar cortical defect. The compressed and noncompressed ACBG recipient sites were radiographed at weekly intervals. Dogs were euthanatized 8 weeks after surgery, and the ACBG recipient sites were harvested for histomorphometric analysis. Optical densitometry was performed on all radiographs. There was no significant difference between compressed and noncompressed ACBG with optical densitometry or histomorphometric analysis for total bone area. We concluded that there was no difference in osteogenic capability between compressed and noncompressed ACBG of equal mass.     

Comparison of bone healing,as assessed by computed tomography,following tibial tuberosity advancement in dogs with and without autogenous cancellous bone grafts

Abstract

AIMS: To objectively compare measures of bone healing, using computed tomography (CT) in dogs following bilateral tibial tuberosity advancement (TTA), between tibiae treated with and without autogenous cancellous bone grafts.

METHODS: Ten dogs with bilateral cranial cruciate ligament disease requiring surgical stabilisation were prospectively recruited to undergo single-session bilateral TTA, with only one, randomly assigned, tibia receiving bone graft in the osteotomy deficit. Bone healing at the osteotomy site was assessed using CT performed 38–70 days post-operatively. CT images were evaluated using both objective measurements of osseous bridging and subjective evaluation by six radiologists. Repeated measures ANOVA was used to compare the objective outcomes between the grafted and non-grafted tibiae.

RESULTS: The mean percentage of the osteotomy deficit bridged at the lateral cortex was greater in grafted (77.6, SD 35.2%) compared to non-grafted (63.0, SD 36.5%) tibiae (p=0.001), but did not differ at the medial cortex (p=0.1). The mean minimum callus width was greater in grafted (7.2, SD 3.3 mm) compared to non-grafted (3.6, SD 2.9 mm) tibiae (p<0.001). There was no difference in mean attenuation (measured in Hounsfield units) of the callus between grafted and non-grafted tibiae (p=0.5). The grafted tibia was deemed to have superior bone healing in 50/60 subjective assessments made by radiologists.

CONCLUSIONS: Superior osseous bridging was detected by CT analysis following TTA using autogenous cancellous bone grafts compared with no graft. This was shown by greater bridging percentage at the lateral cortex and formation of a broader callus. Qualitative assessments made by six radiologists also supported the conclusion that bone healing was improved by use of autogenous cancellous bone graft. CT was a useful method for assessing evidence of bone healing following TTA.

CLINICAL RELEVANCE: These findings justify the application of autogenous cancellous bone graft to augment healing following TTA in dogs.     

The effect of compacted cancellous bone grafting on the healing of subchondral bone defects of the medial femoral condyle in horses

Objective— To compare the quality of second-intention healing and that of compacting sternally harvested cancellous bone into subchondral bone defects of the medial femoral condyle in horses.
Study Design— A controlled experiment using a surgical technique that minimizes soft tissue trauma, customized for consistency among horses.
Animals or Sample Population— Ten horses, aged 2 to 5 years, free of hindlimb lameness and with radiographically normal stifles.
Methods— After a 12.7-mm-diameter × 19-mm-deep defect was created into randomly selected medial femoral condyles, bone and cartilage healing was evaluated over a 6-month period in control horses (  n = 5  ) and horses receiving a compacted cancellous bone graft (  n = 5  ). Healing was evaluated using lameness assessment, radiographic and microradiographic interpretation, arthroscopic appearance, percent bone fill, proteoglycan content, and histology.
Results— Six months after surgery, there was no significant difference between grafted and ungrafted defects with respect to lameness, radiographic score, or percent bone fill. Histologically, grafted defects were characterized by the presence of dead graft and secondary cyst formation in four defects. Ungrafted defects filled with fibrous tissue and no cyst formation were identified.
Conclusions— Grafted defects do not heal better than ungrafted defects, and lameness was not affected by surgical technique.
Clinical Significance— Cartilage healing is similar in grafted and ungrafted defects in the equine medial femoral condyle at 6 months, suggesting that surgical debridement alone of cystic structures remains the treatment of choice.     

Histologic features of the healing of bone graft donor sites in dogs

Healing of cancellous bone graft donor sites in the proximal tibial metaphysis of 12 healthy adult dogs was studied histologically. Cancellous bone was curetted from the metaphysis of the proximal end of the tibia, via a 1-cm diameter circular opening in the medial cortex. A hematoma and fibrovascular tissue filled the bone defect at 2 weeks. At 4 and 8 weeks, endosteal callus, composed initially of cartilage and woven bone and later of lamellar bone, filled the marrow cavity. At 12 weeks, the normal structural arrangement of lamellar bone and hematopoietic marrow was reestablished in the marrow cavity. The medial cortex defect was filled only with lamellar trabecular bone. It was concluded that, in adult dogs, a second cancellous bone graft could be collected from the proximal portion of the tibial metaphysis 12 weeks or more after an initial collection.     

Antigenicity of cortical bone allografts in dogs and effect of ethylene oxide-sterilization.

The aims of the present study were to determine the antigenicity of cortical bone allografts and the effect of ethylene oxide-sterilization (EO-sterilization). Cortical bone allografts from one donor dog were implanted in a muscle pouch in four groups of four dogs each. The grafts were either fresh, EO-sterilized, demineralized or demineralized and EO-sterilized. The immune response against the grafts was determined by measuring the antibody response against surface antigens of donor cells and by the mixed lymphocyte reaction. Dogs receiving EO-sterilized grafts or bone matrix did not demonstrate an immune response. Only two of the four dogs with fresh cortical bone grafts showed a very weak immune response. This suggests a priming of the host by the fresh bone grafts. However, implanting skin grafts from the donor dog subdermally, in one dog of each of the groups, four months after implanting the bone grafts did not induce a secondary immune response. Macroscopic and histologic examination of the bone grafts five months after their implantation consistently revealed graft resorption (activity of osteoclasts) and vascularization of the fresh bone grafts, but not of EO-sterilized fresh grafts. For most EO-sterilized grafts, a strong inflammatory reaction was present in the tissues surrounding the graft and this was not apparent around the non-sterilized grafts. The absence of resorption and the presence of the inflammation seemed to be unwanted effects of the EO-sterilization. The EO-sterilisation did not affect osteoinduction since osteocytes were observed in the EO-sterilized demineralized grafts. Results indicate that cortical bone allografts used in the present study are very weak antigens and that the EO-sterilization procedure used has no effect on osteoinduction, but decreases bone resorption.     

Effects of nonadherent dressing materials on the healing of open wounds in dogs

Four types of nonadherent dressing materials (rayon/polyethylene dressing, cotton nonadherent film dressings, fine mesh gauze petrolatum dressings, and commercial petrolatum emulsion dressings) were applied on small full-thickness skin defects on the backs of 12 Beagles. At 7 days, the wounds treated with the petrolatum-containing dressings had more contraction than wounds dressed with cotton nonadherent film dressings and wounds dressed with rayon/polyethylene dressings. However, by days 14 and 21, there was little difference in the amount of contraction of any of the wounds. At 7, 14, and 21 days, the wounds dressed with petrolatum-containing dressings had less epithelialization than wounds dressed with cotton nonadherent film dressings and rayon/polyethylene dressings. The uniform open mesh of the commercial petrolatum emulsion dressings allowed the best absorption of exudate and bacteria into the secondary overlying bandage.     

Effects of isobutyl-2-cyanoacrylate on bone healing

An in vivo evaluation of isobutyl-2-cyanoacrylate as an osseous adhesive was performed to determine its effect on bone healing. Oblique cortical fractures were induced at the tibial-fibular junctions of adult rabbits. Forty-eight fractures were repaired by the use of isobutyl-2-cyanoacrylate and a full cerclage wire. The opposite control fracture was repaired with a full cerclage wire only. One hundred percent of the glued fractures were not united through the glue interface, although normal periosteal new bone formation (secondary bone healing) took place around the glue. One hundred percent of the nonglued fractures healed by secondary bone union. Reactions were not seen in the bone or soft tissue adjacent to the adhesive material.     

Evaluation of the proximal portion of the femur as an autogenous cancellous bone donor site in dogs.

The proximal portion of the femur was evaluated as a source of autogenous cancellous bone in dogs. Bilateral oval cortical defects were created in the lateral subtrochanteric area of the femur in 16 dogs. Cancellous bone was removed and the weight recorded. Cancellous bone was similarly harvested from the proximal portion of the humerus in 7 of these dogs. Subtrochanteric femoral defects in 11 dogs were randomly assigned to receive cancellous bone graft obtained from the femur (n = 4) or the humerus (n = 7). Subtrochanteric defects in 5 dogs were not grafted. Radiographic assessment of subtrochanteric defects was performed at 4-week intervals, and histologic assessment at 4, 8, 16, and 24 weeks after surgery. Nongrafted donor sites healed by ingrowth of trabecular bone during the first 12 weeks after surgery. By week 24, the lateral cortical wall had reformed, but remodeling was incomplete. Donor sites grafted with cancellous bone healed similarly, but with more rapid healing and more complete remodeling evident by week 24. Although the mean weight of cancellous bone harvested from the proximal portion of the femur (0.82 +/- 0.22 g) was significantly (P less than 0.05) less than that harvested from the proximal portion of the humerus (1.38 +/- 0.29 g), there was no qualitative histologic or radiographic difference in bony healing of grafted defects. We determined that the proximal portion of the femur can be safely used to provide moderate amounts of cancellous bone, and that a second bone graft can be collected from the same subtrochanteric donor site after 12 weeks.     

Effects of orchidectomy on bone metabolism in beagle dogs

The effects of orchidectomy on bone metabolism in male beagle dogs were examined using twelve 2-year-old dogs that were orchidectomized. The dogs' bilateral iliac bones, double-labeled with tetracycline and calcein for the histomorphometry, were obtained from three dogs prior to orchidectomy and at 3, 6, 9, and 12 months afterwards. The serum biochemical constituents related to bone metabolism were examined before and every month after orchidectomy. Between 1 and 6 months after orchidectomy, the value of serum testosterone decreased (1 month), while the levels of parathyroid hormone, calcitonin, total calcium, osteocalcin, and alkaline phosphatase activity increased significantly, indicating a high bone turnover. The mean trabecular thickness and the fraction of labeled osteoid surface decreased significantly 3 months after orchidectomy, but other histomorphometric parameters were unchanged. In the period 7-12 months after orchidectomy, the parathyroid hormone level increased ever and above that of the first 6-month period, while the levels of calcitonin, osteocalcin, alkaline phosphatase activity, and phosphorus decreased. The bone volume, mean trabecular thickness, and the fraction of labeled trabecular surface decreased significantly compared with the pre-orchidectomy values. These findings indicate an imbalance in bone metabolism (i.e. bone resorption > bone formation). These results indicate that a loss of bone volume accompanied the fall in sex hormone levels following orchidectomy and suggest that the orchidectomized dog is available as an animal model for studying osteoporosis caused by hypogonadism and the decline of sex functions in men.     

Effects of periodontal dressings on fibroblasts and gingival wound healing in dogs

In the present study the effects of different commercially available periodontal dressings (Peripac, Barricaid, Fittydent, Reso-Pack and Myzotect-tincture) on fibroblast (V-79-379A) proliferation and survival were tested in vitro. Barricaid, Fittydent and Reso-Pack periodontal dressings have only small inhibitory effects on cell proliferation (83.3 +/- 9%, 71.6 +/- 8.7% and 87.3 +/- 4.5% of control after 48 h, respectively) in comparison with the great inhibitory effect of Myzotect-tincture (2.9 +/- 0.1%) and Peripac (33.7 +/- 11.4%) (p < 0.001). Barricaid was the only dressing where 41% of cells survived after exposure, while the other four dressings killed all the cells in 6 days. In addition, the healing of artificially created gingival wounds covered by Barricaid and Reso-Pack was followed for 7 days in 12 Beagle dogs. Histological evaluation of gingival tissue demonstrated that wounds covered by Reso-Pack showed the best epithelisation and vascularity and the least inflammatory reaction in first 4 days. Later the observed parameters were similar with those of wounds covered by Barricaid or without pack. The present results indicate that Peripac periodontal dressing and Myzotect-tincture showed the highest cytotoxicity to fibroblasts in vitro. From the histological observations in Beagle dogs Reso-Pack has been found to be the most suitable dressing, followed by Barricaid.     

Effects of a hydrolyzed collagen dressing on the healing of open wounds in dogs

OBJECTIVE: To determine the effects of a hydrolyzed bovine collagen dressing (HBCD) on healing of open wounds in healthy dogs. ANIMALS: 9 female Beagles. PROCEDURES: 2 full-thickness skin wounds were made bilaterally on the trunk of each dog. Wounds on 1 side were treated with powdered HBCD covered with a semiocclusive nonadherent bandage. Wounds on the other side (control wounds) were covered with a semiocclusive nonadherent bandage only. Wound healing was subjectively assessed, and percentage increase in tissue perfusion was assessed by use of laser Doppler perfusion imaging (LDPI). Planimetry was performed to determine the percentages of contraction, epithelialization, and total wound healing. Biopsy specimens were examined microscopically to evaluate histologic changes. RESULTS: The HBCD did not induce a strong inflammatory reaction, as reflected by results of LDPI and histologic examination. Moreover, HBCD appeared hydrophilic and provided an environment to keep wounds clean and enhance early epithelialization. After treatment for 7 days, treated wounds had a significantly greater percentage of epithelialization than control wounds (12.13 vs. 7.03%). CONCLUSION AND CLINICAL RELEVANCE: The hydrophilic property of HBCD may cleanse contaminated wounds with the body's homeostatic fluids and enhance early wound epithelialization.     

Effects of a 98% solution of glycerol or sterilization with ethylene oxide on FeLV in bone allografts and effects on bone incorporation of allografts in cats

OBJECTIVES: To compare virucidal effects and bone incorporation properties of cortical bone allografts transplanted into specific-pathogen-free (SPF) cats. Allografts consisted of untreated bone from a SPF cat (negative-control group) and bone from 5 FeLV-infected cats that was subjected to sterilization with ethylene oxide (ETO), preservation with glycerol, or no treatment (positive-control group). SAMPLE POPULATION: Bones from the aforementioned groups and twenty 8-week-old SPF cats (5 cats/group) implanted with an allograft from 1 of the aforementioned groups. PROCEDURE: After implantation, blood samples were collected weekly to monitor FeLV p27 antigen and antibody titers. Quantification of FeLV provirus was performed on blood samples at weeks 0, 4, and 8 and donor bone samples at time of implantation. Cats were euthanatized 8 weeks after transplantation, and graft sites were evaluated. RESULTS: All results for negative-control cats were negative. All ETO group cats had negative results for antigen and provirus in blood, whereas 1 cat had a low antibody titer. Although 3 ETO-treated allografts were positive for provirus, the DNA appeared denatured. One cat in the glycerol group had positive results for all tests in blood samples. All glycerol-preserved allografts were positive when tested for provirus. All results for positive-control group cats were positive. Differences in incorporation of bone grafts were not observed. CONCLUSIONS AND CLINICAL RELEVANCE: Glycerol preservation of FeLV-infected bone allografts did not eliminate transmission of retrovirus to recipients. In contrast, ETO sterilization appeared to denature DNA and prevent infection. Treatments did not affect incorporation of bone grafts in young cats.     

Open drainage and delayed autogenous cancellous bone grafting for treatment of chronic osteomyelitis in dogs and cats

Ten dogs and 1 cat with 12 chronically infected bones were treated surgically by means of saucerization, primary internal or external rigid fixation, and open irrigation drainage. After formation of a healthy granulation bed, autogenous cancellous bone grafting and skin closure were performed. Eleven of 12 bones healed 4 to 7 weeks (mean, 4.8 weeks) after treatment, without recurrence of infection. Eight of 12 chronically infected bones had a polymicrobial infection, with 2 to 6 bacterial species isolated. Gram-negative bacteria were isolated from 7 of the 12 bones, and anaerobic bacteria were isolated twice.     

Recombinant human bone morphogenetic protein-2 in absorbable collagen sponge enhances bone healing of tibial osteotomies in dogs

OBJECTIVE: To compare the efficacy of 2 doses of recombinant human bone morphogenetic protein-2 (rhBMP-2) on tibial osteotomy healing in dogs. STUDY DESIGN: Experimental, randomized complete block (n=7). ANIMALS: Adult female dogs (n=21). METHODS: Right midshaft tibial osteotomies were created and stabilized with a 1-mm gap using type I external fixators. Seven dogs were untreated controls and 14 with osteotomies were treated with either 0.05 or 0.2 mg/mL rhBMP-2 delivered in an absorbable collagen sponge (ACS). At 8 weeks, dogs were euthanatized and bones were mechanically tested and examined by microscopy. RESULTS: Bone healing based on radiographic scoring, was significantly improved in dogs treated with 0.2 mg/mL of rhBMP-2 compared with the other groups; these tibiae were also significantly stronger and stiffer than 0.05 mg/mL rhBMP-2 and control osteotomized tibiae. Histologic scores were significantly better for 0.2 mg/mL rhBMP-2 group than 0.05 mg/mL rhBMP-2 group, but neither was significantly different from control. CONCLUSIONS: rhBMP-2 in ACS at a concentration of 0.2 mg/mL improves healing of tibial osteotomies in dogs compared with untreated controls and 0.05 mg/mL rhBMP-2 based on force plate analysis and radiographic evaluation. This was not confirmed histologically but treated bones had improved mechanical properties at 8 weeks. CLINICAL RELEVANCE: After a long bone fracture, dogs may face a long recovery period before full return of limb function. rhBMP-2, in association with good fracture fixation principles, may enhance bone healing in dogs with diaphyseal fractures.     

Effects of a bioactive glass on healing of closed skin wounds in dogs.

OBJECTIVE: To determine effects of intraincisional bioactive glass on healing of sutured skin wounds in dogs. ANIMALS: 9 purpose-bred mature female Beagles. PROCEDURE: 3 small matched bilateral (treated vs control) full-thickness truncal skin incisions were made and sutured. Treated wounds received intraincisional particulate bioactive glass prior to closure. Laser Doppler perfusion imaging was used to assess percentage change in tissue perfusion 3 and 5 days after incision on 1 set of 2 matched wounds, and skin and subcutaneous tissue-cutaneous trunci breaking strength were assessed at 5 days. The other 2 sets of wounds were used for histologic evaluation at 5 and 21 days, respectively. RESULTS: Subjective signs of gross inflammatory reaction were not detected in treated or control wounds. At 5 days, median subcutaneous tissue-cutaneous trunci breaking strength was significantly higher in treated wounds than in control wounds-(188.75 vs 75.00 g). At 5 days, median scores were significantly higher for neutrophils (1 vs 0), macrophages (2 vs 1), and necrosis (1 vs 0) for treated wounds than for control wounds. At 21 days, median macrophage scores were significantly higher for treated wounds than for control wounds (2 vs 1). CONCLUSIONS AND CLINICAL RELEVANCE: Bioactive glass in soft tissues does not cause a gross inflammatory reaction but causes an increase in histologic signs of inflammation, which decreases with time. Bioactive glass has potential for increasing tissue strength. Increased subcutaneous breaking strength could be beneficial in treating wounds in which early healing strength is needed.     

Correlation of serum alkaline phosphatase activity with the healing process of long bone fractures in dogs

BACKGROUND: Bone healing is monitored mainly by physical and serial radiologic examinations of the fracture site. However, it is sometimes difficult to distinguish a delayed union from a nonunion, and advanced imaging techniques may not be available. Serum biochemical markers of bone formation, such as alkaline phosphatase (ALP) activity, may be clinically useful in evaluating the progress of healing. OBJECTIVE: The purpose of this study was to correlate serial values of serum ALP activity with the process of fracture healing in dogs and to assess its potential as a postsurgical prognostic indicator. METHODS: Changes in serum ALP activity were studied in 83 dogs with closed long bone diaphyseal fractures treated surgically. Physical and radiologic examinations of the fracture site and determination of serum ALP activity and calcium (Ca) and phosphate (P) concentrations were performed on admission (day 0); postoperatively on days 10, 20, and 30; and subsequently on a monthly basis until bone union was completed or signs of nonunion were evident. The dogs were allocated into 3 groups with respect to the fracture healing progress as documented by physical and serial radiologic examination. RESULTS: Group A dogs (n=35) developed a medium-sized callus that led to bone union within 2 months. Group B dogs (n=36) had a hypertrophic callus and delayed union, within 3-5 months. Group C dogs (n=12) had slow progress in fracture healing, with minimal callus formation during a 2-month period. Changes in mean serum ALP activity followed the same pattern in groups A and B, reaching a maximum level on day 10. Group A values returned to normal within 2 months, at which point bone union was complete, whereas group B values remained increased and returned to normal within 3-5 months, thus correlating with delayed union. In Group C, mean serum ALP activities showed no significant changes during the 2-month follow-up period, consistent with failure of bone union (nonunion). Serum P and Ca changes followed a proportional and inverse pattern to ALP changes, respectively. CONCLUSION: Serial determination of serum ALP activity during fracture healing could be an additional tool in predicting fractures at risk of developing a nonunion, helping the clinician to choose the appropriate intervention.