Uterine horn intussusception in immediate post-partum in a Pomeranian Spitz dog.

A 2-year-old Pomeranian-Spitz dog (5.8 kg) was admitted with symptoms of uterine prolapse and lethargy 1 day after whelping three puppies. The prolapsed uterus was corrected, but the next day, prolapse reoccurred. To fix the cervix to the abdominal wall, the dog underwent a surgery operation where intussusception in the left horn of the uterus was found. Ovariohysterectomy was performed as the treatment of choice. The diagnosis of intussusception is challenging, and exploratory laparotomy is the reliable diagnostic approach. Although uterine intussusception is rare, it should be one of the important considerations in dogs within the post-partum period.

The potential prophylactic and therapeutic impacts of niacin on ischemia/reperfusion injury of testis.

INTRODUCTION: The testicular ischemia-reperfusion (I/R) injury is characterized by the excessive aggregation of un-scavenged reactive oxygen species, leading to the heightened levels of oxidative stress. This phenomenon plays a pivotal role in the pathophysiology of testicular torsion damage. OBJECTIVE: The current study aimed to detect the prophylactic and therapeutic effects of niacin on testicular I/R injury. STUDY DESIGN: Twenty-four healthy adult male Sprague Dawley rats were randomly allocated into three groups as follows: (1) sham group, (2) torsion/detorsion (T/D) group, and (3) treatment group which received 200 mg/kg niacin along with testicular T/D. Torsion/detorsion was induced by 2 h of torsion followed by 10 days of reperfusion period. In the treatment group, niacin was injected 30 min before the reperfusion period intraperitoneally and continued for 10 days by oral gavage. RESULTS: T/D was associated with marked decreases in terms of sperm count, viability, and kinematic parameters versus the sham group (P < 0.05), which niacin significantly reverted the kinematic parameters (P < 0.05). I/R injury caused a significant increase in the number of abnormal epididymal sperms compared to the sham group (P < 0.05). Niacin decreased the epididymal sperm abnormality significantly compared to the T/D group (P < 0.05). Tissue abnormalities in T/D group, such as edema, hyperemia, inflammation, and necrosis were completely visible histopathologically, while the histological changes in the niacin-treated group were better than those in the T/D group. Regarding the pathological parametric evaluations, I/R injury significantly reduced the mean testicular biopsy score (MTBS), germinal epithelial cell thickness (GECT), and mean seminiferous tubular diameter (MSTD), and increased the tubular hypoplasia/atrophy (THA) compared to the sham group (P < 0.05), which niacin treatment significantly improved the MTBS and GECT compared to the T/D group (P < 0.05). T/D significantly increased the oxidative stress index (OSI) and lipid peroxidation (MDA) (P < 0.05). Niacin significantly reduced the OSI and MDA levels compared to the T/D group (P < 0.05). DISCUSSION: The current study found that niacin has preventive/therapeutic effects against the elevation of oxidative stress markers and depletion of antioxidants during I/R injury. Following administration of niacin, a reduction in histologic injury was observed in rats. In our study, we showed the antioxidant properties of niacin and its capacity to protect against I/R damage. CONCLUSION: The findings of the present investigation revealed that niacin, as an antioxidant agent, can suppress the oxidative stress induced by testicular I/R injury, and can be used as a supplementary agent in the treatment of those undergoing testicular torsion surgery.

Effect of direct therapeutic ultrasound exposure of ovaries on histopathology, inflammatory response, and oxidative stress in dogs.

BACKGROUND: This research was designed to evaluate the effects of therapeutic ultrasound waves on ovarian germinal tissue and inflammatory cytokines (interleukin-6 (IL-6), IL1ß, tumor necrosis factor-α (TNF-α)), acute phase proteins (serum amyloid A (SAA), C reactive protein (CRP)) and oxidative stress (total antioxidant capacity (TAC), and malondialdehyde (MDA)) in dogs. Twenty-six clinically healthy adult mix-breed female dogs were aligned into three groups. Laparotomy was performed in control (n = 6) and treatment (T5, n = 10; T10, n = 10) groups. The ultrasonic exposure of ovaries in treatment groups was performed during laparotomy by round motions of the therapeutic ultrasonic transducer on both ovaries (1 MHz frequency, 1.5 W/cm2) for 5 min in the T5 group and for 10 min in the T10 group. Blood samples were collected from the jugular vein into a plain glass tube on days 0 (before laparotomy), 3, 6, and 9 after surgery. All control and treatment groups' dogs were ovariectomized for histological evaluation on day 60 after laparotomy or laparotomy + ultrasound exposure. RESULTS: Direct exposure of ovaries with therapeutic ultrasound waves induced inflammation and oxidative stress comparison with the control group. Histopathological evaluation of treated ovaries with ultrasound waves indicated a decreased number of primordial follicles (ovarian reserve) and oocyte preservation scores compared with ovaries in the control group. CONCLUSIONS: These changes may cause subfertility in the long term. It seems that inflammatory response and oxidative stress are factors in the permanent damage of ovarian tissue.

Surgical management of an aspirated bone in a Shih Tzu terrier dog: a case report.

Foreign bodies in the respiratory tract (trachea/bronchus) are very rare conditions; therefore, it is not on top of the list of differential diagnoses in patients with chronic or severe cough, but its occurrence should not be considered unexpected. A 2-year-old male Shih Tzu terrier mix dog (9.20 kg) with a history of dyspnea for 6 days was referred to the Veterinary Teaching Hospital of Shiraz University, Shiraz, Iran. In the clinical examination, respiratory distress and pain during palpation of the neck area were observed. Radiology indicated a triangular radiopaque object (bone) in the trachea of the animal. The size of the foreign body, the weakness of the grasping forceps of the device, and the edges of the bone being stuck in the trachea caused tracheoscopy failure after 30 min of trying. Surgical procedure (tracheotomy) was effective to remove the foreign body using Noyes alligator tissue forceps. Aspirated foreign body is not a common condition. In the case of early recognition and immediate treatment of the aspirated foreign body, the prognosis is usually good.

Effects of concurrent use of royal jelly with hydroxyapatite on bone healing in rabbit model: radiological and histopathological evaluation.

BACKGROUND: Bone grafts have been used to enhance bone fracture healing in orthopedic surgery. Bone grafts enhance bone healing either by mechanical support or acting as a scaffold for bone formation. Fresh autograft is the most effective biomaterial because it is histocompatible with less complication about transmissible disease. Hydroxyapatite is a well-established material for bone repair and very comparable to natural apatite providing a strong biomechanical interlock with host tissue. Royal jelly is the principal food for the honeybee queen. This biomaterial has been demonstrated to have several pharmacological activities, such as antiallergic, antitumor and anti-inflammatory effects. OBJECTIVES: This study was design to evaluate the effect of concurrent using of Royal jelly with hydroxyapatite on bone healing in rabbit model. METHODS: 15 adult rabbits weighting approximately 2 kg had been used. They were divided into three groups randomly. In first group (N = 5) mid radius bone defect created and left empty. The second group (N = 5) filled with hydroxy apatite alone and the last group (N = 5) filled with royal jelly and hydroxy apatite combination. Radiological evaluation performed on days14th, 28th and 42nd after operation. Histopathological evaluation was done on 56th postoperative day. RESULTS: Radiological evaluation showed significant superior bone healing in hydroxyapatite and hydroxyapatite-Royal jelly groups in comparison to control group. Control group was the inferior group between three groups. There were not any significant differences between three groups in histopathological group. CONCLUSION: In conclusion our study showed the best results with using the hydroxyl apatite and Royal-jelly group because they provide not only scaffold for bone healing but also do, they provide some osteoinduction materials for bone healing.

Effects of different intensities of strength and endurance training on some osteometabolic miRNAs, Runx2 and PPARγ in bone marrow of old male wistar rats.

Bone tissue is known as a living dynamic and complex organ in response to physical activity and mechanical loading such as exercise training; thus, the purpose of this study was to determine the effect of different intensities of strength and endurance training on expression of some osteometabolic miRNAs and runt-related transcription factor 2 (Runx2) and peroxisome proliferator-activated receptor γ (PPARγ) in bone marrow of old male Wistar rats. To this end, a total number of 50 male Wistar rats (aged 23 months, 438.27 g) were obtained from Pasteur Institute of Iran. The rats were randomized into five groups (10 rats/per group) including moderate endurance training (MET), high-intensity endurance training (HET), moderate-intensity resistance training (MRT), high-intensity resistance training (HRT), and control (CON). The four training groups completed 8 weeks of a training program, 5 days a week, according to the study protocol. To evaluate miR-133a, miR-103a, miR-204, and other adipogenic and osteogenic genes such as RUNX2 and PPARγ via real-time PCR, total RNA including mRNA and miRNA was isolated from the bone marrow. The statistical analysis was then performed using two-way analysis of variance (ANOVA). No significant differences in miR-133a (p = 0.197), miR-103a (p = 0.302), miR-204 (p = 0.539), RUNX2 (p = 0.960), and PPARγ (P = 0.872) were observed between the intervention groups and the control one. Furthermore, there were no significant differences in bone force (p = 0.641), fracture energy (p = 0.982), stress (p = 0.753), module (p = 0.147), and elongation (p = 0.292) variables between the intervention groups and the control group. Investigating molecular and cellular changes in the bone after such exercises in longer time could provide clearer results about the beneficial or harmful effects of these types of exercises in healthy and passive elderly people.

Propolis extract a new reinforcement material in improving bone healing: An in vivo study.

BACKGROUND: Propolis is known for its antioxidant, immune response modulating, and wound healing effects. In the present study in order to determine the bone healing capacity of the propolis extract, a critical sized, nonunion, radial bone defect model was repaired in rat, using chitosan and demineralized bone matrix (DBM) scaffolds along with propolis extract. MATERIALS AND METHODS: Seventy-two radial bone defects in 36 healthy male rats were randomly divided into 6 groups (n = 12/group). The groups included autograft, defect or untreated group, chitosan, DBM, chitosan and propolis (chitosan-propolis), and DBM and propolis (DBM-propolis). The bone repairing capability was characterized using radiography at 28th, 42nd and 56th postoperative days. Gross morphologic, histopathologic, histomorphometric and biomechanical examinations were performed following euthanasia at the 56th post-operative day. RESULTS: The DBM-propolis group, showed better structural and biomechanical properties compared to the untreated, DBM, chitosan and chitosan-propolis groups. The defect site in the chitosan and untreated groups were mainly restored by fibrous connective tissue while the lesions in the autograft group were mostly filled by cartilage and a lesser amount of woven bone. The woven bone, and the hyaline cartilage were the main constituents of the newly formed tissues in the DBM-propolis group, at the 56th day after injury. CONCLUSION: The results of this study showed that percutaneous injection of diluted aqueous propolis extract in the bone defect (25 mg/defect) can improve bone formation in the critical radial bone defect in rat. Since there was no significant difference between the autograft and DBM-propolis group, probably this therapeutic strategy has high potential in augmentation of autologous bone grafting.

Healing potentials of polymethylmethacrylate bone cement combined with platelet gel in the critical-sized radial bone defect of rats.

Polymethylmethacrylate (PMMA) is the most commonly used filler material that lacks biological properties and osteoconductivity or osteoinductivity. Platelet gel (PG) is a typical source of growth factors, cytokines and molecules efficient for bone formation and remodeling. The aim of this study was to evaluate bone healing and regeneration of bone defect in rat model by combining PMMA with PG. A total of 50 defects were created in the diaphysis of the radii of 25 male Sprague-Dawley rats. These defects were randomly divided into five groups (n = 10 defects for each group) and treated by autograft, plain PMMA, PG and PMMA-PG or left untreated. The rats were examined clinically and radiologically during the experiment and also after euthanasia at the 8th post-operative week, the healed defects were evaluated by gross morphology, histopathology, histomorphometry, computed tomography, scanning electron microscopy and biomechanical testing. PG could function as efficiently as autograft in promoting bone healing of the radial bones. Additionally, bone formation, and densities of cartilaginous and osseous tissues in the defects treated with autograft, PG and PMMA-PG were more satisfactory than the untreated and PMMA treated defects. Compared with the PMMA-PG implant, more PMMA residuals remained in the defect area and induced more intense inflammatory reaction. In conclusion, addition of PG could improve the bone regenerative properties of PMMA bone cement compared with PMMA alone in vivo. Therefore, the PG-PMMA can be proposed as a promising option to increase regenerative potential of PMMA, particularly when it is used as fixator, filler or adhesive in the dentistry, neurosurgery and bone tissue engineering applications.

The effects of gelatin, fibrin-platelet glue and their combination on healing of the experimental critical bone defect in a rat model: radiological, histological, scanning ultrastructural and biomechanical evaluation.

Fibrin-platelet glue (FPG) is a blood derivative, in which platelets and fibrinogen are concentrated in a small plasma volume, by differential centrifugation and precipitation. It can form a three-dimensional and biocompatible fibrin scaffold with a myriad of growth factors and proteins that are released progressively to the local environment and contribute to the accelerated postoperative bone healing. Gelatin (Gel) is a derivative of collagen and can promote cell adhesion and proliferation due to its unique sequence of amino acids, so it is suitable for bone tissue applications. This study examined the effects of Gel, FPG and their combinations as bone scaffold on the healing of surgically created critical-size defects in rat radius. Fifty critical size defects of 5 mm long were bilaterally created in the radial diaphysis of 25 rats. The animals were randomly divided into five equal groups as empty defect, autograft, Gel, FPG and Gel-FPG groups (n = 10 in each group). Radiographs of each forelimb were taken postoperatively on the 1st day and then at the 28th and 56th days post injury to evaluate bone formation, union and remodeling of the defect. After 56 days, the rats were euthanized and their harvested healing bone samples were evaluated by histopathology, scanning electron microscopy (SEM) and biomechanical testing. The results of present study showed that the Gel alone did not significantly affect bone healing and regeneration; however, the Gel treated defects promoted healing more than those that were left untreated (negative control). Furthermore, the FPG-enhanced grafts provided a good scaffold containing numerous growth factors for proliferation of osteoinduction and was effective in improving the structural and functional properties of the newly formed bone more than that of the untreated and also the Gel treated groups. Incorporation of Gel into the FPG scaffold improved healing potential of the FPG scaffold; however, it was still inferior to the autograft (positive control). Although the Gel-FPG scaffolds had best effectiveness during bone regeneration, it still needs to be further enhanced by incorporation of the ceramic and osteoinductive biomaterials.

Comparative Expression Analysis of HSP70, HSP90, IL-4, TNF, KITLG and KIT-receptor Gene between Varicocele-Induced and Non-Varicocele Testes of Dog.

BACKGROUND: This study was designed to create an experimental varicocele model by a simple surgical procedure in dog with minimum invasion and to investigate the effect of varicocele-induced infertility on the expression of six related genes (HSP90, HSP70, IL- 4, TNF, KITLG and KIT receptor). MATERIALS AND METHODS: In this experimental study, the proximal part of the pampiniform plexus of dog testes was partially occluded without abdominal incision which was confirmed by venographic examination. To evaluate varicocele in its acute form, dogs were castrated after 15 days and testes were dissected. Histopathologic evaluation was undertaken and the relative expression of the six genes was assessed by quantitative realtime polymerase chain reaction (PCR). RESULTS: Microscopic changes showed tubule degeneration. The Johnson score was significantly decreased in the varicocele testes when compared with non-varicocele testes. Expressions of HSP90, TNF, KITLG and the KIT-receptor gene were significantly downregulated (P=0.029, 0.047, 0.004 and 0.035 respectively) in varicocele-induced testes while HSP70 was upregulated (P=0.018). IL-4 did not show differential expression (P=0.377). CONCLUSION: We conclude that partial occlusion of the proximal part of the pampiniform plexus induces varicocele in the testis of dog. Differential expression of the mentioned genes may be responsible for the pathophysiology of varicocele and related subfertility.

Dicalcium Phosphate Anhydrous: An Appropriate Bioceramic in Regeneration of Critical-Sized Radial Bone Defects in Rats.

The present study aimed to evaluate and compare the effectiveness of composites of calcium phosphates including ß-tri calcium phosphate (ß-TCP), dicalcium phosphate anhydrous (DCPA, monetite), mono-calcium phosphate monohydrate (MCPM), and hydroxyapatite (HA) with the chitosan-gelatin-platelet gel (CGP) on the healing of experimentally induced critical size radial bone defects in rats after 8 weeks of injury. Eighty bilateral bone defects were created in the radial bones of 40 adult male Sprague-Dawley rats. The defects were either left empty (untreated or defect group), or treated with autograft, CGP, CGP-DCP, CGP-TCP, CGP/ß-TCP/DCPA (CGP-TD), CGP-TD/MCPM (CGP-TDM), and CGP-TDM/HA (CGP-TDMH) scaffolds. The injured forelimbs were evaluated by radiography, gross morphology, three-dimensional computed tomography scanning, histopathology, histomorphometry, scanning electron microscopy, and biomechanical testing. The materials were analyzed using X-ray diffraction to verify the crystalline nature of their structures, and their crystallinity was revealed based on the diffraction peaks achieved from the XRD analysis. The best results were achieved by the CGP-DCP scaffold and the autograft. The CGP-TCP and CGP-TDMH scaffolds were not degraded, while the CGP-DCP, CGP-TDM, CGP-TD, and CGP scaffolds were biodegraded and enhanced bone formation compared with the CGP-TCP and CGP-TDMH groups (P < 0.05). Overall, the CGP-DCP treated defects showed significant improvement in bone formation and union, bone volume, maximum load, and stiffness compared to the CGP group (P < 0.05). It could be concluded that the CGP-DCP scaffold can be considered as a suitable substitute to autograft. In fact, this study demonstrated that DCPA or monetite has high healing potential due to its biocompatibility, biodegradability and biomechanical, osteoconductive and osteoinductive properties of this bioceramic.

Effectiveness of tissue engineered based platelet gel embedded chitosan scaffold on experimentally induced critical sized segmental bone defect model in rat.

BACKGROUND: Healing and regeneration of large bone defects are a challenging problem for reconstructive orthopedic surgeons. PURPOSE: This study investigated the effectiveness of chitosan scaffold (CS), platelet gel (PG) and their combination (CS-PG) on healing process of an experimentally induced critical sized segmental bone defect model in rat. METHODS: Fifty bilateral defects were created in the mid diaphysis of the radial bones of 25 Sprague-Dawley rats. The animals were randomly divided into five equal groups. The bone defects were either left untreated or treated with corticomedullary autograft, CS, PG or CS-PG. Plain radiographs were provided from the radial bones on weeks 2, 5, and 8 after injury. In addition, clinical examinations were done for the healing radial bones. The animals were euthanized after 8 weeks of injury, and their harvested samples were evaluated by gross morphology, histopathology, scanning electron microscopy, CT-scan, and biomechanical testing. RESULTS: Compared with the defect group, the PG and autograft treated bone defects had significantly superior radiological scored values, bone volume and biomechanical performance which had positive correlation with their superior gross pathological, histopathological and ultra-structural features. Compared with the untreated defects, the PG and CS-PG treated defects showed significantly superior structural and functional properties so that PG had the highest value. In addition, CS had low value in bone regeneration. Although combination of CS and PG improved the healing efficacy of the CS, this strategy reduced the ability of PG to increase osteoconduction and osteoinduction during bone regeneration. CONCLUSION: Application of PG alone enhanced bone healing and can be regarded as a promising option for bone tissue engineering in clinical settings. Chitosan was not effective in bone reconstruction surgery and further investigations should be conducted to find a suitable carrier for PG.

Chitosan/gelatin/platelet gel enriched by a combination of hydroxyapatite and beta-tricalcium phosphate in healing of a radial bone defect model in rat.

This study compared the regeneration potentials of the hydroxyapatite (HA) and beta-tricalcium phosphate (ß-TCP) alone or in combination with a HA:TCP ratio of 30:70 in the critical-sized radial bone defects of rats. Bilateral 60 radial bone defects created were randomly divided into six equal groups (n=10 defects/group) including autograft, untreated or defect, chitosan-gelatin-platelet gel (CGP), CGP-HA, CGP-TCP and CGP-HA/TCP. The defects were evaluated by radiography, morphology, histopathology, histomorphometry, CT scan, scanning electron microscopy and biomechanical testing after eight weeks. Compared with the untreated and CGP-HA groups, the CGP and CGP-HA/TCP groups showed significantly higher new bone formation, bone volume, and mechanical properties. The CGP-HA and CGP-TCP scaffolds showed low biodegradability, whereas the CGP scaffolds were completely degraded. Osteoconductivity and osteoinductivity of the CGP and CGP-HA/TCP scaffolds were superior to the CGP-HA ones. The untreated and CGP-HA groups repaired mostly through fibrosis, while there were evidence of higher bone formation in the autograft, CGP and CGP-HA/TCP groups. In conclusion, addition of HA or ß-TCP alone into the CGP scaffolds impaired bone regeneration, while bone regeneration with the CGP and CGP-HA/TCP scaffolds was comparable with the autograft. Therefore, the CGP-HA/TCP scaffold can be a possible option to substitute the autologous bone grafting.

Effectiveness of tissue engineered chitosan-gelatin composite scaffold loaded with human platelet gel in regeneration of critical sized radial bone defect in rat.

Although many strategies have been utilized to accelerate bone regeneration, an appropriate treatment strategy to regenerate a new bone with optimum morphology and mechanical properties has not been invented as yet. This study investigated the healing potential of a composite scaffold consisting of chitosan (CS), gelatin (Gel) and platelet gel (PG), named CS-Gel-PG, on a bilateral critical sized radial bone defect in rat. Eighty radial bone defects were bilaterally created in 40 Sprague-Dawley rats and were randomly divided into eight groups including untreated, autograft, CS, Gel, CS-PG, Gel-PG, CS-Gel, and CS-Gel-PG treated defects. The bone defects were evaluated clinically and radiologically during the study and their bone samples were assessed by gross and histopathology, histomorphometry, CT-scan, scanning electron microscopy, and biomechanical testing after 8weeks of bone injury. The autograft and CS-Gel-PG groups showed significantly higher new bone formation, density of osseous and cartilaginous tissues, bone volume, and mechanical performance than the defect, CS and Gel-PG groups (P˂0.05). In addition, bone volume, density of osseous and cartilaginous tissues, and numbers of osteons in the CS-Gel-PG group were significantly superior to the CS-PG, CS-Gel and Gel groups (P˂0.05). Increased mRNA levels of alkaline phosphatase, runt-related transcription factor 2, osteocalcin, collagen type 1 and CD31, vascular endothelial growth factor as osteogenic and angiogenic differentiation markers were found with the CS-Gel-PG scaffold by quantitative real-time PCR in vitro after 30days of culturing on bone marrow-derived mesenchymal stem cells. In conclusion, the healing potential of CS-Gel scaffold embedded with PG was comparable to autografting and therefore, it can be offered as an appropriate scaffold in bone tissue engineering and regenerative applications.

Comparative study on the healing potential of chitosan, polymethylmethacrylate, and demineralized bone matrix in radial bone defects of rat.

This study aimed to compare the effectiveness of xenogeneic demineralized bone matrix (DBM), chitosan (CS), and polymethylmethacrylate (PMMA) on the regeneration of the critical-sized radial bone defects in rats after eight weeks. Fifty bilateral radial bone defects were randomly divided into five groups including untreated defects and those treated with autograft, CS scaffold, PMMA, and DBM. The defects were evaluated by diagnostic imaging, histopathology, histomorphometry, scanning electron microscopy, and biomechanical testing. Compared with the defect, CS, and PMMA groups, the autograft and DBM treated defects showed significantly higher new bone formation, bone volume, ultimate mechanical strength, and stiffness, but significantly lower inflammatory cells, fibroblasts, fibrocytes, and strain. Moreover, DBM showed significantly superior biocompatibility, biodegradability, osteoconductivity, and osteoinductivity to the CS scaffold and PMMA. In conclusion, both CS and PMMA alone were non-biocompatible polymers with slow biodegradation which retarded bone regeneration, whereas DBM significantly improved bone healing close to the gold method. However CS was not osteoconductive or osteoinductive alone, it can be combined with other biomaterials and molecules considering the excellent properties of this carbohydrate biopolymer for bone healing and regeneration.

Role of platelet gel embedded within gelatin scaffold on healing of experimentally induced critical-sized radial bone defects in rats.

PURPOSE: We investigated the role of human platelet gel (PG) embedded within gelatin (Gel) scaffold on healing of critical-sized radial bone defects in rats. METHODS: Twenty-five Sprague-Dawley rats were randomly divided into five equal groups. In each animal, critical-sized 5-mm bone defects were created in the radial bones of both forelimbs (n = 10/group). The defects were then either left untreated or filled with autograft, Gel, PG or Gel-PG. Before euthanasia, the healing defects were evaluated radiologically and clinically. The animals were euthanized after eight weeks and their radial bones evaluated by radiography, computed tomography (CT) scan, histology, biomechanical testing and ultrastructural evaluations. RESULTS: PG implantation significantly increased cellular differentiation, osteoblastic proliferation and consequently new bone formation so that those defects treated with PG showed superior structural and biomechanical properties to the Gel and PG-Gel-treated defects. The PG-treated defects had radiological, morphological and mechanical properties closely comparable with those of the autograft group. In contrast, in the PG-Gel group, Gel significantly reduced the beneficial effects of PG on bone healing. CONCLUSIONS: Human PG had beneficial effects on bone regeneration, while combination of PG and Gel had no remarkable beneficial effect. Therefore, PG when used alone can be regarded as a promising osteoinductive and osteoconductive option in bone tissue engineering applications.

Comparative study on the role of gelatin, chitosan and their combination as tissue engineered scaffolds on healing and regeneration of critical sized bone defects: an in vivo study.

Gelatin and chitosan are natural polymers that have extensively been used in tissue engineering applications. The present study aimed to evaluate the effectiveness of chitosan and gelatin or combination of the two biopolymers (chitosan-gelatin) as bone scaffold on bone regeneration process in an experimentally induced critical sized radial bone defect model in rats. Fifty radial bone defects were bilaterally created in 25 Wistar rats. The defects were randomly filled with chitosan, gelatin and chitosan-gelatin and autograft or left empty without any treatment (n = 10 in each group). The animals were examined by radiology and clinical evaluation before euthanasia. After 8 weeks, the rats were euthanized and their harvested healing bone samples were evaluated by radiology, CT-scan, biomechanical testing, gross pathology, histopathology, histomorphometry and scanning electron microscopy. Gelatin was biocompatible and biodegradable in vivo and showed superior biodegradation and biocompatibility when compared with chitosan and chitosan-gelatin scaffolds. Implantation of both the gelatin and chitosan-gelatin scaffolds in bone defects significantly increased new bone formation and mechanical properties compared with the untreated defects (P < 0.05). Combination of the gelatin and chitosan considerably increased structural and functional properties of the healing bones when compared to chitosan scaffold (P < 0.05). However, no significant differences were observed between the gelatin and gelatin-chitosan groups in these regards (P > 0.05). In conclusion, application of the gelatin alone or its combination with chitosan had beneficial effects on bone regeneration and could be considered as good options for bone tissue engineering strategies. However, chitosan alone was not able to promote considerable new bone formation in the experimentally induced critical-size radial bone defects.

Histopathological and biomechanical evaluation of bone healing properties of DBM and DBM-G90 in a rabbit model.

OBJECTIVE: The present study was designed to investigate the effects of DBM and DBM-G90 on bone healing in a rabbit model. METHODS: Thirty male white albino rabbits were used in this study. An incision was made in all rabbits under general anesthesia directly over the radius in order to expose it. A 10-mm segmental defect was created on the middle portion of each radius. The defects of 10 rabbits (Group I) were filled with DBM Block and Strip (Zimmer, Inc., Warsaw, IN, USA), the defects of 10 rabbits (Group II) were filled with DBM soaked in G90, and the defects of 10 rabbits (Group III/control) were left empty. The rabbits were euthanized at 60 days postoperatively for histopathological and biomechanical evaluation. RESULTS: At the histopathologic level, the defects of the animals in the DBM and DBM-G90 groups showed more advanced healing criteria than those of the control group. In biomechanical findings, there was a statistically significant difference between the injured bones and contralateral normal bones of the control group in terms of measured strength. There was not a statistically significant difference between the treated bones of the DBM and DBM-G90 groups with contralateral normal bones, nor was there a statistically significant difference between the treated bones of the DBM and DBM-G90 groups with the treated bones of the control group, in terms of other biomechanical tests. CONCLUSION: Based on the histopathological and biomechanical findings, the DBM and DBM-G90 groups demonstrated superior osteogenic potential; however, G90 shows no superiority over DBM on bone healing.

Selection of animal models for pre-clinical strategies in evaluating the fracture healing, bone graft substitutes and bone tissue regeneration and engineering.

In vitro assays can be useful in determining biological mechanism and optimizing scaffold parameters, however translation of the in vitro results to clinics is generally hard. Animal experimentation is a better approximation than in vitro tests, and usage of animal models is often essential in extrapolating the experimental results and translating the information in a human clinical setting. In addition, usage of animal models to study fracture healing is useful to answer questions related to the most effective method to treat humans. There are several factors that should be considered when selecting an animal model. These include availability of the animal, cost, ease of handling and care, size of the animal, acceptability to society, resistance to surgery, infection and disease, biological properties analogous to humans, bone structure and composition, as well as bone modeling and remodeling characteristics. Animal experiments on bone healing have been conducted on small and large animals, including mice, rats, rabbits, dogs, pigs, goats and sheep. This review also describes the molecular events during various steps of fracture healing and explains different means of fracture healing evaluation including biomechanical, histopathological and radiological assessments.