Wounds In Wistar Rats Research Articles

Aplikasi biomaterial aktif dari daging ikan gabus (Channa striata) untuk penyembuhan luka pascaoperasi pada hewan model tikus Wistar

One problem for postoperative patients is tissue recovery takes a long time due to the occurrence of blood clotting disorders and immune system. Snakehead fish was known to contain bioactive ingredients that can potentially speed up the process of tissue repair as a result of injuries. Snakehead fish meat contains a lot of minerals, albumin, and amino acids needed for tissue protein synthesis and increases the antioxidant glutathione. This study was conducted to test the extraction of snakehead fish flesh against skin tissue wound healing in Wistar rats. The study was conducted with a completely randomized design consisting of three treatments with 6 replication, which include (P0) a negative control (without treatment madecassol and snakehead fish meat extract), (P1) madecassol commercial drug treatment, and (P2) treatment of snakehead fish meat extract. Each material was injected intramuscularly in Wistar rats at a dose of 9 ml/kg bb for 7 days. Data analysis uses Anova followed by test Honestly Significant Difference (HSD) with a significance of 95%. Treatment of snakehead meat extract with a dose of 9 ml/kg bb a significant effect on skin tissue repair after injury, better than treatment with madecassol and control with the average value of extensive tissue remaining after recovering from the condition of the wound by 0.2 cm2; 0.33 cm2; and 0.4 cm2, respectively. Snakehead fish meat extract with a dose of 9 ml/kg bb injected intramuscularly can accelerate the recovery of tissue in the skin after the wound in Wistar rats.

EFFECT OF JNK MAPK ON THE REPAIR OF DAMAGED SKELETAL MUSCLE

Regeneration of muscles after injuries, as well as the development of methods that stimulate this process, is an important problem in medicine and biology. The objective of the study was to evaluate the effect of local blocking of mitogen-activated protein kinase (MAPK) activity of the JNK group (c-Jun N-terminal kinase) on the repair of muscle tissue. Materials and methods. The effect of the JNK MAPK SP600125 blocker on the repair of muscle tissue was studied on a model of a skin and muscle wound in Wistar rats. The main group (n = 30) was injected with a drug plate containing SP600125 with a slow release of the active substance, the control group (n = 30) – the plate without the active substance. The number of dividing myosatellites and muscle kidneys in the damage zone was estimated. Results. Experimental studies have shown that when using a drug plate containing a JNK SP600125 blocker with a slow release of the active substance, the number of dividing myosatellites and forming muscle kidneys in the injury zone of the muscle in the main group on the 7th, 14th and 30th days was significantly higher (p < 0.05) than in the control. Conclusion. Local blockade of JNK MAPK in the zone of muscle damage provides the ability to stimulate the repair of damaged skeletal muscle.

Evaluation of nano hydrogel composite based on gelatin/HA/CS suffused with Asiatic acid/ZnO and CuO nanoparticles for second degree burns

In the present work, a hydrogel platform composed of biopolymer gelatin, and glycosaminoglycan's (Hyaluronic acid and Chondroitin sulfate) incorporated with Asiatic acid (a triterpenoid) and nanoparticles (Zinc oxide and Copper oxide) has been designed and developed to find out the efficacy of healing in second degree burn wounds in Wistar rats. The developed hydrogel composite has been characterized by physico-chemical methods such as; SEM, swelling, mechanical strength, degradation and drug release kinetics. Results showed that the morphology of composite scaffolds are porous with maximum water uptake capacity of 1068% and possessed tensile strength of ~0.196 MPa. Anti-microbial evaluation depicted increase in zone of inhibition with hydrogel containing gelatin + ZnO (5.3 ± 0.2 mm in E. coli and 4.9 ± 0.6 mm in S. aureus) and gelatin + CuO (4.8 ± 0.7 mm in E. coli, 3.8 ± 0.3 mm in S. aureus) in comparison to hydrogel composite scaffold. In-vitro cytocompatibility of developed hydrogel composite was assessed in terms of MTT and DNA quantification on L929 fibroblast cells. In-vivo studies for the composite scaffolds were evaluated on Wistar rats after second degree burn wounds were induced and studied for 28 days which showed the significant wound healing activity in comparison to the control (NeuSkin™ and Cotton guaze) in terms of DNA, total protein, hexosamine and hydroxyproline content. Histopathology studies showed the significant progress in re-epithelization, collagen fibers arrangement and angiogenesis in comparison to control. Additionally, a decrease of TNF-α and increase of MMP-2 expression on day 7 of animal experiment support healing. Furthermore, no toxicity was seen with the developed scaffolds suggesting their suitability to use as a wound dressing in second degree burns.

Evaluation of wound healing effect of alginate films containin g Aloe vera ( Aloe barbadensis Miller) gel.

A new alginate dressing with Aloe vera gel was developed and used to study its cutaneous healing effect in vivo by surgical wounds in Wistar rats. The aloe-alginate film was characterized by Fourier-transform infrared spectroscopy, mechanical properties, scanning electron microscopy, hydration rate, and calcium release. The aloe-alginate film demonstrated desirable physical and mechanical characteristics for wound dressing application. The animals were divided in three groups (n = 20): gauze bandage, alginate film, and aloe-alginate film. The histological analysis on the 3rd, 7th, 14th, and 22nd days after the treatment showed that the aloe-alginate films modulated the inflammatory phase and decreased the quantity of macrophages when compared to the alginate film group and the control (gauze bandage). The evaluation of collagen fibers showed a decrease in type III and increase in type I fibers on group treated with aloe-alginate film suggesting an improvement in the progression of the healing process.

Study of the non-clinical healing activities of the extract and gel of Portulaca pilosa L. in skin wounds in wistar rats: A preliminary study

IntroductionPortulaca pilosa L., belonging to the family Portulacaceae, is a common herbaceous plant in the Americas and in the Amazon, is popularly known as love-grown and is traditionally used as an aid in the treatment of burns, buds, insect bites and wound healing. This study aims to evaluate the non-clinical topical healing activity of the P. pilosa gel (GPP) and the propyleneglycol extract of P. pilosa (EPP) in Wistar rats. MethodsFor the healing activity, wistar rats were divided into the following groups: negative control (GVE – vehicle, 150mg/kg), positive control (Fibrinase®- FIB, 100U/kg), Portulaca pilosa gel 10%, (GPP, 150mg/kg), and propylenglycollic extract of Portulaca pilosa (EPP, 150 mg/kg), which were submitted to a surgical procedure to obtain the wounds, and were treated topically for 7days. After treatment, the treated area was removed and a histopathological analysis was performed. ResultsThe EPP when analyzed in HPLC was able to identify the presence of gallic acid. EPP significantly modulated the tissue inflammatory response, presenting low number of inflammatory cells in the histopathological study. Treatment with EPP and GPP significantly stimulated angiogenesis and this response was superior to the fibrinase® group. Treatment with EPP and GPP significantly stimulated the proliferation of fibroblasts. The groups treated with EPP and GPP presented an organization pattern of the epidermis and dermis better than the control group, with a mild inflammatory process, with fibroblast proliferation and increased formation of collagen fibers. ConclusionThus, from the results obtained it can be suggested that the phytochemical marker of the P. pilosa species for healing activity is gallic acid and, together with the macroscopic and microscopic findings triggered by the topical applications of EPP and GPP, it can be concluded that this plant species has topical healing activity, with great potential for use, since this pharmacological action is associated with a possible topical anti-inflammatory activity.

Modulator effect of a polysaccharide-rich extract from Caesalpinia ferrea stem barks in rat cutaneous wound healing: Role of TNF-α, IL-1β, NO, TGF-β

Ethnopharmacological relevanceIn folk medicine stem barks of Caesalpinia ferrea (Caesalpinioideae) are used to treat enterocolitis, rheumatism and wounds and in experimental procedures, its aqueous extracts demonstrated antiulcer, anti-inflammatory, analgesic, and healing effects. Aim of the studyThe healing mechanism of the polyssacharide-rich extract of C. ferrea stem barks (TPL-Cf) was investigated in a model of excisional cutaneous wound in Wistar rats. Materials and methodsExcisional wounds received topical treatment with TPL-Cf (0.025–0.1%) during 21 days. Hypernociception, macroscopical, histological and immunohistochemical parameters were evaluated and analyzed by ANOVA, Bonferroni and Kruskal-Wallis tests, followed by Dunn and Chi-Square tests. ResultsTPL-Cf (0.1%) reduced wound area and hypernociception, and increased wound contraction. TPL-Cf reduced leukocyte infiltration and vascular permeability, and stimulated fibroblasia, angiogenesis, well formed granulation tissue, collagen deposition and epithelial layer formation. TPL-Cf reduced TNF-α expression and the levels of PGE2 (73%-day 5), IL-1 (42%-day 2), MDA (38%-day 5), total protein (53%-day 2; 73%-day 5) and MPO activity (53%-day 2), but increased the expression of i-NOS (days 5 and 7), TGF-β (day 5) and the levels of NO (3.6 fold-day 5). ConclusionThe polysaccharide-rich extract of C. ferra stem barks accelerates wound healing by the control of the inflammatory phase and attenuates hypernociception via modulation of inflammatory mediators (TNF-α, IL-1β, NO, TGF-β).

Nowy model ran przewlekłych w cukrzycy – badanie pilotażowe u szczurów rasy Wistar

Nowy model ran przewlekłych w cukrzycy – badanie pilotażowe u szczurów rasy Wistar

Effect of Laser Photostimulation on the Healing of Third-Degree Burn wounds in Rats

Background: Burns are one of the most common injuries in the world, most importantly during the wars, and the patients usually experience high levels of pain. Complications include both physical and psychological aspects as well as aesthetic concerns. Management comprises special strategies via specific treatment protocols. Treatment stages include: rescue and control of breathing rhythm, hemorrhage management, fracture closure, particle removal, infection control and control of body temperature. It has been suggested that laser therapy with low level energy could have a positive effect on wound healing process. Objectives: The aim of this study was to examine the effect of different wavelengths of Ga-As laser with blue (405 nm) and green (532 nm) wavelengths on the healing of third-degree burn wounds in Wistar rats. Materials and Methods: In this study, 36 male Wistar rats (8-weekold, weighting 250-300 g) were divided into three groups of equal numbers. All animals were anesthetized by intraperitoneal injections. Dorsal hairs were shaved and third-degree skin burns were created on the back of the animals by using a copper stamp. The stamp was warmed up first to 95˚C via a heater. Afterwards, the animals were randomly divided into the blue and green laser and control groups. The length and width of each wound were measured by a caliper and the area of the lesion was calculated on the first, third, seventh, tenth, fourteenth, seventeenth, and twenty-first days of the study. Six samples were taken from each group for pathobiological analysis. Results: A mild anti-inflammatory effect of blue laser was observed during acute phase, which could possibly accelerate wound healing during the first week of repair. Results of our study showed that using low-level laser therapy (LLLT) with green and blue wavelengths may accelerate the wound healing process as compared with the control group. Conclusions: The laser groups have a more rapid rate of wound contraction than the control group during the healing process of burns; however, the effect of LLLT on burn wound healing was neither statistically nor histologically significant. More studies should be performed to investigate other treatment protocols and prescriptions of soft laser to clarify its role in the treatment of wounds and burn injuries.

The use of the andiroba oil (Carapaguianensis) on cutaneous wounds in Wistar rats

The use of the andiroba oil (<i>Carapaguianensis</i>) on cutaneous wounds in Wistar rats

Influence of different energy densities of laser phototherapy on oral wound healing

The aim of the present prospective study was to evaluate the impact of laser phototherapy (LPT) on the healing of oral ulcers. Different power densities were used on oral wounds in Wistar rats (n=72) randomly divided into three groups: control (0 J/cm2), 4 J/cm2 laser, and 20 J/cm2 laser. Ulcers (3 mm in diameter) were made on the dorsum of the tongue with a punch. Irradiation with an indium-gallium-aluminum-phosphide laser (660 nm; output power: 40 mW; spot size: 0.04 cm2) was performed once a day in close contact with the ulcer for 14 consecutive days. A statistically significant acceleration in healing time was found with wounds treated with 4 J/cm2 LPT. Moreover, striking differences were found in the ulcer area, healing percentage, degree of reepithelialization, and collagen deposition. The most significant changes occurred after 5 days of irradiation. Based on the conditions employed in the present study, LPT is capable of accelerating the oral mucosa wound-healing process. Moreover, faster and more organized reepithelialization and tissue healing of the oral mucosa were achieved with an energy density of 4 J/cm2 in comparison to 20 J/cm2.

Comparative study of the effects of laser photobiomodulation and extract of Brassica oleracea on skin wounds in wistar rats: A histomorphometric study

The objective of this study was to investigate the effect of a photobiomodulation laser and Brassica oleracea on tissue morphology in skin wounds. The parameters analyzed were type I and III collagen fibers, and thickness and surface density of the epithelial tissue, as well as how quickly the wound closed. Five skin wounds 12mm in diameter were made on the backs of the animals, which were randomized into four groups (8 animals each). Saline Group: 0.9% saline solution; Ointment Group (extract of Cabbage, B. oleracea, 10% lanolin); Balsam Group (10% glycolic extract of B. oleracea emulsion oil); L60 Group (laser GaAsAl 60J/cm2). The applications were made daily during a 20-day treatment, and every 4 days tissue from different wounds was removed. The reduction in the size of the wounds on the 4th, 8th, 12th and 16th days was significantly greater in the treated groups compared to the control group. At all the time points analyzed, there was a greater proportion of collagen in the Balsam and L60 groups (p<0.05). There was also a greater proliferation of epithelial cells in the L60 and Balsam groups after 20 days of treatment (p<0.05). The healing extract and laser 60j/cm2 exerted a great effect on collagen proliferation in stimulating scar tissue maturation.

Effect of Topical Application of Chlorogenic Acid on Excision Wound Healing in Rats

This study was undertaken to evaluate the therapeutic effects of topical chlorogenic acid on excision wounds in Wistar rats. A 1 % (w/w) chlorogenic acid or silver sulfadiazine ointment was applied topically once a day for 15 days on full-thickness excision wounds created on rats. The 1 % (w/w) chlorogenic acid ointment had potent wound healing capacity as evident from the wound contraction on the 15th post-surgery day, which was similar to that produced by 1 % (w/w) silver sulfadiazine ointment. Increased rates of epithelialization were observed in the treated rats. It also improved cellular proliferation, increased tumor necrosis factor-α levels during the inflammatory phase (12 h, 24 h, 48 h, and 72 h post-wounding) of wound healing, upregulated transforming growth factor-β1 and elevated collagen IV synthesis in the chlorogenic acid-treated group. The results also indicated that chlorogenic acid possesses potent antioxidant activity by increasing superoxide dismutase, catalase, and glutathione, and decreasing lipid peroxidation. In conclusion, these results demonstrate that topical application of chlorogenic acid can accelerate the process of excision wound healing by its ability to increase collagen synthesis through upregulation of key players such as tumor necrosis factor-α and transforming growth factor-β1 in different phases of wound healing as well as by its antioxidant potential.

Healing potential of aStreptomycessp. secondary metabolite, SEM-1-111, on experimental full-thickness excision cutaneous wounds in Wistar rats

The potent biomedical properties of Streptomyces secondary metabolites make them a possible source of promising new targets to use as wound-healing-promoting agents. To investigate the properties of a Streptomyces sp. secondary metabolite, the SEM-1-111 fraction, in the healing full-thickness cutaneous wounds. In this experiment, creams were prepared using a Eucerin ointment base with 5%, 10% or 15% SEM-1-111 added. Circular excision wounds were surgically created on the back of Wistar rats, and the creams were applied daily to the wound area. The negative control group was treated with the vehicle, and the positive control group with a commercial phenytoin 1% cream. We analysed planimetric and histological parameters, and the expression of procollagen-I mRNA. SEM-1-111 substantially influenced wound healing. The cream containing the highest dose of metabolite (15%) significantly decreased wound size during the study, and histological evaluation showed beneficial effects of this concentration in the wound-healing process. Procollagen-I mRNA levels were significantly increased in both the 15% SEM-1-111 group and the phenytoin group compared with the vehicle control on postoperative day 7. Regular topical application of the Streptomyces sp. secondary metabolite SEM-1-111 fraction at a concentration of 15% accelerates the wound-healing process.

Antibacterial microemulsion prevents sepsis and triggers healing of wound in wistar rats

Microemulsions are thermodynamically stable isotropic systems comprising of oil, surfactant and water. Cinnamon oil (Cinnamomum zeylanicum) microemulsion was formulated using non-ionic surfactant Tween 20 and water. With oil to surfactant (v/v) ratio of 1:4, cinnamon oil microemulsion (CMF4) was formulated having droplet diameter of 5.79 and kinetically stable for 240 days. TEM images confirmed the spherical morphology of emulsion droplets. Surfactant concentration demonstrated a negative correlation with droplet diameter and turbidity but positive correlation with stability and viscosity. Formulated microemulsion (CMF4) was evaluated for bactericidal activity against Staphylococcus aureus by dose and time dependent killing experiment. SEM analysis suggested that microemulsion treatment resulted in alteration of permeability and surface features of bacterial cell membrane which lead to lysis and cell death. CMF4 showed no erythema for skin irritation test in wistar rats and triggered wound-healing process in wistar rats. Control group without any treatment, cinnamon oil only and neomycin treated group healed in 20, 16 and 16 days, whereas cinnamon oil formulation (CMF4) treated group was healed in 14 days.

Anti-inflammatory effect of low-intensity laser on the healing of third-degree burn wounds in rats

Third-degree burn wounds are considered severe injuries because they destroy all the skin layers and may affect subcutaneous tissues, fasciae, muscles, and bones. To favor the healing process of the injured tissues, it is very useful to diminish the occurrence of the inflammatory process. The present study was aimed at comparing the effect of different energetic densities of AlGaInP laser on the inflammatory process and in the healing of third-degree burn wounds in Wistar rats. This study was approved by the Ethics Committee, in which 36 adult male rats were selected and suffered the induction of third-degree burn injury. These rats were divided as follows: group 1-control (treated with silver sulfadiazine), group 2-received energy density of 3J/cm(2), and group 3-received energy density of 6J/cm(2). All animals daily received an occlusive bandage with silver sulfadiazine and 8% papain. The laser therapy was performed alternatively three times a week. The animals were evaluated on the 3rd, 7th, 14th, and 21st days after the initial lesion and euthanized for the macroscopic, histologic, and morphometric analysis. A higher production of collagen was observed at 7days and a greater re-epithelialization at 21days in group 3 (6J/cm(2)). Furthermore, the latter when compared to the other groups presented macroscopically a better aspect of the scar at 21days with more granulation tissue and fibrosis. We conclude that the AlGaInP laser used in dosages of 3 and 6J/cm(2) favors the healing of third-degree burn wounds induced in rats.

Application of Jatropha curcas L. seed oil (Euphorbiaceae) and microcurrent on the healing of experimental wounds in Wistar rats

To investigate the effects of Jatropha curcas L. seed oil and microcurrent stimulation on the healing of wounds experimentally induced in Wistar rats. Forty-eight animals were divided into four groups: (A) control; (B) treated with microcurrent (10 µA/2 min); (C) treated with J. curcas seed oil, and (D) treated with J. curcas seed oil plus microcurrent. Tissues samples were obtained two, six, ten and 14 days after injury and submitted to structural and morphometric analyses. The animals of groups A and C showed similar responses in terms of repair area, total number of cells, number of newly formed blood vessels, epithelial thickness, and percentage of area occupied by mature collagen fibers. Significant differences in all parameters analyzed were observed between animals of groups B and D and the control 10 and 14 days after experimentally induced injury. The morphometric data confirmed the structural findings The application of J. curcas seed oil alone was not effective on experimental wound healing when compared to control, but microcurrent application alone or combined with the oil exerted significant differences in the parameters studied. These findings suggest that the positive results were due to microcurrent stimulation.

Comparative Study of the Effects of Gallium-Aluminum-Arsenide Laser Photobiomodulation and Healing Oil on Skin Wounds in Wistar Rats: A Histomorphometric Study

The present study compared the effects of gallium-aluminum-arsenide diode laser and healing oil on fibroblasts, blood vessels, and collagen maturation of skin wounds in Wistar rats. Twenty-four male rats weighing 325 ± 27 g were used. Five wounds, 12 mm in diameter, were made on the animals' backs. The rats were randomly divided into four groups with six animals in each group. saline solution; L30 group: 30 J/cm(2) laser; L60 group: 60 J/cm(2) laser; Oil group: healing oil. Histomorphometric analysis was performed on the scar tissue removed from the different wounds every 4 d for 20 d. On day 4, there were significantly more fibroblasts in the wounds treated with the laser and the healing oil compared to the controls. On day 8, there were significantly more fibroblasts in the oil group compared to the L30 and L60 groups. On the same day, the quantity of vessels was significantly greater in the L60 group compared to the other groups. On day 16, there was a significant increase in the number of blood vessels in the wounds treated with the 60 J/cm(2) laser compared to the other groups. Analysis of the collagen maturation index throughout the experiment showed significantly higher values in the L60 group compared to the other groups at all time points. The healing oil exerted a greater effect on fibroblast proliferation, whereas the 60 J/cm(2) laser was more effective in stimulating angiogenesis and scar-tissue maturation.

Effect of gallium-arsenide laser, gallium-aluminum-arsenide laser and healing ointment on cutaneous wound healing in Wistar rats

This study determined the effects of gallium-aluminum-arsenide laser (GaAlAs), gallium-arsenide laser (GaAs) and Dersani healing ointment on skin wounds in Wistar rats. The parameters analyzed were: type I and III collagen fiber concentrations as well as the rate of wound closure. Five wounds, 12 mm in diameter, were made on the animals' backs. The depth of the surgical incision was controlled by removing the epithelial tissue until the dorsal muscular fascia was exposed. The animals were anesthetized with ketamine and xylazine via intraperitoneal injection. The rats were randomly divided into five groups of 6 animals each, according to the treatment received. Group 1 (L4): GaAs laser (4 J/cm(2)); group 2 (L30): GaAlAs laser (30 J/cm(2)); group 3 (L60): GaAlAs laser (60 J/cm(2)); group 4 (D): Dersani ointment; group 5 (control): 0.9% saline. The applications were made daily over a period of 20 days. Tissue fragments were stained with picrosirius to distinguish type I collagen from type III collagen. The collagen fibers were photo-documented and analyzed using the Quantum software based on the primary color spectrum (red, yellow and blue). Significant results for wound closing rate were obtained for group 1 (L4), 7.37 mm/day. The highest concentration of type III collagen fibers was observed in group 2 (L30; 37.80 + or - 7.10%), which differed from control (29.86 + or - 5.15%) on the 20th day of treatment. The type I collagen fibers of group 1 (L4; 2.67 + or - 2.23%) and group 2 (L30; 2.87 + or - 2.40%) differed significantly from control (1.77 + or - 2.97%) on the 20th day of the experiment.

The use of amniotic membrane in the repair of duodenal wounds in Wistar rats

In the search of a new material to repair duodenal wounds, a trial was conducted to assess the behavior of human amniotic membrane in the repair of the duodenal wall in rats. Fifty Wistar rats weighing between 250 and 350g, male, were submitted to duodenotomy and randomly distributed into two groups. Group A (n=8) had no treatment and was used as the control group. In Group B (n=42) the duodenal wound was treated with a patch of human amniotic membrane. All animals in Group A died. In Group B no changes were observed with regards to death or the formation of duodenal fistula. All animals presented peritoneal adherences in the region on the duodenal wall repair and intestinal obstruction was observed in two animals. Healing of the duodenal wall in the region of the patch took place progressively as the post-operatory period increased, with regeneration of the mucosa and of the smooth muscle layer. From the clinical standpoint, the amniotic membrane proved to be a biological tissue which served as a temporary seal and allowed the wound to heal by second-intention, with re-establishment of the duodenal wall structure.

Lack of a central role for osteoprogenitor cells from the femoral canal in heterotopic ossification of the hip

We have developed an animal model to examine the formation of heterotopic ossification using standardised muscular damage and implantation of a beta-tricalcium phosphate block into a hip capsulotomy wound in Wistar rats. The aim was to investigate how cells originating from drilled femoral canals and damaged muscles influence the formation of heterotopic bone. The femoral canal was either drilled or left untouched and a tricalcium phosphate block, immersed either in saline or a rhBMP-2 solution, was implanted. These implants were removed at three and 21 days after the operation and examined histologically, histomorphometrically and immunohistochemically. Bone formation was seen in all implants in rhBMP-2-immersed, whereas in those immersed in saline the process was minimal, irrespective of drilling of the femoral canals. Bone mineralisation was somewhat greater in the absence of drilling with a mean mineralised volume to mean total volume of 18.2% (sd 4.5) versus 12.7% (sd 2.9, p < 0.019), respectively. Our findings suggest that osteoinductive signalling is an early event in the formation of ectopic bone. If applicable to man the results indicate that careful tissue handling is more important than the prevention of the dissemination of bone cells in order to avoid heterotopic ossification.