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Introduction: This study explored the synergistic effects of low-level laser therapy (LLLT) and adipose-derived stem cells (ADSCs) on cranial bone regeneration in rats, addressing the limitations of autogenous grafts and advancing bone tissue engineering with innovative photobiomodulation (PBM) applications. Methods: Sixty Wistar rats were allocated to 5 separate groups randomly; (1) natural bovine bone mineral (NBBM); (2) NBBM+LLLT; (3) NBBM+allogenic ADSCs; (4) NBBM+allogenic ADSCs+LLLT; (5) Only defects. 8-mm calvarial defects were made in each rat in the surgical procedure. A diode laser was applied with the following parameters (continuous mode, power of 100mW, wavelength of 808nm, and 4 J/cm2 energy density) immediately after the procedure and every other day. Bone samples were obtained and assessed histomorphometrically and histologically after staining with hematoxylin and eosin (H&E). Results: Different volumes of bony material were observed in two weeks; 2.94%±1.00 in group 1, 5.1%±1.92 in group 2, 7.11%±2.82 in group 3, 7.34%±2.31 in group 4, and 2.01%±0.83 in group 5 (P<0.05). On the other hand, foreign body residuals were up by 23% in the groups with scaffolding by the end of 2 weeks. Four weeks of observation led to 6.74 %±1.95, 13.24%±1.98, 15.76%±1.19, 15.92%±3.4, and 3.11%±1.00 bone formation in groups 1 to 5, respectively (P<0.05). Generally, the difference between groups 2-4 was not statistically significant based on different types of bone and the extent of inflammation. Conclusion: Bearing in mind the limitations of our research, it was demonstrated that ADSCs in combination with PBM have promising effects on bone tissue regeneration in sizeable bony defects. Furthermore, this study also showed that PBM usage improved the newly regenerated bone quality.
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Spinal cord injury (SCI) is a serious medical condition. The search for an effective cure remains a persistent challenge. Current treatments, unfortunately, are unable to sufficiently improve neurological function, often leading to lifelong disability. This systematic review and meta-analysis evaluated the effectiveness of stem cell therapy for SCI using canine models. It also explored the optimal protocol for implementing stem cell therapy. A comprehensive search of studies was conducted from 2000 to October 2022. This study focused on five outcomes: motor function score, histopathology, IHC, western blot, and SEP. The results demonstrated a significant improvement in locomotion post-SCI in dogs treated with stem cell therapy. The therapy also led to an average increase of 3.15 points in the Olby score of the treated dogs compared to the control group. These findings highlights stem cell therapy's potential as a promising SCI treatment. The meta-analysis suggests that using bone marrow stem cells, undergoing neural differentiation in vitro, applying a surgical implantation or intrathecal route of administration, associating matrigel in combination with stem cells, and a waiting period of two weeks before starting treatment can enhance SCI treatment effectiveness.
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Modelos Animales de Enfermedad , Traumatismos de la Médula Espinal , Trasplante de Células Madre , Traumatismos de la Médula Espinal/terapia , Animales , Perros , Trasplante de Células Madre/métodos , Recuperación de la Función/fisiologíaRESUMEN
In the absence of vascular obstruction, central venous pressure (CVP) is a hydrostatic pressure in the cranial and caudal vena cava, providing valuable information about cardiac function and intravascular volume status. It is also a component in evaluating volume resuscitation in patients with septic shock and monitoring patients with right heart disease, pericardial disease, or volume depletion. Central venous pressure is calculated in dogs by invasive central venous catheterization, which is considered high-risk and impractical in critically ill patients. This study aimed to investigate the feasibility of using echocardiographic tricuspid E/E' as a noninvasive method to estimate CVP in anesthetized healthy dogs under controlled hypovolemic conditions. Ten male mixed-breed dogs were included in the study after a thorough health assessment. For hypovolemia induction, blood withdrawal was performed, and echocardiographic factors of the tricuspid valve, including peak E and E' velocities, were measured during CVP reduction. Repeated measures analysis of variance and Bonferroni post hoc tests were employed to compare the average difference between measured echocardiographic indices and CVP values derived from catheterization and intermittent measurement methods. Spearman's ρ correlation coefficient was used to evaluate the correlation between echocardiographic indices and CVP. E peak velocity had a significant negative correlation with venous blood pressure phases (r = -0.44, P = .001), indicating a decrease in peak E velocity with progressive CVP reduction. However, tricuspid valve E' peak velocity and E/E' did not correlate with CVP, suggesting that these parameters are not reliable for CVP estimation in dogs.
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Presión Venosa Central , Ecocardiografía , Válvula Tricúspide , Animales , Perros , Presión Venosa Central/fisiología , Masculino , Ecocardiografía/veterinaria , Válvula Tricúspide/diagnóstico por imagen , Estudios de FactibilidadRESUMEN
The jungle cat (Felis chaus) is a member of the genus Felis within the family Felidae, native to south-east Asia, west Asia and north Africa. A 2-year-old male jungle cat was referred with a history of lameness of 3 days duration. At the time of presentation, the animal had non-weight-bearing lameness of the right hindlimb. Examination identified crepitation and instability in the distal diaphyseal region of the right tibial bone. Radiographs confirmed the presence of fractures in the distal diaphyseal region of the right tibia and fibula. Double plating of the tibia was employed for used fixation. The animal was evaluated 2, 4 and 8 weeks post-operatively, and the procedure was considered to have been successful with no observed surgical complications.
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Felis , Fracturas de la Tibia , Masculino , Animales , Cojera Animal , Fracturas de la Tibia/diagnóstico por imagen , Fracturas de la Tibia/cirugía , Fracturas de la Tibia/veterinaria , Peroné/cirugíaRESUMEN
The treatment of type 1 diabetes through islet cell transplantation is a complex process, facing challenges such as allograft rejections and a limited supply of donors. One potential solution is to utilize the liver as an alternative for natural insulin production, as hepatocytes can secrete proteins and respond to glucose levels. Recent research has shown promising results in using mesenchymal stem cells as a potential cure for diabetes. The study utilized a diabetic rat model, confirmed through blood sugar measurement. A plasmid vector was designed with specific genetic components, synthesized by biotech company, and then Inserted vector into a plasmid with resistance genes and bacterial origin. Bone marrow-derived mesenchymal stem cells (BM-MSCs) were cultured and transfected with the plasmid using Lipofectamine 3000. Polymerase chain reaction was employed to confirm successful transfection using specific primers. For the animal study, 30 male Wistar rats were divided into six groups, each comprising five rats. The control group did not receive any treatment, while the second group received MSCs via Portal Vein Injection. The third group received MSCs transfected with a specific construct via Portal Vein Injection. The fourth group was induced to develop diabetes through streptozotocin (STZ) injection, the fifth group developed diabetes and received untransfected MSCs via Portal Vein Injection, and the sixth group received MSCs transfected with the specific construct via Portal Vein Injection. To manage Pain, appropriate pain control was administered to the rats for 3 days after the surgery. Fixed liver tissues obtained from the euthanized rats were utilized for immunohistochemistry. In this study, immunohistochemical techniques were used to examine insulin expression in different groups of rats. The control groups showed high levels of insulin expression, while the diabetic groups exhibited lower expression. However, there was a significant difference between the diabetic groups treated with MSC and transgenic MSC cells. All groups had similar baseline glucose levels, but the diabetic groups showed a significant increase after STZ injection, whereas the control and MSC groups did not. Postintervention, both the control and MSC groups had similar glucose levels to the post-STZ levels. However, diabetes-induced groups experienced a significant decrease in glucose levels, with the transfected MSCs showing a greater decrease than the untransfected MSCs. The study suggested that treatment with MSCs, especially transfected ones, can effectively reduce glucose levels in rats with diabetes. In this research, rat BM-MSCs were utilized to create insulin-producing mesenchymal cells with glucose-sensitive insulin expression. The cells were transferred to the liver of diabetic rats via portal vein injection, leading to an increase in insulin expression. This study proposes a novel approach for cell therapy and delivery in the treatment of type 1 diabetes.
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Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 1 , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas , Ratas , Masculino , Animales , Insulina/metabolismo , Glucosa/metabolismo , Diabetes Mellitus Tipo 1/terapia , Diabetes Mellitus Tipo 1/metabolismo , Vena Porta/metabolismo , Ratas Wistar , Diabetes Mellitus Experimental/inducido químicamente , Diabetes Mellitus Experimental/terapia , Expresión Génica Ectópica , Diferenciación Celular , Glucemia , Células Madre Mesenquimatosas/metabolismo , Dolor/metabolismo , Trasplante de Células Madre Mesenquimatosas/métodosRESUMEN
Due to the increasing prevalence of bone disorders among people especially in average age, the future of treatments for osseous abnormalities has been illuminated by scaffold-based bone tissue engineering. In this study, in vitro and in vivo properties of 58S bioactive glass-based scaffolds for bone tissue engineering (bare (B.SC), Zein-coated (C.SC), and Zein-coated containing Kaempferol (KC.SC)) were evaluated. This is a follow-up study on our previously published paper, where we synthesized 58S bioactive glass-based scaffolds coated with Kaempferol-loaded Zein biopolymer, and characterized from mostly engineering points of view to find the optimum composition. For this aim, in vitro assessments were done to evaluate the osteogenic capacity and biological features of the scaffolds. In the in vivo section, all types of scaffolds with/without bone marrow-derived stem cells (BMSC) were implanted into rat calvaria bone defects, and potential of bone healing was assessed using imaging, staining, and histomorphometric analyses. It was shown that, Zein-coating covered surface cracks leading to better mechanical properties without negative effect on bioactivity and cell attachment. Also, BMSC differentiation proved that the presence of Kaempferol caused higher calcium deposition, increased alkaline phosphatase activity, bone-specific gene upregulation in vitro. Further, in vivo study confirmed positive effect of BMSC-loaded KC.SC on significant new bone formation resulting in complete bone regeneration. Combining physical properties of coated scaffolds with the osteogenic effect of Kaempferol and BMSCs could represent a new strategy for bone regeneration and provide a more effective approach to repairing critical-sized bone defects.
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Células Madre Mesenquimatosas , Zeína , Ratas , Animales , Ingeniería de Tejidos/métodos , Andamios del Tejido , Estudios de Seguimiento , Quempferoles/farmacología , Zeína/farmacología , Osteogénesis , Regeneración Ósea , Vidrio , Diferenciación Celular , CráneoRESUMEN
Magnesium (Mg) plays an important role in controlling bone apatite structure and density and is a potential bioactive material in repairing critical-sized bone defects. In this study, we aimed to evaluate the effect of adding NanoMgO to polycaprolactone/beta-tricalcium phosphate (PCL/ß-TCP) scaffolds on bone regeneration. Novel 3D-printed porous PCL/ß-TCP composite scaffolds containing 10% nanoMgO were fabricated by fused deposition modeling (FDM) and compared with PCL/ß-TCP (1:1) scaffolds (control). The morphology and physicochemical properties of the scaffolds were characterized by ATR-FTIR, XRD, scanning electron microscope-energy dispersive X-ray analysis (SEM-EDX), transmission-electron-microscopy (TEM), water contact angle, and compressive strength tests and correlated to its cytocompatibility and osteogenic capacity in-vitro. To evaluate in-vivo osteogenic capacity, bone-marrow-derived stem cell (BMSC)-loaded scaffolds were implanted into 8 mm rat critical-sized calvarial defects for 12 weeks. The hydrophilic scaffolds showed 50% porosity (pore size = 504 µm). MgO nanoparticles (91.5 ± 27.6 nm) were homogenously dispersed and did not adversely affect BMSCs' viability and differentiation. Magnesium significantly increased elastic modulus, pH, and degradation. New bone formation (NBF) in Micro-CT was 30.16 ± 0.31% and 23.56 ± 1.76% in PCL/ß-TCP/nanoMgO scaffolds with and without BMSCs respectively, and 19.38 ± 2.15% and 15.75 ± 2.24% in PCL/ß-TCP scaffolds with and without BMSCs respectively. Angiogenesis was least remarkable in PCL/ß-TCP compared with other groups (p < .05). Our results suggest that the PCL/ß-TCP/nanoMgO scaffold is a more suitable bone substitute compared to PCL/ß-TCP in critical-sized calvarial defects.
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Nanopartículas , Ingeniería de Tejidos , Ratas , Animales , Andamios del Tejido/química , Óxido de Magnesio/farmacología , Magnesio , Fosfatos de Calcio/farmacología , Fosfatos de Calcio/química , Poliésteres/farmacología , Poliésteres/química , Impresión TridimensionalRESUMEN
This study investigated the effects of hydroxyapatite (HA) and hydroxyapatite/titanium (HA/Ti) coatings on osseointegration and bonding strength at the bone-implant interface. The coatings were made using air plasma spray (APS), and three study groups were examined: 1) Uncoated commercial pure titanium (CP-Ti) rods; 2) HA-coated CP-Ti rods, and 3) Composite of 50 %wt HA + 50 %wt Ti coated CP-Ti rods. The rods were implanted into the distal femurs and proximal tibias of fifteen New Zealand white rabbits, and 8 weeks after the implantation, the samples were harvested. The results of pull-out tests showed that the ultimate strength of HA and HA/Ti coatings were significantly greater than the uncoated samples (P < 0.05). Moreover, even though the histological evaluations showed significantly greater osseointegration of HA/Ti composite coatings compared with HA coatings (P < 0.05), nonetheless, the composite of HA/Ti offers no significant increase in the ultimate strength, stiffness, and bonding strength at the bone-implant interface, compared with the HA group (P > 0.05). Thus, in an eight-week study, there was no linear correlation between the osseointegration and the bonding strength at the bone-implant interface. The results of this work may imply that the extent of osseointegration at the bone-implant interface does not necessarily determine the value of the bonding strength at the bone-implant interface. It is speculated that, in a longer-term study, a greater quality of bone formation may occur during osseointegration, between the implant and its adjacent bone, which can lead to a more enhanced bonding strength, compared with the 8-weeks post-surgery follow up.
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Durapatita , Oseointegración , Conejos , Animales , Titanio , Interfase Hueso-Implante , Materiales Biocompatibles Revestidos/farmacología , Prótesis e Implantes , Propiedades de Superficie , Ensayo de MaterialesRESUMEN
Bone-marrow derived stem cells (BMSCs) can differentiate into several mesenchymal cell lines that are suitable for bone and dental tissue engineering. This study was aimed to assess the efficacy of cell therapy in direct pulp capping (DPC) of canine teeth using autologous BMSCs along with collagen/hydroxyapatite hybrid scaffold in terms of the quantity and quality of calcified bridge formation. The teeth were randomly divided into three groups of DPC with mineral trioxide aggregate (MTA), hydroxyapatite/collagen hybrid scaffold alone and BMSCs with hydroxyapatite/collagen hybrid scaffold. DPC was performed under general anesthesia in cavities prepared on the buccal surfaces of mandibular and maxillary premolars of the same dogs from which, stem cells had been isolated. All cavities were then restored with light-cure resin modified glass ionomer cement. Histomorphometric assessments after 12 weeks showed formation of dentinal bridge following DPC with BMSCs and MTA. The efficacy of MTA for calcified bridge formation following DPC was significantly higher than that of BMSCs plus hybrid scaffold. According to the present study, we concluded DPC using BMSCs and hybrid scaffold did not provide clinically noticeable results in canine patients.
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In this study, polyvinyl alcohol hydrogel chains were crosslinked by polyurethane in order to synthesize a suitable substrate for cartilage lesions. The substrate was fully characterized, and in vitro and in vivo investigations were conducted based on a sheep model. In vitro tests were performed based on the chondrocyte cells with the Alcian Blue and safranin O staining in order to prove the presence of proteoglycan on the surface of the synthesized substrate, which has been secreted by cultures of chondrocytes. Furthermore, the expression of collagen type I, collagen type II, aggrecan, and Sox9 was presented in the chondrocyte cultures on the synthesized substrate through RT-PCR. In addition, the H&E analysis and other related tests demonstrated the formation of neocartilage tissue in a sheep model. The results were found to be promising for cartilage tissue engineering and verified that the isolated chondrocyte cultures on the synthesized substrate retain their original composition.
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Condrocitos , Poliuretanos , Agrecanos/metabolismo , Azul Alcián/metabolismo , Animales , Cartílago , Células Cultivadas , Condrocitos/metabolismo , Colágeno Tipo I/metabolismo , Colágeno Tipo II , Poliuretanos/metabolismo , Proteoglicanos/metabolismo , Ovinos , Ingeniería de Tejidos/métodos , Andamios del TejidoRESUMEN
In vivo bioreactors serve as regenerative niches that improve vascularization and regeneration of bone grafts. This study has evaluated the masseter muscle as a natural bioreactor for ßTCP or PCL/ßTCP scaffolds, in terms of bone regeneration. The effect of pedicle preservation, along with sole, or MSC- or rhBMP2-combined application of scaffolds, has also been studied. Twenty-four mongrel dogs were randomly placed in six groups, including ßTCP, ßTCP/rhBMP2, ßTCP/MSCs, PCL/ßTCP, PCL/ßTCP/rhBMP2, and PCL/ßTCP/MSCs. During the first surgery, the scaffolds were implanted into the masseter muscle for being prefabricated. After 2 months, each group was divided into two subgroups prior to mandibular bone defect reconstruction; one with a preserved vascularized pedicle and one without. After 12 weeks, animals were euthanized, and new bone formation was evaluated using histological analysis. Histological analysis showed that all ß-TCP scaffold groups had resulted in significantly greater rates of new bone formation, either with a pedicle surgical approach or non-pedicle surgical approach, comparing to their parallel groups of ßTCP/PCL scaffolds (p ≤ .05). Pedicled ß-TCP scaffold groups that were treated with either rhBMP2 (48.443% ± 0.250%) or MSCs (46.577% ± 0.601%) demonstrated the highest rates of new bone formation (p ≤ .05). Therefore, masseter muscle can be used as a local in vivo bioreactor with potential clinical advantages in reconstruction of human mandibular defects. In addition, scaffold composition, pedicle preservation, and treatment with MSCs or rhBMP2, influence new bone formation and scaffold degradation rates in the prefabrication technique.
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Músculo Masetero , Andamios del Tejido , Animales , Reactores Biológicos , Regeneración Ósea , Perros , Mandíbula/cirugíaRESUMEN
This study was based on in vivo assessment of bone regeneration capacity of synthesized porous ß-tricalcium phosphate (ß-TCP) nanocomposite granules and aimed to explore the effects of fabricated ß-TCP granules reinforced with layered double hydroxides (LDH) nanoclay compared to ß-TCP granules, in terms of osteoconductivity and biodegradability. Granules with diameters of 2-3 mm were implanted into cavities drilled in rabbit distal femur and were left in situ for up to 3 months. The mechanical study demonstrated that the presence of LDH nanoparticles in ß-TCP granules resulted in a significant increase in compressive modulus from 174.4 to 231.4 MPa, while the porosity was constant at 76%-80%. The results revealed that the obtained granules showed no cytotoxicity. In this study, x-ray radiographic, micro-computed tomography, and histological staining analysis were taken to evaluate the percentage of bone ingrowth and biodegradability of the porous granules. The results exhibited that both granules support bone regeneration and also the amount of new bone formation in the bone defect filled with both granules was almost six times higher than the empty defects. Although no significant difference in bone formation for two different granules was observed, a higher biodegradability was detected in ß-TCP granules in comparison to ß-TCP/LDH granules. Overall, the addition of LDH nanoclay (10%) enhanced the physicochemical and mechanical properties of ß-TCP granules while it is biological and osteoconductity properties have been maintained and its biodegradation rate has been decreased.
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Sustitutos de Huesos , Nanocompuestos , Animales , Regeneración Ósea , Sustitutos de Huesos/farmacología , Fosfatos de Calcio/química , Fosfatos de Calcio/farmacología , Hidróxidos/farmacología , Conejos , Microtomografía por Rayos XRESUMEN
This study aimed to explore the effects of drilling rotational speed and feed-rate on the stability of dental implants through in-vivo and ex-vivo experiments. To this end, a total of 16 identical dental implants were inserted in the mandible of four dogs. The osteotomies were made with two drilling rotational speeds, i.e., 800 and 1500 rpm, and two different feed-rates, i.e., 1 and 2 mm/s. Implant stability quotients (ISQs) were recorded immediately after inserting implants and then each week for four subsequent weeks. Then, all animals were euthanized, and a bone sample containing the implants was extracted from each hemi-mandible for the pull-out test. A two-way ANOVA was performed for ISQs, and pull-out strengths (PoS), and the significance level was set to <0.05. The effect of rotational speed and feed-rate, used in this study, on the primary stability quotients was not significant (P > 0.05). Increasing the rotational speed from 800 to 1500 rpm significantly increased both ISQ and PoS values at the end of the 4th week after the implantation (P = 0.022 and P = 0.001, respectively). Moreover, by decreasing the feed-rate from 2 to 1 mm/s, a significant increase in PoSs of the dental implants was observed four weeks after the implantation (P = 0.019). Results of this study showed that either by increasing drilling rotational speed, here from 800 to 1500 rpm, or by reducing feed-rate, here from 2 to 1 mm/s, the secondary stability would be reinforced. Further investigations are needed to see if and how the conclusions made in this study can be generalized.
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Implantes Dentales , Análisis de Varianza , Animales , Implantación Dental Endoósea , Perros , Mandíbula/cirugía , Oseointegración , OsteotomíaRESUMEN
Separating cells from the body and cultivating them in vitro will alter the function of cells. Therefore, for optimal cell culture in the laboratory, conditions similar to those of their natural growth should be provided. In previous studies, it has been shown that the use of cellular shape at the culture surface can regulate cellular function. In this work, the efficiency of the imprinting method increased by using microfluidic chip design and fabrication. In this method, first, a cell-imprinted substrate of chondrocytes was made using a microfluidic chip. Afterwards, stem cells were cultured on a cell-imprinted substrate using a second microfluidic chip aligned with the substrate. Therefore, stem cells were precisely placed on the chondrocyte patterns on the substrate and their fibroblast-like morphology was changed to chondrocyte's spherical morphology after 14-days culture in the chip without using any chemical growth factor. After chondrogenic differentiation and in vitro assessments (real-time PCR and immunocytotoxicity), differentiated stem cells were transferred on a collagen-hyaluronic acid scaffold and transplanted in articular cartilage defect of the rabbit. After 6 months, the post-transplantation analysis showed that the articular cartilage defect had been successfully regenerated in differentiated stem cell groups in comparison with the controls. In conclusion, this study showed the potency of the imprinting method for inducing chondrogenicity in stem cells, which can be used in clinical trials due to the safety of the procedure.
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Cartílago Articular , Células Madre Mesenquimatosas , Animales , Diferenciación Celular , Células Cultivadas , Condrocitos , Condrogénesis , Dispositivos Laboratorio en un Chip , Conejos , Regeneración , Ingeniería de TejidosRESUMEN
Production of a 3D bone construct with high-yield differentiated cells using an appropriate cell source provides a reliable strategy for different purposes such as therapeutic screening of the drugs. Although adult stem cells can be a good source, their application is limited due to invasive procedure of their isolation and low yield of differentiation. Patient-specific human-induced pluripotent stem cells (hiPSCs) can be an alternative due to their long-term self-renewal capacity and pluripotency after several passages, resolving the requirement of a large number of progenitor cells. In this study, a new biphasic 3D-printed collagen-coated HA/ß-TCP scaffold was fabricated to provide a 3D environment for the cells. The fabricated scaffolds were characterized by the 3D laser scanning digital microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and mechanical test. Then, the osteogenesis potential of the hiPSC-seeded scaffolds was investigated compared to the buccal fat pad stem cell (BFPSC)-seeded scaffolds through in vitro and in vivo studies. In vitro results demonstrated up-regulated expressions of osteogenesis-related genes of RUNX2, ALP, BMP2, and COL1 compared to the BFPSC-seeded scaffolds. In vivo results on calvarial defects in the rats confirmed a higher bone formation in the hiPSC-seeded scaffolds compared to the BFPSC-seeded groups. The immunofluorescence assay also showed higher expression levels of collagen I and osteocalcin proteins in the hiPSC-seeded scaffolds. It can be concluded that using the hiPSC-seeded scaffolds can lead to a high yield of osteogenesis, and the hiPSCs can be used as a superior stem cell source compared to BFPSCs for bone-like construct bioengineering.
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Tejido Adiposo/diagnóstico por imagen , Células Madre Pluripotentes Inducidas/metabolismo , Osteogénesis/fisiología , Impresión Tridimensional/normas , Andamios del Tejido/normas , Tejido Adiposo/fisiopatología , Animales , Diferenciación Celular , Proliferación Celular , Humanos , Masculino , Ratas , Ratas WistarRESUMEN
In this study, we developed an injectable in situ forming hydrogel/microparticle system consisting of two drugs, melatonin and methylprednisolone, to investigate the capability of the system for chondrogenesis in vitro and in vivo. The chemical, mechanical, and rheological properties of the hydrogel/microparticle were investigated. For in vitro evaluation, the adipose-derived stem cells might be mixed with hydrogel/microparticles, then cellular viability was analyzed by acridine orange/propidium iodide and 4',6-diamidino-2-phenylindole staining and also dimethylmethylene blue assay were conducted to find the amount of proteoglycan. The real-time polymerase chain reaction for aggrecan, sex-determining region Y-Box 9, collagen I (COL1), and COL2 gene expression was performed after 14 and 21 days. For evaluation of cartilage regeneration, the samples were implanted in rabbit knees with cartilaginous experimental defects. Defects were created in both knees of three groups of rabbits. Group 1 was the control with no injection, and Groups 2 and 3 were loaded with hydrogel/cell and hydrogel/microparticle/cell; respectively. Then, after 3 and 6 months, histological evaluations of the defected sites were carried out. The amount of glycosaminoglycans after 14 and 21 days increased significantly in hydrogels/microparticles loaded with cells. The expression of marker genes was also significant in hydrogels/microparticles loaded with cells. According to histology analysis, the hydrogels/microparticles loaded with cells showed the best cartilage regeneration. Overall, our study revealed that the developed injectable hydrogel/microparticle can be used for cartilage regeneration.
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Cartílago/fisiología , Liberación de Fármacos , Hidrogeles/química , Inyecciones , Microesferas , Regeneración , Alginatos/química , Animales , Cartílago/patología , Proliferación Celular , Supervivencia Celular , Sistemas de Liberación de Medicamentos , Módulo de Elasticidad , Regulación de la Expresión Génica , Glicosaminoglicanos/metabolismo , Masculino , Ensayo de Materiales , Oxidación-Reducción , Conejos , Reología , Estrés Mecánico , Factores de TiempoRESUMEN
During dental trauma, periodontal ligament (PDL) contributes to the stability of the tooth-PDL-bone structure. When a dental implant is inserted into the bone, the dental implant-bone construct will be more prone to mechanical damage, caused by impact loading, than the tooth-PDL-bone construct. In spite of the prevalence of such traumas, the behavioral differences between these two constructs have not been well-understood yet. The main goal of this study was to compare the momentum transferred to the tooth-PDL-bone and dental implant-bone constructs under impact loading. First, mechanical impact tests were performed on six canine mandibles of intact (N = 3) and implanted (N = 3) specimens using a custom-made drop tower apparatus, from release heights of 1, 2, and 3 cm. Next, computed tomography-based finite element models were developed for both constructs, and the transferred momenta were calculated. The experimental results indicated that, for the release heights of 1, 2, and 3 cm, the linear momenta transferred to the dental implant-bone construct were 33.1, 31.0, and 27.5% greater than those of the tooth-PDL-bone construct, respectively. Moreover, results of finite element simulations were in agreement with those of the experimental tests (error <7.5%). This work tried to elucidate the effects of impact loading on the dental implant-bone and tooth-PDL-bone constructs using both in-vitro tests and validated in-silico simulations. The findings can be employed to modify design of the current generation of dental implants, based on the lessons one can take from the biomechanical behavior of a natural tooth structure.
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Postoperative sensory disturbances of inferior alveolar nerve (IAN) are major challenges in dental procedures. We aimed to investigate the effect of photobiomodulation therapy (PBMT) with 810 nm and 980 nm diode lasers on behavioral and immunological factors in a rat IAN crush model. Seventy-two rats were randomly assigned to the four groups of 810 nm laser (crush injury+810 nm laser; 6 J/cm2, 15 sessions, every 48 h), 980 nm laser (crush injury+980 nm laser; same protocol), control (crush injury without irradiation), and sham surgery (no crush injury and no irradiation). The neurosensory response of IAN was evaluated by Von Frey behavioral test before (baseline) and post-surgery in a period of one month. Changes of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), nuclear factor-kappa B (NF-κB), TNF-α, and IL-1ß, were assessed on days 2 and 30 post injury. Data were analyzed for significant differences by repeated measures and one-way ANOVA (p < .05). One day after surgery, all rats subjected to nerve injury showed significant increase in the withdrawal threshold of von Frey test compared to the baseline (p = .02 for control and p = .03 for laser groups). The threshold gradually returned to the baseline scores in 810 nm, 980 nm, and control groups from days 11, 17, and 29, respectively. There was a significant lower withdrawal threshold in 810 nm and 980 nm laser groups compared to the control group in days 11 to 19 and 9 to 23, respectively. At both time points, the levels of NGF and BDNF were significantly higher in 810 nm laser group compared to the control group. There was a significant difference between laser and control groups regarding NF-κB expression (all p values<.001). TNF-α and IL-1ß were significantly lower in laser groups compared to the control group (all p values < .001). PBMT with 810 and 980 nm diode laser protocol used in this study, promoted the neurosensory recovery of IAN after crush injury in rats. In addition, application of 810 nm diode laser was associated with more improvement in immunological responses compared to that of 980 nm laser.
Asunto(s)
Láseres de Semiconductores , Nervio Mandibular/efectos de la radiación , Animales , Conducta Animal/efectos de la radiación , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Modelos Animales de Enfermedad , Regulación hacia Abajo/efectos de la radiación , Interleucina-1beta/metabolismo , Terapia por Luz de Baja Intensidad , Masculino , Nervio Mandibular/inmunología , Nervio Mandibular/metabolismo , Lesiones del Nervio Mandibular/inmunología , Lesiones del Nervio Mandibular/metabolismo , Lesiones del Nervio Mandibular/radioterapia , FN-kappa B/metabolismo , Factor de Crecimiento Nervioso/metabolismo , Ratas , Ratas Wistar , Factor de Necrosis Tumoral alfa/metabolismo , Regulación hacia Arriba/efectos de la radiaciónRESUMEN
Hypoxia, the result of disrupted vasculature, can be categorized in the main limiting factors for fracture healing. A lack of oxygen can cause cell apoptosis, tissue necrosis, and late tissue healing. Remedying hypoxia by supplying additional oxygen will majorly accelerate bone healing. In this study, biphasic calcium phosphate (BCP) scaffolds were fabricated by robocasting, an additive manufacturing technique. Then, calcium peroxide (CPO) particles, as an oxygen-releasing agent, were coated on the BCP scaffolds. Segmental radial defects with the size of 15 mm were created in rabbits. Uncoated and CPO-coated BCP scaffolds were implanted in the defects. The empty (control) group received no implantation. Repairing of the bone was investigated via X-ray, histological analysis, and biomechanical tests at 3 and 6 months postoperatively, with immunohistochemical examinations at 6 months after operation. According to the radiological observations, formation of new bone was augmented at the interface between the implant and host bone and internal pores of CPO-coated BCP scaffolds compared to uncoated scaffolds. Histomorphometry analysis represented that the amount of newly formed bone in the CPO-coated scaffold was nearly two times higher than the uncoated one. Immunofluorescence staining revealed that osteogenic markers, osteonectin and octeocalcin, were overexpressed in the defects treated with the coated scaffolds at 6 months of postsurgery, demonstrating higher osteogenic differentiation and bone mineralization compared to the uncoated scaffold group. Furthermore, the coated scaffolds had superior biomechanical properties as in the case of 3 months after surgery, the maximal flexural force of the coated scaffolds reached to 134 N, while it was 92 N for uncoated scaffolds. The results could assure a boosted ability of bone repair for CPO-coated BCP scaffolds implanted in the segmental defect of rabbit radius because of oxygen-releasing coating, and this system of oxygen-generating coating/scaffold might be a potential for accelerated repairing of bone defects.