Tricalcium phosphate (TCP) is one of the most widely used bioceramics for constructing bone tissue engineering scaffold. The three-dimensional (3D) printed TCP scaffold has precise and controllable pore structure, while with the limitation of insufficient mechanical properties. In this study, we investigated the effect of sintering temperature on the mechanical properties of 3D-printed TCP scaffolds in detail, due to the important role of the sintering process on the mechanical properties of bioceramic scaffolds. The morphology, mass and volume shrinkage, porosity, mechanical properties and degradation property of the scaffold was studied. The results showed that the scaffold sintered at 1 150℃ had the maximum volume shrinkage, the minimum porosity and optimal mechanical strength, with the compressive strength of (6.52 ± 0.84) MPa and the compressive modulus of (100.08 ± 18.6) MPa, which could meet the requirements of human cancellous bone. In addition, the 1 150℃ sintered scaffold degraded most slowly in the acidic environment compared to the scaffolds sintered at the other temperatures, demonstrating its optimal mechanical stability over long-term implantation. The scaffold can support bone mesenchymal stem cells (BMSCs) adherence and rapid proliferation and has good biocompatibility. In summary, this paper optimizes the sintering process of 3D printed TCP scaffold and improves its mechanical properties, which lays a foundation for its application as a load-bearing bone.
Objective To prepare silver-containing hydroxyapatite coating (hydroxyapatite/Ag, HA/Ag) and investigate its antibacterial property and biocompatibil ity in vitro. Methods Vacuum plasma spraying technique was adopted to prepare HA/Ag coating on titanium alloy substrate (3% Ag). After incubating the HA/Ag and the HA coating under staphylococcus aureus and pseudomonas aeruginosa suspensions of 2% tryptic soy broth (TBS) medium for 2, 4 and 7 days, respectively, the biofilm on the coatings was examined by confocal laser scanning microscope, and the bacterial density and viable bacterial percentage of bacterial biofilm were calculated. Meanwhile, the micro-morphology of bacterial biofilm was observed by SEM, the cytotoxicity was detected via MTT and the biocompatibil ity of biofilm was evaluated by acute aemolysis test. Results Compared with HA coating, the bacterial biofilm’s thickness on the surface of HA/Ag coating witnessed no significant difference at 2 days after culture (Pgt; 0.05), but decreased obviously at 4 and 7 days after culture (P lt; 0.01). The bacterial density of the biofilm increased with time, but there was no significant difference between two coatings (P gt; 0.05) at 2, 4 and 7 days after culture. The viable bacterial percentage of the biofilms on the surface of HA/Ag coating decreased obviously compared with that of HA coating at 2, 4 and 7 days after cultureP lt; 0.01). The MTT notified the cytotoxic grade of both coatings was zero. The acute haemolysis assay showed that the hemolytic rate of HA/Ag and HA coating was 0.19% and 0.12%, respectively. Conclusion With good biocompatibil ity, significant antibacterial property against staphylococcus aureus and pseudomonas aeruginosa, no obvious cytotoxicity and no erythrocyte destruction, the vacuum plasma sprayed HA/Ag coating is a promising candidate for the surface of orthopedic metal implants to improve their osseointegration and antibacterial property.
Objective To explore the biomechanic effects of multi ple freeze-thaw on human allograft tendons. Methods Thirty tendons (24 flexor digitorum superficial is tendons and 6 flexor poll icis longus tendons) were harvested from 3 fresh cadaver donors and were divided into 6 groups randomly (fresh group; 1 cycle, 2 cycle, 3 cycle, 5 cycle, and 10 cycle freeze-thaw groups). There was 4 flexor digitorum superficial is tendons and 1 flexor poll icis longus tendon in each group. The structural and mechanical properties as well as viscoelastic change were estimated. Results The results of the structural and mechanical properties in 1 cycle, 2 cycle, and 3 cycle freeze-thaw groups were similar to that of the fresh group (P gt;0.05). The tendons in 5 cycle and 10 cycle freeze-thaw groups showed a significantly lower ultimate load and maximum stress when compared with those of fresh group (P lt; 0.05), but there was no significant difference in maximum tensile or maximum strain (P gt; 0.05). Moreover, the tendons in 5 cycle and 10 cycle freeze-thaw groups had a significant increase in viscoelastic properties when compared with fresh group (P lt; 0.05). Conclusion In the cryopreservation of tendon allografts, the cycle of freeze-thaw should not exceed 3 times. Multiple cycle freeze-thaw will weaken the biomechanical properties of tendon allografts, which make grafts easier to fatigue or even rupture.
Objective To summarize the influence of microstructure on performance of triply-periodic minimal surface (TPMS) bone scaffolds. Methods The relevant literature on the microstructure of TPMS bone scaffolds both domestically and internationally in recent years was widely reviewed, and the research progress in the imfluence of microstructure on the performance of bone scaffolds was summarized. Results The microstructure characteristics of TPMS bone scaffolds, such as pore shape, porosity, pore size, curvature, specific surface area, and tortuosity, exert a profound influence on bone scaffold performance. By finely adjusting the above parameters, it becomes feasible to substantially optimize the structural mechanical characteristics of the scaffold, thereby effectively preempting the occurrence of stress shielding phenomena. Concurrently, the manipulation of these parameters can also optimize the scaffold’s biological performance, facilitating cell adhesion, proliferation, and growth, while facilitating the ingrowth and permeation of bone tissue. Ultimately, the ideal bone fusion results will obtain. Conclusion The microstructure significantly and substantially influences the performance of TPMS bone scaffolds. By deeply exploring the characteristics of these microstructure effects on the performance of bone scaffolds, the design of bone scaffolds can be further optimized to better match specific implantation regions.
ObjectiveTo investigate the effects of modification of acellular bovine pericardium with 1-ethyl-3-(3-dinethylami-nopropyl) carbodimide (EDC)/N-hydroxysuccininide (NHS) or genipin and find out the best crosslinking reagent. MethodsThe cellular components of the bovine pericardiums were removed. The effects of decellularization were tested by HE staining. The acellular bovine pericardiums were crosslinked with EDC/NHS (EDC/NHS group) or genipin (genipin group). The properties of the crosslinked acellular matrix were evaluated by scanning electron microscope (SEM), matrix thickness, crosslinking index, mechanical property, denaturation temperature, enzymatic degradation, and cytotoxicity test before and after the crosslinking. Acellular bovine pericardium (ABP group) or normal bovine pericardium (control group) were harvested as controls. ResultsSEM showed that collagen fibers were reticulated in bovine pericardial tissues after crosslinked by EDC/NHS or genipin, and relative aperture of the collagen fiber was from 10 to 20 μm. The thickness and denaturation temperature of the scaffolds were increased significantly after crosslinking with EDC/NHS or genipin (P<0.05), while there was no significant difference between EDC/NHS group and genipin group (P>0.05). The difference had no statistical significance in crosslinking index between EDC/NHS group and genipin group (t=0.205, P=0.218). The degradation rate in EDC/NHS group and genipin group was significantly lower than that in ABP group and control group (P<0.05). Elastic modulus and fracture stress in EDC/NHS group and genipin group were significantly lower than those in ABP group (P<0.05), but there was no significant difference among EDC/NHS group, genipin group, and control group (P>0.05). The break elongation in EDC/NHS group and genipin group were significantly increased than those in ABP group and control group (P<0.05). The difference had no statistical significance in stability and mechanical properties between EDC/NHS group and genipin group (P>0.05). Cytotoxicity of genipin crosslinked tissue (grade 1) were much lower than that of EDC/NHS (grade 2) at 5 days. ConclusionAcellular bovine pericardium crosslinked with genipin has better biocompatibility than EDC/NHS.
Decellularized tissue engineering scaffolds appear to have the properties of similar structure and mechanical characteristics to native tissues，good biocompatibility，suitability for cell adhesion，growth and angiogenesis induction，and non-immunogenicity. Genipin has anti-inflammatory，antithrombotic and antioxidative features which can considerably suppress vascular and endothelial inflammatory activation，increase mechanical strength of biological scaffolds，inhibit inflammatory response and decrease degradation rate of biological scaffolds. By cross-linking with decellularized matrices，Genipin can further improve corresponding performance of tissue engineering matrices，which is very helpful to promote the application of tissue engineering into clinical practice of cardiothoracic surgery. This review focuses on recent research process and possible prospects of Genipin cross-linking in tissue engineering in the field of cardiothoracic surgery.
ObjectiveTo observe the changes of visual acuity and fixation properties of eyes with idiopathic macular hole (IMH) before and after surgery. MethodsA prospective clinical study. From September 2019 to December 2020, 25 patients with 25 eyes of IMH diagnosed in Department of Ophthalmology of The Fourth People's Hospital of Shenyang were included in the study. All patients underwent pars plana vitrectomy (PPV) combined with internal limiting membrane stripping. All eyes underwent best corrected visual acuity (BCVA), optical coherence tomography (OCT), and microperimetry before and after surgery. The BCVA examination was carried out using the Snellen visual acuity chart, which was converted into logarithmic minimum resolution angle (logMAR) visual acuity during statistics. The 12° macular sensitivity (MS) and bivariate contour ellipse area (BCEA) were measured by MP-3 microperimetry. The minimum diameter (MIN) and base diameter (BASE) of the macular hole were measured by OCT; the distance between the preferred retinal location (PRL) and the center of the fovea was measured by Image-proplus 6.0 image processing software. At 1 and 3 months follow-up after surgery, the same equipment and methods as before surgery were used to conduct related examinations. The changes of BCVA, PRL distance from the fovea, MS, BCEA, and macular hole shape before and after surgery were compared and observed. One-way analysis of variance was used to compare the indicators before and after surgery. Pearson correlation analysis was used for the correlation between BCVA and preoperative BCVA, PRL and foveal center distance at 3 months after surgery. The correlation between MIN, BCVA, PRL and foveal center before surgery distance, MS, BCEA and BCVA at 3 months after surgery were analyzed by multiple linear regression. ResultsAmong 25 eyes of 25 cases, 1 male had 1 eye, and 24 females had 24 eyes. The macular hole in stage Ⅲ and Ⅳ were 11 eyes and 14 eyes, respectively. MIN and BASE were 537.68±200.09 and 905.48±278.79 μm, respectively. One month after surgery, the hiatus was closed. Before surgery and 1 and 3 months after surgery, the logMAR BCVA of the affected eyes were 0.80±0.17, 0.70±0.21, 0.60±0.25, and the MS were 22.20±3.86, 23.60±3.14, 24.38±2.68 dB, the distances between PRL and the center of the fovea were 537.72±426.05, 402.00±395.06, 236.80±219.54 μm, and BCEA were 7.90±3.43, 6.40±2.67, 4.80±2.32 deg2. Compared with before operation, BCVA (F=7.047, 20.104) and MS (F=1.980, 5.390) were significantly improved at different time after operation, the distance between PRL and fovea center (F=1.265, 9.530), BCEA (F=2.762, 13.617) were decreased, the difference were statistically significant (P＜0.05). The results of correlation analysis showed that BCVA at 3 months after surgery was significantly associated with preoperative MIN (r=0.810), BASE (r=0.664), BCVA before surgery and 1 month after surgery (r=0.854, 0.940), preoperative and surgical MS at 1 month after surgery (r=－0.548, －0.578), distance between PRL and foveal center before surgery and at 1 month after surgery (r=0.833, 0.915), BCEA before surgery and at 1 month after surgery (r=0.636, 0.732) were significantly correlated (P＜0.05). The results of multiple linear regression analysis showed that the distance between PRL and foveal center before surgery and BCVA were risk factors for poor prognosis of BCVA at 3 months after surgery. ConclusionsThe BCVA and MS of eyes with IMH are significantly improved after surgery, and the distance between PRL and foveal center and BCEA decreased. BCVA, PRL and foveal center distance before surgery are risk factors for poor visual acuity after surgery.
ObjectiveThe tissue engineered osteochondral integration of multi-layered scaffold was prepared and the related mechanical properties and biological properties were evaluated to provide a new technique and method for the repair and regeneration of osteochondral defect.MethodsAccording to blend of different components and proportion of acellular cartilage extracellular matrix of pig, nano-hydroxyapatite, and alginate, the osteochondral integration of multi-layered scaffold was prepared by using freeze-drying and physical and chemical cross-linking technology. The cartilage layer was consisted of acellular cartilage extracellular matrix; the middle layer was consisted of acellular cartilage extracellular matrix and alginate; and the bone layer was consisted of nano-hydroxyapatite, alginate, and acellular cartilage extracellular matrix. The biological and mechanics characteristic of the osteochondral integration of multi-layered scaffold were evaluated by morphology observation, scanning electron microscope observation, Micro-CT observation, porosity and pore size determination, water absorption capacity determination, mechanical testing (compression modulus and layer adhesive strength), biocompatibility testing [L929 cell proliferation on scaffold assessed by MTT assay, and growth of green fluorescent protein (GFP)-labeled Sprague Dawley rats’ bone marrow mesenchumal stem cells (BMSCs) on scaffolds].ResultsGross observation and Micro-CT observation showed that the scaffolds were closely integrated with each other without obvious discontinuities and separation. Scanning electron microscope showed that the structure of the bone layer was relatively dense, while the structure of the middle layer and the cartilage layer was relatively loose. The pore structures in the layers were connected to each other and all had the multi-dimensional characteristics. The porosity of cartilage layer, middle layer, and bone layer of the scaffolds were 93.55%±2.90%, 93.55%±4.10%, and 50.28%±3.20%, respectively; the porosity of the bone layer was significantly lower than that of cartilage layer and middle layer (P<0.05), but no significant difference was found between cartilage layer and middle layer (P>0.05). The pore size of the three layers were (239.66±35.28), (153.24±19.78), and (82.72±16.94) μm, respectively, showing significant differences between layers (P<0.05). The hydrophilic of the three layers were (15.14±3.15), (13.65±2.98), and (5.32±1.87) mL/g, respectively; the hydrophilic of the bone layer was significantly lower than that of cartilage layer and middle layer (P<0.05), but no significant difference was found between cartilage layer and middle layer (P>0.05). The compression modulus of the three layers were (51.36±13.25), (47.93±12.74), and (155.18±19.62) kPa, respectively; and compression modulus of the bone layer was significantly higher than that of cartilage layer and middle layer (P<0.05), but no significant difference was found between cartilage layer and middle layer (P>0.05). The osteochondral integration of multi-layered scaffold was tightly bonded with each layer. The layer adhesive strength between the cartilage layer and the middle layer was (18.21±5.16) kPa, and the layer adhesive strength between the middle layer and the bone layer was (16.73±6.38) kPa, showing no significant difference (t=0.637, P=0.537). MTT assay showed that L929 cells grew well on the scaffolds, indicating no scaffold cytotoxicity. GFP-labeled rat BMSCs grew evenly on the scaffolds, indicating scaffold has excellent biocompatibility.ConclusionThe advantages of three layers which have different performance of the tissue engineered osteochondral integration of multi-layered scaffold is achieved double biomimetics of structure and composition, lays a foundation for further research of animal in vivo experiment, meanwhile, as an advanced and potential strategy for osteochondral defect repair.
In order to overcome the influence of stray light and impurity on the image of video laryngoscope, we designed an optical structure by using TracePro, a simulation software, to imitate optical path status. Images are captured by CMOS sensor which has the size of 4.5 mm×18 mm and the pixel size is 1.75 μm×1.75 μm. The sensor is placed in the elbow of the laryngoscope, and the elbow has the size of 9 mm×10 mm. As a result, the video laryngoscope could meet the requirements, including wide viewing angle (80°), short focal length (2.8 mm), long working distance (10 cm), and least impurity. In the test, the image was clear and there was no facula or impurity in the condition of required illumination, and thus stray light and image impurity were eliminated and the image quality was improved.
ObjectiveTo isolate nucleus pulposus cells (NPCs) from the caudal and lumbar intervertebral disc of rat, and to identify the morphology and to compare the characteristics. MethodsThe whole spine was separated from 8-week-old Sprague Dawley rats under the sterile conditions. NPCs of different segments (lumbar group: L1,2-L6, S1; caudal group: C1,2-C17,18) were cultured by adherent cultivation approach. Cellular morphologic change was noted by HE staining and continuous observation under inverted phase contrast microscope. Besides, the aggrecan and collagen type Ⅱexpression were examined by toluidine blue and immunocytochemistry staining respectively. The total protein contents, senescence level, and the cell viability of passage 1-5 (P1-5) were detected. The growth curves of the P1 cells in lumbar and caudal groups were determined by cell counting kit 8. ResultsThe NPCs were isolated and identified successfully. The adherence time of the primary cells (the cell fusion reached 90%) in lumbar group was significantly longer than that in caudal group in primary generation (P<0.05). HE staining showed that cytoplasm was pink with the blue nucleus. Lumbar disc NPCs were spindle. The larger caudal disc NPCs were polygonal or irregular. Toluidine blue staining showed that the proteoglycan was stained as blue. In the cytoplasm of cells, collagen type Ⅱwas stained as brown surround the blue-black nucleus. The cell viability had no significant difference between lumbar and caudal groups and between different passages in the same group (P>0.05). The caudal disc NPCs reached their logarithmic growth phase after 3 days of culture, while the cells in lumbar segments did after 4-5 days of culture. The cell proliferation in caudal segments was more than that in lumbar segments at 3-9 days (P<0.05). The difference in the total protein contents was not significant between cells at P1-5 in 2 groups (P>0.05), and the caudal disc NPCs had higher protein contents than lumbar disc NPCs (P<0.05). There was no significant difference in cell senescence rate between cells at P1, P2, and P3 in 2 groups (P>0.05), but significant difference was shown in senescence rate between 2 groups in cells at P4 and P5 (P<0.05). ConclusionCaudal disc NPCs have a better status, which is more suitable for experiment as a seed cell than the lumbar disc NPCs in the same generation.