Objective To investigate the feasibility of a new kind of porous β tricalcium phosphate (β-TCP) as a scaffold for the bone tissue engineering Methods The inverted phase contrast microscope was used to observe the growth of the marrow mesenchymal stem cells (MSCs) in the experimentalgroup and the control group at 10 days.In the experimental group, the MSCs were cultured with β-TCP(3 mm×3 mm×3 mm) in the 24-hole cultivation board, and in the control to control group, only MSCs were cultivated. The scanning electron microscope was used to observe growth of MSCs at 6 days. Cultivated with β-TCP at 3, 6, 9, 12 days, the MTT assay was used to judge the biocompatibility. The cytotoxicity was analyzed with the method that used the different density(100%， 50%， 10%， 1%，0%) leaching liquor gained from β-TCP to raise MSCs. MSCs were induced into the osteoblasts and were mixed with β-TCP, and the composite was used to repair a large radius bone defect in the rabbit. The specimens were made at 2,6,12 weeks. The histology imageology, and the radionuclide bone scan were used to analyze the bone formation. Results Some MSCs had a good adherence 4 hours after MSCs were inoculated and had a complete adherence at 12 hours. The cells were shaped like polyangle, spindle or converge monolayer after 8-10 days. The cells in the two groups had no difference. The cell adhesion was good, when observed by the inverted phase contrast microscope and the scanning electron microscope at 6 days. MTT showed that the absorbance (A)was not statistically different between the experimental group and the control group (P＞0.05); the different density leaching liquor had no cytotoxicity at the different time points. Histology, X-ray, and CT tomograph showed that itcould repair the large radius bone defect in the rabbit and its in vivo degradationrate was the same as the bone formation rate. Conclusion The new porous β-TCP has a unique three dimensional (3D) stereochemical structure and superordinary physicochemical property, and so it is a good scaffold for the bone tissue engineering.
Objective To review the research progress of articular cartilage scaffold materials and look into the future development prospects. Methods Recent literature about articular cartilage scaffold for tissue engineering was reviewed, and the results from experiments and clinical application about natural and synthetic scaffold materials were analyzed. Results The design of articular cartilage scaffold for tissue engineering is vital to articular cartilage defects repair. The ideal scaffold can promote the progress of the cartilage repair, but the scaffold materials still have their limitations. Conclusion It is necessary to pay more attention to the research of the articular cartilage scaffold, which is significant to the repair of cartilage defects in the future.
ObjectiveTo explore the influence of three central venous catheter biomedical materials (polyurethane, silicone, and polyvinyl chloride) on the proliferation, apoptosis, and cell cycle of Xuanwei Lung Cancer-05 (XWLC-05) cells so as to provide the basis for clinical choice of central venous catheter. MethodsXWLC-05 cells were cultured and subcultured, and the cells at passage 3 were cultured with polyurethane, silicone, and polyvinyl chloride (1.0 cm × 1.0 cm in size), and only cells served as a control. At 24, 48, and 72 hours after cultured, MTT assay was used to detect the cellular proliferation and flow cytometry to detect the cell cycle and apoptosis. At 72 hours after cultured, inverted microscope was used to observe the cell growth. ResultsInverted microscope showed the cells grew well in control group, polyurethane group, and silicone group. In polyvinyl chloride group, the cells decreased, necrosed, and dissolved; residual adherent cells had morphologic deformity and decreased transmittance. At 24 and 48 hours, no significant difference in proliferation, apoptosis, and cell cycle was found among 4 groups (P gt; 0.05). At 72 hours, the proliferations of XWLC-05 cells in three material groups were significantly inhibited when compared with control group (P lt; 0.05), and the cells in polyvinyl chloride group had more significant proliferation inhibition than polyurethane group and silicone group (P lt; 0.05), but there was no signifcant difference in proliferation inhibition between polyurethane group and silicone group (P gt; 0.05). Compared with the control group, three material groups had significant impact on the rate of apoptosis and cell cycle: polyvinyl chloride group was the most remarkable, followed by silicone group, polyurethane group was minimum (P lt; 0.05). ConclusionPolyvinyl chloride can significantly impact the proliferation, apoptosis, and cell cycle of XWLC-05 cells; polyurethane has better biocompatibility than polyvinyl chloride and silicone
Objective To evaluate the biocompatibility of a new bone matrix material (NBM) composed of both organic and inorganic materials for bone tissue engineering. Methods Osteoblasts combined with NBM in vitro were cultured. The morphological characteristics was observed; cell proliferation, protein content and basic alkaline phosphatase(ALP) activity were measured. NBM combined with osteoblasts were implanted into the skeletal muscles of rabbits and the osteogenic potential of NBM was evaluated through contraat microscope, scanning electromicroscope and histological examination. In vitro osteoblasts could attach and proliferate well in the NBM, secreting lots of extracellular matrix; NBM did not cause the inhibition of proliferation and ALP activity of osteoblasts. While in vivo experiment of the NBM with osteoblasts showed that a large number of lymphacytes and phagocytes invading into the inner of the material in the rabbit skeletalmuscle were seen after 4 weeks of implantation and that no new bone formation was observed after 8 weeks. Conclusion This biocompat ibility difference between in vitro and in vivo may be due to the immunogenity of NBM which causes cellular immuno reaction so as to destroy the osteogenic environment. The immunoreaction between the host and the organic-inorganic composite materials in tissue engineering should be paid more attention to.
OBJECTIVE To investigate the feasibility of freeze-dried demineralized bone matrix (FDBM) as scaffold material in bone tissue engineering. METHODS Osteoblasts which were isolated from cranial periosteum of New Zealand rabbits were cultured as the seeding cells, then the cells were cocultured with heterogenous FDBM in vitro. The cell-material complex was observed under phase microscope, light microscope and electronic scanning microscope in order to evaluate the interaction between cells and FDBM. RESULTS Eight hours after coculture, the osteoblasts adhered to FDBM scaffolds. Seven days later, the osteoblasts differentiated and proliferated in FDBM network. Extracellular matrix was secreted and calcium nodes were formed among osteoblasts. CONCLUSION FDBM is a good scaffold material for the bone tissue engineering.
The study aimed to evaluate the safety and function of poly(lactic-acid-co-ε-caprolactone) (PLCL)/fibrinogen nanofibers (P/F-Ns), and provide theoretical basis for the clinical application. The surface morphology, mechanical properties, the hydrophilicity and the fibrinogen content of P/F-Ns were tested by scanning electron microscope, the material testing machine, the contact angle meter and the microplate reader, respectively. The cell adhesion, proliferation and ligament remodeling genes expression of Hig-82 cells on P/F-Ns were conducted through cell counting kit-8 (CCK-8) and real-time quantitative PCR analyses, respectively. The results showed that with the increase of the fibrinogen content, the pore sizes and hydrophilicity of three P/F-Ns increased, but the mechanical properties decreased. Cell adhesion and proliferation tests showed that P/F-N-2 held the best ability to promote cell adhesion and proliferation. The ligament remodeling genes expressions of Hig-82 cells on P/F-N-1, P/F-N-2 and P/F-N-3 were all up-regulated compared to P/F-N-0 on days 3 and 7. All the three P/F-Ns containing fibrinogen (P/F-N-1, P/F-N-2 and P/F-N-3) had better biocompatibility compared to P/F-N-0, and could be efficiently applied to the reconstruction of anterior cruciate ligament.
Objective To study the inflammation response and the biocompatibil ity of valved bovine jugular vein conduit (BJVC) and valved bovine jugular vein patch (VBJV-P) in treating complex congenital heart disease (CHD). Methods From December 2007 to March 2008, 16 patients with complex CHD were treated. Of 16 patients, 6 underwent conjunction right ventricular to pulmonary artery with BJVC and broaden right ventricular outflow tract (RVOT) with VBJV-P (BJVC group), and 10 underwent broaden RVOT with self pericardial patch (control group). In BJVC group, there were 3 males and 3 females, aging (5.6 ± 3.6) years, and including 1 case of type I truncus arteriosus, 1 case of type I truncus arteriosus with ventricular septal defect and patent foramen ovale, 1 case of congenital pulmonary atresia with ventricular septal defect and patent arterial duct, and 3 cases of Fallot’s tetrad. In control group, there were 5 males and 5 females, aging(4.3 ± 3.1) years, all being Fallot’s tetrad. The periphery vein blood of the two groups was collected during operation and after operation, and the levels of cytokine were detected with ELISA method. Meanwhile the cl inical data of the two groups were collected. Results There were no significant differences at levels of TNF-α and IL-6 between BJVC group and control group 1 week after operation (P gt; 0.05), and there was significant difference at level of IL-10 [(25.7 ± 5.0) pg/mL vs (19.5 ± 4.7) pg/ mL, P lt; 0.05]. There were no significant differences at levels of IL-6 and IL-10 within groups both in control group and in BJVC group (P gt; 0.05) between 1 week after operation and the anesthesia inducing period. And there was significant difference at level of TNF-α in BJVC group [(77.0 ± 1.6) pg/mL vs (82.9 ± 1.3) pg/mL, P lt; 0.05] and in control group [(78.6 ± 3.4) pg/mL vs (83.1 ± 1.9) pg/mL, P lt; 0.05] between 1 week after operation and the anesthesia inducing period. There were no statistically significant differences (P gt; 0.05) in leukocyte count and body temperature between BJVC group and control group. The X-ray films showed no abnormal ity in BJVC group and control group before operation and after operation. No hepatic and renal dysfunction occurred in control group; and 2 patients had hepatic dysfunction, which may be caused by antibiotics. Conclusion BJVC has a good biocompatibil ity in treating complexty CHD.
Objective To investigate the biocompatibility of diamond-like carbon(DLC) coated NickelTitanium shape memory alloy with osteoblasts cultured invitro. Methods Rabbit’s osteoblasts were incubated with DLCcoated NickelTitanium shape memory alloy disks and uncoated ones of equal size for 5 days. The control group（without shape memory alloy in culture media） was performed simultaneously. The cultured cells were counted and graphed. The samples from culture media were collected and the concentrations of alkaline phosphatase (ALP) and nickel（Ni2+） were measured from the 1st to 5th day respectively. Results The proliferation of osteoblasts and the concentration of ALP in both DLC-coated group and control gruop was higher than uncoated group. The proliferation of osteoblasts on the 3rd, 4th, and 5th day in both DLC-coatedgroup and control group was significantly higher than that in the uncoated group(P＜0.05). The concentration of ALP in DLC-coated group on the 2nd, 3rd, and 5th day and in the control group on the 3rd, 4th, and 5th day was significantly higher than that in the uncoated group(P＜0.05). The concentration of Ni2+ on the 3rd, 4th, and 5th day was significantly lower than that in the uncoated group(P＜0.05). Conclusion DLC- coated NickelTitanium shape memory alloys appears to have better biocompatibility with osteoblast cultured in vitro compared to uncoated ones.
Objective To observe the biocompatibil ity of self-assembled FGL peptide nano-fibers scaffold with neural stem cells (NSCs). Methods FGL peptide-amphiphile (FGL-PA) was synthesized by sol id-phase peptide synthesistechnique and thereafter It was analyzed and determined by high-performance l iquid chromatography (HPLC) and massspectrometry (MS). The diluted hydrochloric acid was added into FGL-PA solution to reduce the pH value and accordinglyinduce self-assembly. The morphological features of the assembled material were studied by transmission electron microscope (TEM). NSCs were cultured and different concentrations of FGL-PA assembled material were added with the terminal concentrations of 0, 50, 100, 200, 400 mg/L, respectively. CCK-8 kit was used to test the effect of FGL assembled material on prol iferation of NSCs. NSCs were added into differentiation mediums (control group: DMEM/F12 medium containing 2% B27 supplement and 10% FBS; experimental group: DMEM/F12 medium containing 2% B27 supplement, 10% FBS and 100 mg/L FGL-PA, respectively). Immunofluorescence was appl ied to test the effect of FGL-PA assembled material on differentiation of NSCs. Results FGL-PA could be self-assembled to form a gel. TEM showed the self-assembled gel was nano-fibers with diameter of 10-20 nm and length of hundreds nanometers. After NSCs were incubated for 48 hours with different concentrations of FGL-PA assembled material, the result of CCK-8 assay showed that FGL-PA with concentrations of 50, 100 or 200 mg/L could promote the prol iferation of NSCs and absorbance of them was increased (P lt; 0.05). Immunofluorescence analysis notified that the differentiation ratio of neurons from NSCs in control group and experimental group were 46.35% ± 1.27% and 72.85% ± 1.35%, respectively, when NSCs were induced to differentiation for 14 days, showing significant difference between 2 groups (P lt; 0.05). Conclusion FGL-PA can self-assemble to nano-fiber gel, which has good biocompatibil ity and neural bioactivity.
Objective To investigate the influence of different dose levels of hydroxyapatite/tricalcium phosphate (HA/TCP) on the proliferation and alkalinephosphatase (ALP) activity of rabbit osteoblasts. Methods Three different doselevels of HA/TCP (10%, 40%, 70%) were co-cultivated with rabbit osteoblasts respectively. The proliferation and ALP expression capacity of osteoblasts were examined with MTT method and enzyme histochemistry once every 24 hours until 5 days. Three control groups of other materials were treated and examined in the sameway: rabbit osteoblasts as normal control; polyvinylchloride as positive control; titanium alloy as negative control. Results There was remarkable timeeffect relationship in the proliferation of osteoblasts. Ten percent HA/TCP did not affect osteoblasts growth while 40% HA/TCP could slow the cell growth rate down though time-effect relationship still existed. The proliferation of osteoblasts stagnated when co-cultivated with 70% HA/TCP. On the other hand, 10% HA/TCP could cause reversible damage on ALP activity of osteoblasts, whereas when the dose was40%, and the cultivation lasted 6 days the damage was irreversible. Three different dose levels of titanium alloy (10%, 40%, 70%) had no effect on the proliferation or ALP activity of osteoblasts. Conclusion Dosage is an important factor affecting the biocompatibility evaluation of biomaterial. It suggests that dose choosing should be more specified upon each individual biomaterial. It also indicates that ALP may be a good supplementary index of the cell compatibility of material.