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 prepare and study the biocompatibil ity of selectively decellular xenoskin which has the character of the lower antigen, continuous epidermis, and the dermal matrix without any cellular components. Methods The porcine skin was treated with glutaraldehyde solution, trypsin, and detergent solution TritonX-100 to prepare the selectivelydecellular xenoskin. The cytotoxicity was tested according to GB/T16886.5-2003 biological evaluation of medical devices for in vitro cytotoxicity, and the levels of cytotoxicity were evaluated with the United States Pharmacopeia. Subdermal implantation was tested according to GB/T16886.6-1997 biological evaluation of medical devices for local effects after implantation. Seventytwo mature Wistar rats were randomly assigned to groups A, B, and C (n=24). Three kinds of materials were implanted into subcutaneous of rats back. Selectively decellular xenoskin was transplanted into group A, fresh porcine skin was transplanted into group B, and allogeneic skin was transplanted into group C. The samples were collected to make the observation of gross and histology after 1, 2, 4, 8, 12, and 16 weeks. Results The cytotoxicity was proved to be first grade by biocompatibil ity test. The gross and histological observation of subdermal implantation: after implantation, the most severe inflammatory reactions were seen in group B which dispersion was very slow. Inflammatory reactions in groups A and C alleviated gradually. In groups A and C, there was an increased collagen fiber density and angiogenesis at late stage; the transplanted skin was gradually degraded and absorbed. In group B, no obvious degradation and absorption were observed. Conclusion Selectively decellular xenoskin, prepared with glutaraldehyde solution, trypsin, and detergent solution, possesses characteristics of integral skin structure andexcellent biocompatibil ity, so it can be used as a new type substitute to repair the burn wound.
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.
ObjectiveTo observe the morphological characteristic by implanting domestic porous tantalum in rabbit patellar tendon and to evaluate biocompatibility features so as to provide experimental basis for porous tantalum used as interface fixation between tendon and bone.
MethodsA total of 48 adult New Zealand white rabbits, male or female, weighing 2.5-3.0 kg, were selected. Porous tantalum flake (5 mm×5 mm×2 mm) was implanted in the left patellar tendon (experimental group) and the same size porous titanium flake in the right patellar tendon (control group). The animals were sacrificed at 2, 4, 8, and 12 weeks after implantation, then the specimens were harvested for gross observation, HE staining, scanning electron microscope (SEM) observation, and hard slices observation.
ResultsNo animal died after operation. Porous tantalum was bonded closely with host tendon and no inflammatory reaction was found. Loose and thick fibrous capsule was observed at the beginning and became density and thinner in the end by microscope, showing significant difference between different time points in 2 groups (P<0.05), but no significant difference was found between 2 groups at different time points (P>0.05). The SEM observation showed that fibrous tissue attached to the surface and inner walls of porous tantalum at early stage, and extended on the material to reach confluence at late period, but the experimental group was more than the control group. Hard slices observation showed that the collagen fibrils were seen on porous tantalum interface with host tendon, and blood vessels grew into the pores. The control group and the experimental group showed no significant difference.
ConclusionThe domestic porous tantalum has good biocompatibility. Connection and integration can be established between tendon and porous tantalum, and therefore it could be used in reconstruction of tendon-bone fixation device.
Objective To evaluate the biocompatibil ity of manufactured heterogeneous demineral ized bone matrix(DBM) particles and to provide basis for further experimental study and cl inical application. Methods Heterogeneous DBMparticles A (degreased and demineralized) and B (degreased, demineralized and acellular), particle size from 250 to 810 μm, and leaching l iquor were made with a series of physical and chemical methods from pig l imbs cortical bone. The residual calcium and phosphorus contents of bone particles were measured after degreased and demineral ized. The acute toxicity test, skin stimulating test, pyrogeneous test, hemolysis test, cellular toxicity test and muscular embedded test were carried out according standard toxicological method. Results The contents of calcium and phosphorus in cortical bone were (189.09 ± 3.12) mg/g and (124.73 ± 2.87) mg/g, and in demineral ized bone matrix particles were (3.48 ± 0.09) mg/g and (3.46 ± 0.07) mg/ g. The residual calcium content was 1.87%, of phosphorus was 2.69%. The activity of mice was normal in the acute toxicity test. No animal died and no toxicity symptom or adverse effects were shown within 7 days. The mean weight daily increased showed no statistically significant difference (P gt; 0.05) between two groups after 7 days. Skin stimulating reactions were not found in the two experimental groups and negative control group by intradermal stimulation test. The maximal increase of body temperature in two experimental groups were 0.4℃ , which meet the national standard (lt; 0.6 ). The rate of haemolysis to the leaching liquor was 1.14% (A) and 0.93% (B), which was lower than the national standard (lt; 5%). The cell prol iferation rates of two experimental groups when compared with control group showed no statistically significant difference (P gt; 0.05). The toxicity of DBM particlesleaching liquor was graded from 0 to 1, which means the material has no cytotoxicity. All the animals survived well. There was no tissue necrosis, effusion or inflammation at all implantation sites. For the index of HE and Masson staining, there were no effusion around the material and inflammatory cell infiltrate obviously in two experimental groups. Inflammatory cell infiltrate is sl ight in control group 2 weeks postoperatively. The inflammatory cell infiltration was mitigate gradually over time in two experimental groups after 4, 8 and 12 weeks. New bone and collagen fibers formation were observed when the material was degraded and absorpted. Score evaluation of local cellular immune response at different time after operation of two experimental groups showed no statistically significant difference (P gt; 0.05). Conclusion Heterogeneous DBM has no obvious toxicity, skin irritation, pyrogenicity, and no cytotoxicity with a rate of haemolysis lt; 5%, so it has good biocompatibility and partial osteoinductive.
ObjectiveTo study the biocompatibility of bioprosthetic heart valve material with a non-glutaraldehyde-based treatment, and to provide the safety data for the clinical application. MethodsAll the tests were conducted according to GB/T16886 standards. The in vitro cytotoxicity was determined by methyl thiazolyl tetrazolium assay. Fifteen guinea pigs were divided into a test group (n=10) and a control group (n=5) in the skin sensitization test. Three New Zealand white rabbits were used in the intradermal reactivity test. Five sites on both sides of the rabbit back were set as test sites and control sites, respectively. In the acute systemic toxicity test, a total of 20 ICR mice were randomly assigned to 4 groups: a test group (polar medium), a control group (polar medium), a test group (non-polar medium) and a control group (non-polar medium), 5 in each group. Forty SD rats were divided into a test group (n=20) and a control group (n=20) in the subchronic systemic toxicity test. ResultsThe viability of the 100% extracts of the bioprosthetic heart valve material with a non-glutaraldehyde-based treatment was 75.2%. The rate of positive reaction was 0.0%. The total intradermal reactivity test score was 0. There was no statistical difference in the body weight between the test group and control group in the acute systemic toxicity test. There was no statistical difference in the body weight, organ weight, organ weight/body weight ratio, blood routine test or blood biochemistry between the test group and control group in the subchronic systemic toxicity test. ConclusionThe bioprosthetic heart valve material with a non-glutaraldehyde-based treatment has satisfying biocompatibility, which conforms to relevant national standards. The material might be a promising material for application in valve replacement.
Objective To evaluate the biocompatibility and safety of a novel orthopedics materials-graded zirconia(ZrO2)hydroxyapatite(HA) composite biomaterials. Methods First, ultrafine powers of ZrO2 and HA powder were prepared by chemical precipitation method, then graded ZrO2-HA composite was synthesized by dry-laying and sintering method. After the physiological saline and culture medium extracts of the composite were prepared, four experiments were conducted as follows:① The mouse acute toxic test consists of 2 groups(n=10). The extracts were intravenously injected to mice in the first group, and physiological saline to mice in the second group. The dose was 50 g/kg. Their toxicity manifestation, morality and the change of weight were recorded.② The standard curve of proliferation and metabolism of L929 cells was established. ③ The cytotoxinic test consists of 3 groups: materials group (extracts of the materials), positive control group (culture fluid with 0.64% phenol), and negative control group (RPMI-1640 culture fluid). Each of three was cultured with cell suspension, and then the morphology of the cells was observed, the relative proliferation rate (RGR) was calculated, and the toxicity was classified. ④ In vitrohemolytic test was divided into 3 groups: extracts, sterile distilled water (positive control) and 0.9% physiological saline. In each of three, 0.2 ml anticoagulant diluted fresh rabbit blood was added. The percentage of hemolysis was tested. ⑤ The muscle and implantation test were divided into 4 groups(n=3). The composite biomaterials were implanted into pygal muscleson either side and lateral condyles of femurs. After surgery, the rats of four groups were sacrificed at 12 and 24 weeks respectively.Tissue slice and scanning electronic microscopy were performed. Results General acute toxic test: no mouse died within 3 weeks; no toxicity symptom or adverse effects were shown within 3 days. The weight of materials group increased by 3.57±0.49 g, and the control group by 3.62±0.61 g, showing no statistically significant difference（Ρgt;0.05）.The standard curve of L929 cell perliferation and metabolism showed that their existed a positive correlation between the number of L929 cells and the perliferation. ③ Cytotoxinic test: cytosomes in the positive control group diminished and appeared round, there were pyknotic nucleus, the attached cells agglomerated; the toxicity was level Ⅳ. The morphology of cells in materials groupand negative control group was normal, and the number of them increased; the toxicity was level Ⅰand level 0, respectively. The MTT color experiments showed that positive control group was significantly lower than materials group and negative control group, showing statistically significant difference (Plt;0.01); there was no statistically significant difference between materials group and negative group.④ Hemolytic test: in vitrohemolytic rate of negative control group was0, of positive control group was 100%, and of materials group was 1.66%, which accords with the standard that hemolytic rate should be lower than 5% specified in ISO. ⑤ Implant test:No apparent rejection reaction took place after the composite was implanted; the composite bonded with the bones of the receptors firmly, which had good bonedinduced effect. Conclusion Graded ZrO2-HA composite bioceramic has good biocompatibility and is suitable for orthopedic biomaterials.
Biomedical metal materials have always been a major biomedical material with a large and wide range of clinical use due to their excellent properties such as high strength and toughness, fatigue resistance, easy forming, and corrosion resistance. They are also the preferred implant material for hard tissues (bones and teeth that need to withstand higher loads) and interventional stents. And nano-medical metal materials have better corrosion resistance and biocompatibility. This article focuses on the changes and improvements in the properties of several typical medical metal materials surfaces after nanocrystallization, and discusses the current problems and development prospects of nano-medical metal materials.
Objective To evaluate the biocompatibility and in vivo degradation of novel chest wall prosthesis materials and provide some data for their clinical application. MethodsAccording to the standard for the biological evaluation of the medical devices, several tests were performed to evaluate the tissue toxic effects induced by polydioxanone (Group A), chitosan (Group B), and hydroxyapitite/collagen (Group C),which were tested as component materials of the chest wall prosthesis. In the hemolysis test, 0.2 ml of the anticoagulant rabbit blood was added to the component materials and the normal saline (negative control) and to the distilled water(positive control). Five samples were made in each group. Absorbency was measured and the hemolysis rate was determined. In the acute systemic toxicity test, 20 mice were randomly divided into 4 groups (Groups A, B and C, and the normal saline group, n=5). The leaching liquid (50 ml/kg) was injected through the caudal vein, which was observed at 24, 48 and 72 hours. In the pyrogen test, 12 rabbits were randomly divided into 4 groups (Groups A, B, C and the normal saline group, n=3) the leaching liquid(10 ml/kg) was injected through the ear vein,and the body temperature was recorded within 3 hours. In the in vivo degradable test, the component materials (10 mm×10 mm) were implanted in 12 rabbits at 2, 4, 8, 12, 16 and 24 weeks, respectively, after operation. Two rabbitswere sacrificed for the macroscopic and the microscopic examinations. Results The chest wall component materials had no hemolytic reaction, no acute systemic toxicity, and no pyrogen reaction. The results demonstrated that the implanted materials had only a mild inflammatory reaction during the early days of the grafting, which subsided gradually. There was no tissue denaturation, necrosis or pathological hyperplasia when the prosthesis materials were degraded. Conclusion The degradable materials of the chest wall prosthesis have a good biocompatibility and agreat biological safety though their surgical application still requires a further clinical research.
