Abstract To investigate the ectopic new bone formation following implantation of bovine hydroxyapatite Bio-oss together with free periosteum, 12 chabb: ch rabbits were selected. In 10 rabbits, Bio-oss block together with free periosteum was implanted in the gastrocnemius muscle of one leg randomly, and Bio-oss block alone was implanted in the same muscle of the other leg. In the other 2 rabbits, the periosteum was implanted into the gastrocnemius musle of both legs. Histologic examination and quantitative analysis of newbone formation were performed at 3 and 6 weeks postoperatively. The results showed that in the legs implanted bovine hydroxyapatite Bio-oss together with freeperiosteum, new bone formation began at 5th day after implantation. The area ofnew bone composed of 19.0% of the specimens at 3 weeks postoperatively. No boneformation through out the experimental period in Bio-oss block alone implantedlegs and also periosteum implanted legs. We concluded that bovine hydroxyapatite Bio-oss has a good capacity of osteoconduction. New bone can be formed after the implantation of hydroxyapatite combined with free periosteum.
Abstract To restore the bone defect after curettage of bone cyst, hydroxyapatite bioactive microcrystal glass (HBG) was used. From 1990 to 1995, HBG was applied in 17 cases. The bone involved were humerus, femur, tibia and fibula. Among them, 6 were complicated with pathological fracture. After eradication of the focus, the cyst was filled in ZnCl2 powder and irrigated with saline, then particles or segments of HBG were implanted into the cavity. The fracture were fixed with Enders rod. All the extremities were immobilized with plaster splint for about 6 to 8 weeks. Three months later, the lower limbs began to have functional exercises. By X-ray examination, the border between HBG and bone was clear in 2 weeks, after 1 month the clear border become blurred, and 2 months after operation, HBG was intermingled with bone. After 1 year there was neither absorption of bone nor HBG. No recurrence of the aptic lesion occurred in 1 year. HBG was a kind of artificial bone composed of hydroxyapatite and bioactive microcrystal glass, the latter contained silicon.It was characterized by its bioactivity, osteoinductivity and good tissue compatibility. The microcrystal would facilitate the growth of osseous tissues, which caused HBG intermingled with the surrounding bone. The source of HBG was abundant. It might be an ideal artificial bone.
Objective To evaluate the feasibility and the value of the layered cylindric collagenhydroxyapatite composite as a scaffold for the cartilage tissue engineering after an observation of how it absorbs the chondrocytes and affe cts the cell behaviors. Methods The chondrocytes were isolated and multiplied in vitro, and then the chondrocytes were seeded onto the porous collagen／h ydro xyapatite composite scaffold and were cultured in a three-dimensional environme n t for 3 weeks. The effects of the composite scaffold on the cell adhesivity, proliferation, morphological changes, and synthesis of the extracellular matrix were observed by the phase-contrast microscopy, histology, scanning electron micros copy, and immunohistochemistry. Results The pore diameter of the upper layer of the collagen-hydroxyapatite composite scaffold was about 147 μm. and the porosity was 89%; the pore diameter of the bottom layer was about 85 μm and the porosity was 85%. The layered cylindric collagenhydroxyapatite composite scaffold had good hydrophilia. The chondrocytes that adhered to the surface of the scaffold, proliferated and migrated into the scaffold after 24 hours. The chondrocytesattached to the wall of the microholes of the scaffold maintained a rounded morphology and could secrete the extracellular matrix on the porous scaffold. Conclusion The layered cylindric collagenhydroxyapatite composite scaffold has a good cellular compatibility, and it is ber in the mechanical property than the pure collagen. It will be an ideal scaffold for the cartilage tissue enginee ring.
Natural nonorganic bone(NNB) was obtained after the fresh bone of pig was heated to 100℃. The NNB was white and in a shape as its original bone.The tensile strength of the compact bone was 200kg/cm3 and that of the cancellous bone was 25kg/cm3. The ratio of calcium to phosphorus was 10∶6. The main componentwas hydroxyapatite. The material was composed of trabeculae and intertrabecular spaces. Three experiments were performed. Experiment 1: 18 pieces of NNB in a size of 0.5×0.5×0.5cm3 were implanted intothe back muscle of 18 rabbits. At 4, 8 and 12 weeks after the operation, 6 specimens were obtained seperately and were stained by HE, and then examined under microscope. The result showed that the mesenchymal cells had no regeneration and differentiation, and the NNB and the surrounding tissues had no evidence of formation of new bone or chondrosynthesis. This NNB did not produce rejection reaction between tissues but the new blood vessels could easily grow into the space of the NNB. The fibrils had intimate contact with NNB. Experiment 2: The NNB and hydroxyapatite(HA) were mixed to make a cylindroid body with 2mm in diameter and 4mm in length and was implanted in the bilateral tibias of 40 rabbits, respectively. The roentgenography, fluroscent microscopic examination and histological observation were carried out at 2, 3, 4 and 8 weeks after the implantation. Experiment 3: In 50 rabbits, a defect of 2.5cm was made on both radius, and in one sideNNB was implanted and the other side was served as the control. Another 50 rabbitsHA was implanted in the defect in one side and the other side was served as the control. The results showed that in the NNB group at the 16th week, the bone united in 16 of the 30 cases, while in the HA group, in the 30 cases,only 2 had the bone united, while those of the controls no union had occurred.It was suggested that NNB had more formation of new bone than HA did.
ObjectiveTo explore the effect of hydroxyapatite nanoparticle (nHAP) on hepatocellular carcinoma (HCC) and its mechanisms. MethodsThe literatures about the effect of nHAP on HCC were reviewed and summarized. ResultsAs a new nanoparticle, nHAP could suppress the DNA synthesis and subsequent division and proliferation of HCC cells through the inhibition of proliferating cell nuclear antigen (PCNA) and telomerase gene expression and increase of intracellular Ca2+. Moreover, nHAP was able to suppress the differentiation and metastases of HCC cells through the effect on the expressions of Paxillin and P130cas and the decrease of expressions of multiple drug resistance gene protein, microvessel density, and vascular endothelial growth factor. Finally, nHAP induced the apoptosis of HCC tumor cells by the regulation of bcl-2 and bax protein expressions. The combined use of nHAP and chemoembolization drugs could enhance the efficacy, prolong drug duration and reduce toxicity. ConclusionnHAP can inhibit the division, proliferation, differentiation, and metastases, and promote the apoptosis of HCC cells and combined use with chemoembolization drugs can enhance the efficacy and reduce toxicity.
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.
OBJECTIVE To confirm membrane-guided tissue regeneration in the healing course of segmental bone defects and study the mechanism. METHODS Segmental, 1 cm osteoperiosteal defects were produced in both radii of 12 rabbits. One side was covered with hydroxyapatite/polylactic acid(HA/PLA) membrane encapsulated as a tube. The contralateral side served as an untreated control. Healing courses were detected by radiographic and histologic examinations. RESULTS All control sides showed nonunion, whereas there were consistent healing pattern in test sides. CONCLUSION Membrane technique can promote bone regeneration.
Repair of bone defect by compound of bone morphogenic protein (BMP) and its prior bone matrix gelatin (BMG) was compared with repair by BMP with hydroxyapatite(HA). The results showed that the BMP/BMG group was found fibrous callus in the bone defect in 4th week. In 8th week a large quantity of osseos trabecula was found. In 12th week the BMP was absorbed completely and was replaced by newly formed bone. In 16th week the recanalization appeared in the bone cavity. While in the BMP/HA group, although the fibrous callus was appeared in the 4th and 8th weeks, the HA was not absorbed. In the 12th and 16th weeks the change was similar to that in the 8th week and no recanalization of bone marrow cavity. It was suggested the BMP/BMG compound might be an ideal material to repair the bone defect.
The primary results of five patients in whomthe block hydroxyapatite artificial bone (BHAB)used in maxillofacial plastic repair were reported. All incisions healed up with no evidence ofinfection. None of the implants was rejected norhad resorption changes. Satisfactory estheticaleffects were maintained. The results demonst-rated BHAB had a good biocampatibility andcould be used as a bone graft substitute inmaxillofacial plastic repair. This kind of material could be carved and contoured ...
ObjectiveTo evaluate the in vivo biological safety of porous zinc oxide (ZnO)/hydroxyapatite (HA) composite materials.MethodsThe porous ZnO/HA composite materials and porous HA materials were prepared by the spark plasma sintering technology. First, the materials were characterized, including scanning electron microscopy to observe the material structure, in vitro degradation experiments to detect the degradation rate of the materials, and inductively coupled plasma emission spectrometer to detect the concentration of Zn2+ dissolved out of the composite material degradation. Then the two kinds of material extracts were prepared for acute systemic toxicity test. Fifteen male Kunming mice were randomly divided into groups A, B, and C (n=5) and injected intraperitoneally with normal saline, HA extracts, and ZnO/HA extracts, respectively. The body mass of the mice was recorded before injection and at 24, 48, and 72 hours after injection. The liver and kidney tissues were taken at 72 hours for HE staining to evaluate the safety of the composite material. Finally, the biological safety of the material in vivo was evaluated by implantation experiment. The eighteen male New Zealand white rabbits were randomly divided into HA group and ZnO/HA group (n=9); a bilateral radius defect model (1 cm) was established, and the right forelimbs of the two groups were implanted with porous HA materials and porous ZnO/HA composite materials, respectively; the left untreated as a blank control. The general condition of the animals were observed after operation. The rabbit blood was collected at 1 day before operation and at 1 day, 1 week, 4 weeks, and 8 weeks after operation for routine blood test (inflammation-related indicators) and blood biochemistry (liver and kidney function-related indicators). X-ray films were taken at 4, 8, and 12 weeks after operation to observe the repair of bone defects.ResultsMaterial characterization showed that porous ZnO/HA composite materials had interconnected large and small pore structures with a pore size between 50 and 500 μm, which degraded faster than porous HA materials, and continuously and slowly dissolved Zn2+. The acute systemic toxicity test showed that the mice in each group had no abnormal performance after injection, and the body mass increased (P<0.05). HE staining showed that the cells shape and structure of liver and kidney tissue were normal. Animal implantation experiments showed that all rabbits survived until the experiment was completed; routine blood tests showed inflammation in each group (neutrophils, monocytes, and lymphocytes increased) at 1 day after operation, and all returned to normal at 8 weeks (P>0.05); compared with 1 day before operation, the content of inflammatory cells in the HA group increased at 1 day, 1 week, and 4 weeks after operation (P<0.05), and the ZnO/HA group increased at 1 day after operation (P<0.05); blood biochemistry showed that the liver and kidney function indexes were in the normal range; X-ray films showed that the ZnO/HA group had better osseointegration than the HA group at 4 weeks after operation.ConclusionThe porous ZnO/HA composite material has good in vivo biological safety and good bone repair ability, which is a potential bone repair material.