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.
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 ...
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.
OBJECTIVE: To study the effect of collagen/hydroxyapatite(CHA) instead of autogenous bone transplantation on repairing the mandibular defects. METHODS: Ten Chinese experimental minipigs were made 2 cm bone defects in diameter in the mandible. The experimental group was implanted CHA, while the control group was implanted autogenous bone. The basic parameters of bone dynamics were determined by bone metrology. RESULTS: There was remarkable difference between the two groups in the mean distance and mineralization apposition rate of double label bands marked by tetracycline(P lt; 0.05), while the mean osteoid seam width and mineralization lag time had no remarkable difference(P gt; 0.05). It suggested that CHA had good osteogenesis. The collagen in CHA offered the condition of bone mineralization, and the mineralization peak of experimental group was present at 4 weeks earlier than that of control group (8 weeks). CONCLUSION: CHA may be a substitute of autogenous bone transplantation in repairing the mandibular defects, and the second operation for offering the implanting bone is avoidable, therefore, CHA may be an ideal material to repair bone defects.
The hydroxyapatite particles were used to repair 23 cases of depressed deformities of face. The patients were follwed up for 3 to 8 months and the short termresults were satisfactory. The operative procedure was briefly introduced. The advantages and attentions relevant to the operation were discussed.
To observe the collagen-hydroxylaptite composite in the repair of bone defect, ten minipigs were chosen to make a mandibular dafect measuring 2 cm in diameter and the composite was implanted, while the use of autogenous bone graft and the blank wese served as control. On the 4, 8, 12, 24 and 48 weeks after the operation, the animals were sacrificed and the samples were examined under light microscope. The result showed that: no infection or necrosis occurred. The composite coalesced with host bone and the outcome was similar to that of the autogenous bone graft. No foreign body giant cells or vacuum left from osteonecrosis was observed. It was suggested that the composite had the advantage of abundant supply, easy to handle and no harm. The biocompatibility was good and might be hopeful as a bone substitute.
Objective To investigate the antibacterial and osteogenic capabil ities in vivo of hydroxyapatite (HA)/silver (Ag) coating. Methods HA/Ag coating (Ag qual ity percentage was 3%) and HA coating were deposited to external fixator Schanz screws. The tibial fracture model was establ ished in right hindl imb of 18 adult male Beagle dogs (weighing 15-20 kg). Thetibia was stabil ized with an external fixator and 2 Schanz screws of HA coating at proximal tibia (control group, n=18) and HA/Ag coating at distal tibia (experimental group, n=18), and every screw incision was infected with Staphylococcus aureus. Infection in screw holes and the changes of bone-screw interface were observed by wound grading and X-ray films. Results In control group, wounds infection became worse with time (χ2=13.492, P=0.001), while in experimental group, no obvious change was observed (χ2=0.208, P=0.901). The wound grading of experimental group was significantly better than that of the control group at 1, 2, and 3 weeks (P lt; 0.05). Laser scanning confocal microscope showed that there was bacterial adhesion on the surface of screws in 2 groups, viable becteria mainly in control group and non-viable becteria mainly in experimental group. The scanning electron microscope (SEM) observation results of the fractured sclerous tissue section showed that an obvious transparent boundary between screw and bone in control group, but no obvious boundary in experimental group. The osseointegration ratios were 76.23% ± 15.54% in control group and 93.42% ± 5.53% in experimental group, showing significant difference (t=8.843, P=0.000). The SEM observation showed that HA/Ag coating integrated with new bone and the surface of implant was filled with new bone in experimental group; obvious interspace was seen between the HA coating and new bone in control group. Conclusion HA/Ag coating has good antibacterial and osteogenic capabil ities, so it can take effects in preventing infection in screw holes and loosening of implants.
Objective To investigate and compare the osteogenic potential of three kinds of calcium phosphate ceramic as carriers for recombinant human bone morphogenetic protein-2(rhBMP-2) in vivo.Methods BCPceramics (HA,TCP,HA/TCP) impregnated with rhBMP-2 (experimental groups) and without rhBMP-2(control groups) were implanted into 6 muscles pockets on the dorsum of 3month-old Wistar rabbits. The rabbits were sacrificed 2, 4 and 8 weeks after implantation and bone induction was estimated by alkaline phosphatase(ALP) activity measurement. The implants were also examined histologically and histomorphometrically by HE staining and computerized graphical analysis. Results The ALPactivity of implants withrhBMP-2 was higher than that of control groups(P＜0.05), but there was no difference between 2 and 4 weeks in experimental groups. In all experimental groups,theimplants exhibited that new bone formation increased with the lapse of time. The amount of new bone formation is more in -HA/rhBMP-2 group than in the other two group in the 2nd and 4th weeks, but there was no difference between them (P＞0.05).In the 8th week, the amount of bone formation was most in HA/TCP with -rhBMP-2, and was more than that in the 2nd and 4th weeks. Whereas in control groups, there was only fibrous connective tissue. Conclusion HA/TCP- is a good carriers of rhBMP-2 and can be used as bone substitutes clinically.
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.
ObjectiveTo construct bone morphogenetic protein 2 (BMP-2) gelatin/chitosan hydrogel sustained-release system, co-implant with induced pluripotent stem cells (iPS) derived mesenchymal stem cells (MSCs) to hydroxyapatite (HA)/zirconium dioxide (ZrO2) bio porous ceramic foam, co-culture in vitro, and to explore the effect of sustained-release system on osteogenic differentiation of iPS-MSCs.MethodsBMP-2 gelatin/chitosan hydrogel microspheres were prepared by water-in-oil solution. Drug encapsulation efficiency, drug loading, and in vitro sustained release rate of the microspheres were tested. HA/ZrO2 bio porous ceramic foam composite iPS-MSCs and BMP-2 gelatin/chitosan hydrogel sustained release system co-culture system was established as experimental group, and cell scaffold complex without BMP-2 composite gelatin/chitosan hydrogel sustained release system as control group. After 3, 7, 10, and 14 days of co-culture in the two groups, ALP secretion of cells was detected; gene expression levels of core binding factor alpha 1 (Cbfa1), collagen type Ⅰ, and Osterix (OSX) were detected by RT-PCR; the expression of collagen type Ⅰ was observed by immunohistochemical staining at 14 days of culture; and cell creep and adhesion were observed by scanning electron microscopy.ResultsBMP-2 gelatin/chitosan hydrogel sustained-release system had better drug encapsulation efficiency and drug loading, and could prolong the activity time of BMP-2. The secretion of ALP and the relative expression of Cbfa1, collagen type Ⅰ, and OSX genes in the experimental group were significantly higher than those in the control group at different time points in the in vitro co-culture system (P<0.05). Immunohistochemical staining showed that the amount of fluorescence in the experimental group was significantly more than that in the control group, i.e. the expression level of collagen type Ⅰ was higher than that in the control group. The cells could be more evenly distributed on the materials, and the cell morphology was good. Scanning electron microscopy showed that the sustained-release system could adhere to cells well.ConclusioniPS-MSCs have the ability of osteogenic differentiation, which is significantly enhanced by BMP-2 gelatin/chitosan hydrogel sustained-release system. The combination of iPS-MSCs and sustained-release system can adhere to the materials well, and the cell activity is better.