【Abstract】 Objective To investigate the effectiveness of the medical calcium sulfate—OsteoSet bone graft substitute in the treatment of defect after excision of jaw cyst. Methods Between December 2009 and May 2010, 15 cases of jaw cystic lesion were treated，including 9 males and 6 females with an average age of 36.6 years (range, 15-75 years). Orthopantomography (OPT) method was used to measure the cyst size before operation, and the size ranged from 1.5 cm × 1.5 cm to 8.0 cm × 3.0 cm. The range of bone defect was from 1.5 cm × 1.5 cm × 1.5 cm to 8.0 cm × 3.0 cm × 3.0 cm after cyst excision intraoperatively. The patients underwent cyst curettage and OsteoSet bone graft substitutes implantation (2-15 mL). Radiological method was used to evaluate the repair effect of OsteoSet pellets. Results The pathology biopsy was periapical cyst in 7 cases, odontogenic keratocyst in 5 cases, and dentigerous cyst in 3 cases. Fifteen patients were followed up 6-12 months. Thirteen patients achieved wound healing by first intention; 2 cases had longer drainage time (5 and 7 days, respectively), the incision healed after the pressure bandage. Swelling occurred in 1 case after 1 month with no symptom of infection. No postoperative infection and rejection was found. The X-ray examination showed that the materials filled the bone defect well after 1 day of operation. OsteoSet bone graft substitutes were absorbed by one-half after 1 month of operation and totally after 3 months by OPT. The low density area was smaller in the original cysts cavity, and high density in the cysts increased significantly with fuzzy boundaries of cysts. At 6 months after operation， there was no obvious difference in image density between the original cavity and normal bone, and the capsule cavity boundary disappeared, and defect area was full of new bone. Conclusion The medical calcium sulfate—OsteoSet bone graft substitute is an ideal filling material for bone defect.
Objective To evaluate the effectiveness of the submental island flap for repair of oral defects after radical resection of early-stage oral squamous cell carcinoma (OSCC). Methods Between February 2010 and August 2011, 15 cases of early-stage OSCC were treated. Of 15 cases, 9 were male and 6 were female, aged from 48 to 71 years (mean, 63 years). The disease duration was 28-73 days (mean, 35 days). Primary lesions included tongue (3 cases), buccal mucosa (8 cases), retromolar area (2 cases), and floor of mouth mucosa (2 cases). According to TNM classification of International Union Against Cancer (UICC, 2002) of oral cancer and oropharyngeal cancer, 2 cases were classified as T1N0M0 and 13 cases as T2N0M0. The results of the pathologic type were high differentiated squamous cell carcinoma in 11 cases and moderately differentiated squamous cell carcinoma in 4 cases. The defect after resection of the lesion ranged from 5 cm × 3 cm to 8 cm × 6 cm. All the cases underwent radical resection of the primary lesion and immediate reconstruction with submental island flap except 1 case with radial forearm free flap because of no definite venous drainage. The sizes of the submental island flap varied from 6 cm × 4 cm to 9 cm × 6 cm. Results Operation time ranged from 4 hours and 30 minutes to 7 hours and 10 minutes (mean, 5 hours and 53 minutes) in 14 cases undergoing repair with submental island flap. All the flaps survived completely in 13 cases except 1 case having superficial necrosis of the flap, which was cured after conservative treatment. Temporary marginal mandibular nerve palsy occurred in 1 case, and was cured after 3 months; submandibular effusion was observed in 3 cases, and was cured after expectant treatment. The follow-up period ranged from 8 to 15 months (mean, 10.5 months) in 14 cases undergoing repair with submental island flap. Hair growth was seen on the flap and became sparse after 3 months in 2 male cases. The appearance of the face, opening mouth, swallowing, and speech were recovered well in 14 cases, and the donor site had no obvious scar. The follow-up period was 13 months in 1 case undergoing repair with radical free forearm flap, and the appearance and function were recovered well. No local recurrence was found during follow-up. Conclusion The submental island flap has reliable blood supply, and could be harvested simply and rapidly. It can be used to repair oral defects in patients with early-stage OSCC after radical resection.
Objective To construct a new type of self-assembling peptide nanofiber scaffolds—RGDmx, and to study the cell compatibility of the new scaffolds and the proliferation and chondrogenic differentiation of precartilaginous stem cells(PSCs) in scaffolds. Methods PSCs were separated and purified from newborn Sprague Dawley rats by magnetic activated cell sorting and indentified by immunohistochemistry and immunofluorescent staining. The RGDmx were constructed by mixing KLD-12 and KLD-12-PRG at volume ratio of 1 ∶ 1. PSCs at passage 3 were seeded into the KLD-12 scaffold (control group) and RGDmx scaffold (experimental group). The proliferation of PSCs in 2 groups were observed with the method of cell counting kit (CCK) -8 after 1, 3, 7, and 14 days after culture. The RGDmx were constructed by mixing KLD-12-PRG and KLD-12 at different volume ratios of 0, 20%, 40%, 60%, 80%, and 100% and the prol iferation of PSCs was also observed. The complete chondrogenic medium (CCM) was used to induce chondrogenic differentiation of PSCs in different scaffolds. The differentiation of PSCs was observed by toluidine blue staining and RT-PCR assay. Results PSCs were separated and purified successfully, which were identified by immunohistochemistry and immunofluorescent staining methods. The results of CCK-8 showed that the absorbance (A) value in the experimental group increased gradually and reached the highest at 7 days; the A value in the experimental group was significantly higher than that in the control group at 7 days and 14 days (P lt; 0.05). Meanwhile, the A value in the RGDmx scaffold with a volume ratio of 40% was significantly higher than those in others (P lt; 0.05). After 14 days of induction culture with CCM, the toluidine blue staining results were positive in 2 groups; the results of RT-PCR showedthat the expression levels of collagen type II and the aggrecan in the experimental group were significantly higher than those in the control group (P lt; 0.05). Conclusion The self-assembling peptide nanofiber scaffold—RGDmx is an ideal scaffold for tissue engineer because it has good cell compatibility and more effective properties of promoting the differentiation of PSCs to chondrocytes.
Objective To compare the effect between vascularization osteogenesis and membrane guided osteogenesis in the bone repair by the tissue engineered bone with pedicled fascial flap packing autologous red bone marrow (ARBM), so as to provide a reference for the bone defect repair in cl inic. Methods The tissue engineered bone was constructed with ARBM and the osteoinductive absorbing recombinant human materials with recombinant human bone morphogenetic protein 2. Sixty New Zealand rabbits (aged 4-5 months, weighing 2.0-2.5 kg) were randomly divided into group A (n=16), group B (n=22), and group C (n=22). The complete periosteum defect model of 1.5 cm in length was prepared in right ulnar bone, then the tissue engineered bone was implanted in the bone defect area in group A, the tissue engineered bonewith free fascial flap in group B, and the tissue engineered bone with pedicled fascial flap in group C. At 4, 8, 12, and 16 weeks, the tissue of bone defect area was harvested from 4 rabbits of each group for the general, histological, and immunohistochemical staining observations; at 8, 12, and 16 weeks, 2 rabbits of groups B and C, respectively were selected to perform ink perfusion experiment by axillary artery. Results The general observation showed that the periosteum-l ike tissues formed in the fascial flap of groups B and C, chondroid tissues formed in group B, new bone formed in group C, and the fibrous and connective tissues in group A at 4 and 8 weeks; a few porosis was seen in group A, more new bone in group B, and bone stump formation in group C at 12 and 16 weeks. Histological observation showed that there were few new blood vessels and new bone trabeculae in groups A and B, while there were large amounts of new blood vessels and mature bone trabeculae in group C at 4 and 8 weeks. There were a few new blood vessels and new bone trabeculae in group A; more blood vessels, significantly increased mature trabeculae, and the medullary cavity formation in group B; and gradually decreased blood vessels, the mature bone structure formation, and the re-opened medullary cavity in group C at 12 and 16 weeks. The immunohistochemical staining observation showed that the levels of CD105, CD34, and factor VIII were higher in group C than in groups A and B at different time points.The bone morphometry analysis showed that the trabecular volume increased gradually with time in 3 groups after operation; the trabecular volume in group C was significantly more than those in groups A and B at different time points (P lt; 0.05); and there was significant difference between groups A and B (P lt; 0.05) except the volume at 4 weeks (P gt; 0.05). The vascular image analysis showed that the vascular regenerative area ratio in group C was significantly higher than those in groups A and B at different time points (P lt; 0.05). The ink perfusion experiment showed that the osteogenic zone had sparse ink area with no obvious change in group B, while the osteogenic zone had more intensive ink area and reached the peak at 8 weeks, then decreased in group C. Conclusion The tissue engineered bone with pedicled fascial flap packing ARBM has the vascularization osteogenesis effect at early stage, but the effect disappears at late stage gradually when the membrane guided osteogenesis is main.
Objective To investigate the effect of simvastatin on inducing endothel ial progenitor cells (EPCs) homing and promoting bone defect repair, and to explore the mechanism of local implanting simvastatin in promoting bone formation. Methods Simvastatin (50 mg) compounded with polylactic acid (PLA, 200 mg) or only PLA (200 mg) was dissolved in acetone (1 mL) to prepare implanted materials (Simvastatin-PLA material, PLA material). EPCs were harvested from bone marrow of 2 male rabbits and cultured with M199; after identified by immunohistochemistry, the cell suspension of EPCs at the 3rd generation (2 × 106 cells/mL) was prepared and transplanted into 12 female rabbits through auricular veins(2 mL). After 3 days, the models of cranial defect with 15 cm diameter were made in the 12 female rabbits. And the defects were repaired with Simvastatin-PLA materials (experimental group, n=6) and PLA materials (control group, n=6), respectively. The bone repair was observed after 8 weeks of operation by gross appearance, X-ray film, and histology; gelatin-ink perfusion and HE staining were used to show the new vessels formation in the defect. Fluorescence in situ hybridization (FISH) was performed to show the EPCs homing at the defect site. Results All experimental animals of 2 groups survived to the end of the experiment. After 8 weeks in experimental group, new bone formation was observed in the bone defect by gross and histology, and an irregular, hyperdense shadow by X-ray film; no similar changes were observed in control group. FISH showed that the male EPC containing Y chromosome was found in the wall of new vessels in the defect of experimental group, while no male EPC containing Y chromosome was found in control group. The percentage of new bone formation in defect area was 91.63% ± 4.07% in experimental group and 59.45% ± 5.43% in control group, showing significant difference (P lt; 0.05). Conclusion Simvastatin can promote bone defect repair, and its mechanism is probably associated with inducing EPCs homing and enhancing vasculogenesis.
Aiming at the problem of scaffold degradation in bone tissue engineering, we studied the feasibility that controlls bone defect repair effect with the inhomogeneous structure of scaffold. The prediction model of bone defect repair which contains governing equations for bone formation and scaffold degradation was constructed on the basis of analyzing the process and main influence factors of bone repair in bone tissue engineering. The process of bone defect repair and bone structure after repairing can be predicted by combining the model with finite element method (FEM). Bone defect repair effects with homogenous and inhomogeneous scaffold were simulated respectively by using the above method. The simulation results illustrated that repair effect could be impacted by scaffold structure obviously and it can also be controlled via the inhomogeneous structure of scaffold with some feasibility.
ObjectiveTo investigate the feasibility of tissue engineered periosteum (TEP) constructed by porcine small intestinal submucosa (SIS) and bone marrow mesenchymal stem cells (BMSCs) of rabbit to repair the large irregular bone defects in allogenic rabbits. MethodsThe BMSCs were cultivated from the bone marrow of New Zealand white rabbits (aged, 2 weeks-1 month). SIS was fabricated by porcine proximal jejunum. The TEP constructed by SIS scaffold and BMSCs was prepared in vitro. Eighteen 6-month-old New Zealand white rabbits whose scapula was incompletely resected to establish one side large irregular bone defects (3 cm×3 cm) model. The bone defects were repaired with TEP (experimental group,n=9) and SIS (control group,n=9), respectively. At 8 weeks after operation, the rabbits were sacrificed, and the implants were harvested. The general condition of the rabbits was observed; X-ray radiography and score according to Lane-Sandhu criteria, and histological examination (HE staining and Masson staining) were performed. ResultsAfter operation, all animals had normal behavior and diet; the incision healed normally. The X-ray results showed new bone formation with normal bone density in the defect area of experimental group; but no bone formation was observed in control group. The X-ray score was 6.67±0.32 in experimental group and was 0.32±0.04 in control group, showing significant difference (t=19.871,P=0.001). The general observation of the specimens showed bone healing at both ends of the defect, and the defect was filled by new bone in experimental group; no new bone formed in the control group. The histological staining showed new bone tissue where there were a lot of new vessels and medullary cavity, and no macrophages or lymphocytes infiltration was observed in the defect area of experimental group; only some connective tissue was found in the control group. ConclusionTEP constructed by porcine SIS and BMSCs of rabbit can form new bone in allogenic rabbit and has the feasibility to repair the large irregular bone defects.
ObjectiveTo investigate the application of three-dimensional (3-D) printing technique combining with 3-D CT and computer aided-design technique in customized artificial bone fabrication, correcting mandibular asymmetry deformity after mandibular angle ostectomy. MethodsBetween April 2011 and June 2013, 23 female patients with mandibular asymmetry deformity after mandibular angle ostectomy were treated. The mean age was 27 years (range, 22-34 years). The disease duration of mandibular asymmetry deformity was 6-16 months (mean, 12 months). According to the CT data and individualized mandibular angle was simulated based on mirror theory, 3-D printed implants were fabricated as the standard reference for manufacturers to fabricated artificial bone graft, and then mandible repair operation was performed utilizing the customized artificial bone to improve mandibular asymmetry. ResultsThe operation time varied from 40 to 60 minutes (mean, 50 minutes). Primary healing of incisions was obtained in all patients; no infection, hematoma, and difficulty in opening mouth occurred. All 23 patients were followed up 3-10 months (mean, 6.7 months). After operation, all patients obtained satisfactory facial and mandibular symmetry. 3-D CT reconstructive examination results after 3 months of operation showed good integration of the artificial bone. Conclusion3-D printing technique combined with 3-D CT and computer aided design technique can be a viable alternative to the approach of maxillofacial defects repair after mandibular angle ostectomy, which provides a accurate and easy way.
ObjectiveTo explore the application of three-dimensional (3-D) printing technique in repair and reconstruction of maxillofacial bone defect. MethodsThe related literature on the recent advance in the application of 3-D printing technique for repair and reconstructing maxillofacial bone defect was reviewed and summarized in the following aspects:3-D models for teaching, preoperative planning, and practicing; surgical templates for accurate positioning during operation; individual implantable prosthetics for repair and reconstructing the maxillofacial bone defect. Results3-D printing technique is profoundly affecting the treatment level in repair and reconstruction of maxillofacial bone defect. Conclusion3-D printing technique will promote the development of the repair and reconstructing maxillofacial bone defect toward more accurate, personalized, and safer surgery.
ObjectiveTo investigate the effects of micro-fracture and insul in-l ike growth factor 1 (IGF-1) in treatment of articular cartilage defect in rabbits. MethodsTwenty-four New Zealand white rabbits (aged, 4-6 months; weighing, 2.5-3.5 kg) were randomly divided into 4 groups (n=6):micro-fractures and recombinant human IGF-1 (rhIGF-1) treatment group (group A), micro-fracture control group (group B), rhIGF-1 treatment control group (group C), and blank control group (group D). Full thickness articular cartilage defects of 8 mm×6 mm in size were created in the bilateral femoral condyles of all rabbits. The micro-fracture surgery was performed in groups A and B. The 0.1 mL rhIGF-1 (0.01 μg/μL) was injected into the knee cavity in groups A and C at 3 times a week for 4 weeks after operation, while 0.1 mL sal ine was injected in groups B and D at the same time points. At 4, 12, and 24 weeks, the gross, histological, and immunohistochemical observations were performed, and histological score also was processed according to Wakitani's score criteria. The collagen contents in the repair tissues and normal patellofemoral cartilage were detected by the improved hydroxyproline (HPR) method at 24 weeks. Electron microscope was used to observe repair tissues of groups A and B at 24 weeks. Results All animals were survival at the end of experiment. At 24 weeks after operation, defect was repaired with time, and the repair tissue was similar to normal cartilage in group A; the repair tissue was even without boundary with normal cartilage in group B; and the repair tissue was uneven with clear boundary with normal cartilage in groups C and D. Histological staining showed that the repair tissues had no difference with normal cartilage in group A; many oval chondrocytes-l ike cells and l ight-colored matrix were seen in the repair tissues of group B; only a few small spindle-shaped fibroblasts were seen in groups C and D. Moreover, histological scores of group A were significantly better than those of groups B, C, and D (P<0.05) at 4, 12, and 24 weeks. Electron microscope observation showed that a large number of lacuna were seen on the surface of repair tissue in group A, and chondrocytes contained glycogen granules were located in lacunae, and were surrounded with the collagen fibers, which was better than that in group B. Collagen content of the repair tissue in group A was significantly higher than that in groups B, C, and D (P<0.05), but it was significantly lower than that of normal cartilage (P<0.05). Conclusion Combination of micro-fracture and rhIGF-1 for the treatment of full thickness articular cartilage defects could promote the repair of defects by hyaline cartilage.