目的 探讨对Mirizzi综合征实施临床合理有效的手术方法。方法 自1990年1月至2003年12月期间，我院采用经肝放置胆道支撑引流管治疗Ⅱ、Ⅲ型Mirizzi综合征21例,胆道支撑引流管放置6个月以上，并行胆道造影检查。结果 所有患者恢复良好,胆道造影检查见胆道通畅后拔除支撑引流管，随访2～10年,无并发症发生。结论 经肝放置胆道支撑引流管治疗Ⅱ、Ⅲ型Mirizzi综合征,是保持胆道生理功能完整的有效方法。
Objective To investigate the effect of dynamic compression and rotation motion on chondrogenesis of the 3rd passage cell-loaded three-dimensional scaffold in a joint-specific bioreactor in vitro so as to provide theoretical basis of the autologous chondrocyte transplantation in clinical practice. Methods Primary chondrocytes were isolated and cultured from the knee cartilage of 3-4 months old calves. The 3rd passage cells were seeded onto fibrin-polyurethane scaffolds (8 mm × 4 mm). Experiment included 5 groups: unloaded culture for 2 weeks (group A), direct load for 2 weeks (group B), unloaded culture for 4 weeks (group C), direct load for 4 weeks (group D), and unload for 2 weeks followed by load for 2 weeks (group E). The cell-scaffold was incubated in incubator (unload) or in a joint-specific bioreactor (load culture). At different time points, the samples were collected for DNA and glycosaminoglycan (GAG) quantification detect; mRNA expressions of chondrogenic marker genes such as collagen type I, collagen type II, Aggrecan, cartilage oligomeric matrix protein (COMP), and superficial zone protein (SZP) were detected by real-time quantitative PCR; and histology observations were done by toluidine blue staining and immunohistochemistry staining. Results No significant difference was found in DNA content, GAG content, and the ratio of GAG to DNA among 5 groups (P gt; 0.05). After load, there was a large number of GAG in the medium, and the GAG significantly increased with time (P lt; 0.05). The mRNA expression of collagen type I showed no significant difference among 5 groups (P gt; 0.05). The mRNA expression of collagen type II in group B was significantly increased when compared with group A (P lt; 0.01), and groups D and E were significantly higher than group C (P lt; 0.01); the mRNA expression of Aggrecan in groups D and E were significantly increased when compared with group C (P lt; 0.01), and group E was significantly higher than group D (P lt; 0.01); the mRNA expression of COMP in group B was significantly increased when compared with group A (P lt; 0.01), and group E was significantly higher than group C (P lt; 0.01); and the mRNA expression of SZP in group E was significantly increased when compared with groups C and D (P lt; 0.05). The toluidine blue staining and immunohistochemistry staining displayed that synthesis and secretion of GAG could be enhanced after load; no intensity changes of collagen type I and collagen type II were observed, but intensity enhancement of Agrrecan was seen in groups D and E. Conclusion Different dynamic loads can promote chondrogenesis of the 3rd passage chondrocytes. Culture by load after unload may be the best culture for chondrogenesis, while the 3rd passage chondrocytes induced by mechanical load hold less capacity of chondrogenesis.
【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 study the effectiveness of one-stage repairing pharyngeal defect with the tongue flaps after resection of advanced stage hypopharyngeal neoplasm and laryngeal neoplasm. Methods Between June 2006 and March 2011, 20 patients with hypopharyngeal neoplasm (8 cases) and laryngeal neoplasm (12 cases) with advanced stage were treated. There were 19 males and 1 female, aged 47-78 years (mean, 62.8 years). All neoplasms were squamous cell carcinomas. The disease duration was 1-8.5 months (mean, 3.9 months). According to the standards of International Union Against Cancer (UICC, 1987), 12 cases were in stage III and 8 cases were in stage IV. The size of pharyngeal defect was 5 cm × 2 cm to 4 cm × 4 cm after resection of tumor. Defects were repaired by the whole base of the tongue flaps in 16 cases and by the horizontal base of the tongue flaps in 4 cases. The size of the flaps ranged from 5 cm × 2 cm to 4 cm × 4 cm. Postoperative radiotherapy and chemotherapy were regularly performed. Results The 20 tongue flaps were alive. Healing of incision by first intention was achieved in 18 cases and delayed healing in 2 cases because of subcutaneous fluid. The patients were followed up 12-63 months (mean, 36.7 months). The patients had normal feeding ability and tongue function. Of 20 cases, 12 died and 1 of local recurrence was alive with tumor. The 3-year survival rate was 69.2% (9/13). Conclusion One-stage repair of pharyngeal defect with the tongue flaps after resection of hypopharyngeal neoplasm and laryngeal neoplasm can obtain good effectiveness because the tongue flap is easy-to-obtain and easy-to-survive, and has abundant blood supply.
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 expression levels and significance of vascular endothel ial growth factor (VEGF) and microvessel density (MVD) in rabbit radius defects repaired with allogeneic and autogenic bone. Methods Forty adult New Zealand rabbits were chosen, and 10 mm bone defect model was created in the bilateral radii of 28 experimental rabbits. The other 12 rabbits provided allogeneic bone under the standard of American Association of Tissue Bank. In the left side, allogeneic bone were used to repair bone defect (experimental group), equal capacity autogenous il iac bone was used in the right side (control group). Animals were sacrificed at 2, 4, 8, and 12 weeks postoperatively. Immunohistochemical method was used to determine the expression of VEGF, CD34 protein and MVD counting. Bone histomorphometric parameters, including percent trabecular area (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp) were measured by von Kossa staining undecalcified sl ices. The relation was analyzed between VEGF and MVD, histomorphometric parameters. Results The positive signals of VEGF protein were detected in cytoplasm of vascular endothel ial cells, chondrocytes, osteoblasts, fibroblasts and osteoclasts. At 2 weeks, there was no significant difference in VEGF protein expression between experimental group and control group (P gt; 0.05); at 4 and 8 weeks, the expression of VEGF in control group was significantly higher than that in experimental group (P lt; 0.05); and at 12 weeks, there was no significant difference between two groups (P gt; 0.05). There was a positive correlation (P lt; 0.01) between VEGF expression and MVD value in two groups at 2, 4, 8, and 12 weeks postoperatively. There was no significant difference in bone histomorphometric parameters (BV/TV, Tb.Th, Tb.N, Tb.Sp) between two groups at 12 weeks postoperatively (P gt; 0.05), but there was a positive correlation between VEGF expression and parameters of BV/TV, Tb.Th, and Tb.N (P lt; 0.01); and a negative correlation between VEGF and Tb.Sp (P lt; 0.01). Conclusion VEGF can express diversity at different time and positions, and the different expressions indicated various biology significances in the process of the bone heal ing. It can coordinate growth of cartilage and bone and profit vascular reconstruction of allogeneic bone. VEGF may participate in promoting osteogenesis in the course of allogeneic bone transplantation.
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.
Objective To report 4 methods of reconstructing soft tissue defects in oral and maxillofacial regions after tumors resection using cervical pedicle tissue flaps. Methods One hundred seventy-two soft tissue defects were repaired with cervical myocutaneous flaps after resection of oral and facial cancer( 165 cases of squamous cell carcinoma and 7 cases of salivary carcinoma). The clinical stage of the tumors was stage Ⅰ in 21 cases, stage Ⅱ in 116 cases and stage Ⅲin 35 cases. Primary sites of the lesions were the tongue (59 cases), buccal mucosa (55 cases), lower gingiva (26 cases), floor of the mouth (25 cases), parotid gland (4 cases) and oropharynx (3 cases). Infrahyoid myocutaneous flaps were used in 60 cases, platysma flaps in 45 cases, sternocleidomastoid flaps in 59 cases and submental island flaps in 8 cases. The sizes of skin paddle ranged from 2.5 cm×5.0 cm to 5.0 cm ×8.0 cm. Results Among 153 survival flaps, there were55 infrahyoid myocutaneous flaps, 40 platysma flaps, 52 sternocleidomastoid flaps and 6 submental island flaps. There were 11 cases of total flap necrosis and8 cases of partial flap necrosis. The success rates were 91.67%(55/60) for infrahyoid myocutaneous flap, 88.89%(40/45) for platysma flap, 88.14% (52/59) for sternocleidomastoid flap and 75%(6/8) for submental island flap. After a follow-up of 3 11 years(5.7 years on average) among 101 cases local reccurence in 18 cases, cervical reccurence in 4 cases, distance metastasis in 2 cases. The survical rate at 3 years were 83.17%(84/101). Conclusion Cervical pedicle tissue flaps haveclinical value in reconstruction of small and medium-sized soft tissue defects after resection of oral and maxillofacial tumors.