1.Allogenic chondrocytes loaded with liquid or gel biocarrier material in repairing full-thickness rabbit articular cartilage defects
Lei ZHANG ; Hongbo YU ; Xiaokun JIAO ; Xicheng ZHAI ; Nazhao GU ; Yingzhen WANG ; Zhenhua Lü ; Yougu HU ; Zonghua QI
Chinese Journal of Tissue Engineering Research 2006;10(45):190-193,插6
BACKGROUND: It has been successful to repair articular cartilage defects by using solid carrier as cytoskeleton. We tried to transplant liquid or gel carrier materials combined cells into the body of animals, and investigated its feasibility.OBJECTIVE: To investigate the feasibility of homo-transplatation with liquid or gel carrier materials of Pluronic F-127-recombinant human bone morphogenetic protein-2 (rhBMP-2) engineered chondrocytes for the repair of full-thickness rabbit articular cartilage defect.DESIGN: A controlled experiment.SETTINGS: Department of Orthopaedics, Weihai Municipal Hospital;Shandong Institute of Orthopaedics and Traumaology.MATERIALS: The experiments were carried out in the laboratory of Shandong Institute of Orthopaedics and Traumaology from November 2001 to September 2003. Thirty-six healthy adult New Zealand rabbits of 2.5-4.5 kg, either male or female, were divided into four groups according to the method of random number table: Pluronic F-127-rhBMP-2 engineered chondrocytes group, Pluronic F-127-rhBMP group, Pluronic F127 engineered chondrocytes group and blank control group, with 9 rabbits in each group.METHODS: After grouping, the 36 rabbits were made into models of articular cartilage defects. Pluronic F-127-rhBMP-2 was used as a vector of chondrocytes which were obtained from New Zealand rabbits after cultured and amplified in vitro. The mixture of Pluronic F-127, Pluronic F-127-rhBMP-2 and cultured chondrocytes was transplanted into the defects of articular cartilage that had been made previously with φb3.5 mm drill.There was not any treatment in the blank control group. At 4, 8 and 12 weeks postoperatively, the repairing conditions of the defects were evaluated with gross observation and histological observation under light microscope and under electron microscope. The repaire quality was assessed blindly according to the Wakitani scoring standard.MAIN OUTCOME MEASURES: ① Healing of cartilage defects; ② Property and morphology of the chondrocytes, characteristics, number and arrangement of collagens in matrix.RESULTS: ① In the Pluronic F-127-rhBMP-2 engineered chondrocytes group, the transplanted chondrocytes could grow better than those in other groups, the defected areas were completely filled at 4 weeks. The regenerated tissues at 8 and 12 weeks had similar appearance with the surrounding normal cartilage tissue, but vague. Delimitation. The histological examination showed that transparent cartilages formed, and the defects were healed. ② Under electron microscope at 8 and 12 weeks, there were mature transparent cartilages in the repaired tissues, and there were irregularly arranged slight, even and non-periodical collagen Ⅱ in surrounding. In the blank control group, only fibrous repair was observed, the regenerated tissue lacked elasticity with rough surface. ③ Repairing quality score: The scores at each time point in the Pluronic F-127-rhBMP-2 engineered chondrocytes group were significantly different from those in the other groups.Those in the Pluronic F-127-rhBMP-2 engineered chondrocytes group and Pluronic F-127-rhBMP-2 group and Pluronic F-127 engineered chondrocytes group were significantly different from those in the blank control group [4 weeks: (3.93±1.91), (4.56±1.07), (4.78±1.09), (8.44±1.13) points:8 weeks: (2.80±1.45), (3.24±1.00), (3.33±1.00), (8.44±1.13) points; 12 weeks (2.22±1.10), (3.01±0.69), (3.00±0.71), (9.00±0.87) points, P < 0.001],but there were no significant differences between the two groups (P > 0.05).CONCLUSION: The mixture of Pluronic F-127-rhBMP-2 and cultured chondrocytes can repair successfully the cartilage defects of femoral condyle of rabbit knees by means of hyaline cartilage than simple application of Pluronic F-127-rhBMP-2 or Pluronic F-127 engineered chondrocytes.
2.Clinical analysis of pedicle screw fixation in the treatment of type Ⅱ odontoid fracture with bone grafting and non-bone grafting
Hongbiao LI ; Xiuqin SUN ; Wenbo NIE ; Xicheng ZHAI ; Lei QIN ; Baoshuai LIU
Chinese Journal of Primary Medicine and Pharmacy 2018;25(8):1002-1005
Objective To explore the clinical efficacy of bone grafting in the treatment of type Ⅱ odontoid fracture with pedicle screw fixation .Methods Of 14 cases with type Ⅱ odontoid fractures ,8 patients in group A received pedicle screw,lamina autologous bone grafts,6 cases of group B received simple posterior pedicle screw fixation.The operative time ,the amount of surgical bleeding and the postoperative cervical spine flexion and rotationwere observed at 3 months postoperatively ,and the patients were followed up for 5-45 months.Results All patients were followed up for 5-45 months,with an average of 26.5 months.The operation time was (1.83 ±0.5) h in the bone graft group,and (1.58 ±0.9)h in the non-bone graft group,the difference was statistically significant (t=2.842,P>0.05).The blood loss of the bone graft group was (150 ±16)mL,which of the non-bone graft group was (120 ±14) mL,the difference between the two groups was statistically significant (t =3.57,P >0.05).After 3 months,the flexion of the cervical spine of the bone graft group was (31.2 ±4.6)°,which in the non -grafted group was (32.3 ±5.7)°,the difference was statistically significant (t=0.675,P<0.05).The rotation of the bone graft group was (40.6 ±4.5)°,which in the non -graft group was (41.3 ±3.5)°,the difference was statistically significant (t=0.278,P<0.05).Both two groups had no vertebral artery and spinal cord injury ,wound healing. During the follow-up period,the two groups of patients had a good reduction of cervical spine ,no internal fixation lossening,fracture,fracture healing well,group A bone graft fusion.Conclusion For this type of fracture,simple atlantoaxial pedicle screw fixation compared with autologous iliac bone graft fusion treatment ,can save the operation time,reduce the amount of bleeding .
3.Effect of the sonic hedgehog inhibitor GDC-0449 on an in vitro isogenic cellular model simulating odontogenic keratocysts.
Jiemei ZHAI ; Heyu ZHANG ; Jianyun ZHANG ; Ran ZHANG ; Yingying HONG ; Jiafei QU ; Feng CHEN ; Tiejun LI
International Journal of Oral Science 2019;11(1):4-4
Odontogenic keratocysts (OKCs) are common cystic lesions of odontogenic epithelial origin that can occur sporadically or in association with naevoid basal cell carcinoma syndrome (NBCCS). OKCs are locally aggressive, cause marked destruction of the jaw bones and have a propensity to recur. PTCH1 mutations (at ∼80%) are frequently detected in the epithelia of both NBCCS-related and sporadic OKCs, suggesting that PTCH1 inactivation might constitutively activate sonic hedgehog (SHH) signalling and play a major role in disease pathogenesis. Thus, small molecule inhibitors of SHH signalling might represent a new treatment strategy for OKCs. However, studies on the molecular mechanisms associated with OKCs have been hampered by limited epithelial cell yields during OKC explant culture. Here, we constructed an isogenic PTCH1 cellular model of PTCH1 inactivation by introducing a heterozygous mutation, namely, c.403C>T (p.R135X), which has been identified in OKC patients, into a human embryonic stem cell line using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system. This was followed by the induction of epithelial differentiation. Using this in vitro isogenic cellular model, we verified that the PTCH1 heterozygous mutation causes ligand-independent activation of SHH signalling due to PTCH1 haploinsufficiency. This activation was found to be downregulated in a dose-dependent manner by the SHH pathway inhibitor GDC-0449. In addition, through inhibition of activated SHH signalling, the enhanced proliferation observed in these induced cells was suppressed, suggesting that GDC-0449 might represent an effective inhibitor of the SHH pathway for use during OKC treatment.
Anilides
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pharmacology
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Basal Cell Nevus Syndrome
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Hedgehog Proteins
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genetics
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pharmacology
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Humans
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Molecular Targeted Therapy
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Odontogenic Cysts
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genetics
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physiopathology
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therapy
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Odontogenic Tumors
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genetics
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physiopathology
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therapy
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Pyridines
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pharmacology