Objective To investigate the effects of flow shear stress and mass transport on the construction of largescale tissue engineered bone using a perfusion bioreactor. Methods Bone marrow (20 mL) was harvested from the il iac crestof the healthy volunteer, and then hBMSCs were isolated, cultured and identified. The hBMSCs at passage 3 were seeded on the critical-size β-TCP scaffold and cultured in a perfusion bioreactor for 28 days. Different flow shear stress (1 ×, 2 × and 3 ×) and different mass transport (3, 6 and 9 mL/min) were exerted on the cells seeded on the scaffold by changing the viscosity of media or perfusion flow rate. The cell prol iferation and ALP activity of cells seeded on the scaffold were detected, and histology observation and morphology measurement of cell/scaffold complex were conducted. Results When the perfusion flow rabe was 3 mL/min, the cell viabil ity of 2 × group was higher than that of other groups (P lt; 0.05). When the flow shear stress was 3 ×, no significant differences were found among 3, 6 and 9 mL/min in cell viabil ity (P gt; 0.05). When the perfusion flow rate was 3 mL/min, the activity of ALP of 2 × and 3 × groups was higher than that of 1 × group (P lt; 0.05). When the flow shear stress was 3 ×, the activity of ALP of 6 mL/min group was the highest (P lt; 0.05). After 28 days of perfusion culture, the ECM of all the groups distributed throughout the scaffold, and the formation and mineral ization of ECM was improved with the increase of flow shear stress when the perfusion flow rate was 3 mL/min. However, the increase of perfusion flow rate decreased the mineral ization of ECM when the flow shear stress was 3 ×. Conclusion As two important fluid dynamics parameters affecting the construction of large-scale tissue engineered bone, the flow shear stress and the mass transport should be measured duringthe process of constructing large-scale tissue engineered bone so as to maximize their roles.
To investigate the cl inical results and the mechanism of bone heal ing for the repair of bone defects following tumor resection with novel interporous TCP bone graft, and to test the hypothesis of “structural transplantation”. Methods From January 2003 to December 2005, 61 cases of various bone defects following the curettage of the benign bone tumors were treated with interporous TCP, with 33 males and 28 females, including bone fibrous dysplasiain 8 cases, bone cyst in 23 cases, eosinophil ic granuloma in 12 cases, enchondroma in 13 cases, non-ossifying fibroma in 2 cases, and osteoblastoma in 3 cases. Tumor sizes varied from 1.5 cm × 1.0 cm to 7.0 cm × 5.0 cm. The plain X-ray, single photon emission computed tomography (SPECT) and histology examination were obtained at various time points after operation. The in vivo biodegradation rate of the implanted TCP was evaluated based on a semi-quantitive radiographic analyzing method. Histopathology examination was performed in 1 revision case. Results All the patients were followed up for 5 to 24 months after operation. They all had good wound heal ing and bone regeneration. There was neither significant reverse reaction to the transplanted material nor locally inflammatory reaction in all of the cases. The bone defects were repaired gradually from 1 to 6 months after operation (bone heal ing at average 2.6 months after surgery) with a bone heal ing rate up to 96.7%. There was only 1 recurrence case (eosinophil ic granuloma in ischium) 3 months after operation. Given revision operation, this case gained bone heal ing. Radiographically, the interface between the implanted bone and host bone became fuzzy 1 month after implantation, indicating the beginning of new bone formation. Three months later, the absorption of the interporous TCP was noticed from peripheral to the center of the implanted bone evidenced by the vague or fuzzy realm. New bone formation could be seen both in peri pheral and central areas. Six months later, implanted bone and host bone merged together and the bone defect was totally repaired, with 78.9% degradation rate of the implanted TCP. Twelve months later, the majority of the implanted bone was absorbed and bone remodel ing was establ ished. In the cases that were followed up for 24 months, the function of affectedextremity was excellent with good bone remodel ing without recurrence. In 2 cases, SPECT showed that nucl ide uptake could be observed in implanted site and the metabol ic activity was high both in the central as well as the peripheral areas of the graft 1 month after implantation, which was an evidence of osteogenesis. Pathologically, the interporous TCP closely contacted the host bone inside the humerus 1 month after grafting. The interface between the implanted bone and host bone became fuzzy, and vascularized tissue began growing inside the implanted graft as a “l ining” structure. Conclusion The interporous TCP proves to be effective for cl inical reparation of bone defects following tumor resection. The inside three-dimensional porous structure simulates the natural bionic bone structure which is suitable for recruitment related cells in-growth into the scaffold, colonizing and prol iferation companied with the process of vascularize, finally with the new bone formation. The novel interporous TCP may boast both bone conductive and bone inductive activities, as an appeal ing “structural transplantation” bone graft.
Objective To perfect the theory system of minimally invasive treatment for osteonecrosis of the femoral head (ONFH) with β tricalcium phosphate (β-TCP) bioceramic system and evaluate the effectiveness. Methods Eighteen New Zealand white rabbits aged 7-8 months were used to establish an animal model to verify the vascularization of porous β-TCP bioceramic rods. Micro-CT based three-dimensional reconstruction and fluorescence imaging were used to display the new blood vessels at 4, 8, and 12 weeks after operation. The inserting depth, number and diameter of vessels in the encapsulated area were analyzed. Nine pig femoral specimens were randomly divided into 3 groups (n=3): group A was normal femur; group B had cavity (core decompression channel+spherical bone defect in femoral head); in group C, mixed bioceramic granules were implanted to fill the defect in femoral head, and porous β-TCP bioceramic rod was implanted into decompression channel. The stiffness and yield load of specimens were analyzed by biomechanical test. A multicenter retrospective study was conducted to analyze 200 patients (232 hips) with femoral head necrosis treated with bioceramic system in 7 hospitals in China between January 2012 and July 2018. There were 145 males and 55 females, with an average age of 42 years (range, 17-76 years). According to the Association Research Circulation Osseous (ARCO) stage, 150 hips were in stage Ⅱ and 82 hips in stage Ⅲ. Postoperative imaging assessment was carried out regularly, and hip function was evaluated by Harris score. The effectiveness of ARCO stage Ⅱ and Ⅲ was also compared. Results Animal experiments showed that blood vessels could grow into the encapsulated area and penetrate it at 12 weeks. The inserting depth, number and diameter of blood vessels in the encapsulated area gradually increased, and there was significant difference between different time points (P<0.05). Biomechanical tests showed that the stiffness and yield load of specimens in groups B and C were significantly lower than those in group A, while the yield load in group B were significantly lower than that in group C (P<0.05). The stiffness in group C was restored to 41.52%±3.96% in group A, and the yield load was restored to 46.14%±7.85%. Clinical study showed that 200 patients were followed up 6-73 months, with an average of 22.7 months. At last follow-up, 12 patients (16 hips) underwent total hip arthroplasty, and the hip survival rate was 93.10%. According to the imaging evaluation, 184 hips (79.31%) were stable and 48 (20.69%) were worse. Harris score (79.3±17.3) was significantly higher than that before operation (57.3±12.0) (t=18.600, P=0.000). The excellent rate of hip function was 64.22% (149/232). The survival rate of hip joint, imaging score and Harris score of patients in ARCO stage Ⅱ were better than those in ARCO stage Ⅲ (P<0.05). Conclusion β-TCP bioceramic system can guide the abundant blood supply of greater trochanter and femoral neck to the femoral head to promote repair; it can partly restore the mechanical properties of the femoral head and neck in the early stage, providing a new minimally invasive hip-preserving method for patients with ONFH, especially for those in early stage.