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.
Objective To discuss the role of heparan sulfate (HS) in bone formation and bone remodeling and summarize the research progress in the osteogenic mechanism of HS. Methods The domestic and abroad related literature about HS acting on osteoblast cell line in vitro, HS and HS composite scaffold materials acting on the ani-mal bone defect models, and the effect of HS proteoglycans on bone development were summarized and analyzed. Results Many growth factors involved in fracture healing especially heparin-binding growth factors, such as fibroblast growth factors, bone morphogenetic protein, and transforming growth factor β, are connected noncovalently with long HS chains. HS proteoglycans protect these proteins from protease degradation and are directly involved in the regulation of growth factors signaling and bone cell function. HS can promote the differentiation of stem cells into osteoblasts and enhance the differentiation of osteoblasts. In bone matrix, HS plays a significant role in promoting the formation, maintaining the stability, and accelerating the mineralization. Conclusion The osteogenesis of HS is pronounced. HS is likely to become the clinical treatment measures of fracture nonunion or delayed union, and is expected to provide more choices for bone tissue engineering with identification of its long-term safety.
【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 investigate the role and regulatory mechanism of ring finger protein 11 (RNF11) on Akt signaling pathway in the process of osteogenesis of bone marrow mesenchymal stem cells (BMSCs) to provide ideas for further clarifying its osteogenesis mechanism and its use in clinical treatment in the future. Methods BMSCs were isolated and cultured from fresh bone marrow of healthy donors and subcultured. The 4th generation cells were used in experiments after identification by flow cytometry, and osteogenic, chondrogenic, and adipogenic induction. BMSCs were cultured in osteogenic differentiation medium for 0-14 days. The degree of osteogenic differentiation was detected by Alizarin red staining and alkaline phosphatase (ALP) staining, and the protein expression of RNF11 was detected by Western blot. The 4th generation BMSCs were divided into blank control group (group A), empty lentivirus (Lv-NC) group (group B), and knockdown RNF11 (Lv-ShRNF11) group (group C). Osteogenesis was induced and cultured for 0-14 days. The expression of RNF11 protein was detected by Western blot, the degree of osteogenic differentiation was detected by Alizarin red staining and ALP staining, and the relative mRNA expressions of Runx2, osteocalcin (OCN), and osteopontin (OPN) were detected by real-time fluorescence quantitative PCR (qRT-PCR). The protein relative expressions of Akt, Smad1/5/8, and β-catenin signaling pathway were detected by Western blot, expressed as the ratio before and after phosphorylation. In order to study the effect mechanism of RNF11 on Akt signaling pathway, the 4th generation BMSCs were divided into Lv-NC transfection group (group A1), Lv-ShRNF11 transfection group (group B1), and Lv-ShRNF11 transfection supplemented with Akt signaling pathway activator SC79 group (group C1). The protein relative expressions of RNF11 and Akt signaling pathway were detected by Western blot, the related osteogenesis indexes were detected by Alizarin red staining, ALP staining, and qRT-PCR. ResultsThe flow cytometry, and osteogenic, chondrogenic, adipogenic induction culture identification showed that the isolated and cultured cells were BMSCs. The protein relative expression of RNF11 increased gradually with the extension of osteogenic differentiation time (P<0.05); after knockdown RNF11, Alizarin red and ALP stainings showed that the degree of osteogenic differentiation of BMSCs in group C were significantly lower than those in groups A and B, and qRT-PCR detection showed that the relative expression of Runx2, OCN, and OPN mRNA significantly decreased (P<0.05). The protein relative expressions of RNF11 and Akt signaling pathway significantly increased with the extensions of osteogenic differentiation time (P<0.05). After knockdown RNF11, the protein relative expression of Akt signaling pathway in group C was significantly lower than that in groups A and B (P<0.05), while Smad1/5/8 and β-catenin signaling pathway had no significant effect (P>0.05). Compared with group A1, the protein relative expression of RNF11 in groups B1 and C1 significantly decreased (P<0.05). Compared with groups A1 and C1, the protein relative expression of Akt signaling pathway in group B1 was significantly lower (P<0.05); Alizarin red and ALP stainings showed that the degree of osteogenic differentiation of BMSCs in group C1 were slightly lower than that of group A1 (P>0.05), but significantly higher than that of group B1 (P<0.05); qRT-PCR detection showed that the relative expressions of Runx2, OCN, and OPN mRNA in group C1 were slightly lower than those of group A1 (P>0.05), but were significantly higher than those of group B1 (P<0.05). ConclusionRNF11 promotes the differentiation of BMSCs into osteoblasts by positively regulating the activation level of Akt signaling pathway. RNF11 can be used as a potential target to improve the bone repair efficacy of BMSCs and treat bone metabolic diseases.
Icariin（ICA） is one of the main active ingredients in the Berberidaceae family Epimedium. It makes a variety of biological activities, such as promoting bone formation, antibacterial and anti-inflammatory, and regulating immunity. Periodontitis is a chronic inflammatory disease that is present in the soft and hard tissues of the periodontium. The ultimate goals of its treatment are the reconstruction of periodontal tissues and bone defect repairing. At present, conventional treatment of periodontitis fails to achieve the ideal periodontal tissue regeneration. In recent years, the rapid development of tissue engineering technology has brought new ideas for the treatment of periodontal disease and bone defect repairing. Because of its anti-inflammatory and osteogenic effects, ICA has great potential for the treatments of periodontitis and bone defect repairing. This paper summarizes the effect and the molecular mechanism of ICA in the treatment of periodontitis and bone defect repairing, and discusses its application prospect as a drug for periodontal adjuvant therapy. This paper aims to provide a theoretical basis for the research and application of ICA in periodontitis treatment and bone defect repairing.
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.
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.
Based on transversely isotropic theory, a finite element model for three-dimensional solid-liquid coupling defect repair of articular cartilage was established. By studying stress state of host cartilage near the restoration interface, we identified deformation type of cartilage and discussed the cause of restoration interface cracking. The results showed that the host cartilage surface node near the restoration interface underwent compression deformation in the condition of surface layer defect repair. When the middle layer, deep layer or full-thickness defect were repaired, the node underwent tensile deformation. At this point, the radial dimension of cartilage increased, which might cause restoration interface cracking. If elastic modulus of the tissue engineered cartilage (TEC) was lower (0.1 MPa, 0.3 MPa), the host cartilage surface layer and middle layer mainly underwent tensile deformation. While elastic modulus of TEC was higher (0.6 MPa, 0.9 MPa), each layer of host cartilage underwent compression deformation. Therefore, the elastic modulus of TEC could be increased properly for full-thickness defect repair. This article provides a new idea for evaluating the effect of cartilage tissue engineering repair, and has a certain guiding significance for clinical practice.
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.
ObjectiveTo objectively evaluate the effectiveness of the ventricular fold pull-down combined with strip myofascial flap to repair laryngeal defect after early glottic carcinoma operation with glottic morphological parameters and voice parameters. MethodsBetween January 2008 and December 2012, 47 patients with early glottic carcinoma and anterior commissure involvement underwent partial laryngectomy. All patients were male, aged from 60 to 75 years (mean, 68.5 years). The disease duration was 4-11 months (mean, 7.2 months). According to American Joint Committee on Cancer (AJCC) TNM criteria, 28 cases were classified as T1aN0M0, 14 cases as T1bN0M0, and 5 cases as T2N0M0. Laryngeal defect after resection of tumor was repaired by ventricular fold pull-down combined with strip myofascial flap. At 1 day before operation and at 1 year after operation, multilayer spiral CT was used to scan larynx, to measure and compare the anteroposterior diameter of vocal area, the distance between both sides of the vocal process, and the thickness of soft tissue of vocal area, and the effect of combined soft tissue flap was objectively assessed in laryngeal morphology reconstruction. The actual voice parameters[including F0, Jitter, Shimmer, normalized noise energy (NNE), and maximum phonatory time (MPT)] were tested and compared, and the effect of the combined soft tissue flap on postoperative laryngeal pronunciation was evaluated. ResultsPostoperative pathological examination revealed well-differentiated squamous cell carcinoma in 38 cases, and moderately-differentiated squamous cell carcinoma in 9 cases; no tumor was found in the resection margin. Healing of neck incision was obtained in all patients at 7-9 days after operation. Forty-four cases were decannulated at 9-11 days after operation and the remaining 3 cases were decannulated at 3 weeks after operation. Oral feeding usually started in all cases at 3-4 days after operation. All patients were followed up 1 year. At 1 year after operation, the anteroposterior diameter of vocal area was significantly reduced when compared with preoperative one (t=15.161, P=0.000); the distance between both sides of the vocal process and the thickness of soft tissue of vocal area had no significant changes (P > 0.05). Compared with preoperative ones, there were significant differences in Shimmer, NNE, and MPT (P < 0.05), but no significant difference was found in F0 and Jitter (P > 0.05) at 1 year after operation. ConclusionVentricular fold pull-down combined with strip myofascial flap can repair laryngeal defect effectively after partial laryngectomy and maintain the effective airway after operation. It not only has no effect on postoperative laryngeal morphology, but also can be used as new laryngeal voice vibration body.