Objective To review the current condition of growth factors and their application to clinical treatment of acute and chronic wounds. Methods Data from the literature and Medline were analyzed according to their different uses in acute and chronic wounds. Their potential side-effects were studied. Results All data showed that wound healing time in acute and chronic wounds was accelerated and wound healing quality was improved after treatment with growth factors. No sideeffect was observed. Conclusion The efficacy and safety of growth factors in improving wound healing were confirmed. However, some reconsideration aboutpotential problems of growth factors must be made to apply them clinically in the future.
Objective To establish a better method of isolating andculturing ofneural stem cells(NSCs) in neonatal rat brain. Methods Tissue of brain was isolated from neonatal rats. Different medium and culture concentration were used toculture NSCs of neonatal rat. The culture concentration used were 1×10 4, 1×105, 1×106and 1×107/ml respectively. Ingredient of medium was classified into group 1 to 8 respectively according to whether to add 2% B27, epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) as well as the difference in culture concentration. The cells were induced to differentiate asto be confirmed as NSCs, and then were checked by phase contrast microscopy and identified by immunocytochemistry. Results The cells isolated and cultured gathered into neurospheres. The cells were capable of proliferating and maintaining longterm survival in vitro. The cells could be differentiated into neurons and glia.It was to the benefit of the survival of NSCs to add 5% fetal bovine serum(FBS)into the medium at the beginning of the culturing. When 10% FBS was added intothe medium, the neurospheres differentiated quickly. When concentration 1×106/ ml was used, the growth rate of the cells was the highest of all the concentrations. Reasonably higher cell concentration promoted the proliferation of NSCs. It was necessary to add 2% B27, EGF, and bFGF into the medium. The cells had the best growth when 2% B27, 20 ng/ml bFGF and 20 ng/ml EGF were added into the culture medium. EGF and bFGF had cooperative effect. Conclusion A better method of isolating and culturing of NSCs in neonatal rat brain is established and the foundation for future research is laid.
【Abstract】 Objective To explore the interventional effect of platelet lysate (PL) on osteogenic differentiation ofBMSCs by induction in rats in vitro. Methods Twenty-four clean-grade adult Wistar rats, weighing from 250 g to 300 g, maleor female, were included in this study. PL was obtained through three times of centrifugation and repeated freeze-thaw for the blood aspirated from cardiac cavities in 16 Wistar rats. ELISA assay was conducted to detect the concentration of growth factors PDGF, TGF-β1, IGF-1 and VEGF in PL. The BMSCs harvested by flushing femurs of 8 adult Wistar rats were isolated, cultivated and expanded in vitro. The cells at the 4 passage were performed for osteogenic differentiation by induction in three groups of A (5% PL of final concentration in basic induction medium), B (1% PL of final concentration in basic induction medium), and C (no presence of PL in basic induction medium as a control). The morphological changes of the cells were dynamically observed with inverted phase contrast microscope during the whole period. At different time-points, ALP staining (7 days) and ALP/TP (2, 8, 12 days) of the cells were detected to evaluate ALP activity, and the mineral formation in extracellular martrix was examined with Al izarin red staining which provided quantitative analysis of mineral deposits. Results ELISA assay showed that the content of PDGF, TGF-β1, IGF-1 and VEGF in PL reached (300 ± 30), (140 ± 25), (80 ± 35), (70 ± 20) pg/mL, respectively. Morphological observation displayed BMSCs in group A or B gradually turned from spindle-shape to square- or polygon-shape as the morphorlogical type of osteoblast-l ike cells at 7 days. The cells in group A showed slower shape changesbut higher prol iferation than that in group B or C. Moreover, at the 20 days, the cells in group A still displayed dense gro wth and produced obviously decreased amount of mineral deposits in ECM when compared with group B or C. At the 7 days, the cells ofgroup A showed smaller amount of granules positive for ALP staining in cytoplasm when compared with groups B and C, and displayed marked reduction in ALP activity assay at the 2, 8, and 10 days compared with that of groups B and C (P lt; 0.05). At the 20 days, Al izarin red staining showed the number of mineral deposits in groups A, B and C were 7.67 ± 1.10, 12.87 ± 0.81 and 15.59 ± 0.25, respectively, while the area of mineral deposits were (161 778.70 ± 44 550.80), (337 349.70 ± 56 083.24), and (415 921.70 ± 71 725.39) pixels, respectively. The number of mineral deposits and the area of mineral deposits in group A were smaller than those in groups B and C (P lt;0.05). But there was no statistically significant difference between groups B and C (P gt; 0.05). Conclusion PL is a kind of system carrying various growth factors. Exposure of PL inhibits both ALP activity and mineral formation of BMCs in a dose-dependent way under the osteogenic induction environment.
Objective To summarize the research progress of controlled release of angiogenic factors and osteogenic factors in bone tissue engineering. Methods The domestic and abroad literature on the controlled release structure of growth factors during bone regeneration in recent years was extensively reviewed and summarized. Results The sustained-release structure includes direct binding, microsphere-three-dimensional scaffold structure, core-shell structure, layer self-assembly, hydrogel, and gene carrier. A sustained-release system composed of different sustained-release structures combined with different growth factors can promote bone regeneration and angiogenesis. Conclusion Due to its controllability and persistence, the growth factor sustained-release system has become a research hotspot in bone tissue engineering and has broad application prospects.
Objective To review the recent progress of the researches in the field of cartilage tissue engineering, and to discuss the challenges in construction of tissue engineered cartilage. Methods Literature related with cartilage tissue engineering was reviewed and analyzed. Results Some techniques have been appl ied in cl inical. As far as the seeding cells, induced pluripotent stem cells have attracted much more attention. Current strategies of scaffold designing are trying to imitate both component and structure of natural extracellular matrix. Cartilage regeneration through the autologous cell homing technique el iminate the transplantation of exotic cells and has become the hot topic. Conclusion Successful treatment of the damaged cartilage using tissue engineering method will depend on the advances of stem cell technology development, biomimetic scaffolds fabrication and proper appl ication of growth factors.
OBJECTIVE To compare the osteogenesis of recombination artificial bones, which are bovine deproteined bone (bDPB) and bovine bone morphogenetic protein (bBMP), combined with tumor necrosis factor alpha (TNF alpha), basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) respectively. METHODS One hundred trephined skull bone defects in fifty rabbits were divided into four groups, which implanted with bDPB/bBMP/TNF alpha, bDPB/bBMP/bFGF, bDPB/bBMP/EGF, and bDPB/bBMP respectively. X-ray and histological changes were observed in the 1st, 2nd, 4th, 6th, 8th weeks after implantation. The content of 35S and 45Ca and ash weight were measured at 10 and 42 days after operation. RESULTS The osteogenesis of bDPB/bBMP/TNF alpha group was ber than that of bDPB/bBMP/bFGF group(P lt; 0.01), while bDPB/bBMP/bFGF group was ber than that of bDPB/bBMP/EGF(P lt; 0.01). No significant statistical difference were found between bDPB/bBMP/EGF and bDPB/bBMP(P gt; 0.05). CONCLUSION TNF alpha combined with bBMP and carrier can stimulate bone formation and increase the volume of new bone in vivo. It suggests that bDPB/bBMP/TNF alpha is a valuable biomaterial of bone graft.
OBJECTIVE: To observe the effects of hyaluronic acid (HA) and basic fibroblast growth factor (bFGF) on the proliferation of the cells from medial collateral ligament (MCL) and anterior cruciate ligament (ACL) cells. METHODS: The MCL cells and ACL cells of mature New Zealand white rabbit were cultured, while HA, bFGF or HA and bFGF were added to the cell culture media, the cellular proliferation was assayed by MTT method. RESULTS: HA only had no effect on the preoliferation of ACL cells, but had a small stimulatory effect on the proliferation of MCL cells. The addition of 1 ng/ml bFGF enhanced the proliferation of both MCL and ACL cells significantly, and this enhancement was maximal in the concentration of 50 ng/ml. However, the enhancement of proliferation of MCL and ACL cells could be achieved when the combination of HA in concentration of 100 micrograms/ml and bFGF in concentration of 100 ng/ml. CONCLUSION: It is evident that bFGF can enhance the proliferation of the ligament cells. HA can maintain the normal growth of ACL cells with no effect on the proliferation of the cells, while HA has a small stimulatory effect on the proliferation of MCL cells. However, when bFGF is coordinated with HA, more improvement of cellular proliferation can be achieved. HA can be used as a potential carrier for bFGF to enhance the healing of ligamentous tissue injuries.
Objective To review the research progress of the seed cells, scaffolds, growth factors, and the prospects for clinical application of the intervertebral disc regeneration. Methods The recent literature concerning the regeneration strategies and tissue engineering for treatment of degenerative intervertebral disc disease was extensively reviewed and summarized. Results Seed cells based on mesenchymal stem cells (MSCs) and multiple-designed biomimetic scaffolds are the hot topic in the field of intervertebral disc regeneration. It needs to be further investigated how to effectively combine the interactions of seed cells, scaffolds, and growth factors and to play their regulation function. Conclusion The biological regeneration of intervertebral disc would have a very broad prospects for clinical application in future.
ObjectiveTo summarize the application status and progress of the strategies to augment tendon-to-bone healing.
MethodsThe present researches focused on augmentation of tendon-to-bone healing were extensively reviewed.
ResultsThe present strategies to augment healing of tendon-to-bone by enhancing the location environment, and increasing the cell numbers and relative growth factor. The mainly strategies include using calcium phosphate materials, biocompatible scaffolds and glue, growth factors, cell matrix, platelet-rich plasma, and periosteum. Although periosteum have been used in clinical and got some possitive effects, the others still not be used in clinical and needs further studies.
ConclusionThere are many strategies to enhance the ability of tendon-to-bone healing, which got some positive results, but results of studies were varied. Thus, further fundamental research and clinical studies are required to achieve the best effects.
To compare the platelet enrichment ratio of platelet-rich plasma (PRP) prepared by different centrifuge methods and to compare the concentration of growth factors released from autologous platelet-rich gel (APG) with the whole blood. Methods Thirteen diabetic patients with refractory skin lesions were enrolled in APG treatment. ① Three kinds of centrifuge methods were selected for PRP by 11 diabetic patients: A(n=6): 529 × g for 4 minutes in the first centrifugeand 854 × g for 6 minutes in the second centrifuge; B (n=5): 313 × g for 4 minutes in the first centrifuge and 1 252 × g for 6 minutes in the second centrifuge; C (n=5): 176 × g for 5 minutes in the first centrifuge and 1 252 × g for 5 minutes in the second centrifuge. Platelet counted on the whole blood and PRP was determined. The APG, produced by combining the PRPwith thrombin and calcium gluconate (10 ∶ 1) was used by patients. ② PDGF-BB, TGF-β1, VEGF, EGF, and IGF-1 were measured in the APG and the whole blood using the enzyme-l inked immunosorbent assay method. Results ① The average platelet concentration was higher in group B [(1 363.80 ± 919.74) × 109/ L] than in groups A[(779.67 ± 352.39) × 109/ L)] and C[(765.00 ± 278.78) × 109/ L] and the platelet recovery rate was 75.2% ± 21.0% in group B. ② The concentration of growth factors all increased with the increasing platelet number. On average, for the whole blood as compared with APG, the PDGF-BB concentration increased from (145.94 ± 133.24) pg/mL to (503.81 ± 197.86) pg/mL (P lt; 0.05); TGF-β1 concentration increased from (3.31 ± 2.27) ng/mL to (5.67 ± 4.80) ng/mL (P lt; 0.05); IGF-1concentration increased from (14.54 ± 35.34) ng/mL to (110.56 ± 84.36) ng/mL (P lt; 0.05); and EGF concentration increased from (160.73 ± 71.10) pg/mL to (265.95 ± 138.43) pg/mL (P lt; 0.05). No increase was found for VEGF(P gt; 0.05). ③ There was positive correlation between the platelet concentration and PDGF-BB and TGF-β1 (r = 0.627, r = 0.437, P lt; 0.05). ④ Thirteen diabetic repractory dermal ulcers received APG treatment for 18 times, 9 ulcers (69.2%) and 10 sinuses (88.3%) were cured at the end of 12-week treatment. Conclusion The method ofgroup B is the best centrifuge method. A variety of growth factors are detected and released from the platelets at significant levels in APG. There is positive correlation between the platelet concentration and PDGF-BB and TGF-β1 .
