1.Osteogenic ability of cryopreserved bone marrow stromal cells complex in vivo
Yuqian ZHENG ; Fang YUAN ; Fuhua YAN ; Xin ZHAO ; Minkui LIN
Chinese Journal of Tissue Engineering Research 2011;15(12):2275-2278
BACKGROUND: Our previous studies have demonstrated that cryopreserved bone marrow stromal cells (BMSCs) still maintain high survival rate, cell proliferation and osteogenic differentiation potentials after thawing. However, this result needs confirmed in vivo environment. OBJECTIVE: To explore the effects of cryopreserved BMSCs and collagenic membrane BME-10X complex on type Ⅰ collagen synthesis in vivo. METHODS: Beagle dog BMSCs were cultured in vitro and cryopreserved for 12 months, which were thawed and prepared complexes with collagenic membrane. The complexes were cultured with mineralization induction medium or normal medium for 5 days, followed by implanting into nude mice. The specimens were harvested and analyzed by gross observation, histopathological and immunohistochemistry at 4 weeks after implantation. The collagenic membrane cultured with mineralization induction medium served as controls. RESULTS AND CONCLUSION: In the control group, the boundary of collagenic membrane was distinctly, without cell growth around boundary or intra collagenic membrane, additionally, there was little type Ⅰ collagen. In the non-induction group, cells grew into collagenic membrane, trabes-like collagen formed, and type Ⅰ collagen distribution increased at 4 weeks. In the induction group, scaffold degraded, more cells grew, and plenty of collagen formed osteoid-like tissues. The distribution of typeⅠcollagen was obviously increased than that of other groups. The findings demonstrated that cryopreserved BMSCs possess strong osteogenic differentiation potentials after proliferation and induction combined with collagenic membranes in vitro.
2.Effect of platelet-rich plasma, activated by different concentrations of thrombin, on the repair of cranial defects
Minkui LIN ; Xiaoling CHEN ; Xin ZHAO ; Fuhua YAN
Chinese Journal of Tissue Engineering Research 2010;14(2):209-213
BACKGROUND: The biological functions of platelet-rich plasma (PRP) are affected by multiple factors, such as individual difference, PRP concentration, PRP carder, PRP-activated methods and so on. OBJECTIVE: To evaluate the effect of PRP, activated by different concentrations of thrombin, on the repair of cranial defects. METHODS: Whole blood of the central artery of rabbit ears was extracted to prepare PRP, which was then diluted so that the final platetet count was about 5 times of the whole blood. Four whole-thickness layer of cranial defects at an 8-mm diameter were created in 16 New Zealand rabbits and randomly grafted with β-tdcalcium phosphate (β-TCP) and PRP, activated by 60 U/mL. thrombin; β-TCP and PRP, activated by 1 000 U/mL thrombin; β-TCP and PRP; β-TCP alone. At 1 and 3 months following implantation, X-ray analysis and microscopic observation were performed to onserve cranial repair, the area percent of new bone formation was calculated. RESULTS AND CONCLUSION: At one month post-surgery, the edge of defects was clear in each group, with varying degrees of new bone formation surrounding the defects, β-TCP particles partially degraded and the degradation lesion was replaced by new bone, only a small amount of bone lacunae was seen, fiber wrapped around the defect center β-TCP, only a small number of specimens showed new bone formation; X-ray showed a clear boundary and uniform defect density; the percentage of new bone formation in the PRP groups were higher than β-TCP groups (P < 0.05). However, there was no significant difference between PRP group groups (P > 0.05). At 3 months post-surgery, the defect boundary was unclear in each group, the new bone formation increased, the β-TCP particles surrounding defects partially or all degraded and were replaced by new bones, some regions appeared trabecular bone, bone lacuna in new bone was increased, the central defect of the majority of specimens exhibited new bone formation; X-ray showed defect boundary was unclear in each group, defect surrounding density was higher than the center defect, and bone mineral density was equivalent to other normal parts; the percentage of new bone formation in the PRP groups was significantly higher than that in the β-TCP groups (P < 0.05), PRP +β-TCP group was higher than the other 3 groups (P <0.05), there was no significant difference between two thrombin groups (P > 0.05). It is indicated that although PRP improves the repair of cranial defects, 60 and 1 000 U/mL of thrombin has no effects on PRP rapairing cranial defects in New Zealand white rabbits, compared with PRP+β-TCP group, possible the absence of the optimal concentration of thrombin.
3.Human leptin gene overexpression enhances osteogenesis of bone marrow stromal cells from rats
Baoyu ZHENG ; Jun JIANG ; Yuling CHEN ; Minkui LIN ; Kai LUO ; Fuhua YAN
Basic & Clinical Medicine 2017;37(2):169-175
Objective To investigate the effects of human leptin ( hLEP) gene transfection on rat bone marrow stro-mal cells ( rBMSCs) .Methods rBMSCs were cultured and transfected with adenoviruses encoding hLEP ( Ad5-hLEP-EGFP) in vitro as experimental group while rBMSCs transfected with Ad 5-EGFP and non-transfected were control groups.The proliferation was detected by MTT and the expression of collagen type Ⅰ(Col-Ⅰ) and alkaline phosphatase ( ALP) were assessed by real-time PCR.The ability of mineralized nodule forming was also examined by Alizarin red staining .The combination of transfected rBMSCs and β-tricalcium phosphate (β-TCP ) was con-structed and the osteogenic ability of the construction was evaluated in nude mice .Results hLEP could be trans-fected into rBMSCs successfully by adenovirous .After transfection , the proliferation was not affected while Col-Ⅰand ALP expressions were more pronounced in rBMSCs transfected with Ad 5-hLEP-EGFP ( P<0.05 ) .Alizarin red staining showed the ability of mineralized nodule forming was also up-regulated in Ad5-hLEP-EGFP group (P<0.05).In addition, the transfected rBMSCs adhered to β-TCP and survived well and the combination showed more new bone like tissue formation in nude mice compared to control groups .Conclusions rBMSCs transfected with hLEP might be potently used in bone or periodontal tissue regeneration .
4. Application of CiteSpace software in the visualization analysis of work-related musculoskeletal disorders
Peifang LIU ; Xuyan XU ; Yan CHEN ; Fujiang WANG ; Lihua HE ; Minkui LIN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2019;37(10):768-772
Objective:
To investigate the visual application of the CiteSpace software in the field of work