The Effects Of PQQ And BFGF On Proliferation Of Schwann Cells On Microfluidic Chips

Objective: This study is to explore PQQ and bFGF in promoting Schwann cellsproliferation in three-dimensional and flow micro-environment which made bymicrofluidic chips, and to compare the similarities and differences with traditional cellculture platform.Methods:(1)Extracted sciatic nerve from3-5d SD rats and stripped the epineurium andfiber membrane under anatomy microscope, then cultured and amplified the SCsthrough the double enzyme digestion method.The cell morphology and quantity wereobserved under inverted microscope and SCs type was identified by S-100proteinimmunohistochemical method.(2)Used CAD software to design and make microfluidic chips and examinedtheir availability.(3)Cultured the purified Schwann cells mixed with collagen in single channelmicrofluidic chip, joined the PQQ biological factor, then observed and counted theSCs with fluorescence microscopy and Graphpad Prism5software, recorded cellgrowth and proliferation rate on1week,compared with the ordinary culture in2Denvironment.(4) Cultured the purified Schwann cells mixed with collagen in11channelconcentration gradient microfluidic chip, added PQQ and bFGF biological factor,then counted and observed the SCs with fluorescence microscopy and GraphpadPrism5software, recorded cell growth and proliferation rate on1,2,3,4weeks,compared with the ordinary culture in2D environment.Results: (1) The schwann cells extracted from3-5d SD rats grew well, and the cellsshape was a bipolar spindle,there was tip to tip or side by side arrangement betweencells,the cells nucleus were oval.The purity of SCs were more than95%and theresults of S-100protein immunohistochemical method identified as schwann cells ofrats.(2) Microfluidic chips were well sealed,the concentration value after testingconsistent with the theoretical values, which can be applied to the screening ofcytokines.(3)When PQQ accelerated the proliferation of Schwann cells,100nmol/L Grouphad the maximum proliferation effect.(4)When PQQ unite bFGF accelerated the proliferation of Schwann cells, theGroup of30ng/ml PQQ and7ng/ml bFGF had the maximum proliferation effect.Conclusions:(1)A certain concentration of PQQ can promote the proliferation of rat Schwanncells, but the high concentration inhibited the proliferation;(2)A certain concentration of PQQ and bFGF can work together to produce thelargest effect on the proliferation of rat Schwann cells;(3)Microfluidic chip technology can be used to peripheral nerve tissueengineering, it has broad application prospects. Objective:To investigate the diagnosis and therapy of infection after total hiparthroplasty.Methods: We retrospectively collected42infected cases after total hip arthroplasty injoint department of the First Affiliated Hospital of Dalian Medical University,betweenOct.2006to Oct.2011, including28male patients,14female patients,and the averageage was66.44.2.Then compared the temperature, WBC,ESR and CPR differencesbefore and after the therapy, and made a bacterial culture test on joint puncturefluid.Finally,10patients were treated with antibiotic therapy,4patients were treatedwith joint debridement therapy combine with antibiotic therapy,20patients weretreated with two-stage revision operation, the other8patients were treated byarthrodesis operation.After treatment, all patients were followed up with3-6months,and evaluated the joint function by Harris scores.Results: This retrospective clinical tests studied42infected patients after THA,according to Conventry classification criteria, I stage infection was24cases and itsproportion in all infected patients was57.14%;II stage infection was13cases and theproportion was30.95%; III stage infection was5cases and the proportion was11.91%. More than half of infected patients’ temperature became elevated, theaverage temperature was（37.320.21）℃, after treatment, the average temperaturedropped to（36.50.19）℃, there was significant difference between them (P <0.05);Before therapy WBC was（7.710.27）×109,after treatment WBC was（7.620.34）×109,there was no significant difference between the targets before and after treatment （P＞0.05）;before therapy ESR was（58.334.34）mm/h and after treatment ESRwas（10.763.11）mm/h, there was statistically significant difference between them（P＜0.01);Before therapy CPR was（75.211.98）mg/L, after treatment CPR was（6.021.01）mg/L, there was statistically significant difference between the targetsbefore and after treatment（P＜0.01).Joint puncture fluid bacterial culture test showedthat17patients infected with Staphylococcus epidermidis infection,3patientsinfected with Staphylococcus aureus infection,3cases of patients infected withStaphylococcus epidermidis combined Staphylococcus aureus infection, and1patients infected with Pseudomonas aeruginosa infection, the positive rate was57.14%. After treatment, all infected cases after total hip arthroplasty were cured,Harris scores in antibiotic therapy group were （85.632.83) points, in jointdebridement therapy combine with antibiotic therapy group were（81.123.13）points, in two-stage revision operation group were（89.453.99) points, there weresignificantly statistical significant differences compared with pre-treatment scores(P <0.01). Eight patients were treated with arthrodesis operation, but all ipsilateraljoint function was lost.Conclusions: The therapy of infected patients after total hip arthroplasty should selectthe appropriate method based on their actual prevalence，two-stage revision operationwas an ideal method for treatment.