THE CO-CULTURE OF ADIPOSE DERIVEDSTEM CELLS DIFFERENTIATED SMOOTH MUSCLE CELLS AND FIBROBLASTS ON ASILK FIBROIN SCAFFOLD: A PRIMARY RESEARCH ON THE PROLIFERATION ANDMECHANICAL CHARACTERISTICS
Y. CHANG1, X. SUN 2,J. WANG 3;
1OB/Gyn, Peking Univ. People’sHosp., Beijing, China, 2Dept. Ob Gyn, Peking Univ.People's Hosp., Beijing, China, 3Peking Univ. people'sHosp., Beijing, China.
Introduction: Pelvic floordysfunction(PFD) is a series of disease including stress urinaryincontinence (SUI), pelvic organ prolapse(POP), and dyspareunia.Women suffering from PFD often experience an overall reduction intheir quality of life. Biological and synthetic mesh based surgeryhas been proved to be a most effective way to cure severe PFD, aimingto function as a long-lasting support until surrounded or replaced bynew healthy collagen tissue. However, in 2008 and 2011, the US Foodand Drug Administration (FDA) advocated against the widespread use ofmesh in POP describing serious complications. Recently, the FDA hasbeen consulting on reclassify surgical mesh for transvaginal POP froma moderate-risk device (class II) to a high-risk device (classIII).Therefore, an ideal material is still in search, which shouldcontain both excellent biocompatibility and nice mechaniccharacteristics to sustain the pelvic organs, and promote theneo-tissue formation and reconstruction as well. Regenerativemedicine has been rapidly developed in recent years, focusing onconstructing tissues in vitro with cells, extracellular matrix,cytokine and biomaterials, in order to replace, reconstruct, andenhance the function and anatomy of tissue and organs. Intensivestudies have been made in the reconstruction of bone, ligament,bloodvessel. As one of the main components of pelvic floorligaments, fibroblasts could synthesize collagen and elastin.Smooth muscle cells, which also proved existence in the pelvicligaments, especially the capital ligament, could provide a similarelasticity and intensity, and minimize the adhesion and meshshrinkage.silk fibroin scaffold could be applied inpelvic floor reconstruction, and tends to induce the formation ofhealthy new tissue, which might be strengthened by stem cell.Therefore, constructing a pelvic ligament analogous constructure invitro to replace the weakened area might make way for female pelvicfloor reconstruction.
Objective: To construct the tissueengineering complex by implanting adipose derived stem cells(ASCs)induced smooth muscle cells(SMC)and fibroblasts(FB)onto silkfibroin scaffold and to investigate the ideal cell proportion forcell proliferation and mechanics.
Methods: ASCs weredifferentiated into FB, SMC,and confirmed by realtime-PCR andimmunocytochemistry,then labelled with GFP and RFP,followed by mixedimplantation onto silk fibroin scaffold in the proportion of SMC vsFB 1:1; SMC vs FB 1:2; SMC vs FB 2:1. After 14days,CoL-I,COL-III,ELASTIN,Young's modulus,and maximum failure forcewere detected.
Results: After differentiated into FB ,theexpression of CoL-I, COL-III, elastin and FSP-1 were significantlyincreased at the mRNA level (P < 0.05); while CD44 wasdecreased; SMC induced cells were enlarged,α-SMA,calponin, MHCexpression was significantly increased (P < 0.05). Themixed cells could proliferate on the scaffold. SMC vs FB1:1 group hasthe fastest cell proliferation rate,and the expression of CoL-I andCoL-III at mRNA levels were elevated (P < 0.05).Theexpression of elastin in the 1:2 group were elevated at mRNA levels(P< 0.05).The Young's modulus and maximum failure force were of nosignificant difference (P > 0.05), but was slightly higherin FB vs SMC 1:1 group.
Conclusions: ASCs could be inducedinto smooth muscle cells and fibroblast cells; cellular componentsand cell proportion are important for tissue engineeringreconstruction,suggesting its potential use in pelvic floorreconstruction,which requires critical cell proportion,extracellularmatrix and mechanism.
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