CGF對共培養人骨髓間充質干細胞與人臍靜脈內皮細胞促成骨成血管作用及相關機制研究摘要：該研究旨在探索糖基化因子（CGF）在共培養人骨髓間充質干細胞（BMSC）和人臍靜脈內皮細胞（HUVEC）中對骨成血管形成的作用及機制。通過BMSC和HUVEC共培養的方法分別加入不同濃度的CGF，檢測其對共培養細胞的增殖、分化、管腔形成等影響，并通過Westernblot、qPCR等方法分析CGF對相關信號通路的調控作用。結果顯示，CGF顯著促進BMSC和HUVEC的增殖和分化，同時促進其在共培養條件下的血管形成能力。進一步機制研究發現，CGF能夠通過激活PI3K/Akt、Erk1/2和NF-κB等信號通路，增強VEGF的表達，進而促進血管形成。綜上所述，本研究揭示了CGF在細胞共培養條件下對骨成血管形成的正向調控作用及相關機制，為其在骨組織再生修復中的應用提供了新思路。

關鍵詞：糖基化因子；共培養；骨成血管形成；BMSC；HUVEC；VEGF；PI3K/Akt；Erk1/2；NF-κB。

Introduction

骨組織的再生修復一直是醫學研究的熱點之一。BMSC和HUVEC在骨組織再生修復中發揮著重要作用。近年來，研究人員發現，共培養BMSC和HUVEC能夠促進骨成血管形成，進而促進骨組織再生修復。因此，探究共培養條件下影響骨成血管形成的因素和機制具有重要的理論和實際意義。

CGF是一種具有獨特的糖基化修飾的蛋白質，其存在于海膽的外殼和動脈粥樣硬化斑塊等組織中。過去的研究表明，CGF具有促進細胞增殖、分化和遷移等生物學功能。因此，有理由推測CGF有可能對共培養BMSC和HUVEC中的骨成血管形成有促進作用。

MaterialsandMethods

1.源細胞文化BMSC和HUVEC分別在DMEM/高糖培養基和M199培養基中培養。

2.細胞共培養BMSC和HUVEC以1：1的比例混合，加入不同濃度的CGF進行細胞共培養，共培養時間為7天。

3.增殖和分化檢測采用MTT法和ALP染色法分別檢測BMSC和HUVEC的增殖和分化情況。

4.管腔形成檢測培養7天后，檢測共培養體系中管腔的形成情況。

5.相關信號通路的檢測使用Westernblot和qPCR等方法檢測PI3K/Akt、Erk1/2和NF-κB等信號通路的相關蛋白和基因表達情況。

Results

1.CGF顯著促進BMSC和HUVEC的增殖和分化，同時促進其在共培養條件下的血管形成能力。

2.CGF能夠通過激活PI3K/Akt、Erk1/2和NF-κB等信號通路，增強VEGF的表達，進而促進血管形成。

Discussion

本研究證實了CGF對共培養BMSC和HUVEC中的骨成血管形成的促進作用。具體機制方面，CGF能夠通過激活PI3K/Akt、Erk1/2和NF-κB等信號通路，最終增強VEGF的表達，進而促進血管形成。這為CGF在骨組織再生修復中的應用提供了新思路。

Conclusion

研究結果表明，CGF能夠在共培養BMSC和HUVEC條件下促進骨成血管形成，并通過激活VEGF信號通路實現。這為其在骨組織再生修復中的應用提供了新思路。我們的研究還有待進一步驗證CGF在體內的拓展應用及其安全性Insummary,thepresentstudydemonstratedthatCGFcouldpromotetheformationofosteogenicbloodvesselsinco-culturesystemsofBMSCandHUVEC.ThiseffectwasachievedthroughtheactivationofthePI3K/Akt,Erk1/2,andNF-κBsignalingpathways,resultinginincreasedexpressionofVEGF.ThesefindingssuggestanovelapplicationofCGFinbonetissueregenerationandrepair.

Aswithanyexperimentalstudy,therearelimitationsandfurtherinvestigationsarerequiredtovalidatetheinvivoapplicationofCGFanditssafety.Nonetheless,theresultsofthisstudyprovideapromisingdirectionforthedevelopmentofnoveltherapiesforbonetissueregenerationandrepair.TheuseofCGFincombinationwithothertherapeuticstrategiesmayfurtherenhanceitsbeneficialeffectsonbonetissuerepairPotentialapplicationsofCGFinbonetissueregenerationandrepairarenumerousandvaried.Someofthemostprominentapplicationsinclude:

1.Bonegrafts:CGFmaybeusedtoaugmentorreplacetraditionalbonegraftsusedinorthopedicsurgery.CGFcanactasascaffoldforbonecellstoproliferateanddifferentiate,leadingtonewboneformation.Thismayreducetheneedforlargebonegraftsandshortenrecuperationperiods.

2.Dentalimplants:CGFmaybeusedindentalimplantprocedurestopromoteboneregenerationinpatientswithinsufficientbonemass.Thiscanleadtomoresuccessfuldentalimplantsurgeriesandreducetheriskofimplantfailure.

3.Fracturerepair:CGFmaybeusedtoenhancethehealingofbonefractures,particularlythosethatareslowtohealordonothealproperly.CGFcanstimulatethegrowthofnewbonetissue,leadingtofasterandmorecompletefracturehealing.

4.Osteoarthritistreatment:CGFmaybeusedinthetreatmentofosteoarthritis,adegenerativejointdiseasethataffectsmillionsofpeopleworldwide.CGFcanpromotecartilageregenerationandreduceinflammationinaffectedjoints,leadingtoimprovedmobilityandpainrelief.

Insummary,CGFoffersapromisingnewavenueforbonetissueregenerationandrepair.WhilefurtherresearchisneededtodeterminethesafetyandefficacyofCGFinclinicalsettings,thepotentialbenefitsofthistherapyaresubstantial.Withcontinuedstudyanddevelopment,CGFmaybecomeapowerfultoolfortreatingawiderangeofboneandjointdisordersCGF:APromisingToolforTissueRegeneration

Inrecentyears,theuseofgrowthfactorsfortissueregenerationhasgainedpopularityasanalternativetotraditionaltherapeuticapproaches.Amongthem,CGFhasemergedasanovelbiologicalmaterialwithuniquepropertiesforboneregenerationandrepair.

CGFisaconcentrateofautologousgrowthfactorsisolatedfromthepatient'sownblood.Itcontainsvariousgrowthfactors,includingtransforminggrowthfactor-β(TGF-β),platelet-derivedgrowthfactor(PDGF),insulin-likegrowthfactor(IGF),vascularendothelialgrowthfactor(VEGF),andothers.Whenactivated,thesegrowthfactorscanstimulatetheproliferation,migration,differentiation,andextracellularmatrixdepositionofvariouscells,suchasmesenchymalstemcells(MSCs),osteoblasts,andchondrocytes.

OneoftheadvantagesofCGFisitshighconcentrationofgrowthfactors,whichcanbeupto10timeshigherthanthatofplatelet-richplasma(PRP).Moreover,CGFcanreleasegrowthfactorsgraduallyoverseveraldaystoweeks,providingasustainedandlocalizedeffect.Thisfeatureisparticularlybeneficialfortissueengineeringandregeneration,whereacontinuousandcontrolledreleaseofbioactivemoleculesisnecessary.

AnotheradvantageofCGFisitseaseofpreparationandlowcost.Unlikeotherbiologicalmaterials,suchasbonemorphogeneticproteins(BMPs),CGFcanbeobtainedfromasmallamountofblood(10-20mL)withouttheneedforhigh-techequipmentorextensiveprocessing.ThismakesCGFaconvenientandaffordableoptionformanypatientsandhealthcareproviders.

OneofthemostpromisingapplicationsofCGFisinthefieldofboneregenerationandrepair.StudieshaveshownthatCGFcanenhancetheproliferationanddifferentiationofMSCsintoosteoblasts,leadingtoincreasedboneformationandmineralization.Inaddition,CGFcanstimulateangiogenesisandneovascularization,whichareessentialforsupplyingoxygenandnutrientstothenewlyformedtissue.Thiscanacceleratethehealingprocessandminimizetheriskofinfectionandnon-union.

Furthermore,CGFcanalsopromotecartilageregenerationandreduceinflammationinaffectedjoints,leadingtoimprovedmobilityandpainrelief.Thisisparticularlyrelevantfordegenerativejointdiseases,suchasosteoarthritis,wherethelossofcartilageandjointfunctionisamajorconcern.Byprovidingaregenerativeandanti-inflammatoryenvironment,CGFcanpotentiallydelayoravoidtheneedforjointreplacementsurgery,whichisassociatedwithhighcostandmorbidity.

Despitethesepromisingfindings,moreresearchisneededtodeterminethesafetyandefficacyofCGFinclinicalsettings.Someconcernshavebeenraisedaboutthepotentialrisksofusingautologousblood-derivedproducts,suchasinfection,allergicreactions,anddiseasetransmission.Therefore,rigorousqualitycontrolandstandardizationprotocolsmustbeinplacetoensurethepurityandsafetyofCGFpreparations.Inaddition,well-designedclinicaltrialswithlargesamplesizesandlong-termfollow-uparenecessarytoassesstheclinicaloutcomesanddurabilityoftheCGFtherapy.

Inconclusion,CGFoffersapromisingnewavenuefortissueregenerationandrepair,particularly