Oral mucositis(OM)remains a painful complication of anticancer chemotherapy(CT),tending to progress in severity in the presence of Fusobacterium nucleatum(Fn).Yet,no effective therapy exists to suppress OM since in vi...Oral mucositis(OM)remains a painful complication of anticancer chemotherapy(CT),tending to progress in severity in the presence of Fusobacterium nucleatum(Fn).Yet,no effective therapy exists to suppress OM since in vitro models mimicking CT-induced OM are lacking,halting the discovery of new drugs.Here,we developed an integrated millifluidic in vitro tissue culture system for OM disease modeling.This bioengineered system integrates magnetically bioassembled oral epithelium sheets with millifluidics for CT-based 5-fluorouracil perfusion and Fn infection to model CT-induced OM.After modeling OM with all pro-inflammatory hallmarks,we were able to suppress OM with our in-house plant-produced epidermal growth factor(P-EGF),a well-known re-epithelialization cue.Thus,this the first instance where a milifluidic system enabled OM modeling in the presence of CT drug perfusion and Fn infection.This bioengineered system is a novel tool for drug discovery as it propelled P-EGF as a promising therapy for OM.展开更多
Mucoepidermoid carcinoma(MEC)is a rare malignancy of the salivary gland(SG)that poses significant treatment challenges.This highlights the need for in vitro cancer modeling platforms towards anti-cancer drug screening...Mucoepidermoid carcinoma(MEC)is a rare malignancy of the salivary gland(SG)that poses significant treatment challenges.This highlights the need for in vitro cancer modeling platforms towards anti-cancer drug screening applications.Emerging organ-on-a-chip(OoC)microfluidic technologies represent promising new approach methodologies(NAMS)and a real alternative to animal testing.While tissue-specific decellularized extracellular matrix(ECM)can recapitulate in vivo-like microenvironments,its application in SG-on-a-chip(SGoC)is still underexplored.This study developed an injectable porcine decellularized submandibular gland(dSMG)hydrogel for bioengineering an SG MEC tissue chip.dSMG was prepared using a chemical and enzymatic decellularization process with 0.1%or 1%sodium dodecyl sulfate(SDS).Both treatments effectively removed DNA content while preserving key ECM components,including collagens,glycoproteins,and mucins.Proteomic analysis revealed that 1%SDS-treated dSMG contained a greater abundance of ECM components involved in matrix assembly and cell-ECM interactions compared to the 0.1%group.The 1%SDS-treated dSMG was subsequently digested with a pepsin-based buffer to form hydrogels.At 5 mg/mL,dSMG hydrogel exhibited nanofibrous architecture,thermo-responsive gelation,injectability into microfluidic devices,and minimal batch-to-batch biological variations.In static conditions,dSMG hydrogel significantly enhanced SG cell viability and mitochondria-dependent proliferation compared to Matrigel.Under gravity-driven flow,dSMG hydrogel promoted a ductal phenotype on human SG MEC cells,unlike on Matrigel.Additionally,dSMG hydrogel supported cholinergic-specific signaling and functional activity.These findings demonstrate the potential of dSMG hydrogel as a physiologically relevant matrix for SG cancer modeling towards drug screening applications in SGoC microfluidic systems.展开更多
基金funded by the National Research Council of Thailand(NRCT)and Chulalongkorn University with project number:N42A670176 to JNF(main PI)and to RC(Co-I)supported by the International Association for Dental,Oral and Craniofacial Research(IADR)2022 Innovation in Oral Care Award funded by GlaxoSmithKline to JNF,CHLH,KST,SY and WP+5 种基金supported by Thailand Science Research and Innovation Fund Chulalongkorn University to JNFRC.Center of Excellence and Innovation for Oral Health and Healthy Longevity is funded by the Ratchadaphiseksomphot Endowment Fund,Chulalongkorn University(Grant number:CE68_003_3200_001)supported by a Chulalongkorn University scholarship from Graduate Scholarship Program for ASEAN or Non-ASEAN Countriessupported by the 90th Anniversary of Chulalongkorn University Scholarship under Ratchadapisek Somphot Endowment Fund(Grant number:GCUGR1125662098M)the IADR-Southeast Asia Mentor-Mentee Funding provided to TTTT under the mentorship of JNF and OMsupported by the Second Century Fund(C2F),Chulalongkorn University provided to TVP.
文摘Oral mucositis(OM)remains a painful complication of anticancer chemotherapy(CT),tending to progress in severity in the presence of Fusobacterium nucleatum(Fn).Yet,no effective therapy exists to suppress OM since in vitro models mimicking CT-induced OM are lacking,halting the discovery of new drugs.Here,we developed an integrated millifluidic in vitro tissue culture system for OM disease modeling.This bioengineered system integrates magnetically bioassembled oral epithelium sheets with millifluidics for CT-based 5-fluorouracil perfusion and Fn infection to model CT-induced OM.After modeling OM with all pro-inflammatory hallmarks,we were able to suppress OM with our in-house plant-produced epidermal growth factor(P-EGF),a well-known re-epithelialization cue.Thus,this the first instance where a milifluidic system enabled OM modeling in the presence of CT drug perfusion and Fn infection.This bioengineered system is a novel tool for drug discovery as it propelled P-EGF as a promising therapy for OM.
文摘Mucoepidermoid carcinoma(MEC)is a rare malignancy of the salivary gland(SG)that poses significant treatment challenges.This highlights the need for in vitro cancer modeling platforms towards anti-cancer drug screening applications.Emerging organ-on-a-chip(OoC)microfluidic technologies represent promising new approach methodologies(NAMS)and a real alternative to animal testing.While tissue-specific decellularized extracellular matrix(ECM)can recapitulate in vivo-like microenvironments,its application in SG-on-a-chip(SGoC)is still underexplored.This study developed an injectable porcine decellularized submandibular gland(dSMG)hydrogel for bioengineering an SG MEC tissue chip.dSMG was prepared using a chemical and enzymatic decellularization process with 0.1%or 1%sodium dodecyl sulfate(SDS).Both treatments effectively removed DNA content while preserving key ECM components,including collagens,glycoproteins,and mucins.Proteomic analysis revealed that 1%SDS-treated dSMG contained a greater abundance of ECM components involved in matrix assembly and cell-ECM interactions compared to the 0.1%group.The 1%SDS-treated dSMG was subsequently digested with a pepsin-based buffer to form hydrogels.At 5 mg/mL,dSMG hydrogel exhibited nanofibrous architecture,thermo-responsive gelation,injectability into microfluidic devices,and minimal batch-to-batch biological variations.In static conditions,dSMG hydrogel significantly enhanced SG cell viability and mitochondria-dependent proliferation compared to Matrigel.Under gravity-driven flow,dSMG hydrogel promoted a ductal phenotype on human SG MEC cells,unlike on Matrigel.Additionally,dSMG hydrogel supported cholinergic-specific signaling and functional activity.These findings demonstrate the potential of dSMG hydrogel as a physiologically relevant matrix for SG cancer modeling towards drug screening applications in SGoC microfluidic systems.
