title: A three-dimensional assemblage of gingiva-derived mesenchymal stem cells and NO-releasing microspheres for improved differentiation.
abstract: Stem cell therapy is an attractive approach to bone tissue regeneration. Nitric oxide (NO) has been reported to facilitate osteogenic differentiation of stem cells. To enhance osteogenic differentiation of gingiva-derived mesenchymal stem cells (GMSCs), we designed a method for in situ delivery of exogenous NO to these cells. A NO donor, polyethylenimine/NONOate, was incorporated into poly(lactic-co-glycolic acid) microspheres to deliver NO to the cells for an extended period of time under in vitro culture conditions. A hybrid aggregate of GMSCs and NO-releasing microspheres was prepared by the hanging drop technique. Confocal microscopy revealed homogeneous arrangement of the stem cells and microspheres in heterospheroids. Western blot analysis and live-dead imaging showed no significant change in cell viability. Importantly, the in situ delivery of NO within the heterospheroids enhanced osteogenic differentiation indicated by a 1.2-fold increase in alkaline phosphatase activity and an approximately 10% increase in alizarin red staining. In addition, a low dose of NO promoted proliferation of the GMSCs in this 3D system. Thus, delivery of the NO-releasing microsphers to induce differentiation of stem cells within this three dimensional system may be one of possible strategies to direct differentiation of a stem cell-based therapeutic agent toward a specific lineage.
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk 38541, Republic of Korea.
authors.forename: Shobha
authors.initials: S
authors.lastname: Regmi
authors.affiliation: College of Pharmacy, Pusan National University, Busan 46241, Republic of Korea.
authors.forename: Jiafu
authors.initials: J
authors.lastname: Cao
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk 38541, Republic of Korea.
authors.forename: Shiva
authors.initials: S
authors.lastname: Pathak
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk 38541, Republic of Korea.
authors.forename: Biki
authors.initials: B
authors.lastname: Gupta
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk 38541, Republic of Korea.
authors.forename: Bijay
authors.initials: B
authors.lastname: Kumar Poudel
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk 38541, Republic of Korea.
authors.forename: Pham Thanh
authors.initials: PT
authors.lastname: Tung
authors.affiliation: College of Pharmacy, Keimyung University, Daegu 42601, Republic of Korea.
authors.forename: Simmyung
authors.initials: S
authors.lastname: Yook
authors.affiliation: Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea.
authors.forename: Jun-Beom
authors.initials: JB
authors.lastname: Park
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk 38541, Republic of Korea.
authors.forename: Chul Soon
authors.initials: CS
authors.lastname: Yong
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk 38541, Republic of Korea.
authors.forename: Jong Oh
authors.initials: JO
authors.lastname: Kim
authors.affiliation: College of Pharmacy, Pusan National University, Busan 46241, Republic of Korea. Electronic address: jinwook@pusan.ac.kr.
authors.forename: Jin-Wook
authors.initials: JW
authors.lastname: Yoo
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk 38541, Republic of Korea. Electronic address: jeeheon@me.com.
authors.forename: Jee-Heon
authors.initials: JH
authors.lastname: Jeong
mesh_terms: D001391:Azo Compounds / Q000737:chemistry; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D005881:Gingiva / Q000166:cytology*; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D008863:Microspheres; D009569:Nitric Oxide / Q000737:chemistry / Q000494:pharmacology*; D010012:Osteogenesis / Q000187:drug effects; D011094:Polyethyleneimine / Q000737:chemistry; D011100:Polyglycolic Acid / Q000737:chemistry; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer
chemical_list: D001391:Azo Compounds; C452539:diazeniumdiolate; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D009569:Nitric Oxide; D019344:Lactic Acid; D011094:Polyethyleneimine
keywords: Heterospheroid; Mesenchymal stem cell; Nitric oxide; Osteogenic differentiation; Poly(lactic-co-glycolic acid) microsphere