Kang I Ko1, Leila S Coimbra2, Chen Tian1, Jazia Alblowi3, Rayyan A Kayal3, Thomas A Einhorn4, Louis C Gerstenfeld4, Robert J Pignolo5,6, Dana T Graves1. 1. Department of Periodontics, University of Pennsylvania, 240 S 40th St, Levy 122 Philadelphia, PA19104, USA. 2. Department of Physiology and Pathology, Araraquara Dental School, State University of São Paulo, Araraquara, São Paulo , Brazil. 3. Department of Oral Basic and Clinical Sciences, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia. 4. Department of Orthopaedic Surgery, School of Medicine, Boston University, Boston, MA, USA. 5. Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. 6. Department of Orthopaedic Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Abstract
AIMS/HYPOTHESIS: Diabetes interferes with bone formation and impairs fracture healing, an important complication in humans and animal models. The aim of this study was to examine the impact of diabetes on mesenchymal stem cells (MSCs) during fracture repair. METHODS: Fracture of the long bones was induced in a streptozotocin-induced type 1 diabetic mouse model with or without insulin or a specific TNFα inhibitor, pegsunercept. MSCs were detected with cluster designation-271 (also known as p75 neurotrophin receptor) or stem cell antigen-1 (Sca-1) antibodies in areas of new endochondral bone formation in the calluses. MSC apoptosis was measured by TUNEL assay and proliferation was measured by Ki67 antibody. In vitro apoptosis and proliferation were examined in C3H10T1/2 and human-bone-marrow-derived MSCs following transfection with FOXO1 small interfering (si)RNA. RESULTS: Diabetes significantly increased TNFα levels and reduced MSC numbers in new bone area. MSC numbers were restored to normal levels with insulin or pegsunercept treatment. Inhibition of TNFα significantly reduced MSC loss by increasing MSC proliferation and decreasing MSC apoptosis in diabetic animals, but had no effect on MSCs in normoglycaemic animals. In vitro experiments established that TNFα alone was sufficient to induce apoptosis and inhibit proliferation of MSCs. Furthermore, silencing forkhead box protein O1 (FOXO1) prevented TNFα-induced MSC apoptosis and reduced proliferation by regulating apoptotic and cell cycle genes. CONCLUSIONS/ INTERPRETATION: Diabetes-enhanced TNFα significantly reduced MSC numbers in new bone areas during fracture healing. Mechanistically, diabetes-enhanced TNFα reduced MSC proliferation and increased MSC apoptosis. Reducing the activity of TNFα in vivo may help to preserve endogenous MSCs and maximise regenerative potential in diabetic patients.
AIMS/HYPOTHESIS: Diabetes interferes with bone formation and impairs fracture healing, an important complication in humans and animal models. The aim of this study was to examine the impact of diabetes on mesenchymal stem cells (MSCs) during fracture repair. METHODS:Fracture of the long bones was induced in a streptozotocin-induced type 1 diabeticmouse model with or without insulin or a specific TNFα inhibitor, pegsunercept. MSCs were detected with cluster designation-271 (also known as p75 neurotrophin receptor) or stem cell antigen-1 (Sca-1) antibodies in areas of new endochondral bone formation in the calluses. MSC apoptosis was measured by TUNEL assay and proliferation was measured by Ki67 antibody. In vitro apoptosis and proliferation were examined in C3H10T1/2 and human-bone-marrow-derived MSCs following transfection with FOXO1 small interfering (si)RNA. RESULTS:Diabetes significantly increased TNFα levels and reduced MSC numbers in new bone area. MSC numbers were restored to normal levels with insulin or pegsunercept treatment. Inhibition of TNFα significantly reduced MSC loss by increasing MSC proliferation and decreasing MSC apoptosis in diabetic animals, but had no effect on MSCs in normoglycaemic animals. In vitro experiments established that TNFα alone was sufficient to induce apoptosis and inhibit proliferation of MSCs. Furthermore, silencing forkhead box protein O1 (FOXO1) prevented TNFα-induced MSC apoptosis and reduced proliferation by regulating apoptotic and cell cycle genes. CONCLUSIONS/ INTERPRETATION:Diabetes-enhanced TNFα significantly reduced MSC numbers in new bone areas during fracture healing. Mechanistically, diabetes-enhanced TNFα reduced MSC proliferation and increased MSC apoptosis. Reducing the activity of TNFα in vivo may help to preserve endogenous MSCs and maximise regenerative potential in diabeticpatients.
Authors: M F Siqueira; J Li; L Chehab; T Desta; T Chino; N Krothpali; Y Behl; M Alikhani; J Yang; C Braasch; D T Graves Journal: Diabetologia Date: 2009-11-10 Impact factor: 10.122
Authors: Jazia Alblowi; Chen Tian; Michelle F Siqueira; Rayyan A Kayal; Erin McKenzie; Yugal Behl; Louis Gerstenfeld; Thomas A Einhorn; Dana T Graves Journal: Bone Date: 2012-12-20 Impact factor: 4.398
Authors: Joshua M Hare; Joel E Fishman; Gary Gerstenblith; Darcy L DiFede Velazquez; Juan P Zambrano; Viky Y Suncion; Melissa Tracy; Eduard Ghersin; Peter V Johnston; Jeffrey A Brinker; Elayne Breton; Janice Davis-Sproul; Ivonne H Schulman; John Byrnes; Adam M Mendizabal; Maureen H Lowery; Didier Rouy; Peter Altman; Cheryl Wong Po Foo; Phillip Ruiz; Alexandra Amador; Jose Da Silva; Ian K McNiece; Alan W Heldman; Richard George; Albert Lardo Journal: JAMA Date: 2012-12-12 Impact factor: 56.272
Authors: Jazia Alblowi; Rayyan A Kayal; Michelle Siqueira; Michelle Siqueria; Erin McKenzie; Nanarao Krothapalli; Jody McLean; Jason Conn; Barbara Nikolajczyk; Thomas A Einhorn; Louis Gerstenfeld; Dana T Graves Journal: Am J Pathol Date: 2009-09-10 Impact factor: 4.307
Authors: Naiomy D Rios-Arce; Andrew Dagenais; Derrick Feenstra; Brandon Coughlin; Ho Jun Kang; Susanne Mohr; Laura R McCabe; Narayanan Parameswaran Journal: J Cell Physiol Date: 2019-09-19 Impact factor: 6.384
Authors: Jason C Lim; Kang I Ko; Marcelo Mattos; Miao Fang; Citong Zhang; Daniel Feinberg; Hisham Sindi; Shuai Li; Jazia Alblowi; Rayyan A Kayal; Thomas A Einhorn; Louis C Gerstenfeld; Dana T Graves Journal: Bone Date: 2017-03-08 Impact factor: 4.398
Authors: Jing Zhang; Katherine J Motyl; Regina Irwin; Ormond A MacDougald; Robert A Britton; Laura R McCabe Journal: Endocrinology Date: 2015-07-02 Impact factor: 4.736
Authors: Ping Wang; Yang Song; Michael D Weir; Jinyu Sun; Liang Zhao; Carl G Simon; Hockin H K Xu Journal: Dent Mater Date: 2015-12-29 Impact factor: 5.304
Authors: L S Coimbra; J P Steffens; S Alsadun; M L Albiero; C Rossa; R J Pignolo; L C Spolidorio; D T Graves Journal: J Dent Res Date: 2015-07-28 Impact factor: 6.116
Authors: Philip M Roper; Pegah Abbasnia; Aleksandra Vuchkovska; Roman M Natoli; John J Callaci Journal: J Orthop Res Date: 2016-07-29 Impact factor: 3.494