Role of TNF alpha and PLF in bone remodeling in a rat model of repetitive reaching and grasping.
We have previously developed a voluntary rat model of highly repetitive reaching that provides an opportunity to study effects of non-weight bearing muscular loads on bone and mechanisms of naturally occurring inflammation on upper limb tissues in vivo. In this study, we investigated the relationship between inflammatory cytokines and matricellular proteins (Periostin-like-factor, PLF, and connective tissue growth factor, CTGF) using our model. We also examined the relationship between inflammatory cytokines, PLF and bone formation processes. Rats underwent initial training for 5 weeks, and then performed a high repetition high force (HRHF) task (12 reaches/min, 60% maximum grip force, 2 h/day, 3 days/week) for 6 weeks. We then examined the effect of training or task performance with or without treatment with a rat specific TNFalpha antibody on inflammatory cytokines, osteocalcin (a bone formation marker), PLF, CTGF, and behavioral indicators of pain or discomfort. The HRHF task decreased grip strength and induced forepaw mechanical hypersensitivity in both trained control and 6-week HRHF animals. Two weeks of anti-TNFalpha treatment improved grip strength in both groups, but did not ameliorate forepaw hypersensitivity. Moreover, anti-TNFalpha treatment attenuated task-induced increases in inflammatory cytokines (TNFalpha, IL-1alpha, and MIP2 in serum; TNFalpha in forelimb bone and muscles) and serum osteocalcin in 6-week HRHF animals. PLF levels in forelimb bones and flexor digitorum muscles increased significantly in 6-week HRHF animals, increases attenuated by anti-TNFalpha treatment. CTGF levels were unaffected by task performance or anti-TNFalpha treatment in 6-week HRHF muscles. In primary osteoblast cultures, TNFalpha, MIP2 and MIP3a treatment increased PLF levels in a dose dependent manner. Also in primary osteoblast cultures, increased PLF promoted proliferation and differentiation, the latter assessed by measuring Runx2, alkaline phosphatase (ALP) and osteocalcin mRNA levels; ALP activity; as well as calcium deposition and mineralization. Increased PLF also promoted cell adhesion in MC3T3-E1 osteoblast-like cell cultures. Thus, tissue loading in vivo resulted in increased TNFalpha, which increased PLF, which then induced anabolic bone formation, the latter results confirmed in vitro.
Rani
Shobha
S
Department of Anatomy and Cell Biology, Temple Medical School, Philadelphia, Pennsylvania 19140, USA.
Barbe
Mary F
MF
Barr
Ann E
AE
Litivn
Judith
J
D016475:3T3 Cells; D000818:Animals; D001522:Behavior, Animal / Q000502:physiology*; D015415:Biomarkers / Q000378:metabolism; D016723:Bone Remodeling / Q000502:physiology*; D001842:Bone and Bones / Q000166:cytology / Q000502:physiology; D002448:Cell Adhesion / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D018925:Chemokines / Q000378:metabolism; D055513:Connective Tissue Growth Factor / Q000235:genetics / Q000378:metabolism; D012090:Cumulative Trauma Disorders / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D005260:Female; D005552:Forelimb / Q000033:anatomy & histology / Q000502:physiology; D018737:Hand Strength / Q000502:physiology*; D007249:Inflammation / Q000378:metabolism; D051379:Mice; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000166:cytology / Q000502:physiology; D010006:Osteoblasts / Q000166:cytology / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012677:Sensation / Q000502:physiology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism*
D015415:Biomarkers; D015815:Cell Adhesion Molecules; D018925:Chemokines; D016207:Cytokines; D009124:Muscle Proteins; D014409:Tumor Necrosis Factor-alpha; C520508:periostin-like factor, rat; D055513:Connective Tissue Growth Factor
Cancer Res. 2002 Sep 15;62(18):5358-64
12235007
Anat Rec A Discov Mol Cell Evol Biol. 2005 Dec;287(2):1205-12
16240445
Int J Biol Markers. 2008 Jul-Sep;23(3):182-6
18949745
Brain Res. 2008 Jul 7;1218:103-13
18511022
Mol Biol Cell. 1993 Jun;4(6):637-45
8374172
J Bone Miner Res. 1999 Jul;14(7):1239-49
10404027
J Cell Commun Signal. 2009 Dec;3(3-4):163-5
19779848
Mol Cell Endocrinol. 2010 Feb 5;315(1-2):195-200
19879324
Exp Eye Res. 2009 Apr;88(4):694-703
19101543
J Orthop Res. 2008 Oct;26(10):1320-6
18464247
Circ Res. 2000 May 12;86(9):939-45
10807865
J Cell Biol. 1995 Aug;130(3):503-6
7542656
J Cell Biochem. 2004 Aug 1;92(5):1044-61
15258926
Spine (Phila Pa 1976). 2007 Feb 15;32(4):413-6
17304130
Growth Factors. 2008 Apr;26(2):80-91
18428027
J Cell Physiol. 1990 Jun;143(3):420-30
1694181
Atherosclerosis. 2010 Feb;208(2):358-65
19695571
Calcif Tissue Int. 2003 Dec;73(6):584-93
14517717
J Histochem Cytochem. 2009 Nov;57(11):1061-73
19620321
Breast Cancer Res Treat. 2003 Feb;77(3):245-52
12602924
J Bone Miner Res. 2003 Nov;18(11):2023-32
14606516
J Clin Invest. 2003 Mar;111(6):821-31
12639988
Exp Cell Res. 2002 Sep 10;279(1):91-9
12213217
Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3632
7724610
J Neurotrauma. 2003 Jul;20(7):681-95
12908929
Cytokine Growth Factor Rev. 2001 Mar;12(1):9-18
11312114
J Immunol. 1987 Apr 1;138(7):2137-42
3494060
J Biol Chem. 2009 Dec 18;284(51):35987-95
19854828
Bone. 2009 Nov;45(5):1010-6
19651258
Mol Cell Biol. 1993 Sep;13(9):5276-89
8355682
Int J Biochem Cell Biol. 2009 Apr;41(4):771-83
18775791
Cancer Res. 2006 Jul 15;66(14):6928-35
16849536
Curr Opin Cell Biol. 2002 Oct;14(5):608-16
12231357
Bone. 2005 Feb;36(2):232-42
15780949
Mol Cell Biochem. 2009 Feb;322(1-2):25-36
18985280
J Orthop Res. 2010 Mar;28(3):298-307
19743505
J Biol Chem. 2006 Feb 17;281(7):4326-33
16373342
Pain. 2000 Apr;85(3):333-343
10781907
J Immunol. 2000 May 15;164(10):5352-61
10799898
J Clin Pathol. 2008 Apr;61(4):494-8
17938160
Neurosci Lett. 2005 Jul 1-8;382(1-2):148-52
15911139
J Orthop Res. 2003 Jan;21(1):167-76
12507595
Cytokine Growth Factor Rev. 2008 Apr;19(2):133-44
18358427
Cardiovasc Pathol. 2006 Jan-Feb;15(1):24-32
16414453
Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3540-5
10097072
Cytokine. 2004 May 7;26(3):138-44
15135808
Wound Repair Regen. 2008 Sep-Oct;16(5):585-601
19128254
Cancer Cell. 2004 Apr;5(4):329-39
15093540
Ann N Y Acad Sci. 2008 Mar;1123:30-40
18375575
Differentiation. 2008 Sep;76(7):809-19
18294225
Nat Med. 2007 Aug;13(8):962-9
17632525
J Histochem Cytochem. 2008 Apr;56(4):329-45
18040074
Int J Cancer. 2008 Jun 15;122(12):2707-18
18381746
J Biol Chem. 2008 Aug 22;283(34):23084-92
18567580
Histol Histopathol. 2007 Oct;22(10):1167-74
17616943
J Immunol. 2002 Jun 1;168(11):5621-8
12023359
J Cell Physiol. 2009 Mar;218(3):584-92
19006175
J Cell Commun Signal. 2010 Mar;4(1):1-4
19798591
Exerc Sport Sci Rev. 2003 Oct;31(4):188-94
14571958
J Immunol. 1991 Sep 1;147(5):1694-700
1715367
Neuroscience. 2009 Jan 23;158(2):922-31
19032977
J Neuroimmunol. 2005 Oct;167(1-2):13-22
16026858
J Orthop Sports Phys Ther. 2004 May;34(5):244-53
15189016
Prostate. 2009 Sep 15;69(13):1398-403
19479898
Neurosci Lett. 2009 Apr 17;454(1):33-7
19429049
J Cell Physiol. 2007 Mar;210(3):843-52
17133352
J Orthop Res. 2004 May;22(3):520-5
15099630
J Biol Chem. 2001 Jan 5;276(1):563-8
11032840
Spine (Phila Pa 1976). 2000 Dec 1;25(23):2975-80
11145807
J Cell Physiol. 2007 Jan;210(1):26-37
17034042
Bone. 2009 Mar;44(3):502-12
19095091
J Cell Biochem. 2000 Feb;77(1):103-15
10679821
Annu Rev Immunol. 1996;14:649-83
8717528
Arterioscler Thromb Vasc Biol. 2005 Jan;25(1):77-83
15514205
J Orthop Res. 2010 May;28(5):657-63
19890995
J Biol Chem. 1997 Dec 5;272(49):31092-9
9388261
Physiol Genomics. 2003 Aug 15;14(3):261-71
12799472
Pain. 2003 May;103(1-2):175-86
12749972
J Invest Dermatol. 1996 Apr;106(4):729-33
8618012
Mol Cell Biol. 1993 Nov;13(11):7191-8
8413306