title: Role of TNF alpha and PLF in bone remodeling in a rat model of repetitive reaching and grasping.
abstract: 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.
authors.affiliation: Department of Anatomy and Cell Biology, Temple Medical School, Philadelphia, Pennsylvania 19140, USA.
authors.forename: Shobha
authors.initials: S
authors.lastname: Rani
authors.forename: Mary F
authors.initials: MF
authors.lastname: Barbe
authors.forename: Ann E
authors.initials: AE
authors.lastname: Barr
authors.forename: Judith
authors.initials: J
authors.lastname: Litivn
mesh_terms: 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*
chemical_list: 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
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